<%BANNER%>

Pharaoh Ant Consumption of Fluids Used in Hospital Environments


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P H A R A O H A N T C O N S U M P T I O N O F F L U I D S U S E D I N H O S P I T A L E N V I R O N M E N T S B y R O X A N N E G R A C E B U R R U S A T H E S I S P R E S E N T E D T O T H E G R A D U A T E S C H O O L O F T H E U N I V E R S I T Y O F F L O R I D A I N P A R T I A L F U L F I L L M E N T O F T H E R E Q U I R E M E N T S F O R T H E D E G R E E O F M A S T E R O F S C I E N C E U N I V E R S I T Y O F F L O R I D A 2 0 0 4

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C o p y r i g h t 2 0 0 4 b y R o x a n n e G r a c e B u r r u s

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I d e d i c a t e t h i s t h e s i s t o G o d T h e R i s e n L o r d a n d C r e a t o r o f a l l t h i n g s

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i v A C K N O W L E D G M E N T S I a m d e e p l y i n d e b t e d t o f a m i l y a n d f r i e n d s w h o s e a d v i c e m o r a l s u p p o r t a n d a s s i s t a n c e i n a m u l t i t u d e o f w a y s h a v e p r o v e n s o v i t a l l y i m p o r t a n t a n d u p l i f t i n g F i r s t I a m g r a t e f u l t o G o d f o r a n i n q u i s i t i v e m i n d a d e l i g h t i n H i s c r e a t i o n a n d a n u n e n d i n g c u r i o s i t y a b o u t h o w t h i n g s c a m e t o b e t h e w a y t h e y a r e E v e n a s a c h i l d I e n j o y e d e x p l o r i n g o u t d o o r s I t h a n k m y p a r e n t s f o r t h e i r g u i d a n c e s u p p o r t a n d e n c o u r a g e m e n t o v e r t h e p a s t s e v e r a l y e a r s i n s o m a n y w a y s I n a d d i t i o n t o b e i n g g r e a t p a r e n t s t h e y a r e g o o d f r i e n d s w h o s e c o m p a n y I e n j o y a n d w h o s e c o u n s e l I a p p r e c i a t e T h e y a r e r e a l l y s t r o n g i n t h e c o m m o n s e n s e d e p a r t m e n t w h i c h h a s b e e n g o o d b a l l a s t a g a i n s t t h e i d e a l i s t i c a t t i t u d e I t a c k l e l i f e ’ s c h a l l e n g e s w i t h T w o y e a r s a g o w h e n c i r c u m s t a n c e s p r e v e n t e d m e f r o m c o l l e c t i n g m y h o u s e h o l d g o o d s d u r i n g t h e u n i v e r s i t y ’ s s p r i n g b r e a k m y p a r e n t s d r o v e f r o m F l o r i d a t o M a s s a c h u s e t t s p a c k e d m y b e l o n g i n g s a n d c a t s i n t o a r e n t a l t r u c k a n d d e l i v e r e d i t t o m e i n G a i n e s v i l l e F L – a l l w i t h i n o n e w e e k T h a t k i n d o f g e n e r o s i t y a n d k i n d n e s s w i l l a l w a y s b e r e m e m b e r e d a n d g r e a t l y a p p r e c i a t e d I t h a n k m y s i s t e r K a r e n w h o s e u n w a v e r i n g s u p p o r t a n d s e l f l e s s a t t i t u d e h a s g i v e n m e t h e e n e r g y t o p e r s e v e r e m o r e t h a n s h e k n o w s S h e h a s b e e n a r o l e m o d e l o f e n c o u r a g e m e n t a t e v e r y s t e p o f t h e w a y I a m f o r t u n a t e t o c o u n t h e r a s a f r i e n d a s w e l l a s a s i b l i n g

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v S a r a h S m i t h h a s b e e n b o t h f r i e n d a n d m e n t o r i n m a n y w a y s A s a b i o l o g i s t a n d n a t u r a l i s t h e r k n o w l e d g e a n d a p p r e c i a t i o n f o r l i f e a n d l i v i n g o r g a n i s m s h a v e b e e n i n s p i r i n g S h e p u t u s e f u l t o o l s i n m y p a t h s o t h a t I e x p e r i e n c e d t h e j o y o f d i s c o v e r y O t h e r f r i e n d s h a v e f i r m l y e n t r e n c h e d t h e m s e l v e s i n m y l i f e M a r s h a A s q u i t h M i k e M a g e e D a v e C o t t o n D o u g N i c h o l s A l i n e R o b e r J o h n C r e w a n d M a r k A l a r i e s p o k e w o r d s o f w i s d o m a t j u s t t h e r i g h t t i m e o n s o m a n y o c c a s i o n s a n d h e l p e d m e s t a y t h e c o u r s e d u r i n g d i f f i c u l t t i m e s T h e y p r o v i d e d i d e a s a n d i n s i g h t s t h a t p r o v e d i n v a l u a b l e i n b o t h m y p e r s o n a l l i f e a n d i n m y r e s e a r c h W h i l e a t t h e U n i v e r s i t y o f M a s s a c h u s e t t s A m h e r s t I h a d t h e g o o d f o r t u n e t o m e e t R e g C o l e r H i s e n t h u s i a s m a n d v i v a c i t y a r e t o p p e d o n l y b y h i s k e e n i n t e r e s t a n d k n o w l e d g e o f e n t o m o l o g y A s m y f i r s t g r a d u a t e a d v i s o r h e t a u g h t m e r e s e a r c h s k i l l s t h a t w i l l p r o v e i n v a l u a b l e f o r t h e r e s t o f m y l i f e T h e k n o w l e d g e t h a t h e i m p a r t e d i s e s p e c i a l l y r e m a r k a b l e w h e n o n e c o n s i d e r s t h e s h o r t a m o u n t o f t i m e w e w e r e a b l e t o w o r k t o g e t h e r M y t e n u r e a t t h e U n i v e r s i t y o f F l o r i d a b r o u g h t n e w f r i e n d s a n d m e n t o r s i n t o m y l i f e G e n e G e r b e r g h a s m e n t o r e d m e f r o m h i s s t r o n g b a c k g r o u n d o f a l i f e t i m e o f w o r k i n t h e f i e l d o f m e d i c a l e n t o m o l o g y H i s a d v i c e h a s a l r e a d y p r o v e n v a l u a b l e i n m a n y r e s p e c t s a n d I a m s u r e i t w i l l c o n t i n u e t o d o s o H e t o o k t h i s f l e d g l i n g e n t o m o l o g i s t u n d e r h i s w i n g s a n d e n s u r e d t h a t I s t a y e d o n t r a c k d u r i n g m y t i m e h e r e i n F l o r i d a K n o w i n g h i m h a s b e e n a r e a l p l e a s u r e ; I h a v e e n j o y e d a n d b e n e f i t e d f r o m h i s a n e c d o t e s o f d i f f e r e n t e n t o m o l o g i s t s h e w o r k e d w i t h H i s a u t o b i o g r a p h y w i l l b e i n t e r e s t i n g t o r e a d F a i t h O i h a s s p e n t a g r e a t d e a l o f t i m e h e l p i n g m e w i t h s t a t i s t i c s a n d f i n e t u n i n g t h e m a n u s c r i p t I n a d d i t i o n s h e t a u g h t m e u s e f u l r e s e a r c h s k i l l s t h a t h a v e b e e n t i m e

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v i s a v i n g t o o l s I a p p r e c i a t e h e r h e l p a n d a d v i c e I a m a b e t t e r r e s e a r c h e r d u e t o h e r i n f l u e n c e D a v i d O i h a s a l s o s p e n t a g r e a t d e a l o f t i m e h e l p i n g w i t h t h e t h e s i s a n d w i t h s t a t i s t i c s H i s c o n t r i b u t i o n s i m p r o v e d t h e e x p e r i m e n t a l d e s i g n t h e a n a l y s i s o f d a t a a n d t h e m a n u s c r i p t H i s k n o w l e d g e o f a n t b e h a v i o r a n d o f t h e l i t e r a t u r e h a s b e e n i n s p i r i n g T h e k n o w l e d g e a n d r e s e a r c h s k i l l s h e i m p a r t e d t o m e h a v e a l r e a d y m a d e m e a b e t t e r s c i e n t i s t a n d w i l l c o n t i n u e t o p r o v e h e l p f u l P h i l K o e h l e r h a s h e l p e d m e p u t t o g e t h e r n i c e p r e s e n t a t i o n s M u c h o f w h a t I h a v e l e a r n e d f r o m w o r k i n g w i t h h i m a n d i n h i s l a b e x t e n d s b e y o n d t e x t b o o k k n o w l e d g e a n d w i l l p r o v e u s e f u l i n m a n y s i t u a t i o n s I t h a n k t h e s t a f f i n t h e e n t o m o l o g y d e p a r t m e n t e s p e c i a l l y D e b b i e H a l l a n d M y r n a L i t c h f i e l d T h e y h a v e b e e n w o n d e r f u l i n e v e r y r e s p e c t F i n a l l y I t h a n k m y f e l l o w T r i G a t o r a t h l e t e s a n d c o a c h e s T r a i n i n g f o r a n d p a r t i c i p a t i n g i n t r i a t h l o n s h a v e b e c o m e a n i n t e g r a l p a r t o f m y l i f e C o m p l e t i o n o f l o n g e v e n t s h a s h e l p e d c l e a r m y f o c u s a n d s t e e l m y m i n d T h i s h a s i m p r o v e d m y r e s e a r c h a n d h e l p e d m e e n j o y t h e j o u r n e y

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v i i T A B L E O F C O N T E N T S p a g e A C K N O W L E D G M E N T S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i v L I S T O F T A B L E S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x L I S T O F F I G U R E S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x i A B S T R A C T . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x i i i C H A P T E R 1 I N T R O D U C T I O N . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2 L I T E R A T U R E R E V I E W . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 G e n e r a l A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 T r a m p A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 P h a r a o h A n t . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 O r i g i n a n d D i s t r i b u t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 B i o l o g y . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 F o r a g i n g a n d F o o d . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 M e d i c a l I m p o r t a n c e . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 C o n t r o l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 3 N O C H O I C E L I Q U I D T E S T . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 3 I n t r o d u c t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 3 M a t e r i a l s a n d M e t h o d s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 5 A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 5 F o r a g i n g A r e n a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 5 E x p e r i m e n t a l F l u i d s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 7 E x p e r i m e n t a l D e s i g n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 9 F l u i d M e a s u r e m e n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1 S t a t i s t i c a l A n a l y s i s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2 R e s u l t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 3 D i s c u s s i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 5 F l u i d M e a s u r e m e n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 6 W a t e r C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 6 D e x t r o s e C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 8

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v i i i P l a s m a a n d B l o o d C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 9 S a l i n e ( = 0 9 % N a C l ) C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 1 E n s u r e C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 1 4 C H O I C E L I Q U I D T E S T . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 8 I n t r o d u c t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 8 M a t e r i a l s a n d M e t h o d s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 0 A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 0 F o r a g i n g A r e n a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 0 E x p e r i m e n t a l F l u i d s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 2 E x p e r i m e n t a l D e s i g n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 2 F l u i d M e a s u r e m e n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 3 S t a t i s t i c a l A n a l y s i s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 5 R e s u l t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 5 N u m b e r o f A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 5 F l u i d M e a s u r e m e n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 6 C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 7 D i s c u s s i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 7 N u m b e r o f A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 7 F l u i d M e a s u r e m e n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 8 C o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 9 D e x t r o s e . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 0 E n s u r e . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 0 P l a s m a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 1 N a C l . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 2 B l o o d . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 3 5 N O C H O I C E D E H Y D R A T E D F L U I D T E S T . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 0 I n t r o d u c t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 0 M a t e r i a l s a n d M e t h o d s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 1 A n t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 1 F o r a g i n g A r e n a a n d N e s t . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2 H o s p i t a l S h e e t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 2 D e h y d r a t i o n o f F l u i d s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 4 E x p e r i m e n t a l D e s i g n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 5 T i m e d O b s e r v a t i o n s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 5 S t a t i s t i c a l A n a l y s i s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 6 R e s u l t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 6 D i s c u s s i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 8 6 S U M M A R Y A N D C O N C L U S I O N S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 7 A P P E N D I X A A N T M A I N T E N A N C E . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 0

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i x B S E L E C T I O N O F A N T S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3 C C O N S U M P T I O N F O R M U L A S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 5 L I S T O F R E F E R E N C E S . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 9 B I O G R A P H I C A L S K E T C H . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 0 7

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x L I S T O F T A B L E S T a b l e p a g e 3 1 M e a n ( l i q u i d n o c h o i c e ) i n i t i a l w e i g h t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 4 3 2 M e a n ( l i q u i d n o c h o i c e ) c o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 5 4 1 M e a n n u m b e r o f a n t s r e m a i n i n g . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 4 4 2 M e a n ( c h o i c e t e s t ) i n i t i a l w e i g h t s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 5 4 3 M e a n ( c h o i c e t e s t ) c o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 6

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x i L I S T O F F I G U R E S F i g u r e p a g e 3 1 N o c h o i c e l i q u i d a r e n a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 6 3 2 M e a n ( l i q u i d n o c h o i c e ) e v a p o r a t i o n ( 2 4 h ) i n c o n t r o l a r e n a s . . . . . . . . . . . . . . . . 3 6 3 3 M e a n ( l i q u i d n o c h o i c e ) c o n s u m p t i o n ( 2 4 h ) i n t e s t a r e n a s . . . . . . . . . . . . . . . . . . . 3 7 4 1 C h o i c e l i q u i d a r e n a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 1 4 2 M e a n n u m b e r o f a n t s r e m a i n i n g i n f l u i d a f t e r 2 4 h . . . . . . . . . . . . . . . . . . . . . . . . . 5 7 4 3 M e a n ( c h o i c e t e s t ) e v a p o r a t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 8 4 4 M e a n ( c h o i c e t e s t ) c o n s u m p t i o n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 9 5 1 D e h y d r a t e d f l u i d a r e n a . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6 3 5 2 M e a n n u m b e r o f a n t s p e r 1 5 m i n t i m e i n t e r v a l s o v e r e n t i r e 2 h . . . . . . . . . . . . . . 7 2 5 3 M e a n n u m b e r o f a n t s p e r d e h y d r a t e d f l u i d o v e r e n t i r e 2 h . . . . . . . . . . . . . . . . . . 7 3 5 4 M e a n n u m b e r o f a n t s p e r f l u i d a t 1 5 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 4 5 5 M e a n n u m b e r o f a n t s p e r f l u i d a t 3 0 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 5 5 6 M e a n n u m b e r o f a n t s p e r f l u i d a t 4 5 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 6 5 7 M e a n n u m b e r o f a n t s p e r f l u i d a t 6 0 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 7 5 8 M e a n n u m b e r o f a n t s p e r f l u i d a t 7 5 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 8 5 9 M e a n n u m b e r o f a n t s p e r f l u i d a t 9 0 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 9 5 1 0 M e a n n u m b e r o f a n t s p e r f l u i d a t 1 0 5 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 0 5 1 1 M e a n n u m b e r o f a n t s p e r f l u i d a t 1 2 0 m i n . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 1 5 1 2 M e a n n u m b e r o f a n t s f e e d i n g o n E n s u r e . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 2 5 1 3 M e a n n u m b e r o f a n t s f e e d i n g o n 5 % d e x t r o s e . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 3

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x i i 5 1 4 M e a n n u m b e r o f a n t s f e e d i n g o n s a l i n e . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 4 5 1 5 M e a n n u m b e r o f a n t s f e e d i n g o n h u m a n b l o o d c e l l s . . . . . . . . . . . . . . . . . . . . . . . 8 5 5 1 6 M e a n n u m b e r o f a n t s f e e d i n g o n h u m a n p l a s m a . . . . . . . . . . . . . . . . . . . . . . . . . . 8 6

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x i i i A b s t r a c t o f T h e s i s P r e s e n t e d t o t h e G r a d u a t e S c h o o l o f t h e U n i v e r s i t y o f F l o r i d a i n P a r t i a l F u l f i l l m e n t o f t h e R e q u i r e m e n t s f o r t h e D e g r e e o f M a s t e r o f S c i e n c e P H A R A O H A N T C O N S U M P T I O N O F F L U I D S U S E D I N H O S P I T A L E N V I R O N M E N T S B y R o x a n n e G r a c e B u r r u s M a y 2 0 0 4 C h a i r : P h i l i p G K o e h l e r C o c h a i r : D a v i d H O i M a j o r D e p a r t m e n t : E n t o m o l o g y a n d N e m a t o l o g y M o n o m o r i u m p h a r a o n i s ( L ) t h e P h a r a o h a n t i s a p e s t s p e c i e s w h i c h i n h a b i t s b u i l d i n g s t h r o u g h o u t t h e w o r l d I t s p o p u l a t i o n s a r e d i f f i c u l t t o c o n t r o l W h e n i n h o s p i t a l s a n d o t h e r h e a l t h c a r e e n v i r o n m e n t s t h e s e a n t s a r e o f t e n f o u n d n e s t i n g i n p l a c e s a s d i v e r s e a s o p e r a t i n g r o o m s l i n e n c l o s e t s b a b y i n c u b a t o r s a n d i n s i d e u n o p e n e d p a c k a g e s o f i n t r a v e n o u s t u b i n g P h a r a o h a n t s h a v e b e e n o b s e r v e d f e e d i n g u p o n h u m a n s e c r e t i o n s a n d o n i n t a c t e p i d e r m i s I n a d d i t i o n t h e y h a v e b e e n r e p o r t e d t o c o n s u m e p a t i e n t c a r e f l u i d s s u c h a s d e x t r o s e a n d s a l i n e s o l u t i o n s S m a l l P h a r a o h a n t c o l o n i e s t h a t w e r e s t a r v e d f o r 2 d c o n s u m e d s i g n i f i c a n t a m o u n t s o f e i t h e r b l o o d 5 % d e x t r o s e E n s u r e d i e t a r y s u p p l e m e n t 0 9 % N a C l s o l u t i o n ( s a l i n e ) o r p l a s m a w h e n g i v e n 2 4 h a c c e s s t o e a c h f l u i d s e p a r a t e l y C o m p a r i s o n o f c o n s u m p t i o n s b a s e d o n t h e s e n o c h o i c e e x p o s u r e s i n d i c a t e d t h a t s i g n i f i c a n t l y m o r e 5 % d e x t r o s e w a s c o n s u m e d a m o n g a l l t h e o t h e r f l u i d s S i m u l t a n e o u s o r c h o i c e e x p o s u r e s o f

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x i v t h e s a m e f l u i d s a g a i n s h o w e d t h e m o s t s i g n i f i c a n t c o n s u m p t i o n o c c u r r e d o n 5 % d e x t r o s e C o u n t s o f f e e d i n g P h a r a o h a n t s w h i c h w e r e s t a r v e d f o r 3 d a n d g i v e n i n d i v i d u a l a c c e s s t o t h e f l u i d s d r i e d o n h o s p i t a l l i n e n s w e r e h i g h e s t o n t h e E n s u r e d i e t a r y s u p p l e m e n t f o l l o w e d b y t h e 5 % d e x t r o s e

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1 C H A P T E R 1 I N T R O D U C T I O N M o n o m o r i u m p h a r a o n i s ( L ) c o m m o n l y k n o w n a s t h e P h a r a o h a n t h a s a c l o s e a s s o c i a t i o n w i t h h u m a n h a b i t a t s ( W h e e l e r 1 9 1 0 ) a n d i s p r o b a b l y o n e o f t h e w o r l d ’ s m o s t s t u d i e d a n t s I t i s a t r o p i c a l a n t t h a t n e e d s b o t h h i g h h u m i d i t y a n d h i g h t e m p e r a t u r e s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) I n t e m p e r a t e z o n e s i t l i v e s i n b u i l d i n g s s u c h a s h o s p i t a l s w h i c h a r e h e a t e d a n d h a v e i n d o o r p l u m b i n g t h r o u g h o u t t h e s t r u c t u r e ( E d w a r d s a n d S h o r t 1 9 9 0 ) B e a t s o n ( 1 9 7 2 ) r e p o r t e d t h a t P h a r a o h a n t s c a n c a r r y s e v e r a l p a t h o g e n s i n c l u d i n g S t r e p t o c o c c u s S t a p h y l o c o c c u s E s c h e r i c h i a c o l i B a c i l l u s P s e u d o m o n a s a n d o t h e r s T h i s d i s c o v e r y s p a r k e d a b r i e f i n t e r e s t i n t h e p a t h o g e n i c v e c t o r c a p a c i t y o f P h a r a o h a n t s t h a t l a s t e d i n t o t h e 1 9 8 0 ’ s M o r e r e c e n t l y i n d e p e n d e n t o b s e r v a t i o n s b y E d w a r d s a n d S h o r t ( 1 9 9 0 ) a n d ( E d w a r d s a n d S h o r t 1 9 9 0 E i c h l e r 1 9 9 0 ) o f P h a r a o h a n t s f e e d i n g o n h u m a n p a t i e n t s r e k i n d l e d i n t e r e s t i n t h i s s p e c i e s A d d i t i o n a l l y a l i m i t e d n u m b e r o f t r a m p a n t s p e c i e s i n c l u d i n g P h a r a o h a n t s h a v e b e e n i m p l i c a t e d a s v e c t o r s o f h o s p i t a l a c q u i r e d d i s e a s e s ( F o w l e r 1 9 9 0 ) a n d w e r e a s s o c i a t e d w i t h a h i g h e r i n c i d e n c e o f n o s o c o m i a l i n f e c t i o n s ( R i c h a r d s e t a l 1 9 9 9 ) T h e a b i l i t y o f P h a r a o h a n t s t o m o v e b e t w e e n p a t i e n t s r e c e i v i n g m e d i c a l t r e a t m e n t a n d h o s p i t a l a r e a s t h a t m a y h a r b o r p a t h o g e n s s u c h a s s e w a g e p l u m b i n g f o o d p r e p a r a t i o n a r e a s a n d e v e n d i s e a s e d p a t i e n t s s u g g e s t s t h a t P h a r a o h a n t s h a v e t h e p o t e n t i a l o f s p r e a d i n g p a t h o g e n i c d i s e a s e s t h r o u g h o u t a h o s p i t a l E i c h l e r ( 1 9 9 0 ) c o n s i d e r e d P h a r a o h a n t s t o b e o n e o f t h e m o s t d a n g e r o u s i n s e c t s i n h e a l t h c a r e e n v i r o n m e n t s

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2 T h e d i s e a s e v e c t o r i n g p o t e n t i a l o f P h a r a o h a n t s l i k e t h a t o f o t h e r a r t h r o p o d s m o r e t r a d i t i o n a l l y a s s o c i a t e d w i t h d i s e a s e t r a n s m i s s i o n s u c h a s c o c k r o a c h e s a n d f l i e s m a y b e d i r e c t l y l i n k e d t o t h e i r f o o d p r e f e r e n c e s a n d f o r a g i n g b e h a v i o r T h e r e f o r e P h a r a o h a n t c o n s u m p t i o n o f l i q u i d s u b s t a n c e s t h a t a r e s p e c i f i c a l l y u s e d i n p a t i e n t c a r e n e e d e d t o b e e x a m i n e d m o r e c l o s e l y T h e o b j e c t i v e o f t h i s r e s e a r c h w a s t o d e t e r m i n e w h i c h f l u i d ( s ) u s e d i n p a t i e n t c a r e a t t r a c t P h a r a o h a n t s T w o e x p e r i m e n t s w e r e c o n d u c t e d t o d e t e r m i n e i f P h a r a o h a n t s c o n s u m e ( 1 ) l i q u i d f l u i d s u s e d i n h o s p i t a l s ( C h a p t e r 3 ) a n d ( 2 ) d e h y d r a t e d h o s p i t a l f l u i d s ( C h a p t e r 5 ) A t h i r d e x p e r i m e n t ( C h a p t e r 4 ) c o m p a r e d t h e a t t r a c t i o n a m o n g t h e l i q u i d h o s p i t a l f l u i d s w h e n p r e s e n t e d s i m u l t a n e o u s l y t o P h a r a o h a n t s

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3 C H A P T E R 2 L I T E R A T U R E R E V I E W G e n e r a l A n t s T h e r e a r e o v e r 9 5 0 0 k n o w n s p e c i e s o f a n t s i n t h e w o r l d ( B o l t o n 1 9 9 5 ) A n t s a r e e u s o c i a l i n s e c t s w h i c h l i v e i n c o l o n i e s c o m p r i s e d o f m u l t i p l e c a s t e s u s u a l l y m o r p h o l o g i c a l b u t s o m e t i m e s t e m p o r a l ( O s t e r a n d W i l s o n 1 9 7 8 ) a n d m u l t i p l e d e v e l o p m e n t a l s t a g e s E u s o c i a l b e h a v i o r i n a n t s i s s e e n i n t h e d i v i s i o n o f l a b o r / f u n c t i o n a m o n g s t d i f f e r e n t c a s t e s i n t h e p r e s e n c e o f o v e r l a p p i n g g e n e r a t i o n s a n d i n t h e t e n d i n g o f i m m a t u r e b r o o d b y n o n p a r e n t a l a d u l t s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) T r a m p A n t s A n t s w h i c h a r e h a r m f u l t o h u m a n a c t i v i t y a r e c l a s s i f i e d a s p e s t s a n d a r e o f t e n c o n s i d e r e d t o b e t r a m p s p e c i e s T r a m p s p e c i e s h a v e m a n y t r a i t s t h a t e n a b l e d t h e m t o b e s y n a n t h r o p i c T h e y a r e a t t r a c t e d t o c h a n g i n g e n v i r o n m e n t s t h a t a r e l i n k e d s t r o n g l y t o h u m a n a c t i v i t y ( P a s s e r a 1 9 9 4 ) a n d o f t e n n e s t i n s i d e b u i l d i n g s ( E b e l i n g 1 9 7 8 ) M a n y h a v e b e e n t r a n s p o r t e d t o n e w c o u n t r i e s a r o u n d t h e w o r l d v i a h u m a n c o m m e r c e ( R i l e y 1 8 8 9 ) T r a m p s p e c i e s t e n d t o m i g r a t e q u i c k l y i n r e s p o n s e t o c h a n g e s i n e n v i r o n m e n t a l c o n d i t i o n s o r f o o d a v a i l a b i l i t y o r t o o v e r p o p u l a t e d c o l o n y s i z e ( P a s s e r a 1 9 9 4 ) M a n y a r e p o l y d o m o u s c o l o n i e s t h a t l a c k a g g r e s s i o n b e t w e e n n e i g h b o r i n g n e s t s s o t h a t w o r k e r s q u e e n s a n d b r o o d a r e f r e e l y e x c h a n g e d b e t w e e n n e s t s ( P a s s e r a 1 9 9 4 ) W i t h i n e a c h n e s t t r a m p s p e c i e s a r e o f t e n p o l y g y n e t h a t i s t h e y h a v e m a n y f e r t i l i z e d q u e e n s ( E b e l i n g 1 9 7 8 P a s s e r a 1 9 9 4 ) S e x u a l c a s t e s o f t e n e n g a g e i n i n t r a n i d a l m a t i n g r a t h e r t h a n h a v i n g n u p t i a l f l i g h t s e v e n w h e n w i n g s a r e p r e s e n t ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) C o l o n y

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4 r e p r o d u c t i o n t a k e s p l a c e b y s o c i o t o m y o r b u d d i n g w h e n w o r k e r s a n d n e w l y m a t e d q u e e n s l e a v e t o s t a r t a n e w n e s t n e a r b y ( P a s s e r a 1 9 9 4 ) T r a m p a n t s a r e u s u a l l y m o n o m o r p h i c w i t h o n l y o n e m o r p h o l o g i c a l w o r k e r c a s t e a n d a r e a l w a y s v e r y s m a l l ( P a s s e r a 1 9 9 4 ) P h a r a o h A n t M o n o m o r i u m p h a r a o n i s ( L ) ( F o r m i c i d a e : M y r m i c i n a e ) t h e P h a r a o h a n t i s a w o r l d w i d e p e s t ( W h e e l e r 1 9 1 0 H l l d o b l e r a n d W i l s o n 1 9 9 0 ) t h a t i s c l o s e l y a s s o c i a t e d w i t h h u m a n s a n d i s c o n s i d e r e d a t r a m p a n t ( P a s s e r a 1 9 9 4 ) P h a r a o h a n t c o l o n i e s e x h i b i t a t e m p o r a l d i v i s i o n o f l a b o r ( S u d d 1 9 5 7 b ) i n w h i c h y o u n g e r w o r k e r s s p e n d m o s t o f t h e i r t i m e t e n d i n g t h e b r o o d a n d q u e e n s w h i l e o l d e r w o r k e r s p r i m a r i l y f o r a g e I n t h e a b s e n c e o f o n e c a s t e m e m b e r s o f a n o t h e r c a s t e w i l l p e r f o r m t h e a d d i t i o n a l t a s k s P h a r a o h a n t s f e e d o n a w i d e v a r i e t y o f f o o d s ( B e l l e v o y e 1 8 8 9 P e a c o c k e t a l 1 9 5 0 S u d d 1 9 6 0 G r a n o v s k y a n d H o w e l l 1 9 8 3 E d w a r d s 1 9 8 6 E d w a r d s a n d A b r a h a m 1 9 9 0 ) T h e y a l s o f e e d o n h u m a n s w h e r e t h e y h a v e b e e n o b s e r v e d r e c r u i t i n g t o b o d y a n d w o u n d s e c r e t i o n s ( B e a t s o n 1 9 7 2 E d w a r d s a n d B a k e r 1 9 8 1 a G r a n o v s k y a n d H o w e l l 1 9 8 3 E d w a r d s a n d S h o r t 1 9 9 0 E i c h l e r 1 9 9 0 ) I n a d d i t i o n t h e y h a v e b e e n o b s e r v e d r e c r u i t i n g t o o r f e e d i n g o n t h e u n b r o k e n s k i n o f b a b i e s ’ c h e e k s l i p s n e c k f o l d s a n d b a c k s o f e a r s h a s b e e n r e p o r t e d ( S t e i n b r i n k 1 9 7 8 E i c h l e r 1 9 9 0 ) P h a r a o h a n t s a r e e s p e c i a l l y w e l l a d a p t e d t o t a k i n g a d v a n t a g e o f a n y a v a i l a b l e s p a c e f o r n e s t i n g s i t e s a n d a r e p r i m a r i l y f o u n d n e s t i n g i n s i d e w a l l s a n d c r e v i c e s ( W h e e l e r 1 9 1 0 ) H o w e v e r c o l o n i e s h a v e b e e n d i s c o v e r e d i n e v e n m o r e u n u s u a l l o c a t i o n s s u c h a s i n l i n e n s b e t w e e n s h e e t s o f p a p e r ( E b e l i n g 1 9 7 8 ) i n i n t r a v e n o u s t u b i n g p a c k a g e s i n s u r g i c a l a n d r e s u s c i t a t i o n e q u i p m e n t ( B e a t s o n 1 9 7 3 ) a n d i n b a b y i n c u b a t o r s ( S t e i n b r i n k 1 9 7 8 E i c h l e r 1 9 9 0 )

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5 O r i g i n a n d D i s t r i b u t i o n T h e P h a r a o h a n t w a s o r i g i n a l l y d e s i g n a t e d a s F o r m i c a p h a r a o n i s L i n n e a u s 1 7 5 8 a n d h a d s i x d i f f e r e n t n a m e s u n t i l i t w a s e v e n t u a l l y n a m e d M o n o m o r i u m p h a r a o n i s ( L ) ( B o l t o n 1 9 9 4 ) T h e P h a r a o h a n t ’ s o r i g i n i s u n c e r t a i n w i t h m a n y r e s e a r c h e r s c o n t e n d i n g t h a t t h e a n t o r i g i n a t e d i n t h e t r o p i c a l r e g i o n s o f A f r i c a ( E b e l i n g 1 9 7 8 ) H o w e v e r E m e r y ( 1 9 2 2 ) W i l s o n a n d T a y l o r ( 1 9 6 7 ) a n d B o l t o n ( 1 9 8 7 ) a g r e e d t h a t I n d i a i s t h e p r o b a b l e o r i g i n o f t h e P h a r a o h a n t ( A r n o l d 1 9 1 6 ) s t a t e d S o u t h A m e r i c a w a s t h e o r i g i n a l l o c a t i o n o f t h i s a n t T h e P h a r a o h a n t i s c o m m o n a n d w i d e l y p r e v a l e n t i n a l l t h e s e a r e a s a n d i s e a s i l y s p r e a d b y h u m a n c o m m e r c e H e n c e i t s t r u e o r i g i n m a y n e v e r b e d e t e r m i n e d T h e P h a r a o h a n t i s a t r a m p s p e c i e s t h a t i s s t r o n g l y a s s o c i a t e d w i t h h u m a n s ( E d w a r d s 1 9 9 1 ) a n d w a s p r o c l a i m e d a s t h e “ m o s t w i d e l y d i s t r i b u t e d o f a l l a n t p e s t s ” b y E d w a r d s a n d S h o r t ( 1 9 9 0 ) T h e y l i s t e d i t s d o c u m e n t e d p r e s e n c e t o t h e U n i t e d S t a t e s C a n a d a S o u t h A m e r i c a E g y p t N o r t h A f r i c a E u r o p e R u s s i a A u s t r a l i a a n d J a p a n T h e P h a r a o h a n t i s a l s o p r e s e n t i n t h e H a w a i i a n I s l a n d s ( R e i m e r e t a l 1 9 9 0 ) B i o l o g y P h a r a o h a n t w o r k e r s a r e m o n o m o r p h i c a n d a r e v e r y s m a l l w i t h a l e n g t h o f 1 5 2 0 m m T h e y a r e r e d d i s h t o o r a n g e b r o w n i n c o l o r h a v e a t w o s e g m e n t e d p e t i o l e w i t h e a c h s e g m e n t h a v i n g a n o d e a n d t h e i r g a s t e r s h a v e a w h i t e s t r i p e o n t h e a n t e r i o r p o r t i o n o f i t s d o r s a l s u r f a c e ( H a a c k 1 9 8 7 H a r r i s 1 9 9 1 ) E a c h a n t e n n a h a s 1 2 s e g m e n t s w i t h t h e f i n a l 3 s e g m e n t s f o r m i n g a v i s i b l e c l u b A d u l t m a l e s a r e w i n g e d b l a c k i n c o l o r a n d s l i g h t l y l a r g e r t h a n t h e w o r k e r s b u t s m a l l e r t h a n q u e e n s I n n a t u r a l c o l o n i e s m a l e s a p p e a r i n f r e q u e n t l y a l t h o u g h m a l e e g g s a r e l a i d c o n t i n u o u s l y ( P e a c o c k e t a l 1 9 5 4 ) A t 4 m m i n l e n g t h q u e e n s a r e t h e l a r g e s t c a s t e o f t h e P h a r a o h a n t V i r g i n q u e e n s c a n b e

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6 d i s t i n g u i s h e d f r o m e g g l a y i n g q u e e n s b y t h e p r e s e n c e o f t h e i r w i n g s w h i c h a r e b i t t e n o f f a f t e r m a t i n g R e p r o d u c t i v e m a l e s a n d f e m a l e s e m e r g e i n t h e n e s t w i t h w i n g s b u t t h e y d o n o t h a v e n u p t i a l f l i g h t s I n s t e a d t h e y m a t e w i t h i n t h e n e s t ( P e a c o c k a n d B a x t e r 1 9 5 0 ) W i n g m u s c l e s i n t h e t h o r a x o f m a t e d q u e e n s h y s t o l i z e s o t h a t t h e t h o r a x i s m o d i f i e d t o c o n t a i n a t h o r a c i c c r o p t h a t i s s u p p l e m e n t a l t o t h e g a s t e r ’ s c r o p ( C a e t a n o 1 9 9 0 ) W i t h i n t h e n e s t b r o o d i s n o r m a l l y s e p a r a t e d i n t o d i s t i n c t p i l e s t h a t a r e s o r t e d b y a g e ( P e a c o c k a n d B a x t e r 1 9 4 9 ) P h a r a o h a n t e g g s a r e r o u n d a n d s t i c k t o g e t h e r ( H a r r i s 1 9 9 1 ) L a r v a e a r e i m m o b i l e a n d m u s t b e t r a n s p o r t e d a n d c a r e d f o r b y a d u l t w o r k e r s S e x u a l b r o o d d e v e l o p s f a s t e r t h a n w o r k e r b r o o d b u t d i f f e r e n t i a t i o n b e t w e e n t h e t w o b r o o d c a s t e s i s d i f f i c u l t u n t i l t h e g u t c o l o r o f w o r k e r l a r v a e c h a n g e s t o b l a c k o r t h e s e x u a l l a r v a e b e c o m e s n o t i c e a b l y l a r g e r t h a n t h e w o r k e r l a r v a e ( P e a c o c k a n d B a x t e r 1 9 5 0 ) L i f e s p a n s o f P h a r a o h a n t s d i f f e r a m o n g t h e c a s t e s W o r k e r s l i v e ~ 9 1 0 w e e k s m a l e s n o r m a l l y l i v e ~ 3 w e e k s a n d q u e e n s l i v e ~ 3 9 w e e k s ( P e a c o c k a n d B a x t e r 1 9 5 0 ) a l t h o u g h t h e y c a n l i v e a s l o n g a s 5 2 w e e k s ( E d w a r d s 1 9 8 7 ) P h a r a o h a n t c o l o n i e s c a n v a r y t r e m e n d o u s l y i n s i z e ( P e a c o c k e t a l 1 9 5 5 a ) a n d c a n c o n t a i n t h o u s a n d s o f i n d i v i d u a l s i n c l u d i n g m a n y f e r t i l e q u e e n s Q u e e n f e c u n d i t y i s d i r e c t l y r e l a t e d t o t h e n u m b e r o f l a s t i n s t a r l a r v a e p r e s e n t i n t h e c o l o n y ( B r g e s e n a n d J e n s e n 1 9 9 5 ) N a t u r a l P h a r a o h a n t c o l o n i e s t e n d t o h a v e a 1 : 1 : 1 r a t i o o f w o r k e r s : e g g s : l a r v a e ( P e a c o c k e t a l 1 9 5 5 b ) P h a r a o h a n t c o l o n i e s a r e p o l y d o m o u s a n d e x c h a n g e q u e e n s w o r k e r s a n d b r o o d a m o n g n e s t s ( E d w a r d s 1 9 9 1 ) M i g r a t i n g g r o u p s o f a n t s o f t e n f o r m a t e m p o r a r y n e s t w h i l e s e a r c h i n g f o r a m o r e p e r m a n e n t n e s t s i t e ( E d w a r d s a n d B a k e r 1 9 8 1 a ) N e w n e s t s a r e

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7 f o r m e d v i a b u d d i n g ( H e d g e s 1 9 9 7 ) s i n c e P h a r a o h a n t s d o n o t h a v e n u p t i a l f l i g h t s ( E b e l i n g 1 9 7 8 ) P e a c o c k a n d B a x t e r ( 1 9 4 9 ) o r i g i n a l l y s t a t e d t h a t P h a r a o h a n t s p r e f e r r e d t e m p e r a t u r e s ~ 2 7 C a n d 8 0 % R H H a r r i s ( 1 9 9 1 ) d e m o n s t r a t e d t h a t P h a r a o h a n t s p r e f e r r e d a l o w e r r e l a t i v e h u m i d i t y o f ~ 6 5 % I n t e m p e r a t e r e g i o n s P h a r a o h a n t s n e s t p r i m a r i l y i n d o o r s ( E b e l i n g 1 9 7 8 ) F o r a g i n g a n d F o o d P h a r a o h a n t s e n g a g e i n e x t e n s i v e f o r a g i n g a n d a r e m o s t a c t i v e a t n i g h t { H e d g e s 1 9 9 7 # 7 2 1 } T h e y l i k e l y u s e “ g u i d e l i n e o r i e n t a t i o n ” ( K l o t z a n d R e i d 1 9 9 2 ) t o f o l l o w e d g e s a n d c r a c k s o f s t r u c t u r e s t o f i n d f o o d s o u r c e s T h e y w i l l a l s o f o r a g e v i a d u c t s a n d w a l l s p a c e s a l u m i n u m w i n d o w a n d d o o r f r a m e s a n d t h e y c a n f o r a g e u p t o d i s t a n c e s o f 4 5 m ( V a i l a n d W i l l i a m s 1 9 9 4 ) P h a r a o h a n t s a r e w e l l k n o w n f o r t h e i r c o n s u m p t i o n o f a w i d e v a r i e t y o f f o o d s ( S m i t h 1 9 6 5 E d w a r d s a n d B a k e r 1 9 8 1 a ) S o m e r e s e a r c h e r s c o n t e n d t h a t t h e P h a r a o h a n t i s p r i m a r i l y a t t r a c t e d t o s u g a r s ( G r a n o v s k y a n d H o w e l l 1 9 8 3 ) w h i l e o t h e r s t h i n k t h a t t h e y p r i m a r i l y c o n s u m e p r o t e i n s ( E d w a r d s a n d A b r a h a m 1 9 9 0 ) O t h e r r e s e a r c h i n d i c a t e d t h a t P h a r a o h a n t c o l o n i e s c o m p e n s a t e d f o r w h a t w a s l a c k i n g n u t r i t i o n a l l y w h e r e b y l a b o r a t o r y c o l o n i e s t h a t w e r e r e a r e d o n c a r b o h y d r a t e s e x h i b i t e d m o r e i n t e r e s t i n p r o t e i n b a i t s a n d v i c e v e r s a ( E d w a r d s a n d A b r a h a m 1 9 9 0 ) T h u s t h e f o o d p r e f e r e n c e o f P h a r a o h a n t s i s v a r i a b l e a n d d y n a m i c c h a n g i n g w i t h n u t r i t i o n a l n e e d s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) a n d t i m e ( E d w a r d s 1 9 8 6 ) M e d i c a l I m p o r t a n c e T h e P h a r a o h a n t i s a t r o p i c a l a n t a n d n e e d s h i g h h u m i d i t y a n d h i g h t e m p e r a t u r e s t o s u r v i v e ( P e a c o c k a n d B a x t e r 1 9 5 0 P e a c o c k e t a l 1 9 5 5 c ) I n t e m p e r a t e z o n e s i t i s

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8 d e p e n d e n t u p o n h e a t e d b u i l d i n g s a n d i s a c o m m o n p e s t s p e c i e s f o u n d i n h o s p i t a l s a n d o t h e r h e a l t h c a r e f a c i l i t i e s w o r l d w i d e A l t h o u g h h o s p i t a l i n f e s t a t i o n s o f P h a r a o h a n t s w e r e a l r e a d y r e c o g n i z e d a s a n i n c o n v e n i e n c e t h e d i s c o v e r y o f s e v e r a l p a t h o g e n i c o r g a n i s m s i n c l u d i n g S t r e p t o c o c c u s s p p S t a p h y l o c o c c u s s p p E s c h e r i c h i a c o l i B a c i l l u s s p p P s u e d o m o n a s s p p K l e b s i e l l a s p p a n d C l o s t r i d i u m s p p o n h o s p i t a l p o p u l a t i o n s o f P h a r a o h a n t s ( B e a t s o n 1 9 7 2 ) c r e a t e d i n t e r e s t i n t h e p o t e n t i a l f o r P h a r a o h a n t s t o t r a n s m i t d i s e a s e s t o h u m a n s I n B r a z i l t h e p o t e n t i a l f o r P h a r a o h a n t s t o v e c t o r h o s p i t a l a c q u i r e d ( n o s o c o m i a l ) i n f e c t i o n s m a y h a v e b e e n e s p e c i a l l y h i g h a s t h i s c o u n t r y h a d s o m e o f t h e h i g h e s t r a t e s o f i n t r a h o s p i t a l i n f e c t i o n s I n t h e s e h o s p i t a l s i t w a s d e t e r m i n e d t h a t o n l y t r a m p a n t s p e c i e s i n c l u d i n g P h a r a o h a n t s w e r e n e a r p a t i e n t s ( B u e n o a n d F o w l e r 1 9 9 4 ) B e s i d e s b e i n g p o t e n t i a l v e c t o r s P h a r a o h a n t s c a n c a u s e s k i n i r r i t a t i o n s a n d l e s i o n s ( E i c h l e r 1 9 9 0 ) T h e i r s m a l l s i z e e n a b l e s t h e m t o c r a w l t h r o u g h g a u z e c o v e r i n g b a n d a g e d w o u n d s a n d a n t s a s s m a l l a s 2 0 m m i n l e n g t h c a n c a u s e i n t e n s e p a i n w h e n f o r a g e r s w a l k o n o p e n w o u n d s ( E i c h l e r 1 9 9 0 ) P h a r a o h a n t s a l s o h a v e b e e n f o u n d f e e d i n g d i r e c t l y o n o p e n w o u n d s o f b u r n u n i t p a t i e n t s ( A n o n 1 9 8 6 ) o n o r u n d e r b a n d a g e s c o v e r i n g w o u n d s ( C a r t w r i g h t a n d C l i f f o r d 1 9 7 3 ) o n p r e m a t u r e n e w b o r n s i n i n c u b a t o r s ( E d w a r d s a n d S h o r t 1 9 9 0 ) o n b e d r i d d e n e l d e r l y o r p o s t o p e r a t i v e p a t i e n t s ( e s p e c i a l l y p a t i e n t s w h o w e r e l e a k i n g b o d y f l u i d s ) ( A n o n 1 9 7 4 ) a n d i n i n t r a v e n o u s t u b e s t h a t s u p p l y f l u i d s t o p a t i e n t s ( B e a t s o n 1 9 7 3 E d w a r d s a n d B a k e r 1 9 8 1 b ) C o n t r o l O n c e e n s c o n c e d i n a s t r u c t u r e P h a r a o h a n t p o p u l a t i o n s a r e e x t r e m e l y d i f f i c u l t t o e r a d i c a t e p a r t i a l l y d u e t o c r y p t i c h a b i t s i n c l u d i n g a t e n d e n c y t o l i v e i n s i d e w a l l s a n d u n d e r f o u n d a t i o n s I t i s o f t e n d i f f i c u l t t o l o c a t e P h a r a o h a n t n e s t s w i t h o u t d a m a g i n g t h e

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9 b u i l d i n g ’ s s t r u c t u r e ( V a i l 1 9 9 6 ) M a n y d i f f e r e n t m e t h o d s o f P h a r a o h a n t e l i m i n a t i o n h a v e b e e n a t t e m p t e d w i t h v a r y i n g d e g r e e s o f s u c c e s s T h e e f f i c a c y o f p e s t i c i d e s p r a y s a n d d u s t s s t o m a c h p o i s o n s b a i t s c o n t a i n i n g t o x i c a n t s a n d b a i t s c o n t a i n i n g i n s e c t g r o w t h r e g u l a t o r s ( I G R s ) h a v e a l l b e e n i n v e s t i g a t e d I n t h e 1 9 9 0 s s p r a y s a n d d u s t s w e r e u s u a l l y i n e f f e c t i v e a g a i n s t P h a r a o h a n t s b e c a u s e t h e y a f f e c t e d o n l y a s m a l l p e r c e n t a g e o f w o r k e r s ( W i l l i a m s 1 9 9 0 ) d u e t o t h e f a c t t h a t l e s s t h a n 2 5 % o f w o r k e r s f o r a g e a n d m a n y o f t h e c h e m i c a l s w e r e i n t h e p y r e t h r o i d c l a s s a n d h i g h l y r e p e l l e n t T h e i r u s e w a s e v e n h y p o t h e s i z e d t o r e s u l t i n b u d d i n g ( W i l l i a m s a n d V a i l 1 9 9 4 ) s i n c e P h a r a o h a n t c o l o n i e s r e s p o n d r a p i d l y t o d i s t u r b a n c e ( P e a c o c k e t a l 1 9 5 5 b P a s s e r a 1 9 9 4 ) E i c h l e r ( 1 9 9 0 ) b e l i e v e d t h a t i n s e c t i c i d e s p r a y i n g o f P h a r a o h a n t i n f e s t a t i o n s t o b e a f o r m o f m a l p r a c t i c e I n a d d i t i o n t h e e f f e c t i v e n e s s o f s p r a y s w a s b e l i e v e d t o h a v e b e e n r e d u c e d i f m u l t i p l e n e s t s a r e p r e s e n t i n t h e g e n e r a l a r e a ( W i l l i a m s 1 9 9 0 ) S t o m a c h p o i s o n s w o r k t o o q u i c k l y a n d r e d u c e w o r k e r n u m b e r s s o m u c h t h a t p o i s o n s a r e n o t d i s t r i b u t e d t o q u e e n s a n d b r o o d ( E d w a r d s 1 9 7 5 ) C o l o n i e s c a n b e p e r p e t u a t e d b y a s f e w a s 1 0 0 w o r k e r s a n d 5 0 b r o o d ( P e a c o c k e t a l 1 9 5 5 a ) o r e v e n w i t h a s f e w a s 5 w o r k e r s 3 0 e g g s 1 9 l a r v a e a n d 3 p u p a e ( V a i l 1 9 9 6 ) T h e r e f o r e t o e l i m i n a t e P h a r a o h a n t p o p u l a t i o n s i t i s e s s e n t i a l f o r t o x i c a n t s t o r e a c h t h e q u e e n s a n d b r o o d . B a i t s s h o u l d b e c o m p r i s e d o f a s l o w a c t i n g n o n r e p e l l e n t t o x i c a n t i n c o r p o r a t e d i n t o a n a t t r a c t i v e b a i t m a t r i x s o t h a t t h e b a i t i s c o n s u m e d a n d s p r e a d t h r o u g h t h e c o l o n y v i a t r o p h a l l a x i s E i c h l e r ( 1 9 9 0 ) r e c o m m e n d e d m e a t b a s e d b a i t m a t r i c e s a n d s o d i u m a r s e n a t e o r c h l o r d e c o n e F i n a l l y h e r e c o m m e n d e d p r e b a i t i n g t o d e t e r m i n e w h a t f o o d a t t r a c t s t h e P h a r a o h a n t s a t t h e t i m e s i n c e t h e y a r e k n o w n t o c h a n g e f o o d p r e f e r e n c e s h e

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1 0 s u g g e s t e d p l a c i n g b a i t s n e a r l o c a t i o n s t h a t t h e y w o u l d l i k e l y f o r a g e s u c h a s s i n k s H e a l s o s u g g e s t e d d e n y i n g a n t s a c c e s s t o a l t e r n a t i v e f o o d s o u r c e s s o t h a t t h e b a i t w o u l d b e t h e i r o n l y f o o d s o u r c e K l o t z e t a l ( 1 9 9 6 ) n o t e d t h a t b o r i c a c i d s w e r e u s e d a g a i n s t a n t s a s e a r l y a s t h e l a t e 1 8 0 0 s ( R i l e y 1 8 8 9 ) a n d a r e s t i l l i m p o r t a n t i n a n t c o n t r o l T h e y c i t e d a r e c o m m e n d a t i o n b y W r i g h t a n d S t o u t ( 1 9 7 8 ) f o r 2 % b o r i c a c i d i n e i t h e r a l i q u i d o r s o l i d b a i t K l o t z e t a l ( 1 9 9 6 ) t e s t e d t h e e f f i c a c y o f 1 % b o r i c a c i d i n s u c r o s e s o l u t i o n a n d 0 9 % h y d r a m e t h y l n o n w i t h b o t h l i m i t e d a n d c o n t i n u o u s a v a i l a b i l i t y i n t h e l a b o r a t o r y T h e y d e t e r m i n e d t h a t c o l o n y g r o w t h w a s r e d u c e d b u t n o t e l i m i n a t e d a f t e r 3 d e x p o s u r e t o b o r i c a c i d b a i t I n c o n t r a s t c o n t i n u o u s f e e d i n g o f b o r i c a c i d b a i t r e s u l t e d i n e l i m i n a t i o n o f a l l c o l o n i e s C o l o n i e s e x p o s e d t o h y d r a m e t h y l n o n h a d 9 0 % r e d u c t i o n o f w o r k e r s b y 3 w k a n d 9 0 % r e d u c t i o n o f b r o o d b y 4 w k ; q u e e n s w e r e e l i m i n a t e d i n 2 w k T h e y c o n c l u d e d t h a t b o r i c a c i d s u c r o s e s o l u t i o n s m i g h t p r o v i d e a n a l t e r n a t i v e t o o t h e r c o m m e r c i a l l y a v a i l a b l e P h a r a o h a n t b a i t s M a n y t o x i c a n t s s u c h a s M i r e x a r e n o l o n g e r u s e d d u e t o c a r c i n o g e n i c i t y ( E i c h l e r 1 9 9 0 ) I n s e c t g r o w t h r e g u l a t o r s ( I G R s ) w h i c h m i m i c j u v e n i l e h o r m o n e s h a v e b e e n d e v e l o p e d ( N a t i o n 2 0 0 3 ) I G R s o f t e n c o n t a i n e i t h e r t h e a c t i v e i n g r e d i e n t m e t h o p r e n e o r p y r i p r o x y f e n I n s e c t g r o w t h r e g u l a t o r s a r e s l o w e r a c t i n g t h a n s t o m a c h p o i s o n s b e c a u s e t h e y a f f e c t o n l y q u e e n a n d b r o o d : t h e y i n d u c e s t e r i l i t y i n q u e e n a n d d i s r u p t b r o o d d e v e l o p m e n t ( E d w a r d s 1 9 7 5 ) T h e w o r k e r s r e m a i n a c t i v e f o r s e v e r a l w e e k s b u t a r e n o t a b l e t o s t a r t n e w c o l o n i e s d u e t o d e a t h o f q u e e n s a n d b r o o d O i e t a l ( 1 9 9 6 ) c o m p a r e d t h e e f f i c a c y o f s e v e r a l d i f f e r e n t a p p l i c a t i o n s t o d e t e r m i n e i f i n d o o r P h a r a o h a n t i n f e s t a t i o n s c o u l d b e c o n t r o l l e d b y e x t e r i o r t r e a t m e n t s

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1 1 s i n c e P h a r a o h a n t s h a v e b e e n o b s e r v e d f o r a g i n g o n e x t e r i o r w a l l s u r f a c e s a n d o u t s i d e b u i l d i n g s ( H a a c k 1 9 9 1 O i e t a l 1 9 9 4 V a i l a n d W i l l i a m s 1 9 9 4 ) T h e y t e s t e d s e v e r a l m e t h o d s r e g i s t e r e d f o r a n t c o n t r o l : a c o n t a c t i n s e c t i c i d e ( c y f l u t h r i n ) a g r a n u l a r c o n t a c t i n s e c t i c i d e ( d i a z i n o n ) a g r a n u l a r b a i t c o n t a i n i n g a c o n t a c t i n s e c t i c i d e ( p r o p o x u r ) a n d a g r a n u l a r b a i t c o n t a i n i n g a d e l a y e d a c t i o n t o x i c a n t ( h y d r a m e t h y l n o n ) T h e y d e t e r m i n e d t h a t e x t e r i o r a p p l i c a t i o n o f t h e d e l a y e d a c t i o n t o x i c a n t h y d r a m e t h y l n o n r e s u l t e d i n s u c c e s s f u l c o n t r o l i n d o o r P h a r a o h a n t i n f e s t a t i o n s w h i l e a l l o t h e r t e s t e d t r e a t m e n t s r e s u l t e d i n i n c r e a s e d f o r a g i n g T h e y c o n c l u d e d t h a t e x t e r i o r b a i t i n g c o u l d r e d u c e t h e n e e d f o r i n d o o r p e s t i c i d e a p p l i c a t i o n s I n 2 0 0 3 a n e w c o m p o u n d f i p r o n i l ( T e r m i d o r B A S F R e s e a r c h T r i a n g l e N C ) r e c e i v e d r e g i s t r a t i o n f o r a n t a p p l i c a t i o n s I t w a s a n o l d i d e a o f s p r a y i n g t h e p e r i m e t e r o f a b u i l d i n g f o r a n t c o n t r o l w i t h a n e w c o m p o u n d t h a t h a d i m p o r t a n t b a i t l i k e c h a r a c t e r i s t i c s F i p r o n i l i s a n o n r e p e l l e n t s l o w a c t i n g t o x i c a n t S u c c e s s f u l c o n t r o l o f P h a r a o h a n t s f o r a g i n g o u t s i d e a n d i n s i d e h o m e s t r e a t e d o n t h e p e r i m e t e r o n l y w i t h f i p r o n i l h a s b e e n d e m o n s t r a t e d ( F O i a n d J J o n o v i c h p e r s o n a l c o m m u n i c a t i o n ) a c c o r d i n g t o l a b e l d i r e c t i o n s ( 2 5 c m u p a n d 2 5 c m o u t ) V a i l e t a l ( 1 9 9 6 ) e v a l u a t e d t h e e f f i c a c y o f d i f f e r e n t b a i t f o r m u l a t i o n s o f t h e I G R p y r i p r o x y f e n a s a n e x t e r i o r p e r i m e t e r t r e a t m e n t a g a i n s t P h a r a o h a n t i n f e s t a t i o n s T h e y n o t e d t h a t w h i l e p y r i p r o x y f e n h a d a l r e a d y b e e n e v a l u a t e d a s a n e f f e c t i v e c o n t r o l a g e n t a g a i n s t m o s t u r b a n i n s e c t s t e s t e d p r e v i o u s l y a n d d e s p i t e t h e d e m o n s t r a t i o n o f i t s e f f e c t i v e n e s s a g a i n s t P h a r a o h a n t s i n t h e l a b o r a t o r y ( V a i l a n d W i l l i a m s 1 9 9 5 ) n o I G R s w e r e c o m m e r c i a l l y a v a i l a b l e f o r u s e a g a i n s t P h a r a o h a n t s T h e y d e t e r m i n e d t h a t t h e b a i t s f o r m u l a t e d w i t h v a r i o u s a m o u n t s o f p y r i p r o x y f e n i n t w o d i f f e r e n t b a i t m a t r i c e s w e r e a l l s u c c e s s f u l i n c o n t r o l l i n g t h e a n t p o p u l a t i o n s a l t h o u g h c o m p l e t e e l i m i n a t i o n d i d n o t o c c u r

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1 2 u n t i l 1 6 w k a f t e r t r e a t m e n t T h e y o b s e r v e d t h a t b r o o d r a p i d l y d e c l i n e d i n t r e a t e d c o l o n i e s D y e s u s e d i n b a i t f o r m u l a t i o n s w a s v i s i b l e i n l a r v a e q u e e n s a n d w o r k e r s i n d i c a t i n g t h o r o u g h d i s t r i b u t i o n o f b a i t s t h r o u g h o u t t h e c o l o n i e s F i n a l l y t h e y c o n c l u d e d t h a t a p p l i c a t i o n o f a n I G R b a i t 1 w k b e f o r e a p p l i c a t i o n o f a m e t a b o l i c i n h i b i t o r m i g h t p r o v i d e b o t h q u i c k m o r t a l i t y a n d l o n g t e r m c o n t r o l a n d t h e y s t a t e d t h a t v e r t i c a l p l a c e m e n t o f b a i t s a i m e d a t P h a r a o h a n t s m i g h t p r e v e n t o t h e r a n t s p e c i e s f r o m c o n s u m i n g b a i t s O i e t a l ( 2 0 0 0 ) i n v e s t i g a t e d t h e d i s t r i b u t i o n o f b a i t s c o n t a i n i n g t h e I G R p y r i p r o x y f e n a n d t h e m e t a b o l i c i n h i b i t o r h y d r a m e t h y l n o n a m o n g m u l t i p l e P h a r a o h a n t c o l o n i e s T h e y d e t e r m i n e d t h a t t h e I G R d i d n o t k i l l a d u l t w o r k e r a n t s q u i c k l y s o t h a t m o r e w o r k e r a n t s l i v e d l o n g e n o u g h t o d i s t r i b u t e t h e b a i t t o a l l n e s t s i t e s I n c o n t r a s t h y d r a m e t h y l n o n c a u s e d r a p i d m o r t a l i t y i n a d u l t a n t s a n d a n t s f r o m n e a r b y n e s t s m i g r a t e d i n t o s i t e s w h o s e p o p u l a t i o n s d e c l i n e d d u e t o m e t a b o l i c i n h i b i t o r m o r t a l i t y T h e y f u r t h e r d e t e r m i n e d t h a t t h e I G R p y r i p r o x y f e n r e s u l t e d i n a s l o w g r a d u a l d e c l i n e o f b o t h w o r k e r s a n d q u e e n s i n a l l n e s t s i t e s a n d l a s t e d a l o n g t i m e i n c o n t r a s t t o t h e m e t a b o l i c i n h i b i t o r h y d r a m e t h y l n o n w h i c h w o r k e d m o r e q u i c k l y b u t d i d n o t l a s t a s l o n g F i n a l l y t h e y p o s t u l a t e d t h a t l o n g e r s u r v i v o r s h i p o f w o r k e r s i n c r e a s e d t h e a m o u n t o f t r o p h a l l a x i s b e t w e e n a n t s o f d i f f e r e n t n e s t s i t e s a n d i n c r e a s e d t h e d i s t r i b u t i o n o f t h e I G R p y r i p r o x y f e n a m o n g m u l t i p l e n e s t s t h e r e b y r e d u c i n g t h e p o s s i b i l i t y o f r e i n f e s t a t i o n T h e y c o n c l u d e d t h a t t h e m o d e o f a c t i o n a n d s p e e d o f a c t i o n o f b a i t t o x i c a n t s s h o u l d b e e v a l u a t e d w h e n s e l e c t i n g a n t b a i t s f o r p o l y d o m o u s c o l o n i e s s u c h a s t h e P h a r a o h a n t

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1 3 C H A P T E R 3 N O C H O I C E L I Q U I D T E S T I n t r o d u c t i o n P h a r a o h a n t s ’ c o n s u m p t i o n o f a w i d e v a r i e t y o f s u b s t a n c e s ( B e l l e v o y e 1 8 8 9 G r a n o v s k y a n d H o w e l l 1 9 8 3 E d w a r d s a n d A b r a h a m 1 9 9 0 ) a n d t h e i r a b u n d a n t p r e s e n c e i n h o s p i t a l s i s w e l l d o c u m e n t e d ( G r e e n e t a l 1 9 5 4 B e a t s o n 1 9 7 2 E d w a r d s a n d B a k e r 1 9 8 1 b ) A l t h o u g h P h a r a o h a n t s a r e p r i m a r i l y s a m p l e d i n k i t c h e n s a n d b a t h r o o m s b e c a u s e i t i s a s s u m e d t h a t t h e r e w i l l b e a n a b u n d a n c e o f w a t e r a n d f o o d a v a i l a b l e t o s u s t a i n a n t c o l o n i e s V a i l a n d W i l l i a m s ( 1 9 9 4 ) f o u n d t h a t t h e y a r e c a p a b l e o f f o r a g i n g u p t o 4 5 m P h a r a o h a n t s h a v e b e e n f o u n d i n h o s p i t a l s a n d h e a l t h c a r e f a c i l i t i e s ( E i c h l e r 1 9 9 0 F o w l e r 1 9 9 0 ) T h e y h a v e b e e n d i s c o v e r e d i n s i d e a p e r m a n e n t l y i n s t a l l e d t r a c h e o t o m y t u b e a n d i n t r a v e n o u s t u b e s ( W e i d n e r 1 9 8 2 ) P h a r a o h a n t s h a v e b e e n o b s e r v e d f e e d i n g u p o n p a t i e n t s w i t h i n d i f f e r e n t h o s p i t a l a r e a s I n n e o n a t a l u n i t s w h e r e n e w b o r n s a r e p l a c e d i n s i d e s p e c i a l i n c u b a t i n g u n i t s t h e y h a v e b e e n o b s e r v e d f e e d i n g o n i n t a c t s k i n o f n e w b o r n i n f a n t s s u c h a s c h e e k s l i p s n e c k f o l d s a n d b a c k s o f e a r s ( S t e i n b r i n k 1 9 7 8 ) a s c i t e d i n ( E i c h l e r 1 9 9 0 ) T h e y h a v e b e e n o b s e r v e d f e e d i n g o n b u r n p a t i e n t o p e n w o u n d s ( W e i d n e r 1 9 8 2 ) T h e i r s m a l l s i z e e n a b l e s t h e m t o c r a w l t h r o u g h g a u z e ; t h u s t h e y h a v e a l s o b e e n f o u n d u n d e r b a n d a g e s o f c o v e r e d w o u n d s ( B e a t s o n 1 9 7 2 ) F i n a l l y P h a r a o h a n t i n f e s t a t i o n s h a v e b e e n l o c a t e d i n b e d s o f b e d r i d d e n e l d e r l y o r p o s t o p e r a t i v e p a t i e n t s ( e s p e c i a l l y p a t i e n t s w h o w e r e l e a k i n g b o d y f l u i d s ) ( A n o n 1 9 7 4 ) P h a r a o h a n t s c a n a l s o b e a s s o c i a t e d w i t h o t h e r f l u i d s p r e s e n t i n a h o s p i t a l e n v i r o n m e n t S u p p u r a t i n g w o u n d s e x u d e p l a s m a w h i c h i s t h e n o n c e l l u l a r p o r t i o n o f

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1 4 w h o l e b l o o d A d d i t i o n a l f l u i d s c a n a l s o b e p r e s e n t s u c h a s s a l i n e o r d e x t r o s e s o l u t i o n s t h a t a r e a d m i n i s t e r e d e i t h e r i n t r a v e n o u s l y o r t o p i c a l l y ( A A B B 2 0 0 2 W e n z e l e t a l 2 0 0 2 ) B o t h t y p e s o f a d m i n i s t r a t i o n o f s a l i n e a n d d e x t r o s e d e c r e a s e m o i s t u r e l o s s i n t h e a b s e n c e o f e p i t h e l i a l t i s s u e a n d i n s i t u a t i o n s o f l i m i t e d f l u i d c o n s u m p t i o n ( A A B B 2 0 0 2 ) ; t o p i c a l u s e a l s o w i l l s t e r i l i z e w o u n d s a n d d e c r e a s e t h e p o t e n t i a l f o r i n f e c t i o n a t i n j u r y s i t e P h a r a o h a n t s m i g h t b e a t t r a c t e d t o t h e s e f l u i d s o r t o s u b s t a n c e s p r e s e n t i n p a t i e n t b l o o d o r p l a s m a I n a d d i t i o n t o s a l i n e a n d d e x t r o s e i n t r a v e n o u s a n d t o p i c a l a d m i n i s t r a t i o n s m a n y p a t i e n t s a l s o r e c e i v e l i q u i d d i e t a r y s u p p l e m e n t s a f t e r s u r g e r y v i a t u b i n g t h a t i s i n s e r t e d i n t o p a t i e n t ’ s s t o m a c h ( W e n z e l e t a l 2 0 0 2 ) T h e s e f l u i d s p r o v i d e a n o t h e r p o t e n t i a l l y a t t r a c t i v e f o o d s o u r c e t o P h a r a o h a n t s D e x t r o s e s a l i n e a n d l i q u i d d i e t a r y s u p p l e m e n t s a r e a d m i n i s t e r e d d i r e c t l y i n t o p a t i e n t v i a t u b i n g A n e c d o t a l r e p o r t s f r o m n u r s e s ( S C o b b p e r s o n a l c o m m u n i c a t i o n ) i n d i c a t e t h a t m e d i c a l p r a c t i c e s s u c h a s t u b e c l e a r i n g ( i e p r o c e d u r e o f s q u i r t i n g l i q u i d o u t o f t u b e p r i o r t o i n s e r t i o n o f t u b e i n t o p a t i e n t ) o f t e n r e s u l t i n p l a c e m e n t o f l i q u i d s o n o r n e a r t h e p a t i e n t b e d T h u s e a c h o f t h e s e l i q u i d s i s a v a i l a b l e n o t o n l y a t i n s e r t i o n p o i n t i n p a t i e n t ’ s b o d y b u t m i g h t a l s o b e a v a i l a b l e i n a b u n d a n c e t h r o u g h o u t a h o s p i t a l r o o m p a r t i c u l a r l y c l o s e t o t h e p a t i e n t ’ s b e d P h a r a o h a n t s o c c u r i n h o s p i t a l r o o m s a n d h a v e b e e n d o c u m e n t e d t o f e e d u p o n p a t i e n t s b u t n o p r e v i o u s s t u d y h a s b e e n u n d e r t a k e n t o d e t e r m i n e c o n s u m p t i o n o f p a t i e n t c a r e f l u i d s a n d h u m a n f l u i d s E v e n t h o u g h a n t s w e r e d o c u m e n t e d a s f e e d i n g o n w o u n d s n o e x a m i n a t i o n h a s i n v e s t i g a t e d c o n s u m p t i o n o f h u m a n b o d y f l u i d s T h u s m y o b j e c t i v e w a s t o d e t e r m i n e i f P h a r a o h a n t s c o n s u m e f l u i d s u s e d s p e c i f i c a l l y f o r p a t i e n t c a r e i n h o s p i t a l s

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1 5 M a t e r i a l s a n d M e t h o d s A n t s P h a r a o h a n t t e s t c o l o n i e s w e r e e x t r a c t e d f r o m l a r g e c o l o n i e s h o u s e d a n d r e a r e d a t t h e U r b a n E n t o m o l o g y L a b o r a t o r y U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F L R e a r i n g t e m p e r a t u r e s r a n g e d f r o m 2 1 9 C – 2 9 4 C w i t h r e l a t i v e h u m i d i t y b e t w e e n 2 0 5 3 % ( A p p e n d i x A ) C o l o n i e s c o n s i s t e d o f 2 5 0 w o r k e r s 3 d e a l a t e q u e e n s a n d a p p r o x i m a t e l y 5 0 m g b r o o d H a l f o f t h e w o r k e r s w e r e s e l e c t e d f r o m t h e i n s i d e o f n e s t i n g c e l l s t o e n s u r e t h e p r e s e n c e o f n u r s e s a n d 1 2 5 w o r k e r s w e r e s e l e c t e d f r o m o u t s i d e o f t h e n e s t i n g c e l l s a t f o r a g i n g l o c a t i o n s s u c h a s e x t e r i o r s u r f a c e o f w a t e r v i a l s t o e n s u r e t h e p r e s e n c e o f f o r a g e r s ( H a a c k 1 9 8 7 H a a c k a n d V i n s o n 1 9 9 0 H a a c k e t a l 1 9 9 5 ) B r o o d i n c l u d e d e g g s a l l l a r v a l s t a g e s a n d p u p a e S e e A p p e n d i x B f o r f u l l d e t a i l s o n s e l e c t i o n o f a n t s a n d p r e p a r a t i o n o f e x p e r i m e n t a l t e s t c o l o n i e s F o r a g i n g A r e n a F o r a g i n g a r e n a s c o n t a i n e d o n e n e s t i n g c e l l o n e w a t e r v i a l a n d o n e m i c r o c a p i l l a r y t u b e ( F i g 3 1 ) F o r a g i n g a r e n a s c o n s i s t e d o f c l e a r p l a s t i c b o x e s ( 2 7 b y 1 9 b y 1 0 c m d e e p ; P i o n e e r P l a s t i c s D i x o n K Y ) w h i c h w e r e c o a t e d w i t h F l u o n ( A s a h i G l a s s F l u o r o p o l y m e r s D o w n i n g t o n P A ) o n i n t e r i o r s i d e s t o p r e v e n t a n t e s c a p e N e s t i n g c e l l s c o n s i s t e d o f s t e r i l e c o v e r e d p l a s t i c P e t r i d i s h e s ( 5 0 m m d i a m e t e r b y 1 5 m m d e e p ; F a l c o n M o d e l N o 3 5 1 0 0 7 B e c t o n D i c k i n s o n a n d C o F r a n k l i n L a k e s N J ) f i l l e d h a l f w a y w i t h C a s t o n e d e n t a l p l a s t e r ( M o d e l N o 9 9 0 4 5 D e n t s p l y I n t e r n a t i o n a l Y o r k P A ) A c i r c u l a r i n d e n t a t i o n w a s m a d e i n p l a s t e r b y i n s e r t i n g t h e c a p o f a 1 5 m l v i a l ( M o d e l N o 5 5 9 T h o r n t o n P l a s t i c s S a l t L a k e C i t y U T ) i n t o f r e s h l y m i x e d p l a s t e r i n t h e

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1 6 2 7 c m 1 9 c m 3 0 l c a p i l l a r y t u b e w i t h f l u i d F i g u r e 3 1 N o c h o i c e l i q u i d a r e n a w i t h w a t e r v i a l a n d n e s t i n g c e l l p l a c e d i n d i a g o n a l l y o p p o s i t e c o r n e r s o f t e s t a r e n a A m i c r o c a p i l l a r y t u b e c o n t a i n i n g o n e f l u i d w a s p l a c e d i n t h e c e n t e r o f a r e n a B o t h t e s t a r e n a s ( w i t h a n t s ) a n d c o n t r o l a r e n a s ( n o a n t s ) w e r e p l a c e d a d j a c e n t t o e a c h o t h e r t o c o m p r i s e t e s t p a i r T e s t a r e n a s c o n t a i n e d 1 2 5 i n s i d e a n t s 1 2 5 o u t s i d e a n t s 3 q u e e n s a n d ~ 5 0 m g b r o o d ( i n c l u d i n g p u p a e e g g s a n d a l l s t a g e s o f l a r v a e ) C o n t r o l a r e n a s l a c k e d a n t s W a t e r V i a l ( 1 5 m l ) N e s t i n g c e l l ( 5 0 m m d i a m e t e r ; 1 5 m m d e e p )

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1 7 b a s e o f a P e t r i d i s h T h e v i a l c a p w a s r e m o v e d a f t e r p l a s t e r w a s t h o r o u g h l y d r i e d ( 2 4 h ) a n d h a r d e n e d l e a v i n g a n i n d e n t a t i o n ( 3 3 m m d i a m e t e r b y 2 m m d e e p ) a n d r o u g h s l o p i n g e d g e ( P e a c o c k a n d B a x t e r 1 9 4 9 ) f o r a n t s t o r e a r b r o o d O n e e n t r a n c e h o l e ( ~ 5 m m d i a m e t e r ) w a s m e l t e d i n t o t h e u p p e r e d g e o f t h e P e t r i d i s h b a s e v i a i n s e r t i o n o f t h e m e t a l t i p o f a h o t g l u e g u n ( s a n s g l u e ) j u s t a b o v e l e v e l o f h a r d e n e d p l a s t e r I m m e d i a t e l y p r i o r t o p l a c i n g a n t s i n a r e n a s P e t r i d i s h b a s e s w e r e i m m e r s e d i n a c o n t a i n e r o f t a p w a t e r f o r 3 0 m i n t o s a t u r a t e t h e p l a s t e r S a t u r a t e d b a s e s w e r e r e m o v e d f r o m w a t e r a n d e x c e s s w a t e r w a s b l o t t e d f r o m a l l s u r f a c e s A P e t r i d i s h l i d w a s p l a c e d o n t o p o f t h e b a s e a n d t h e e n t i r e n e s t i n g c e l l w a s p l a c e d i n t o t h e f o r a g i n g a r e n a w i t h t h e e n t r a n c e h o l e f a c i n g t h e o p p o s i t e d i a g o n a l c o r n e r o f t h e a r e n a W a t e r w a s p r o v i d e d a d l i b i t u m i n a 1 5 m l v i a l ( M o d e l N o 5 5 9 T h o r n t o n P l a s t i c s S a l t L a k e C i t y U T ) w i t h t h r e e l a r g e c o t t o n b a l l s i n s e r t e d i n t o t h e t o p t o p r e v e n t l e a k a g e a n d t o p r o v i d e e a s y a c c e s s f o r a n t s A w a t e r v i a l w a s p l a c e d i n t h e o p p o s i t e d i a g o n a l c o r n e r o f t h e a r e n a a c r o s s f r o m t h e n e s t i n g c e l l ( F i g 3 1 ) O n e t e s t a r e n a c o n t a i n i n g a n t s a n d o n e c o n t r o l a r e n a w i t h o u t a n t s w e r e p l a c e d a d j a c e n t t o e a c h o t h e r a n d c o m p r i s e d a p a i r e d e x p e r i m e n t a l u n i t f o r e a c h r e p l i c a t i o n C o n t r o l a r e n a d a t a w e r e u s e d t o q u a n t i f y a n d a d j u s t f o r e v a p o r a t i o n o f f l u i d s A l l a r e n a s w e r e c o v e r e d w i t h c l e a r p l a s t i c l i d s t o m a i n t a i n h u m i d i t y a n d t e m p e r a t u r e A i r f l o w w a s m a i n t a i n e d i n t h e r o o m b y t h e b u i l d i n g ’ s v e n t i l a t i o n s y s t e m d u r i n g t e s t p e r i o d E x p e r i m e n t a l F l u i d s F i v e f l u i d s c o m m o n l y u s e d i n h o s p i t a l e n v i r o n m e n t s f o r p a t i e n t c a r e w e r e u s e d i n t h i s n o c h o i c e t e s t T e s t f l u i d s c a n b e p r o v i d e d t o p a t i e n t s v i a i n s e r t i o n o f t u b i n g e i t h e r i n t r a v e n o u s l y ( i e i n j e c t e d d i r e c t l y i n t o b l o o d s t r e a m ) o r i n t o i n t r a p e r i t o n e a l l y ( i e i n t o

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1 8 s t o m a c h ) F l u i d s c o n s i s t e d o f h u m a n b l o o d c e l l s h u m a n p l a s m a s a l i n e ( 0 9 % s o d i u m c h l o r i d e ) ; 5 % d e x t r o s e ( D g l u c o s e ) a n d E n s u r e ( V a n i l l a P l u s R o s s P r o d u c t s D i v i s i o n A b b o t t L a b o r a t o r i e s C o l u m b u s O H ) B l o o d c e l l s a n d p l a s m a ( a l l o t h e r b l o o d c o m p o n e n t s e x c e p t b l o o d c e l l s ) w e r e o b t a i n e d b y c e n t r i f u g a l s e p a r a t i o n o f d o n a t e d w h o l e b l o o d a n d w e r e t e s t e d a s i n d i v i d u a l f l u i d s s i n c e t h e s e t w o m a j o r c o n s t i t u e n t s o f w h o l e b l o o d d i f f e r g r e a t l y i n c o m p o s i t i o n a n d s t r u c t u r e B l o o d c e l l s c a n b r e a k d o w n w i t h i n 5 t o 7 d ( A A B B 2 0 0 2 ) w i t h o u t a d d i t i o n o f a n t i c o a g u l a n t s d u e t o e n z y m a t i c a c t i v i t y o f i n t r a c e l l u l a r b l o o d p r o t e i n s a n d c e l l d e a t h W h o l e b l o o d s a m p l e s c o n t a i n e d h e p a r i n a n t i c o a g u l a n t w h i c h p r e v e n t s c l o t t i n g a n d i n c r e a s e s s h e l f l i f e o f b l o o d c e l l s t o ~ 4 2 d P l a s m a a n d b l o o d c e l l s w e r e s t o r e d t o g e t h e r a n d s h a r e d a s h e l f l i f e o f ~ 4 2 d w h i c h i s t y p i c a l f o r w h o l e b l o o d u s e d i n t r a u m a a n d e m e r g e n c y s i t u a t i o n s ( M e m m l e r e t a l 1 9 9 6 A A B B 2 0 0 2 ) W h o l e b l o o d w a s o b t a i n e d o n i t s e x p i r a t i o n d a t e a f t e r 3 0 d i n r e f r i g e r a t e d s t o r a g e a t L i f e S o u t h B l o o d B a n k ( G a i n e s v i l l e F L ) T h e r e f o r e a l l w h o l e b l o o d w a s a g e d 3 0 d a n d c o n t a i n e d h e p a r i n a n t i c o a g u l a n t W h o l e b l o o d w a s c e n t r i f u g e d t o s e p a r a t e b l o o d c e l l s ( b o t t o m l a y e r ) f r o m p l a s m a ( t o p l a y e r ) P l a s m a w a s p i p e t t e d o u t a n d p l a c e d i n s e p a r a t e t e s t t u b e s P l a s t i c c a p s w e r e i n s e r t e d i n t o t e s t t u b e o p e n i n g s t o p r e v e n t l e a k a g e B l o o d a n d p l a s m a s a m p l e s w e r e p l a c e d i n t o a r e s e a l a b l e p l a s t i c b a g ( 0 9 5 L Z i p l o c S C J o h n s o n & S o n I n c R a c i n e W I ) w i t h t w o p a p e r t o w e l s f o l d e d i n s i d e t o a b s o r b f l u i d i n c a s e o f s p i l l a g e R e s e a l a b l e p l a s t i c b a g s w e r e p l a c e d i n s i d e a t h i c k w a l l e d S t y r o f o a m c o o l e r I c e ( i n s e p a r a t e r e s e a l a b l e p l a s t i c b a g ) w a s p r e s e n t i n t h e c o o l e r t o k e e p b l o o d c e l l s a n d p l a s m a s a m p l e s c o o l d u r i n g t r a n s p o r t B l o o d c e l l s a n d p l a s m a w e r e r e f r i g e r a t e d a n d u s e d w i t h i n o n e w e e k o f a c q u i s i t i o n B l o o d c e l l s a n d p l a s m a w e r e b o t h u s e d w i t h a p p r o p r i a t e b i o h a z a r d

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1 9 p r o c e d u r e s S h a n d s H o s p i t a l B u r n U n i t p r o v i d e d s e a l e d u n o p e n e d b a g s o f 5 % d e x t r o s e a n d 0 9 % N a C l D e x t r o s e a n d s a l i n e w e r e p l a c e d a n d m a i n t a i n e d i n a r e f r i g e r a t o r ( a l o n g w i t h b l o o d c e l l s a n d p l a s m a ) a t ~ 4 1 C b e f o r e u s e D e x t r o s e a n d s a l i n e p o r t i o n s w e r e s q u i r t e d i n t o d r y c l e a n t e s t t u b e s ( 1 5 0 m m b y 1 0 m m i n n e r d i a m e t e r N o 9 8 0 0 P y r e x L a b w a r e C o r n i n g I n c C o r n i n g N Y ) f o r u s e i n e x p e r i m e n t s B a g s w e r e s e a l e d i m m e d i a t e l y a f t e r u s e a n d r e m a i n e d o u t o f t h e r e f r i g e r a t o r f o r n o l o n g e r t h a n 5 m i n w h i l e r e m o v i n g f l u i d f o r t e s t i n g E n s u r e ( E n s u r e P l u s V a n i l l a ) a c o m m o n l y u s e d f l a v o r i n h o s p i t a l e n v i r o n m e n t s ( S C o b b p e r s o n a l c o m m u n i c a t i o n ) w a s u s e d i n e x p e r i m e n t s b e c a u s e P h a r a o h a n t s h a v e b e e n o b s e r v e d f e e d i n g u p o n d i e t a r y s u p p l e m e n t s i n h o s p i t a l s a n d o t h e r h e a l t h c a r e e n v i r o n m e n t s b y p e s t m a n a g e m e n t p e r s o n n e l ( W W r i g h t J a c k s o n v i l l e P M C S e m i n a r p e r s o n a l c o m m u n i c a t i o n ) E n s u r e i s a d i e t a r y s u p p l e m e n t t h a t c o n t a i n s s u b s t a n t i a l p r o p o r t i o n s o f p r o t e i n s ( 2 6 % ) s u g a r s ( 1 7 % ) a n d l i p i d s ( 1 7 % ) E x p e r i m e n t a l D e s i g n T h e e x p e r i m e n t w a s d e s i g n e d a s a r a n d o m i z e d c o m p l e t e b l o c k b l o c k i n g o n c o l o n y E a c h b l o c k c o n s i s t e d o f P h a r a o h a n t c o l o n i e s ( 1 2 5 i n s i d e w o r k e r s 1 2 5 o u t s i d e w o r k e r s 3 d e a l a t e q u e e n s a n d ~ 5 0 m g b r o o d ) o b t a i n e d f r o m t h e s a m e p a r e n t c o l o n y a n d r a n d o m l y a s s i g n e d t o e a c h t r e a t m e n t ( f l u i d t y p e ) A n t s w e r e s e l e c t e d f r o m n i n e d i f f e r e n t p a r e n t c o l o n i e s A s i n g l e f l u i d w a s o f f e r e d t o e a c h t e s t c o l o n y t o d e t e r m i n e t h e c o n s u m p t i o n a n d p r e f e r e n c e o f f l u i d s u n d e r n o c h o i c e e x p o s u r e O n e t e s t a r e n a a n d o n e c o n t r o l a r e n a w e r e l o c a t e d a d j a c e n t t o e a c h o t h e r T h e r e w e r e 1 7 r e p l i c a t i o n s f o r a t o t a l o f 8 5 t e s t a r e n a s a n d 8 5 c o n t r o l a r e n a s F l u i d s w e r e t e s t e d o v e r a s i x w e e k p e r i o d f r o m

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2 0 J a n u a r y 2 4 2 0 0 3 t o M a r c h 1 0 2 0 0 3 ; r o o m t e m p e r a t u r e a n d h u m i d i t y w e r e b e t w e e n 2 5 7 + 3 7 C a n d 2 0 5 3 % r e s p e c t i v e l y T h e t e m p e r a t u r e a n d r e l a t i v e h u m i d i t y o f t e s t a r e n a s a n d c o n t r o l a r e n a s i n c o n t r a s t t o r o o m t e m p e r a t u r e a n d h u m i d i t y w a s d e t e r m i n e d i n a s e p a r a t e s t u d y o n o n e d a y ( J u n e 2 0 0 3 ) w h e n r o o m t e m p e r a t u r e w a s 2 6 4 C a n d r o o m h u m i d i t y w a s 3 8 % T h i s w a s a c c o m p l i s h e d b y p l a c i n g a d i g i t a l t h e r m o m e t e r a n d h y g r o m e t e r i n s i d e e a c h o f 1 3 p a i r e d t e s t a n d c o n t r o l a r e n a s f o r 2 m i n a l e n g t h o f t i m e s u f f i c i e n t f o r r e a d i n g s t o s t a b i l i z e b u t n o t l o n g e n o u g h f o r a n t s t o i n v a d e e l e c t r o n i c e q u i p m e n t C o l o n i e s i n s i d e t e s t a r e n a s w e r e i d e n t i c a l i n c o m p o s i t i o n t o t h o s e u s e d i n t h i s n o c h o i c e c o n s u m p t i o n t e s t W a t e r c o n s u m p t i o n w a s d e t e r m i n e d i n a s e p a r a t e s t u d y o v e r a t h r e e w e e k p e r i o d i n N o v e m b e r 2 0 0 3 d u r i n g w h i c h r o o m t e m p e r a t u r e a n d h u m i d i t y w e r e m a i n t a i n e d b e t w e e n 2 8 1 + 1 4 C a n d 4 3 4 8 % A n t s w e r e p r o v i d e d w i t h w a t e r i n m i c r o c a p i l l a r y t u b e s ( 3 0 m l v o l u m e 9 5 m m b y 1 0 m m i n n e r d i a m e t e r ; M o d e l N o 2 1 3 8 0 9 C F i s h e r S c i e n t i f i c A t l a n t a G A ) i n t h e s a m e m a n n e r t h a t t e s t f l u i d s w e r e p r o v i d e d W a t e r c o n t r o l t e s t s w e r e r e p l i c a t e d u s i n g t h e s a m e 9 c o l o n i e s i n t h e s a m e s e q u e n c e u s e d f o r t e s t f l u i d s w i t h a t o t a l o f 1 7 t e s t a r e n a s a n d 1 7 c o n t r o l a r e n a s P h a r a o h a n t c o l o n i e s w e r e s t a r v e d f o r 4 8 h b e f o r e p l a c e m e n t o f f l u i d s i n t o a r e n a s ; w a t e r w a s a v a i l a b l e a d l i b i t u m i n a l l a r e n a s i n c l u d i n g w a t e r c o n t r o l s v i a a 1 5 m l c o t t o n s t o p p e r e d v i a l d u r i n g s t a r v a t i o n p e r i o d ( H a a c k a n d V i n s o n 1 9 9 0 H a a c k e t a l 1 9 9 5 ) A f t e r 4 8 h o f s t a r v a t i o n a m i c r o c a p i l l a r y t u b e c o n t a i n i n g o n e t e s t f l u i d w a s a d d e d t o e a c h a r e n a A n t s w e r e a l l o w e d t o f e e d f o r 2 4 h T h e n f l u i d l o s s w a s m e a s u r e d g r a v i m e t r i c a l l y i n e a c h a r e n a

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2 1 F l u i d M e a s u r e m e n t s E a c h e m p t y 3 0 L ( 9 5 m m l o n g ) m i c r o c a p i l l a r y t u b e w a s w e i g h e d v i a a d i g i t a l s c a l e t o + 0 1 m g a n d t h e m a s s o f t h e e m p t y m i c r o c a p i l l a r y t u b e w a s r e c o r d e d E a c h t u b e c a m e p r e m a r k e d w i t h a g r e e n f i l l l i n e l o c a t e d 6 7 m m f r o m t i p o f t u b e E a c h m i c r o c a p i l l a r y t u b e w a s f i l l e d t o t h e 6 7 m m m a r k a n d h e l d ~ 2 1 2 L o f t e s t f l u i d M a s s o f t h e m i c r o c a p i l l a r y t u b e w i t h f l u i d w a s w e i g h e d a n d r e c o r d e d T h e a m o u n t o f f l u i d p r o v i d e d t o a l l c o n t r o l a n d t e s t a r e n a s w a s c a l c u l a t e d f o r e a c h a r e n a b y s u b t r a c t i n g t h e m a s s o f t h e e m p t y m i c r o c a p i l l a r y t u b e f r o m t h e m a s s o f t h e t u b e c o n t a i n i n g i n i t i a l m a s s o f p r o v i d e d f l u i d ( W e i a n d J o h n s o n 1 9 9 6 ) T h u s i n i t i a l p r o v i d e d a m o u n t s o f e a c h f l u i d w e r e o b t a i n e d f o r c o n t r o l a r e n a s a n d f o r t e s t a r e n a s I n a d d i t i o n t o i n i t i a l w e i g h t s o f f o o d p r o v i d e d t o a r e n a s d a t a c o l l e c t e d d u r i n g t h e e x p e r i m e n t i n c l u d e d p o s t e v a p o r a t i o n w e i g h t s o f f l u i d l o a d e d t u b e s f o r c o n t r o l a r e n a s F i n a l p o s t e v a p o r a t i o n w e i g h t s o f c o n t r o l a r e n a f l u i d s w e r e o b t a i n e d b y s u b t r a c t i o n o f t h e a p p r o p r i a t e e m p t y m i c r o c a p i l l a r y t u b e m a s s ( W e i a n d J o h n s o n 1 9 9 6 ) f r o m t h e i n i t i a l w e i g h t o f t h e m i c r o c a p i l l a r y t u b e l o a d e d w i t h f o o d F i n a l l y d a t a c o l l e c t e d d u r i n g t h e e x p e r i m e n t i n c l u d e d p o s t c o n s u m p t i o n w e i g h t s o f f l u i d l o a d e d t u b e s f o r t e s t a r e n a s F i n a l p o s t c o n s u m p t i o n w e i g h t s o f t e s t a r e n a f l u i d s w e r e o b t a i n e d b y s u b t r a c t i o n o f t h e a p p r o p r i a t e e m p t y m i c r o c a p i l l a r y t u b e m a s s ( W e i a n d J o h n s o n 1 9 9 6 ) F l u i d l o s s i n c o n t r o l a r e n a s w a s c a l c u l a t e d b y s u b t r a c t i n g t h e a m o u n t o f f l u i d i n i t i a l l y p r o v i d e d f r o m t h e a m o u n t o f f l u i d r e m a i n i n g ; i n c o n t r o l a r e n a s f l u i d l o s s w a s d u e e n t i r e l y t o e v a p o r a t i o n F l u i d l o s s i n t e s t a r e n a s w a s c o m p r i s e d o f b o t h c o n s u m p t i o n a n d e v a p o r a t i o n ; t h e r e f o r e c o n s u m e d q u a n t i t i e s w e r e d e t e r m i n e d b y a d j u s t i n g f o r

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2 2 e v a p o r a t i o n P r o v i s i o n o f f l u i d s v i a m i c r o c a p i l l a r y t u b e s e n a b l e d f l u i d s t o r e t a i n c o n s t a n t s u r f a c e a r e a s a n d g r e a t l y m i n i m i z e e v a p o r a t i o n A l t h o u g h s e v e r a l c o n s u m p t i o n f o r m u l a s w e r e c o n s i d e r e d ( A p p e n d i x C ) b e c a u s e i t i s a s t a n d a r d m e t h o d o f m e a s u r i n g c o n s u m p t i o n i n n u t r i t i o n a l e c o l o g y l i t e r a t u r e a s l i g h t l y m o d i f i e d v e r s i o n o f W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a w a s u s e d t o c o r r e c t f o r e v a p o r a t i o n r a t e s o f b o t h i n i t i a l a n d f i n a l f l u i d q u a n t i t i e s i n t e s t a n d c o n t r o l a r e n a s a s f o l l o w s : C = ( 1 – [ Y / G ] ) ( W [ L + ( Y / H ) L ] ) w h e r e C = c o r r e c t e d c o n s u m p t i o n W = m a s s o f f l u i d p r o v i d e d ( g ) i n t e s t a r e n a L = m a s s o f u n e a t e n f l u i d a n d a n t s r e m a i n i n g i n f l u i d ( g ) i n t e s t a r e n a G = m a s s o f f l u i d p r o v i d e d ( g ) i n c o n t r o l a r e n a H = m a s s o f f l u i d r e m a i n i n g ( g ) i n c o n t r o l a r e n a Y = G – H = w e i g h t l o s s o f c o n t r o l f l u i d Y / G = w e i g h t l o s s o f c o n t r o l f l u i d / i n i t i a l m a s s o f c o n t r o l f l u i d ( = a i n W a l d b a u e r 1 9 6 8 ) Y / H = w e i g h t l o s s o f c o n t r o l f l u i d / f i n a l m a s s o f c o n t r o l f l u i d ( = b i n W a l d b a u e r 1 9 6 8 ) S t a t i s t i c a l A n a l y s i s E v a p o r a t i o n o f p a t i e n t c a r e f l u i d s i n c o n t r o l a r e n a s w a s a n a l y z e d v i a a o n e t a i l e d S t u d e n t ’ s t t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) t o d e t e r m i n e i f t h e a m o u n t o f e v a p o r a t i o n w a s s i g n i f i c a n t l y d i f f e r e n t t h a n z e r o ( H o : = 0 ) ( F i g 3 2 ) E v a p o r a t i o n o f e a c h f l u i d w a s a l s o c o m p a r e d v i a a n a l y s i s o f v a r i a n c e a n d m e a n s w e r e s e p a r a t e d u s i n g T u k e y ’ s H S D ( P = 0 0 5 ) I n a s e p a r a t e s t u d y m e a n s o f h u m i d i t y a n d t e m p e r a t u r e i n s i d e c o n t r o l a r e n a s

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2 3 w e r e c o m p a r e d v i a a n a l y s i s o f v a r i a n c e ; m e a n s w e r e s e p a r a t e d u s i n g T u k e y ’ s H S D ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) N e g a t i v e c o n s u m p t i o n v a l u e s ( 9 o f 1 0 1 v a l u e s ) w e r e t r u n c a t e d t o z e r o C o n s u m p t i o n o f p a t i e n t c a r e f l u i d s a n d w a t e r w a s a n a l y z e d v i a a o n e t a i l e d S t u d e n t ’ s t t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) t o d e t e r m i n e w h e t h e r c o n s u m p t i o n w a s s i g n i f i c a n t l y d i f f e r e n t t h a n z e r o ( H o : = 0 ) ( T a b l e 3 2 ) H o m o g e n e i t y o f v a r i a n c e s f o r c o n s u m p t i o n a m o n g t h e d i f f e r e n t f l u i d s w a s t e s t e d w i t h L e v e n e ’ s T e s t V a r i a n c e s w e r e n o t h o m o g e n e o u s s o d a t a w e r e l o g a r i t h m i c a l l y t r a n s f o r m e d ( l o g 1 0 ( x + 1 ) ) T r a n s f o r m e d f l u i d c o n s u m p t i o n w a s c o m p a r e d v i a a n a l y s i s o f v a r i a n c e a n d m e a n s w e r e s e p a r a t e d u s i n g T u k e y ’ s H S D ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) R e s u l t s P h a r a o h a n t w o r k e r s f o r a g e d f r o m t h e n e s t i n g c e l l t o t h e m i c r o c a p i l l a r y t u b e s i n o r d e r t o o b t a i n f o o d a n d w a t e r N o q u e e n s l a r v a e o r p u p a e w e r e o b s e r v e d i n t h e m i c r o c a p i l l a r y t u b e s c o n t a i n i n g p a t i e n t c a r e f l u i d s o r w a t e r W o r k e r s w e r e o b s e r v e d i n m i c r o c a p i l l a r y t u b e s w i t h i n 5 m i n o f p l a c i n g f l u i d s i n t e s t a r e n a s T h e y g e n e r a l l y e n t e r e d a n d f e d o n e a t a t i m e d u e t o t h e n a r r o w t u b e d i a m e t e r ( 1 m m I D ) a n d e x i t e d b y b a c k i n g o u t O c c a s i o n a l l y a n e x t r e m e l y s m a l l w o r k e r w a s a b l e t o t u r n a r o u n d w i t h i n a m i c r o c a p i l l a r y t u b e I n g e n e r a l a n t s w a i t e d a r o u n d t h e o p e n i n g o f t h e m i c r o c a p i l l a r y t u b e s o m e t i m e s e n t e r i n g i m m e d i a t e l y b e h i n d a n o t h e r a n t W h e n t h i s o c c u r r e d a l l a n t s h a d t o e x i t i n r e v e r s e o r d e r t o p e r m i t f i r s t a n t t o e x i t t h e t u b e I n r e l a t i v e l y f e w i n s t a n c e s a n t s b e c a m e s t u c k i n t h e f l u i d a n d w e r e u n a b l e t o e x t r i c a t e t h e m s e l v e s B e c a u s e s t u c k a n t s b e c a m e e m b e d d e d t h e f l u i d s p r e a d o u t a n d e x t e n d e d p a s t t h e s t u c k a n t s o t h a t o t h e r a n t s w e r e s t i l l a b l e t o f e e d u p o n t h e f l u i d N u m b e r s o f a n t s r e m a i n i n g i n f l u i d s w e r e n o t

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2 4 r e c o r d e d A l t h o u g h s o m e a n t s d i e d d u r i n g t h e s t a r v a t i o n a n d t h e 2 4 h t e s t p e r i o d m o r t a l i t y w a s n o t c o u n t e d a n d w a s c o n s i d e r e d a n a t u r a l e v e n t t h a t d i d n o t s i g n i f i c a n t l y a f f e c t m e a n c o n s u m p t i o n p e r a r e n a o f a n y s p e c i f i c f l u i d A v o l u m e o f ~ 2 1 2 l o f e a c h p a t i e n t c a r e f l u i d w a s p r o v i d e d B e c a u s e t h e t e s t e d f l u i d s h a v e d i f f e r i n g s p e c i f i c g r a v i t i e s a n d b e c a u s e o f m i n o r v a r i a t i o n s i n t h e v o l u m e s p r o v i d e d i n i t i a l w e i g h t s w e r e s i g n i f i c a n t l y d i f f e r e n t ( T a b l e 3 1 ) E n s u r e t h e m o s t v i s c o u s f l u i d h a d s i g n i f i c a n t l y m o r e m a s s ( ~ 7 6 m g ) p l a c e d i n m i c r o c a p i l l a r y t u b e s t h a n o t h e r f l u i d s S a l i n e h a d t h e l e a s t m a s s o f f l u i d p r o v i d e d ( ~ 6 8 m g ) M e a n d a t a f o r i n i t i a l a m o u n t s o f w a t e r p r o v i d e d i n a s e p a r a t e s t u d y w a s m e a s u r e d ( T a b l e 3 1 ) b u t w a s n o t a n a l y z e d w i t h o t h e r f l u i d s E v a p o r a t i o n o f t h e t e s t e d p a t i e n t c a r e f l u i d s f r o m t h e m i c r o c a p i l l a r y t u b e s w a s d i s t i n c t l y m e a s u r a b l e d e s p i t e t h e h i g h e r t h a n r o o m h u m i d i t y i n c o v e r e d a r e n a s c o n t a i n i n g c o t t o n s t o p p e r e d w a t e r v i a l s F l u i d l o s s i n c o n t r o l a r e n a s d u e t o e v a p o r a t i o n o n l y r a n g e d f r o m 7 1 3 m g a n d d i d n o t d i f f e r s i g n i f i c a n t l y a m o n g t h e p a t i e n t c a r e f l u i d s ( F = 2 1 0 d f = 1 8 0 ; P = 0 0 8 7 9 ) ( T a b l e 3 1 F i g 3 2 ) I n c o n t r a s t t h e r e w e r e s i g n i f i c a n t d i f f e r e n c e s i n f l u i d l o s s o f t e s t a r e n a s ( T a b l e 3 1 ) w h i c h w a s d u e t o c o m b i n e d e v a p o r a t i o n a n d c o n s u m p t i o n T o t a l f l u i d l o s s o f d e x t r o s e ( 4 2 8 8 2 3 6 m g ) i n t e s t a r e n a s w a s s i g n i f i c a n t l y h i g h e r t h a n t h a t o f a l l o t h e r f l u i d s ; s a l i n e s o l u t i o n l o s t s i g n i f i c a n t l y l e s s f l u i d ( 1 6 3 7 3 6 0 m g ) t h a n a l l o t h e r f l u i d s T e m p e r a t u r e o b s e r v a t i o n s o f 1 3 p a i r e d t e s t u n i t s i n M a y 2 0 0 3 i n d i c a t e d t h a t t e m p e r a t u r e i n s i d e c o v e r e d t e s t a r e n a s ( 2 7 3 6 + 0 3 1 C ) w a s s t a t i s t i c a l l y s i g n i f i c a n t l y h i g h e r t h a n i n s i d e c o v e r e d c o n t r o l a r e n a s ( 2 6 3 8 + 0 3 4 C ) a n d r o o m t e m p e r a t u r e ( 2 6 4 0 C ) b y a b o u t 1 o C ( F = 4 5 8 ; d f = 1 2 5 ; P = 0 0 4 2 8 ) T h e m e a n t e m p e r a t u r e o f

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2 5 c o n t r o l a r e n a s w a s n o t s i g n i f i c a n t l y d i f f e r e n t t h a n r o o m t e m p e r a t u r e H u m i d i t y w a s n o t s i g n i f i c a n t l y d i f f e r e n t b e t w e e n t h e i n s i d e s o f c o v e r e d t e s t ( 5 1 6 9 + 0 8 8 % R H ) a n d c o n t r o l a r e n a s ( 5 1 7 7 + 1 3 4 % R H ) ( F = 0 0 0 ; d f = 1 2 5 ; P = 0 9 6 2 1 ) a l t h o u g h t h e y w e r e b o t h s i g n i f i c a n t l y h i g h e r t h a n r o o m h u m i d i t y ( 3 8 % R H ) P h a r a o h a n t s c o n s u m e d s i g n i f i c a n t q u a n t i t i e s o f a l l p a t i e n t c a r e f l u i d s ( T a b l e 3 2 ) C o n s u m p t i o n o f 5 % d e x t r o s e ( 3 7 9 5 + 5 2 1 m g ) w a s s i g n i f i c a n t l y h i g h e r t h a n t h e c o n s u m p t i o n o f a l l o t h e r f l u i d s ( F = 1 4 8 6 ; d f = 4 8 0 ; P < 0 0 0 0 1 ; S A S I n s t i t u t e 2 0 0 0 ) C o n s u m p t i o n o f h u m a n p l a s m a ( 1 7 6 3 + 3 2 6 m g ) h u m a n b l o o d c e l l s ( 1 0 3 3 + 1 6 7 m g ) s a l i n e s o l u t i o n ( 9 9 4 + 3 6 0 m g ) a n d E n s u r e d i e t a r y s u p p l e m e n t ( 7 3 9 + 1 5 3 m g ) ( i e a l l f l u i d s e x c e p t d e x t r o s e ) w e r e n o t c o n s u m e d s i g n i f i c a n t l y d i f f e r e n t l y C o n s u m p t i o n o f b a s e l i n e w a t e r c o n t r o l s ( 2 5 4 + 0 5 4 m g ) w a s s i g n i f i c a n t l y g r e a t e r t h a n z e r o ( H o : = 0 ) D i s c u s s i o n P h a r a o h a n t s m u s t f o r a g e t o o b t a i n e n e r g y a n d p r o t e i n t o f e e d q u e e n a n d l a r v a e A d u l t w o r k e r s c o n s u m e l a r v a l p o s t p h a r y n g e a l r e g u r g i t a t i o n s a n d a n a l s e c r e t i o n s t o h e l p m e e t t h e i r o w n m e t a b o l i c a n d n u t r i t i o n a l n e e d s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) S i n c e p u p a e d o n o t f e e d d e v e l o p m e n t o f P h a r a o h a n t s m u s t b e n u t r i t i o n a l l y s u p p o r t e d e n t i r e l y d u r i n g l a r v a l s t a g e s A d u l t P h a r a o h a n t s i n g e s t t o c r o p o n l y p a r t i c l e s s m a l l e r t h a n 1 8 m d i a m e t e r ( P e t t i 1 9 9 8 ) L a r g e r p a r t i c l e s a r e f i l t e r e d i n t o t h e i n f r a b u c c a l p o c k e t a n d t r a n s p o r t e d b a c k t o t h e i m m o b i l e l a r v a e i n t h e n e s t ( P e t t i 1 9 9 8 ) w h e r e t h e l a r v a e c a n d i g e s t t h e s e s o l i d p a r t i c l e s a n d l a t e r r e g u r g i t a t e f l u i d s b a c k t o t h e w o r k e r s a n d q u e e n s p r i m a r i l y t h r o u g h t h e l a b i a l g l a n d s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) S i n c e l a r v a e p r o v i d e d i g e s t e d f o o d t o w o r k e r s a n d p r o v i d e u p t o 9 3 % o f s u b s t a n c e s t h a t q u e e n s d e p e n d u p o n ( B r g e s e n a n d J e n s e n 1 9 9 5 ) c o l o n y v i g o r d e p e n d s o n h e a l t h y g r o w i n g o f l a r v a e L a r v a e

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2 6 a r e e s p e c i a l l y i m p o r t a n t i n P h a r a o h a n t c o l o n i e s b e c a u s e t h e a b i l i t y o f q u e e n s t o f o u n d n e w c o l o n i e s i s p o s i t i v e l y l i n k e d t o l a r v a l p r e s e n c e ( B r g e s e n a n d J e n s e n 1 9 9 5 ) P r o t e i n i s a n e c e s s a r y c o n s t i t u e n t o f c u t i c l e a n d o r g a n d e v e l o p m e n t ; i f p r o t e i n a v a i l a b i l i t y i s l i m i t e d m a n y s p e c i e s o f a n t s w i l l c o n s u m e b r o o d ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) P e a c o c k ( 1 9 5 0 ) a t t r i b u t e d b r o o d m o r t a l i t y t o v a r i o u s c a u s e s i n c l u d i n g c a n n i b a l i s m o f s e x u a l b r o o d I n g e n e r a l t h e p r i o r i t y f o r p r o t e i n f e e d i n g i s q u e e n s l a r v a e a n d w o r k e r s ( B e r n d t 1 9 7 7 B r g e s e n a n d J e n s e n 1 9 9 5 ) W o r k e r s a r e l a s t i n t h e p r o t e i n n u t r i t i o n a l h i e r a r c h y s i n c e q u e e n s a n d b r o o d n e e d l a r g e a m o u n t s f o r e g g l a y i n g a n d d e v e l o p i n g r e s p e c t i v e l y ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) F l u i d M e a s u r e m e n t s T h e a m o u n t o f f l u i d p r o v i d e d c a n i m p a c t t h e r a t e o f e v a p o r a t i o n o f a n y g i v e n f l u i d s i n c e e v a p o r a t i o n r a t e s a r e d e p e n d e n t u p o n t h e v o l u m e o f f l u i d p r e s e n t a s w e l l a s u p o n t h e s u r f a c e a r e a e x p o s e d t o a i r D e s p i t e s o m e f l u c t u a t i o n i n t e m p e r a t u r e a n d h u m i d i t y t h e s u r f a c e a r e a f r o m w h i c h e v a p o r a t i o n c o u l d h a v e o c c u r r e d w a s l i m i t e d t o t w o 1 m m d i a m e t e r o p e n i n g s a n d a s a r e s u l t t h e r e w a s n o s i g n i f i c a n t d i f f e r e n c e b e t w e e n e v a p o r a t i o n o f t h e v a r i o u s l i q u i d s t e s t e d F i n a l l y a n t s d i d n o t e a t u n t i l d e p l e t i o n o f p r o v i d e d f l u i d s d u r i n g t h e 2 4 h s t u d y T h u s w h i l e W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a i s t h e s t a n d a r d f o r m e a s u r i n g c o n s u m p t i o n i n n u t r i t i o n a l e c o l o g y s t u d i e s i t w o u l d p r o b a b l y h a v e b e e n a s a c c e p t a b l e t o u s e a s t r a i g h t s u b t r a c t i o n o f e v a p o r a t i o n a s a n a d j u s t m e n t f o r c o n s u m p t i o n i n t h i s e x p e r i m e n t a l s e t u p W a t e r C o n s u m p t i o n W a t e r i s a n e s s e n t i a l c o m p o n e n t o f a n t s u r v i v a l a n d r e p r o d u c t i o n ( C h a p m a n 1 9 9 8 ) F o r a g i n g i s a m u l t i s t e p p r o c e s s f o r P h a r a o h a n t c o l o n i e s ( S u d d 1 9 5 7 b 1 9 6 0 )

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2 7 A f t e r d i s c o v e r y o f w a t e r o r f o o d f o r a g e r s l a y f a r a n a l p h e r o m o n e t r a i l s a s t h e y q u i c k l y r e t u r n t o t h e n e s t ( J e a n s o n e t a l 2 0 0 3 ) a n d r e c r u i t a d d i t i o n a l w o r k e r s I n t h i s e x p e r i m e n t w o r k e r s d i s c o v e r e d t h e w a t e r v i a l s w i t h i n 1 5 m i n o f p l a c e m e n t i n t o a r e n a a t t h e b e g i n n i n g o f t h e 2 4 h s t a r v a t i o n p e r i o d a n d e s t a b l i s h e d a t r a i l f r o m t h e w a t e r v i a l t o t h e n e s t i n g c e l l T h e y m a i n t a i n e d t h e s e t r a i l s d u r i n g t h e e n t i r e s t a r v a t i o n p e r i o d s o t h a t r e c r u i t m e n t t o w a t e r v i a l s w a s e s t a b l i s h e d p r i o r t o p l a c e m e n t o f f l u i d l o a d e d m i c r o c a p i l l a r y t u b e s B e c a u s e f a r a n a l i s a s h o r t l i v e d p h e r o m o n e w i t h a h a l f l i f e o f ~ 9 m i n o n c l e a n p l a s t i c s u r f a c e s ( J e a n s o n e t a l 2 0 0 3 ) P h a r a o h a n t s m u s t c o n t i n u o u s l y d e p o s i t f a r a n a l o n t r a i l s t o m a i n t a i n r e c r u i t m e n t t o d e s i r a b l e r e s o u r c e s A t t h e e n d o f t h e s t a r v a t i o n p e r i o d a m i c r o c a p i l l a r y t u b e c o n t a i n i n g f l u i d w a s p l a c e d i n a r e n a ; w o r k e r s d i s c o v e r e d t h e n e w f l u i d s o u r c e a l m o s t i m m e d i a t e l y I n t h e w a t e r b a s e l i n e s t u d y w h i c h c o n s i s t e d o f p l a c e m e n t o f w a t e r l o a d e d m i c r o c a p i l l a r y t u b e s i n t o t h e a r e n a a l o n g w i t h a d l i b i t u m w a t e r i n 1 5 m l v i a l t h e c h o i c e a v a i l a b l e t o a n t s w a s c o n s u m p t i o n o f w a t e r v i a d i f f e r e n t m e c h a n i s m s e i t h e r v i a t h e m i c r o c a p i l l a r y t u b e c e n t e r e d i n t h e a r e n a o r v i a t h e v i a l p l a c e d i n t h e o p p o s i n g c o r n e r o f t h e a r e n a r a t h e r t h a n b e t w e e n d i f f e r e n t f l u i d s W a t e r c o n s u m p t i o n i n t h e b a s e l i n e s t u d y w a s s i g n i f i c a n t b u t a p p e a r e d t o b e m u c h l e s s t h a n c o n s u m p t i o n o f a l l o t h e r f l u i d s i n d i c a t i n g t h a t w h i l e P h a r a o h a n t s c o n s u m e d w a t e r i n m i c r o c a p i l l a r y t u b e s t h e y p r o b a b l y o b t a i n e d w a t e r p r i m a r i l y f r o m w a t e r v i a l s O b s e r v a t i o n s i n a l l t e s t a r e n a s o f m u l t i p l e a n t s f e e d i n g a t w a t e r v i a l s o f o n l y 1 o r 2 a n t s f e e d i n g a t m i c r o c a p i l l a r y t u b e s a n d m a i n t e n a n c e o f r e c r u i t m e n t t r a i l s t o w a t e r v i a l s s u p p o r t t h i s p o s s i b i l i t y

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2 8 D e x t r o s e C o n s u m p t i o n S i m p l e s u g a r s s u c h a s d e x t r o s e a r e i m m e d i a t e l y d i g e s t e d w i t h o u t n e e d t o b r e a k d e x t r o s e d o w n i n t o s i m p l e r c o m p o n e n t s a s i s n e e d e d f o r m o r e c o m p l e x s u g a r s s u c h a s s u c r o s e f r u c t o s e a n d o t h e r s u g a r s I n f a c t i t i s p o s s i b l e t h a t d e x t r o s e i s d i g e s t e d m o r e r a p i d l y t h a n g l u c o s e t h a t c o n t a i n s b o t h L g l u c o s e a n d D g l u c o s e b e c a u s e L g l u c o s e m u s t b e c o n v e r t e d t o D g l u c o s e f i r s t f o r d i g e s t i o n D e x t r o s e i s a c t u a l l y c o m p o s e d e n t i r e l y o f D g l u c o s e m o l e c u l e s t h e m o s t e a s i l y d i g e s t e d e n a n t i o m e r o f g l u c o s e C o n s u m p t i o n o f d e x t r o s e i n e a c h a r e n a w a s s i g n i f i c a n t l y h i g h e r t h a n c o n s u m p t i o n o f E n s u r e t h e o n l y o t h e r h i g h s u g a r f l u i d p r o v i d e d i n t h i s e x p e r i m e n t F o r a g i n g s t u d i e s r e v e a l e d t h a t P h a r a o h a n t s a r e a t t r a c t e d t o s u g a r f o o d s o u r c e s ( S u d d 1 9 6 0 H a a c k a n d V i n s o n 1 9 9 0 V a i l a n d W i l l i a m s 1 9 9 4 V a i l 1 9 9 6 ) I t i s p o s s i b l e t h a t f o r a g i n g n e e d s o f P h a r a o h a n t s c h a n g e w i t h s e a s o n a l r e q u i r e m e n t s i n a m a n n e r s i m i l a r t o t h a t o f S o l e n o p s i s i n v i c t a ( L ) ( P o r t e r a n d T s c h i n k e l 1 9 8 7 ) D e x t r o s e i s p r o b a b l y m o r e a t t r a c t i v e t o P h a r a o h a n t s t h a n E n s u r e f o r s e v e r a l r e a s o n s : ( 1 ) s u g a r i s t h e o n l y n u t r i e n t p r e s e n t ( 2 ) s u g a r i s p r e s e n t a s D g l u c o s e t h e s i m p l e s t a n d m o s t e a s i l y d i g e s t e d a n d m e t a b o l i z e d f o r m p o s s i b l e a n d ( 3 ) c o n c e n t r a t i o n o f s u g a r i n 5 % d e x t r o s e s o l u t i o n h a s a l o w e r v i s c o s i t y t o a l l o w f o r e a s i e r e x t r a c t i o n b y a d u l t w o r k e r s a n d m a y h e l p s a t i s f y m o i s t u r e r e q u i r e m e n t s a s w e l l a s e n e r g y r e q u i r e m e n t s S i g n i f i c a n t c o n s u m p t i o n ( H o : = 0 ) o f d e x t r o s e a g r e e s w i t h r e s e a r c h o f o t h e r s c i e n t i s t s w h o p r e v i o u s l y r e p o r t e d t h a t a n t s f e e d r e a d i l y a n d f r e q u e n t l y o n s u g a r s M a n y r e s e a r c h e r s u s e d s u g a r o r h o n e y i n e i t h e r l i q u i d s o l u t i o n s o r a s s o l i d s i n b a i t a n d f o r a g i n g s t u d i e s f o r P h a r a o h a n t s R i l e y ( 1 8 8 9 ) s u g g e s t e d c o a t i n g p i e c e s o f c r o c k e r y w i t h s u g a r s o l u t i o n t o a t t r a c t P h a r a o h a n t s a n d B e l l e v o y e ( 1 8 8 9 ) r e p o r t e d P h a r a o h a n t s c o n s u m i n g

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2 9 s w e e t s i n h i s a p a r t m e n t a n d n e i g h b o r i n g b a k e r y T h e r e f o r e i t w a s n o t u n e x p e c t e d t h a t c o n s u m p t i o n o f 5 % d e x t r o s e w a s s i g n i f i c a n t l y h i g h e r t h a n c o n s u m p t i o n o f o t h e r f l u i d s P l a s m a a n d B l o o d C o n s u m p t i o n P h a r a o h a n t s m u s t f o r a g e f o r p r o t e i n t o m e e t m e t a b o l i c n e e d s o f t h e q u e e n f o r e g g l a y i n g a n d t o e n a b l e l a r v a e t o c o m p l e t e t h e i r d e v e l o p m e n t H u m a n w h o l e b l o o d c o n t a i n s a s i g n i f i c a n t a m o u n t o f p r o t e i n ( ~ 8 % ) B l o o d c e l l s c o n t a i n p r o t e i n t h a t i s b o u n d i n b i l i p i d c e l l u l a r m e m b r a n e s a n d i n t e r c e l l u l a r s t r u c t u r e s I n c o n t r a s t p l a s m a c o n t a i n s m o s t o f w h o l e b l o o d ’ s p r o t e i n a s f r e e f l o a t i n g p a r t i c l e s i n a s o l u t i o n t h a t i s 9 0 % w a t e r B l o o d c e l l s s o l i d i f i e d i n t h e m i c r o c a p i l l a r y t u b e s d u r i n g t e s t r e s u l t i n g i n l o s s o f c a p i l l a r y a c t i o n a n d g r e a t l y i n c r e a s e d s u r f a c e a r e a o f e x p o s e d f l u i d A l t h o u g h h e p a r i n s l o w s c o a g u l a t i o n o f b l o o d c e l l s b l o o d c e l l s s o l i d i f i e d o v e r t i m e P i p e t t i n g b l o o d c e l l s o u t o f t e s t t u b e s b e c a m e i n c r e a s i n g l y d i f f i c u l t a s s a m p l e s a g e d A l l s a m p l e s w e r e u s e d w i t h i n 7 d t o r e d u c e s o l i d i f i c a t i o n W h i l e h e p a r i n a n t i c o a g u l a n t w a s p r e s e n t i n w h o l e b l o o d s a m p l e s i n i t i a l r e m o v a l o f p l a s m a v i a p i p e t t e p r o b a b l y i n c l u d e d r e m o v a l o f h e p a r i n s i n c e p l a s m a c o n t a i n s e v e r y t h i n g i n w h o l e b l o o d e x c e p t b l o o d c e l l s ( A A B B 2 0 0 2 ) T h e r e f o r e b l o o d c e l l s a s t e s t e d p r o b a b l y c o n t a i n e d l i t t l e a n t i c o a g u l a n t d u r i n g t e s t p e r i o d ; t h u s c o a g u l a t i o n a n d s o l i d i f i c a t i o n w a s m o r e l i k e l y i n b l o o d c e l l s t h a n i n p l a s m a A d d i t i o n a l l y t h e r e w a s v a r i a b i l i t y i n c o a g u l a t i o n l e v e l s b e t w e e n d o n a t e d s a m p l e s W h o l e b l o o d s a m p l e s w e r e n o t a n a l y z e d f o r h e m a t o c r i t p e r c e n t a g e s ( p e r c e n t a g e o f b l o o d c e l l s p r e s e n t i n w h o l e b l o o d v a r i e s f r o m p e r s o n t o p e r s o n a n d e v e n f r o m d a y t o d a y : i e h e m a t o c r i t c a n i n c r e a s e w h e n d o n o r i s d e h y d r a t e d o r d e c r e a s e i f d o n o r h a s c o n s u m e d l a r g e q u a n t i t i e s o f s a l t i n d i e t ) p r i o r t o u s e i n t h i s e x p e r i m e n t

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3 0 T h e e f f e c t o f h e p a r i n u p o n P h a r a o h a n t c o n s u m p t i o n w a s n o t s t u d i e d i n t h i s e x p e r i m e n t T h e r e i s e v i d e n c e t h a t h e p a r i n m a y n e g a t i v e l y a f f e c t f e e d i n g a b i l i t y o f m o s q u i t o e s ( B o y d a n d K a y 2 0 0 0 ) b u t m o s q u i t o e s a r e c a p i l l a r y f e e d e r s s p e c i a l i z e d t o f e e d d i r e c t l y f r o m v e r t e b r a t e h o s t b l o o d s t r e a m ( S e r v i c e 2 0 0 0 M u l l e n a n d D u r d e n 2 0 0 2 ) a n d h e p a r i n m o s t l y a f f e c t s t h e s u c c e s s f u l b l o o d m e a l a c q u i s i t i o n i n m o s q u i t o ( B o y d a n d K a y 2 0 0 0 ) I n c o n t r a s t P h a r a o h a n t s a r e o m n i v o r o u s f e e d e r s t h a t f e e d u p o n e x t e r i o r o p e n w o u n d s r a t h e r t h a n p e n e t r a t i n g b e l o w v e r t e b r a t e e p i d e r m i s i n t o s e a l e d v e i n s a n d a r t e r i e s F i n a l l y a n t c o l o n i e s u s e d i n t h i s e x p e r i m e n t w e r e s m a l l t h u s p r o t e i n r e q u i r e m e n t s m a y h a v e b e e n l o w e r r e s u l t i n g i n l e s s f e e d i n g o f h i g h p r o t e i n f l u i d s I n r e g a r d s t o b o t h p l a s m a a n d b l o o d c e l l s t h i s e x p e r i m e n t r e p r e s e n t s t h e f i r s t t i m e t h a t b l o o d e i t h e r a s w h o l e b l o o d o r a s i t s c o n s t i t u e n t s h a s b e e n f e d t o P h a r a o h a n t s a l t h o u g h G r a c e e t a l ( 1 9 8 6 ) f e d h u m a n s p u t u m t o I r i d o m y r m e x ( L i n e p i t h e m a ) h u m i l i s ( M a y r ) d u r i n g a f o r e n s i c m u r d e r i n v e s t i g a t i o n W h i l e n o b o d y h a s e v e r f e d h u m a n p l a s m a o r h u m a n b l o o d ( o r a n y a n i m a l p l a s m a o r b l o o d ) t o P h a r a o h a n t s m a n y r e s e a r c h e r s h a v e d e t e r m i n e d t h a t P h a r a o h a n t s w i l l f e e d o n h i g h p r o t e i n f o o d s s u c h a s r a w l i v e r l i v e r p o w d e r f l i e s a n d c o c k r o a c h e s ( B e l l e v o y e 1 8 8 9 S u d d 1 9 6 0 E d w a r d s 1 9 8 6 H a a c k a n d V i n s o n 1 9 9 0 ) T h e r e f o r e s i g n i f i c a n t c o n s u m p t i o n o f h u m a n p l a s m a a h i g h p r o t e i n f l u i d c o m e s a s n o s u r p r i s e a n d I s u s p e c t t h a t p r e s e n c e o f p l a s m a i n o p e n w o u n d s a n d a t b u r n s i t e s m i g h t b e a n i m p o r t a n t f a c t o r i n P h a r a o h a n t a t t r a c t i o n t o p a t i e n t s i n h o s p i t a l s I n c o n t r a s t c o n s u m p t i o n o f b l o o d c e l l s w a s n o t e x p e c t e d t o b e l a r g e s i n c e b l o o d c e l l s a r e n o t a s p r e v a l e n t a t o p e n w o u n d / b u r n s i t e s a s p l a s m a B u r n s t e n d t o o o z e p l a s m a r a t h e r t h a n t o b l e e d d u e t o b o d y ’ s i m m u n e s y s t e m r e s p o n s e w h i c h f a c i l i t a t e s d e l i v e r y o f w h i t e

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3 1 b l o o d c e l l s a n d e n z y m e s ( p r o t e i n s ) t o d a m a g e d e p i t h e l i u m t o f i g h t i n f e c t i o n a n d c l o s e o p e n w o u n d s b y d e v e l o p m e n t o f s c a r t i s s u e S a l i n e ( = 0 9 % N a C l ) C o n s u m p t i o n E l e c t r o l y t e s s u c h a s s o d i u m ( N a + ) a n d c h l o r i n e ( C l ) i o n s ( b o t h p r e s e n t i n s a l i n e s o l u t i o n ) a r e n u t r i e n t s t h a t P h a r a o h a n t s n e e d i n t h e i r d i e t ( C h a p m a n 1 9 9 8 ) V i n s o n ( 1 9 7 0 ) c i t e d B u t l e r ’ s ( 1 9 4 0 ) r e p o r t t h a t s o m e d i l u t e s o l u t i o n s o f N a C l w e r e p r e f e r r e d o v e r w a t e r H o w e v e r s i g n i f i c a n t c o n s u m p t i o n o f s a l i n e s o l u t i o n i n e a c h t e s t a r e n a o v e r a 2 4 h p e r i o d a f t e r o n l y 3 d o f s t a r v a t i o n w a s n o t e x p e c t e d S a l i n e w a s i n c l u d e d b e c a u s e s a l i n e s o l u t i o n i s o f t e n u s e d i n p a t i e n t c a r e i n h o s p i t a l e n v i r o n m e n t s b o t h t o r e p l a c e l o s t f l u i d s a n d t o s t e r i l i z e w o u n d s e s p e c i a l l y i n b u r n u n i t s S a l i n e c o n s u m p t i o n i n t h i s e x p e r i m e n t a g r e e d w i t h W i l l i a m s ( 1 9 9 0 ) w h e n h e c o m p a r e d a c c e p t a b i l i t y o f s a l i n e t o a c c e p t a b i l i t y o f s e v e r a l s u g a r s ( h e u s e d r a w h o n e y a s t e s t s t a n d a r d ) I t i s i n t e r e s t i n g t o n o t e t h a t t h e a c c e p t a n c e r a t i o h e o b t a i n e d f o r s a l i n e c o n s u m p t i o n w a s n e a r l y i d e n t i c a l t o t h o s e f o r c o n s u m p t i o n o f 1 0 % d e x t r o s e a n d 1 0 % g l u c o s e b o t h s i m p l e s u g a r s H e c o n t r a s t e d s a l i n e ’ s a c c e p t a n c e r a t i o ( 0 0 6 9 + 0 0 1 5 ) a g a i n s t r a w h o n e y ’ s a c c e p t a n c e r a t i o n ( 1 0 0 + 0 0 0 0 ) t o d e t e r m i n e t h a t w h i l e s a l i n e w a s a c c e p t e d a s a f o o d s o u r c e i t w a s m u c h l e s s p r e f e r a b l e t h a n m o s t s u g a r s E n s u r e C o n s u m p t i o n E n s u r e w a s t h e o n l y t e s t f l u i d t h a t c o u l d b e c o n s i d e r e d “ c o m p l e t e ” s i n c e i t c o n t a i n s a l l t h r e e m a j o r n u t r i e n t s : 2 6 % p r o t e i n s 1 7 % s u g a r s a n d 1 7 % l i p i d s T h i s f l u i d w a s e x p e c t e d t o b e h i g h l y a t t r a c t i v e t o P h a r a o h a n t s a n d c o n s u m p t i o n o f t h i s c o m p l e t e f o o d w a s a n t i c i p a t e d t o b e h i g h e r t h a n t h a t o f o t h e r f l u i d s C o n s u m p t i o n o f E n s u r e m a y h a v e b e e n a f f e c t e d b y i t s n u t r i t i o n a l c o m p l e t e n e s s V a i l r e p o r t e d t h a t a b a i t c o n t a i n i n g

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3 2 a l l t h r e e n u t r i e n t s ( c a r b o h y d r a t e s l i p i d s a n d p r o t e i n s ) w o u l d a t t r a c t a n y f o r a g i n g P h a r a o h a n t s A l t h o u g h c o n s u m p t i o n o f E n s u r e w a s l e s s t h a n t h a t o f o t h e r f l u i d s t h a t m a y r e f l e c t i t s c o m p l e t e n e s s s i n c e P h a r a o h a n t s w o u l d n o t n e e d t o e a t a s m u c h t o m e e t t h e i r n u t r i t i o n a l r e q u i r e m e n t s I n c o n t r a s t t h e y w o u l d n e e d t o c o n s u m e l a r g e q u a n t i t i e s o f a l o w d e n s i t y s o l e n u t r i e n t f o o d t o o b t a i n t h e s a m e e n e r g y a n d n u t r i t i o n T h i s e x p e r i m e n t r e p r e s e n t s t h e f i r s t t i m e t h a t E n s u r e ( o r a n y d i e t a r y s u p p l e m e n t ) h a s b e e n p r o v i d e d t o P h a r a o h a n t s f o r c o n s u m p t i o n T h e s e r e s u l t s p r o v i d e s o m e i n t e r e s t i n g i n f o r m a t i o n a b o u t c o n s u m p t i o n o f c o m p l e t e f o o d s v s s i m p l e r f o o d s I t i s p o s s i b l e t h a t a n t p r e f e r e n c e f o r E n s u r e m i g h t h a v e b e e n l i m i t e d f o r o n e o r m o r e r e a s o n s : I o b s e r v e d t h a t E n s u r e d r i e d a n d p a r t i a l l y s o l i d i f i e d i n s i d e m i c r o c a p i l l a r y t u b e s w h i l e o t h e r f l u i d s r e m a i n e d i n o r i g i n a l l i q u i d f o r m S o l i d i f i c a t i o n o f E n s u r e w a s m o s t c l o s e l y m a t c h e d b y s o l i d i f i c a t i o n o f b l o o d c e l l s b u t t h e m a n n e r o f d r y i n g w a s o b s e r v e d t o b e d i f f e r e n t S p e c i f i c a l l y b l o o d c e l l s l o s t m i c r o c a p i l l a r y f u n c t i o n a n d s p r e a d o u t i n a t h i n l a y e r a l o n g m i c r o c a p i l l a r y t u b e b o t t o m s a f t e r s o l i d i f i c a t i o n a n d d e s i c c a t i o n I n c o n t r a s t E n s u r e d r i e d i n m i c r o c a p i l l a r y t u b e s i n s u c h a m a n n e r a s t o p a r t i a l l y b l o c k t h e t u b e m a k i n g a n t p e n e t r a t i o n a n d e x t r a c t i o n o f f l u i d p o t e n t i a l l y m o r e d i f f i c u l t L o w c o n s u m p t i o n o f p l a s m a b l o o d c e l l s a n d E n s u r e m i g h t h a v e b e e n d u e t o t h e p r e s e n c e o f h e p a r i n a n t i c o a g u l a n t o r t o h i g h l i p i d c o n t e n t i n h i g h p r o t e i n f l u i d s H e p a r i n w a s p r e s e n t i n w h o l e b l o o d s a m p l e s p r i o r t o c e n t r i f u g a t i o n s o p r o p o r t i o n s o f h e p a r i n i n s e p a r a t e d b l o o d c e l l s a n d p l a s m a w e r e u n k n o w n a n d t h e e f f e c t o f h e p a r i n o n f e e d i n g w a s n o t s t u d i e d P r o t e i n c o m p r i s e s a p p r o x i m a t e l y 8 % o f p l a s m a 8 % o f b l o o d c e l l s ( A A B B 2 0 0 2 ) a n d 1 7 % o f E n s u r e ( n u t r i t i o n l a b e l ) P r e v i o u s r e s e a r c h h a s s h o w n t h a t P h a r a o h a n t s d e p l e t e p r o t e i n s t o r e s a s r a p i d l y a s c a r b o h y d r a t e s t o r e s ( H a a c k a n d V i n s o n 1 9 9 0

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3 3 H a a c k e t a l 1 9 9 5 ) b u t p r o t e i n s u s e d i n t h o s e e x p e r i m e n t s c o n s i s t e d o f a p o w d e r e d e g g y o l k p a s t e a n d e g g y o l k s h a v e a h i g h f a t c o n t e n t o f ~ 7 4 % i n c o n t r a s t t o t h e i r p r o t e i n c o n t e n t o f ~ 2 0 % ( U S D A 2 0 0 2 ) E n s u r e h a s a m u c h l o w e r f a t c o n t e n t ( ~ 1 7 % ) a n d a s l i g h t l y h i g h e r p r o t e i n c o n t e n t ( ~ 2 6 % ) t h a n e g g y o l k s P h a r a o h a n t s h a v e f e d u p o n l i p i d f o o d s w i t h n o s t a r v a t i o n ( W i l l i a m s 1 9 9 0 ) s o t h e e f f e c t o f l i p i d s u p o n c o n s u m p t i o n o f E n s u r e i s u n k n o w n I t i s c l e a r t h a t i n t h i s s t u d y d e x t r o s e c o n s u m p t i o n e x c e e d e d t h a t o f a l l o t h e r f l u i d s S u m p t e r a n d B e e k m a n ( 2 0 0 3 ) r e p o r t e d P h a r a o h a n t s p r e f e r e n t i a l l y r e c r u i t t o f o o d s w i t h h i g h e s t s u c r o s e c o n c e n t r a t i o n K n o w i n g t h a t P h a r a o h a n t s a r e a t t r a c t e d t o a n d w i l l c o n s u m e f l u i d s u s e d i n p a t i e n t c a r e e s p e c i a l l y d e x t r o s e h o s p i t a l s t a f f s h o u l d n o t s p r e a d t h e s e f l u i d s i n h o s p i t a l ; i f t h e s e f l u i d s a r e p r e s e n t ( e s p e c i a l l y n e a r p a t i e n t ) t h e y s h o u l d b e c l e a n e d u p q u i c k l y t o d e c r e a s e p o t e n t i a l f o r P h a r a o h a n t a t t r a c t i o n t o p a t i e n t s w i t h i n h o s p i t a l a n d o t h e r h e a l t h c a r e e n v i r o n m e n t s

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T a b l e 3 1 M e a n ( l i q u i d n o c h o i c e ) i n i t i a l w e i g h t s o f f l u i d q u a n t i t y p r o v i d e d ( m g ) a n d m e a n f l u i d l o s s ( m g ) d u r i n g t e s t p e r i o d ( 2 4 h ) f o r e a c h o f f i v e d i f f e r e n t p a t i e n t c a r e f l u i d s u s e d i n h o s p i t a l e n v i r o n m e n t s T e s t a r e n a s w i t h a n t s ( n = 2 5 0 ) ; c o n t r o l a r e n a s l a c k e d a n t s M e a n ( m g ) S E M _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ C o n t r o l a r e n a s T e s t a r e n a s P a t i e n t c a r e _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ F l u i d I n i t i a l w ta F l u i d L o s s ( E v a p )a b I n i t i a l w ta F l u i d L o s sa b c B l o o d 7 2 7 7 0 2 9 b 1 2 7 1 + 1 9 0 a 7 2 8 8 + 0 2 8 b 2 1 9 7 + 2 3 6 a b c 5 % D e x t r o s e 6 9 7 5 + 0 2 6 c d 9 2 5 + 2 1 4 a 6 9 7 1 + 0 2 6 c 4 2 8 8 + 5 1 6 a E n s u r e 7 7 3 6 + 0 4 1 a 8 5 9 + 0 5 2 a 7 6 5 9 + 0 3 7 a 1 5 3 4 + 1 5 6 b c 0 9 % N a C l 6 8 6 8 + 0 3 3 d 7 5 4 + 1 0 0 a 6 8 3 9 + 0 4 7 d 1 6 3 7 + 3 6 0 c P l a s m a 7 0 3 8 + 0 2 4 c 1 3 5 3 + 2 3 4 a 7 0 6 4 + 0 2 4 c 2 8 4 2 + 3 6 0 a b W a t e rd 6 9 4 8 + 0 1 2 3 2 1 + 0 1 9 6 9 5 4 + 0 1 6 5 6 4 + 0 5 9 a M e a n s w i t h i n a c o l u m n f o l l o w e d b y t h e s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ) b F l u i d L o s s i n c o n t r o l a r e n a s i s t o e v a p o r a t i o n o n l y c F l u i d l o s s i n t e s t a r e n a s d u e t o e v a p o r a t i o n a n d c o n s u m p t i o n c o m b i n e d d W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s 3 4

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T a b l e 3 2 M e a n ( l i q u i d n o c h o i c e ) c o n s u m p t i o n ( 2 4 h ) f o r e a c h o f f i v e d i f f e r e n t p a t i e n t c a r e f l u i d s u s e d i n h o s p i t a l e n v i r o n m e n t s b y s t a r v e d ( 3 d ) P h a r a o h a n t c o l o n i e s ( n = 2 5 0 ) P a t i e n t c a r e T e s t a r e n a s d f t P F l u i d M e a n c o n s u m p t i o n S E M ( m g ) B l o o d 1 0 3 3 + 1 6 7 b 1 6 6 1 7 < 0 0 0 0 1 5 % D e x t r o s e 3 7 9 5 + 5 2 1 a 1 6 7 2 8 < 0 0 0 0 1 E n s u r e 7 3 9 + 1 5 3 b 1 6 4 8 3 < 0 0 0 0 1 0 9 % N a C l 9 9 4 + 3 6 0 b 1 6 2 7 6 0 0 0 9 5 P l a s m a 1 7 6 3 + 3 2 6 b 1 6 5 4 1 < 0 0 0 0 1 W a t e ra 0 4 8 + 0 0 7 1 6 7 2 5 < 0 0 0 0 1 M e a n c o n s u m p t i o n w i t h i n a r o w ( H0: = 0 ) a n a l y z e d v i a S t u d e n t ’ s t t e s t ( S A S I n s t i t u t e 2 0 0 0 ) M e a n c o n s u m p t i o n o f f l u i d s ( m g ) a n d r a n k ( n u m b e r ) l i s t e d i n c o l u m n s a n a l y z e d v i a T u k e y ’ s H S D t e s t ( P = 0 0 5 ) M e a n s w i t h i n a c o l u m n f o l l o w e d b y s a m e l e t t e r s a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( F = 1 4 8 6 ; d f = 4 8 0 ; P < 0 0 0 0 1 ) ; d a t a w e r e l o g a r i t h m i c t r a n s f o r m e d b e f o r e a n a l y s i s a W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s 3 5

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3 2 1 1 3 5 3 a 7 5 4 a 8 5 9 a 9 2 5 a 1 2 7 1 a 0 2 4 6 8 1 0 1 2 1 4 1 6 W a t e r B l o o d 5 % D e x t r o s e E n s u r e 0 9 % N a C l P l a s m aE v a p o r a t i o n ( m g ) F i g u r e 3 2 M e a n ( l i q u i d n o c h o i c e ) e v a p o r a t i o n ( 2 4 h ) i n c o n t r o l a r e n a s f o r e a c h t e s t e d p a t i e n t c a r e f l u i d u s e d i n h o s p i t a l e n v i r o n m e n t s E v a p o r a t i o n m e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( T u k e y ’ s t t e s t ; F = 2 1 0 ; d f = 4 8 0 ; P < 0 0 0 0 1 ) W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s 3 6

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7 3 9 b 9 9 4 b 1 0 3 3 b 1 7 6 3 b 3 7 9 5 a 0 4 80 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0W a t e r 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C l E n s u r e C o n s u m p t i o n ( m g ) F i g u r e 3 3 M e a n ( l i q u i d n o c h o i c e ) c o n s u m p t i o n ( 2 4 h ) i n t e s t a r e n a s f o r e a c h t e s t e d p a t i e n t c a r e f l u i d u s e d i n h o s p i t a l b y s t a r v e d ( 3 d ) P h a r a o h a n t c o l o n i e s ( n = 2 5 0 ) M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( T u k e y ’ s t t e s t ; F = 1 3 9 7 ; d f = 4 8 0 ; P < 0 0 0 0 1 ) ; d a t a w e r e l o g a r i t h m i c t r a n s f o r m e d b e f o r e a n a l y s i s W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s . 3 7

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3 8 C H A P T E R 4 C H O I C E L I Q U I D T E S T I n t r o d u c t i o n M a n y s o c i a l i n s e c t s a r e o m n i v o r e s c o n s u m i n g a w i d e v a r i e t y o f f o o d s f r o m m u l t i p l e n u t r i t i o n a l c l a s s i f i c a t i o n s T h i s i s e s p e c i a l l y t r u e o f h y m e n o p t e r a n c o l o n i e s W i t h i n t h e h y m e n o p t e r a o m n i v o r o u s d i e t s a r e f r e q u e n t l y s e e n i n m a n y s p e c i e s o f a n t s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) T h i s o m n i v o r o u s d i e t d i f f e r e n t i a t e s t h e m f r o m o t h e r s o c i a l i n s e c t s s u c h a s t e r m i t e s w h i c h a r e s p e c i a l i z e d t o f e e d u p o n f o o d s t h a t a r e l i m i t e d i n t h e i r a v a i l a b i l i t y e i t h e r n u t r i t i o n a l l y ( d e a d w o o d i n t h e c a s e o f t e r m i t e s ) o r s e a s o n a l l y ( p o l l e n a n d n e c t a r i n t h e c a s e o f b e e s ) ( W h e e l e r 1 9 1 0 ) L i k e m o s t o t h e r t r a m p a n t s p e c i e s P h a r a o h a n t s f e e d o n a w i d e v a r i e t y o f e n e r g y r i c h f o o d s ( W h e e l e r 1 9 1 0 ) t h a t i n c l u d e i n s e c t s n e c t a r f r u i t d e a d a n i m a l s s w e e t s p a s t r i e s m e a t j e l l y ( P e a c o c k e t a l 1 9 5 0 S u d d 1 9 6 0 ) D i s t u r b i n g l y P h a r a o h a n t s a l s o f e e d o n h u m a n s ( B e a t s o n 1 9 7 2 E d w a r d s a n d S h o r t 1 9 9 0 E i c h l e r 1 9 9 0 ) N u t r i t i o n a l n e e d s o f s o c i a l i n s e c t s a r e c o m p l e x a n d v a r i e d d u e t o t h e s t r u c t u r e o f t h e i r c o l o n i e s A n t c o l o n i e s c o n t a i n d i f f e r e n t c a s t e s a n d d e v e l o p m e n t s t a g e s : w o r k e r s a n d r e p r o d u c t i v e s e g g s l a r v a e a n d p u p a e ; a l l s t a g e s e x c e p t e g g s a n d p u p a e h a v e n u t r i t i o n a l n e e d s a n d c o n s u m e f o o d N u t r i t i o n a l n e e d s d i f f e r a m o n g c a s t e s a n d s t a g e s : f o r e x a m p l e q u e e n s n e e d m o r e p r o t e i n f o r e g g l a y i n g l a r v a e n e e d p r o t e i n t o c o m p l e t e d e v e l o p m e n t a n d w o r k e r s r e q u i r e m o r e c a r b o h y d r a t e s t o s u p p l y e n e r g y f o r f o r a g i n g a n d t e n d i n g o f b r o o d a n d q u e e n s ( H l l d o b l e r a n d W i l s o n 1 9 9 0 ) T h e n u t r i t i o n a l n e e d s o f t h e c o l o n y r e f l e c t t h e c o m b i n a t i o n o f a l l i n d i v i d u a l m e m b e r s ’ n u t r i t i o n a l n e e d s w i t h i n t h e

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3 9 c o l o n y C a s s i l l a n d T s c h i n k e l ( 1 9 9 9 ) r e p o r t e d t h a t t h e o v e r l a p p i n g n u t r i t i o n a l n e e d s o f d i f f e r e n t c a s t e m e m b e r s d e t e r m i n e s f o r a g i n g a n d n u r s i n g b e h a v i o r b y w o r k e r a n t s w i t h i n t h e c o l o n y A n i n t e r e s t i n g a s p e c t o f P h a r a o h a n t s i s t h e i r a b i l i t y t o c h a n g e t h e i r c o n s u m p t i o n t o a d j u s t f o r t h e n u t r i t i o n a l n e e d o f t h e c o l o n y E d w a r d s a n d A b r a h a m ( 1 9 9 0 ) d e m o n s t r a t e d t h a t P h a r a o h a n t s a l t e r n a t e b e t w e e n h i g h p r o t e i n a n d h i g h c a r b o h y d r a t e f o o d s T h e y c o n c l u d e d t h a t f o o d s c o n t a i n i n g b o t h o f t h e s e n u t r i e n t s w o u l d p r o v e m o r e a t t r a c t i v e a s P h a r a o h a n t b a i t m a t r i c e s s i n c e i n c l u s i o n o f b o t h s i m u l t a n e o u s l y w o u l d p r o v i d e t h e a n t s w i t h b o t h h i g h l y r e c r u i t e d n u t r i e n t s T h e y a l s o n o t e d t h a t P h a r a o h a n t c o l o n i e s w h i c h h a v e s a t i a t e d u p o n o n e n u t r i e n t ( e g c a r b o h y d r a t e s ) m i g h t r e c r u i t m o r e t o t h e o t h e r m a j o r n u t r i e n t s u c h a s p r o t e i n B e c a u s e P h a r a o h a n t s v a r y t h e i r d i e t t h e i r f o r a g i n g a n d f e e d i n g b e h a v i o r i s a n e s p e c i a l l y i m p o r t a n t c o n s i d e r a t i o n w h e n a l t e r n a t i v e f o o d s a r e a v a i l a b l e f o r c o n s u m p t i o n I n h o s p i t a l s a w i d e v a r i e t y o f f o o d i s a v a i l a b l e t h r o u g h o u t t h e h o s p i t a l i n k i t c h e n s a n d c a f e t e r i a s i n f o o d c a r t s w h e e l e d t h r o u g h t h e h o s p i t a l a n d i n s n a c k v e n d i n g m a c h i n e s I n a d d i t i o n P h a r a o h a n t s h a v e a c c e s s t o u n i q u e n u t r i t i o n a l f l u i d s s u c h a s h u m a n b o d y f l u i d s a n d f l u i d s u s e d i n p a t i e n t c a r e a v a i l a b l e t o t h e m i n h e a l t h c a r e e n v i r o n m e n t s P h a r a o h a n t s h a v e b e e n o b s e r v e d f e e d i n g a r o u n d w o u n d s a n d b o d y o p e n i n g s w h i c h m i g h t h a v e b e e n l e a k i n g b o d y f l u i d ( E i c h l e r 1 9 9 0 ) P h a r a o h a n t c o n s u m p t i o n o f f l u i d s a s s o c i a t e d w i t h h o s p i t a l p a t i e n t s w a s c o n f i r m e d i n a n o c h o i c e b i o a s s a y ( C h a p t e r 3 ) B e c a u s e p a t i e n t c a r e f l u i d s a r e u s e d t h r o u g h o u t t h e h o s p i t a l f u r t h e r e x a m i n a t i o n w a s d e e m e d n e c e s s a r y t o d e t e r m i n e h o w t h e s i m u l t a n e o u s a v a i l a b i l i t y o f t h e s e p a t i e n t c a r e f l u i d s m i g h t a f f e c t P h a r a o h a n t c o n s u m p t i o n

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4 0 S p e c i f i c a l l y t h e o b j e c t i v e w a s t o d e t e r m i n e t h e p r e f e r e n c e o f s m a l l c o l o n i e s o f P h a r a o h a n t s f o r f i v e l i q u i d p a t i e n t c a r e f l u i d s u s i n g a c h o i c e t e s t M a t e r i a l s a n d M e t h o d s A n t s P h a r a o h a n t t e s t c o l o n i e s w e r e e x t r a c t e d f r o m l a r g e p a r e n t c o l o n i e s h o u s e d a n d r e a r e d a t t h e U r b a n E n t o m o l o g y L a b o r a t o r y U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F L R e a r i n g t e m p e r a t u r e s r a n g e d f r o m 2 1 9 C – 2 9 4 C w i t h r e l a t i v e h u m i d i t y b e t w e e n 2 0 5 3 % ( A p p e n d i x A ) C o l o n i e s c o n s i s t e d o f 2 5 0 w o r k e r s 3 d e a l a t e q u e e n s a n d a p p r o x i m a t e l y 5 0 m g b r o o d O n e h u n d r e d t w e n t y f i v e w o r k e r s w e r e s e l e c t e d f r o m c e l l s t o e n s u r e t h e p r e s e n c e o f n u r s e s a n d 1 2 5 w o r k e r s w e r e s e l e c t e d f r o m o u t s i d e c e l l s t o e n s u r e t h e p r e s e n c e o f f o r a g e r s ( H a a c k 1 9 8 7 H a a c k a n d V i n s o n 1 9 9 0 H a a c k e t a l 1 9 9 5 ) B r o o d i n c l u d e d e g g s a l l l a r v a l s t a g e s a n d p u p a e S e e A p p e n d i x B f o r f u l l d e t a i l s o n s e l e c t i o n o f a n t s a n d p r e p a r a t i o n o f e x p e r i m e n t a l t e s t c o l o n i e s F o r a g i n g A r e n a F o r a g i n g a r e n a s c o n t a i n e d o n e n e s t i n g c e l l o n e w a t e r v i a l a n d f i v e m i c r o c a p i l l a r y t u b e s w i t h f l u i d s ( F i g u r e 4 1 ) T h e s e a r e n a s w e r e i d e n t i c a l t o t h o s e u s e d i n t h e p r e v i o u s n o c h o i c e l i q u i d c o n s u m p t i o n t e s t ( C h a p t e r 3 ) e x c e p t f o r t h e a d d i t i o n o f f o u r m i c r o c a p i l l a r y t u b e s t o p e r m i t f l u i d a v a i l a b i l i t y A d e t a i l e d d e s c r i p t i o n o f t h e f o r a g i n g a r e n a w a s p r o v i d e d i n C h a p t e r 3

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4 1 2 7 c m 1 9 c m F i v e 3 0 l c a p i l l a r y t u b e s w i t h f l u i d s N e s t i n g c e l l ( 5 0 m m d i a m e t e r ; 1 5 m m d e e p ) W a t e r V i a l ( 1 5 m l ) F i g u r e 4 1 A r e n a s w i t h w a t e r v i a l a n d n e s t i n g c e l l s p l a c e d i n d i a g o n a l l y o p p o s i t e c o r n e r s C a p i l l a r y t u b e s p l a c e d i n c e n t e r T e s t a r e n a s ( w i t h a n t s ) a n d c o n t r o l a r e n a s ( n o a n t s ) w e r e p l a c e d a d j a c e n t t o e a c h o t h e r a n d c o m p r i s e d a t e s t p a i r T e s t a r e n a s c o n t a i n e d 1 2 5 i n s i d e a n t s 1 2 5 o u t s i d e a n t s 3 q u e e n s a n d ~ 5 0 m g b r o o d ( i n c l u d i n g a l l s t a g e s o f b r o o d )

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4 2 W a t e r w a s p r o v i d e d a d l i b i t u m i n a 1 5 m l v i a l ( M o d e l N o 5 5 9 T h o r n t o n P l a s t i c s S a l t L a k e C i t y U T ) w i t h t h r e e l a r g e c o t t o n b a l l s i n s e r t e d i n t o t o p t o p r e v e n t l e a k a g e a n d t o p r o v i d e e a s y a c c e s s f o r a n t s A w a t e r v i a l w a s p l a c e d i n t h e o p p o s i t e d i a g o n a l c o r n e r o f a r e n a a c r o s s f r o m t h e n e s t i n g c e l l E a c h r e p l i c a t e c o n s i s t e d o f o n e t e s t a r e n a p l a c e d a d j a c e n t t o o n e c o n t r o l a r e n a T e s t a r e n a s c o n t a i n e d a n t s w h i l e c o n t r o l a r e n a s l a c k e d a n t s C o n t r o l a r e n a d a t a w a s u s e d t o q u a n t i f y a n d a d j u s t f o r e v a p o r a t i o n o f f l u i d s E a c h a r e n a w a s c o v e r e d w i t h a c l e a r p l a s t i c l i d t o m a i n t a i n h u m i d i t y a n d t e m p e r a t u r e E x p e r i m e n t a l F l u i d s T h e s a m e f i v e f l u i d s u s e d i n t h e n o c h o i c e t e s t i n C h a p t e r 3 w e r e u s e d i n t h i s c h o i c e t e s t F l u i d s c o n s i s t e d o f h u m a n b l o o d c e l l s h u m a n p l a s m a s a l i n e ( 0 9 % s o d i u m c h l o r i d e ) ; 5 % d e x t r o s e ( D g l u c o s e ) a n d E n s u r e ( V a n i l l a P l u s R o s s P r o d u c t s D i v i s i o n A b b o t t L a b o r a t o r i e s C o l u m b u s O H ) F l u i d s w e r e c h o s e n t o r e p l i c a t e h o s p i t a l c o n d i t i o n s a n d p o t e n t i a l f l u i d s o u r c e s a s c l o s e l y a s p o s s i b l e T h e d e t a i l e d d e s c r i p t i o n s o f e a c h f l u i d a n d h a n d l i n g p r o c e d u r e s u s e d i n t h i s s t u d y a r e i d e n t i c a l t o t h a t p r o v i d e d i n C h a p t e r 3 E x p e r i m e n t a l D e s i g n T h e e x p e r i m e n t w a s d e s i g n e d a s a r a n d o m i z e d c o m p l e t e b l o c k b l o c k i n g o n c o l o n y E a c h b l o c k c o n s i s t e d o f o n e P h a r a o h a n t c o l o n y ( 1 2 5 i n s i d e w o r k e r s 1 2 5 o u t s i d e w o r k e r s 3 d e a l a t e q u e e n s a n d ~ 5 0 m g b r o o d ) o b t a i n e d f r o m a s i n g l e p a r e n t c o l o n y F i v e f l u i d s w e r e s i m u l t a n e o u s l y o f f e r e d t o e a c h c o l o n y t o d e t e r m i n e t h e c o n s u m p t i o n a m o n g t h e t e s t f l u i d s u n d e r a c h o i c e e x p o s u r e A s i n t h e p r e v i o u s n o c h o i c e e x p e r i m e n t ( C h a p t e r 3 ) a n e x p e r i m e n t a l u n i t c o n s i s t e d o f a p a i r e d t e s t a n d c o n t r o l a r e n a l o c a t e d a d j a c e n t t o o n e a n o t h e r T h e c h o i c e t e s t w a s r e p l i c a t e d 2 1 t i m e s A n t s w e r e s e l e c t e d f r o m 6 d i f f e r e n t p a r e n t c o l o n i e s F l u i d s w e r e t e s t e d o v e r a t w o w e e k p e r i o d f r o m M a y

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4 3 2 1 2 0 0 3 t o J u n e 3 2 0 0 3 ; r o o m t e m p e r a t u r e a n d r e l a t i v e h u m i d i t y w e r e m a i n t a i n e d b e t w e e n 2 4 9 2 8 6 C a n d 3 5 5 7 % R H P h a r a o h a n t c o l o n i e s w e r e s t a r v e d f o r 4 8 h ; w a t e r w a s a v a i l a b l e a d l i b i t u m i n a l l a r e n a s i n c l u d i n g w a t e r c o n t r o l s v i a a 1 5 m l c o t t o n s t o p p e r e d v i a l d u r i n g s t a r v a t i o n p e r i o d ( H a a c k a n d V i n s o n 1 9 9 0 H a a c k e t a l 1 9 9 5 ) A f t e r 4 8 h o f s t a r v a t i o n f i v e m i c r o c a p i l l a r y t u b e s e a c h c o n t a i n i n g o n e t e s t f l u i d w e r e a d d e d t o e a c h a r e n a a n d t h e w a t e r v i a l w a s n o t r e m o v e d T h u s w a t e r w a s a l w a y s a v a i l a b l e i n a l l a r e n a s F l u i d s w e r e o f f e r e d s i m u l t a n e o u s l y t o e a c h c o l o n y t o d e t e r m i n e c o n s u m p t i o n p r e f e r e n c e a m o n g p r o v i d e d f l u i d s M o r t a l i t y a f t e r s t a r v a t i o n p e r i o d a n d p r i o r t o 2 4 h f e e d i n g p e r i o d i n t e s t a r e n a s w a s o b s e r v e d b u t n o t r e c o r d e d D e a d a n t s w e r e n e i t h e r c o u n t e d n o r r e m o v e d M o r t a l i t y r a t e s d u r i n g s t a r v a t i o n p e r i o d s w e r e n o t t a k e n i n t o c o n s i d e r a t i o n w h e n r e c o r d i n g c o n s u m p t i o n f r o m t e s t a r e n a s F l u i d M e a s u r e m e n t s F l u i d p l a c e m e n t i n t o a r e n a s w a s i d e n t i c a l t o t h e p r e v i o u s n o c h o i c e t e s t ( C h a p t e r 3 ) e x c e p t t h a t f i v e d i f f e r e n t f l u i d s w e r e s i m u l t a n e o u s l y p r o v i d e d e a c h i n i t s o w n m i c r o c a p i l l a r y t u b e I n b r i e f e a c h m i c r o c a p i l l a r y t u b e ( M o d e l N o 2 1 3 8 0 9 C F i s h e r S c i e n t i f i c P i t t s b u r g h P A ) w a s w e i g h e d b e f o r e a n d a f t e r t h e l i q u i d w a s l o a d e d T h e a m o u n t o f l i q u i d p r o v i d e d t o a l l c o n t r o l a n d t e s t a r e n a s w a s c a l c u l a t e d f o r e a c h a r e n a b y s u b t r a c t i n g t h e m a s s o f t h e e m p t y m i c r o c a p i l l a r y t u b e f r o m t h e m a s s o f t h e t u b e c o n t a i n i n g i n i t i a l m a s s o f p r o v i d e d f l u i d ( W e i a n d J o h n s o n 1 9 9 6 ) A n t s w e r e a l l o w e d t o f e e d f o r 2 4 h M i c r o c a p i l l a r y t u b e s w e r e t h e n r e w e i g h e d i n b o t h c o n t r o l a n d t e s t a r e n a s D u e t o f r e q u e n t p r e s e n c e o f a n t s s t i l l i n m i c r o c a p i l l a r y t u b e s d u r i n g f i n a l g r a v i m e t r i c m e a s u r e m e n t o n a d i g i t a l s c a l e w e i g h t s o f p o s t c o n s u m p t i o n t e s t

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4 4 f l u i d a f t e r 2 4 h o u r t e s t p e r i o d i n c l u d e d t h e m a s s o f a n t s r e m a i n i n g i n m i c r o c a p i l l a r y t u b e s F i n a l a n t b i o m a s s m e a s u r e m e n t s w e r e i n c l u d e d i n f i n a l m a s s w e i g h t s r e c o r d e d a n d w e r e s u b t r a c t e d l a t e r u s i n g a n a v e r a g e m a s s v a l u e o f 0 0 0 0 2 g p e r P h a r a o h a n t w o r k e r W o r k e r a n t m a s s w a s d e t e r m i n e d b y w e i g h i n g s e v e r a l i n d i v i d u a l w o r k e r a n t s t o d e t e r m i n e a v e r a g e m a s s p e r a n t F l u i d l o s s i n c o n t r o l a r e n a s w a s c a l c u l a t e d b y s u b t r a c t i n g a m o u n t o f f l u i d i n i t i a l l y p r o v i d e d f r o m a m o u n t o f f l u i d r e m a i n i n g r e s u l t i n g i n t h e a m o u n t o f e v a p o r a t i o n t h a t o c c u r r e d i n t h e 2 4 h t e s t p e r i o d F l u i d l o s s i n t e s t a r e n a s w a s c o m p r i s e d o f c o n s u m p t i o n a n d e v a p o r a t i o n ; t h e r e f o r e t o d e t e r m i n e c o n s u m e d q u a n t i t i e s t h e a m o u n t o f f l u i d l o s t t o e v a p o r a t i o n w a s s u b t r a c t e d M a n y r e s e a r c h e r s h a v e d e s i g n e d f o r m u l a s t o a d j u s t c o n s u m p t i o n f o r f l u i d l o s s a n d W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a w a s s e l e c t e d ( A p p e n d i x C ) B e c a u s e m u l t i p l e a n t s r e m a i n e d i n t e s t a r e n a f l u i d s W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a w a s m o d i f i e d t o a c c o u n t f o r m a s s o f a n t s r e m a i n i n g i n f l u i d T h u s c o r r e c t e d c o n s u m p t i o n w a s d e t e r m i n e d a s f o l l o w s : C = ( 1 – [ Y / G ] ) ( W [ ( L – A ) + ( Y / H ) ( L – A ) ] ) w h e r e C = c o r r e c t e d c o n s u m p t i o n W = m a s s o f f l u i d p r o v i d e d ( g ) i n t e s t a r e n a L = m a s s o f u n e a t e n f l u i d a n d a n t s ( g ) i n t e s t a r e n a A = n ( 0 0 0 0 2 g / a n t ) m a s s o f a n t s s t u c k i n t u b e n = n u m b e r o f a n t s s t u c k i n t u b e t h u s ( L A ) = m a s s o f u n e a t e n f l u i d G = m a s s o f f l u i d p r o v i d e d ( g ) i n c o n t r o l a r e n a H = m a s s o f f l u i d r e m a i n i n g ( g ) i n c o n t r o l a r e n a

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4 5 Y = G – H = w e i g h t l o s s ( g ) o f c o n t r o l f l u i d Y / G = w e i g h t l o s s o f c o n t r o l f l u i d / i n i t i a l m a s s o f c o n t r o l f l u i d ( = a i n W a l d b a u e r 1 9 6 8 ) Y / H = w e i g h t l o s s o f c o n t r o l f l u i d / f i n a l m a s s o f c o n t r o l f l u i d ( = b i n W a l d b a u e r 1 9 6 8 ) S t a t i s t i c a l A n a l y s i s D e a d o r a l i v e a n t s r e m a i n i n g i n m i c r o c a p i l l a r y t u b e s a t t h e c o n c l u s i o n o f t h e 2 4 h t e s t p e r i o d w e r e c o m p a r e d a m o n g t e s t f l u i d s M e a n n u m b e r ( n ) o f a n t s r e m a i n i n g i n t e s t f l u i d s w a s c o m p a r e d v i a a n a l y s i s o f v a r i a n c e a n d T u k e y ’ s H S D t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) F l u i d l o s s f r o m e v a p o r a t i o n i n t h e c o n t r o l a r e n a s a n d f l u i d l o s s f r o m e v a p o r a t i o n a n d c o n s u m p t i o n i n t h e t e s t a r e n a s a m o n g t y p e s o f h o s p i t a l f l u i d s w e r e c o m p a r e d b y a n a l y s i s o f v a r i a n c e a n d T u k e y ’ s H S D ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) V a r i a n c e s w e r e t e s t e d u s i n g L e v e n e ’ s H o m o g e n e i t y o f V a r i a n c e s t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) N e g a t i v e c o n s u m p t i o n v a l u e s ( 1 5 o f 1 0 0 v a l u e s ) w e r e t r u n c a t e d t o z e r o C o n s u m p t i o n o f p a t i e n t c a r e f l u i d s a n d w a t e r w a s a n a l y z e d v i a a o n e t a i l e d S t u d e n t ’ s t t e s t t o d e t e r m i n e i f t h e r e w a s s i g n i f i c a n t c o n s u m p t i o n ( H o : = 0 ) C o n s u m p t i o n d a t a w e r e c o m p a r e d v i a a n a l y s i s o f v a r i a n c e a n d m e a n s w e r e s e p a r a t e d u s i n g T u k e y ’ s H S D t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) H o m o g e n e i t y o f v a r i a n c e s f o r c o n s u m p t i o n a m o n g t h e d i f f e r e n t f l u i d s w a s t e s t e d w i t h L e v e n e ’ s T e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) R e s u l t s N u m b e r o f A n t s A n t s w e r e o b s e r v e d e n t e r i n g m i c r o c a p i l l a r y t u b e s w i t h i n 5 m i n o f p l a c i n g f l u i d s i n t e s t a r e n a s O b s e r v a t i o n s o f a n t b e h a v i o r i n t h i s e x p e r i m e n t w e r e s i m i l a r t o t h o s e s e e n i n C h a p t e r 3

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4 6 M e a n n u m b e r o f a n t s r e m a i n i n g i n E n s u r e ( 5 7 7 + 0 9 6 ) a t t h e c o n c l u s i o n o f t h e t e s t p e r i o d w a s s i g n i f i c a n t l y h i g h e r ( F = 1 2 4 0 ; d f = 4 9 5 ; P < 0 0 0 0 1 ) t h a n i n t h e o t h e r f l u i d s ( T a b l e 4 1 ) N u m b e r s o f a n t s r e m a i n i n g i n a l l o t h e r f l u i d s w e r e l o w a n d d i d n o t s i g n i f i c a n t l y d i f f e r r a n g i n g f r o m 0 7 1 t o 1 9 4 ( F i g 4 2 ) F l u i d M e a s u r e m e n t s I n i t i a l q u a n t i t i e s o f f l u i d s p r o v i d e d t o t e s t a n d c o n t r o l a r e n a s d i f f e r e d s i g n i f i c a n t l y ( F = 2 5 6 9 d f = 4 9 5 ; P < 0 0 0 0 1 ) a m o n g t h e t e s t e d p a t i e n t c a r e f l u i d s T h e m a s s o f E n s u r e ( ~ 7 7 m g ) p l a c e d i n t o c o n t r o l a r e n a s w a s h i g h e r t h a n t h a t o f a l l o t h e r f l u i d s T h e m a s s o f b l o o d ( ~ 7 6 m g ) w a s s i g n i f i c a n t l y h i g h e r t h a n t h a t o f p l a s m a d e x t r o s e a n d s a l i n e T h e m a s s o f p r o v i d e d s a l i n e ( ~ 6 9 m g ) w a s l e s s t h a n t h a t o f a l l o t h e r f l u i d s i n c o n t r o l a r e n a s b u t n o t i n t e s t a r e n a s ( T a b l e 4 2 ) I n t e s t a r e n a s i n i t i a l a m o u n t s o f f l u i d s p r o v i d e d w e r e a l s o s i g n i f i c a n t l y d i f f e r e n t ( F = 1 3 3 3 9 ; d f = 4 9 5 ; P < 0 0 0 0 1 ) A s i n c o n t r o l a r e n a s s i g n i f i c a n t l y m o r e E n s u r e b y m a s s w a s p l a c e d i n t o a r e n a s t h a n a l l o t h e r f l u i d s t h e m a s s o f b l o o d p r o v i d e d w a s s i g n i f i c a n t l y h i g h e r t h a n t h a t o f p l a s m a d e x t r o s e a n d s a l i n e a n d t h e m a s s o f p r o v i d e d s a l i n e w a s s i g n i f i c a n t l y l e s s t h a n t h a t o f a l l o t h e r f l u i d s ( T a b l e 4 2 ) E v a p o r a t i o n i n c o n t r o l a r e n a s w a s s i g n i f i c a n t l y d i f f e r e n t b e t w e e n f l u i d s ( F = 4 8 5 ; d f = 4 9 5 ; P = 0 0 0 1 3 ) P l a s m a h a d t h e h i g h e s t f l u i d l o s s d u e t o e v a p o r a t i o n w h i l e s a l i n e a n d d e x t r o s e h a d t h e l e a s t a m o u n t o f f l u i d l o s s d u e t o e v a p o r a t i o n ( T a b l e 4 2 ) F l u i d l o s s o f t e s t e d p a t i e n t c a r e f l u i d s i n t e s t a r e n a s w a s s i g n i f i c a n t l y d i f f e r e n t d u e t o c o m b i n e d e v a p o r a t i o n a n d c o n s u m p t i o n ( F = 2 9 0 1 ; d f = 4 9 5 ; P < 0 0 0 0 1 ) W h e n a n t s w e r e p r e s e n t s i g n i f i c a n t l y m o r e d e x t r o s e ( ~ 7 7 m g ) w a s l o s t t h a n a n y o t h e r f l u i d E n s u r e l o s t m o r e f l u i d t h a n p l a s m a s a l i n e a n d b l o o d S a l i n e a n d b l o o d h a d t h e s m a l l e s t a m o u n t o f f l u i d l o s s ( T a b l e 4 – 2 ) F l u i d l o s s i n c o n t r o l a r e n a s w a s d u e s o l e l y t o

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4 7 e v a p o r a t i o n a n d r a n g e d f r o m 2 5 9 0 1 9 m g f o r d e x t r o s e t o 4 8 7 + 0 6 8 m g f o r p l a s m a I n c o n t r a s t f l u i d l o s s i n t e s t a r e n a s r a n g e d f r o m 5 2 0 0 5 8 m g f o r b l o o d t o 5 0 8 0 3 4 6 m g f o r d e x t r o s e ( F i g 4 3 T a b l e 4 2 ) T e m p e r a t u r e s i n t e s t a r e n a s ( 2 7 3 6 + 0 3 1 C ) w e r e s t a t i s t i c a l l y s i g n i f i c a n t l y h i g h e r ( F = 4 5 8 ; d f = 1 2 4 ; P = 0 0 4 2 8 ) t h a n i n c o n t r o l a r e n a s ( 2 6 3 8 + 0 3 4 C ) p r o b a b l y d u e t o t h e p r e s e n c e o f a n t s C o n s u m p t i o n T h e r e w e r e s i g n i f i c a n t d i f f e r e n c e s ( F = 2 9 9 7 ; d f = 4 9 5 ; P < 0 0 0 0 1 ) i n c o n s u m p t i o n o f t h e f l u i d s b y P h a r a o h a n t s C o n s u m p t i o n o f 5 % d e x t r o s e ( 4 9 1 4 + 3 5 1 m g ) w a s s i g n i f i c a n t l y h i g h e r t h a n c o n s u m p t i o n o f a l l o t h e r f l u i d s C o n s u m p t i o n o f E n s u r e ( 1 6 0 5 + 6 2 3 m g ) w a s s i g n i f i c a n t l y g r e a t e r t h a n c o n s u m p t i o n o f p l a s m a ( 9 3 1 + 3 1 7 m g ) s a l i n e ( 3 0 9 + 1 1 4 m g ) a n d b l o o d c e l l s ( 2 4 6 + 0 4 8 m g ) ( T a b l e 4 2 F i g 4 3 ) B a s e l i n e m e a n c o n s u m p t i o n o f w a t e r ( 0 4 8 + 0 0 7 m g ) w a s l o w e r t h a n t h e c o n s u m p t i o n o f a l l t e s t f l u i d s N o n e o f t h e f l u i d s i n a n y o f t h e m i c r o c a p i l l a r y t u b e s i n e i t h e r t h e t e s t o r c o n t r o l a r e n a s w e r e c o m p l e t e l y d e p l e t e d D i s c u s s i o n N u m b e r o f A n t s N u m b e r s o f a n t s r e m a i n i n g i n m i c r o c a p i l l a r y t u b e s a t c o n c l u s i o n o f t e s t p e r i o d d i d n o t c o r r e l a t e t o h e a v i e s t c o n s u m p t i o n o r t o o b s e r v e d f o r a g i n g b e h a v i o r A n t s r e m a i n i n g i n m i c r o c a p i l l a r y t u b e s a p p e a r e d t o b e f e e d i n g o n r e m a i n i n g f l u i d F o r a g i n g a c t i v i t y o b s e r v e d d u r i n g t h e f i r s t h o u r o f t h e t e s t p e r i o d w a s o f l a r g e s t r e c r u i t m e n t t o t h e m i c r o c a p i l l a r y t u b e s c o n t a i n i n g 5 % d e x t r o s e I n c o n t r a s t t h e E n s u r e h a d t h e h i g h e s t n u m b e r o f a n t s r e m a i n i n g i n t h e t u b e s a f t e r 2 4 h T h e m e a n n u m b e r o f a n t s r e m a i n i n g i n m i c r o c a p i l l a r y t u b e s a f t e r 2 4 h w i t h i n e a c h a r e n a c o m p r i s e d 6 % o f c o l o n y w o r k e r s ( n = 2 5 0 ) ( F i g 4 1 )

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4 8 F l u i d M e a s u r e m e n t s T h e s i g n i f i c a n t d i f f e r e n c e s i n m a s s b e t w e e n d i f f e r e n t f l u i d s p r o b a b l y r e f l e c t s t h e d i f f e r e n t d e n s i t i e s o f t h e f l u i d s u s e d i n t h i s e x p e r i m e n t s i n c e a l l m i c r o c a p i l l a r y t u b e s w e r e f i l l e d t o t h e s a m e v o l u m e o f ~ 2 1 2 l H i g h e r i n i t i a l m a s s e s f o r b o t h E n s u r e a n d b l o o d w e r e a s s o c i a t e d w i t h t h e h i g h e r v i s c o s i t y o b s e r v e d i n t h e s e t w o f l u i d s S i m i l a r l y s a l i n e s o l u t i o n c o n t a i n e d o n l y 0 9 % e l e c t r o l y t e s m a k i n g i t t h e l e a s t v i s c o u s f l u i d u s e d D i f f e r i n g a m o u n t s o f p r o v i d e d f l u i d d i d n o t a f f e c t c o n s u m p t i o n s i n c e n o f l u i d w a s d e p l e t e d D i f f e r i n g a m o u n t s o f p r o v i d e d f l u i d ( T a b l e 4 2 ) m i g h t h a v e r e s u l t e d i n d i f f e r e n t r a t e s o f f l u i d l o s s f o r d i f f e r e n t f l u i d s A l t h o u g h E n s u r e h a d s i g n i f i c a n t l y m o r e f l u i d m a s s p r o v i d e d i n b o t h c o n t r o l a n d t e s t a r e n a s t h a n o t h e r f l u i d s e v a p o r a t i o n o f E n s u r e i n c o n t r o l a r e n a s d i d n o t d i f f e r f r o m t h a t o f o t h e r t e s t f l u i d s F l u i d l o s s i n t e s t a r e n a s ( T a b l e 4 2 ) w a s c o m p r i s e d o f b o t h c o n s u m p t i o n a n d e v a p o r a t i o n ; t h e r e f o r e t h e a m o u n t o f f l u i d l o s t t o e v a p o r a t i o n h a d t o b e s u b t r a c t e d i n o r d e r t o d e t e r m i n e c o n s u m p t i o n v a l u e s A l t h o u g h t e m p e r a t u r e s w i t h i n t e s t a r e n a s w e r e s t a t i s t i c a l l y s i g n i f i c a n t l y h i g h e r t h a n i n c o n t r o l a r e n a s b o t h m e a n s w e r e w i t h i n 1 C o f t e m p e r a t u r e s r e p o r t e d a s i d e a l f o r P h a r a o h a n t s ( P e a c o c k a n d B a x t e r 1 9 4 9 P e a c o c k e t a l 1 9 5 5 c ) H e n c e r e l a t i v e t o t e m p e r a t u r e f o r a g i n g w a s p r o b a b l y o p t i m a l i n t h i s e x p e r i m e n t H u m i d i t y d i d n o t d i f f e r b e t w e e n c o n t r o l a n d t e s t a r e n a s ; h o w e v e r t h e h u m i d i t y l e v e l s i n s i d e a r e n a s o f ~ 5 2 % w e r e w e l l b e l o w t h e 8 0 % r e c o m m e n d e d b y ( P e a c o c k a n d B a x t e r 1 9 4 9 P e a c o c k e t a l 1 9 5 5 c ) a n d a l s o l o w e r t h a n t h e 6 5 % h u m i d i t y r e c o m m e n d e d b y ( H a a c k 1 9 8 7 ) a s o p t i m a l f o r t h e P h a r a o h a n t L o w h u m i d i t y p r o b a b l y i n c r e a s e d t h e m o i s t u r e n e e d s o f t h e t e s t c o l o n i e s a n d p r o b a b l y i n c r e a s e d t h e r a t e o f e v a p o r a t i o n o f a l l f l u i d s I f l o w h u m i d i t y

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4 9 r e s u l t e d i n a n i n c r e a s e d m o i s t u r e r e q u i r e m e n t t h e n t h e c o n s u m p t i o n o f f l u i d s m a y h a v e i n c r e a s e d C o n s u m p t i o n ( M a i l l e u x e t a l 2 0 0 0 ) d e t e r m i n e d t h a t a n t f o r a g i n g b e h a v i o r i s a r e s p o n s e t o t h e n u t r i t i o n a l d e m a n d s o f t h e a n t s a n d t o t h e c h a r a c t e r i s t i c s o f t h e f o o d r e s o u r c e s s u c h a s t h e d e n s i t y a n d s p a t i a l d i s t r i b u t i o n d i s t a n c e t o f o o d a n d t h e q u a l i t y o f f o o d C o n s u m p t i o n o f h i g h c a r b o h y d r a t e a n d h i g h p r o t e i n f l u i d s w a s e x p e c t e d s i n c e a n u t r i t i o n a l n e e d w a s a s s u m e d t o b e i n d u c e d b y a 3 d s t a r v a t i o n p e r i o d F o o d c h a r a c t e r i s t i c s a f f e c t i n g a n t f o r a g i n g b e h a v i o r v a r i e d o n l y i n t h e n u t r i t i o n a l c o m p o s i t i o n a n d q u a l i t y o f t h e d i f f e r e n t f l u i d s o f f e r e d s i n c e a l l m i c r o c a p i l l a r y t u b e s w e r e p l a c e d r a n d o m l y i n r e p l i c a t i o n s ( M a i l l e u x e t a l 2 0 0 0 ) I n t h i s e x p e r i m e n t a l l f l u i d s w e r e c o n s u m e d i n s i g n i f i c a n t p r o p o r t i o n s i n d i c a t i n g t h a t t h e a n t s c o n s u m e d a w i d e v a r i e t y o f f o o d s w h e n p r o v i d e d w i t h a c h o i c e T h i s a g r e e s w i t h o b s e r v a t i o n s t h a t P h a r a o h a n t s c h a n g e f o o d s e l e c t i o n s b a s e d o n s a t i e t y a n d p r e f e r e n c e a n d t h a t t h e y w i l l e v e n a l t e r n a t e b e t w e e n h i g h c a r b o h y d r a t e a n d h i g h p r o t e i n f o o d s o u r c e s ( G r a n o v s k y a n d H o w e l l 1 9 8 3 E d w a r d s a n d A b r a h a m 1 9 9 0 ) C o n s u m p t i o n a l s o c a n b e i n f l u e n c e d b y c o l o n y n e e d s a n d c a n c h a n g e d e p e n d i n g o n a v a r i e t y o f f a c t o r s s u c h a s c o l o n y s i z e p r o p o r t i o n o f l a r v a e t o w o r k e r s a n d n u m b e r s o f e g g l a y i n g q u e e n s F o r e x a m p l e e g g l a y i n g b y q u e e n s a n d l a r g e r n u m b e r s o f l a r v a e b o t h i n c r e a s e t h e c o l o n y ’ s n e e d f o r p r o t e i n s I n t h e r e d i m p o r t e d f i r e a n t S o l e n o p s i s i n v i c t a ( B u r e n ) e v e n w o r k e r s i z e o r c r o p c o n t e n t s c a n h a v e a n i m p a c t o n f o o d f l o w w i t h i n a c o l o n y ’ s m e m b e r s ( C a s s i l l a n d T s c h i n k e l 1 9 9 9 )

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5 0 D e x t r o s e P h a r a o h a n t s h a v e b e e n r e p o r t e d t o f o r a g i n g m o s t s t r o n g l y t o f l u i d s w i t h h i g h e s t s u g a r c o n c e n t r a t i o n s ( S u m p t e r a n d B e e k m a n 2 0 0 3 ) s u c h a s s u c r o s e s o l u t i o n s W h i l e t h e P h a r a o h a n t s c o n s u m e d a l l f l u i d s 5 % d e x t r o s e w a s c o n s u m e d s i g n i f i c a n t l y m o r e t h a n a l l o t h e r l i q u i d s ( F i g 4 3 ) M e a n c o n s u m p t i o n o f 5 % d e x t r o s e ( 4 9 1 4 + 3 5 1 m g ) w a s c o n s u m e d 3 t i m e s m o r e t h a n E n s u r e t h a n t h e n e x t m o s t c o n s u m e d f l u i d ( T a b l e 4 3 ) T h e s e r e s u l t s w e r e s i m i l a r t o t h o s e o b t a i n e d i n t h e n o c h o i c e t e s t ( C h a p t e r 3 ) H a a c k e t a l ( 1 9 9 5 ) r e p o r t e d a s i m i l a r l y h i g h P h a r a o h a n t r e c r u i t m e n t t o s i m p l e c a r b o h y d r a t e s : h e o b s e r v e d t h a t ~ 1 0 0 % o f w o r k e r s f o r a g e d a t c a r b o h y d r a t e s o u r c e w i t h i n ~ 3 h o f i n t r o d u c t i o n o f f o o d i n t o a r e n a w h i l e o n l y 2 % f o r a g e d t o p r o t e i n s w i t h i n 2 4 h a n d ~ 8 0 % f o r a g e d t o l i p i d s a f t e r 2 4 h I n h o s p i t a l s 5 % d e x t r o s e i s o f t e n s q u i r t e d o n t o f l o o r s w h e n n u r s e s o r d o c t o r s c l e a r i n t r a v e n o u s t u b i n g p r i o r t o i n s e r t i n g t u b i n g i n t o p a t i e n t s T h e a m o u n t o f t i m e t h a t m i g h t e l a p s e b e t w e e n s p i l l a g e a n d c l e a n u p i s u n k n o w n ; i f d e x t r o s e i s p r e s e n t i n t h e r o o m s t h i s e x p e r i m e n t c l e a r l y i n d i c a t e s t h a t P h a r a o h a n t s w i l l r e c r u i t t o t h i s f l u i d E n s u r e W h e n g i v e n a c h o i c e o f f l u i d s a f t e r 3 d s t a r v a t i o n P h a r a o h a n t s p r e f e r r e d h i g h s u g a r f l u i d s m o s t ; t h e h i g h c o n c e n t r a t i o n o f s u g a r s i n E n s u r e p r o b a b l y h e l p e d m a k e i t t h e s e c o n d m o s t p r e f e r r e d f l u i d i n t h i s e x p e r i m e n t d e s p i t e i t s r e l a t i v e l y h i g h e v a p o r a t i o n r a t e ( F i g 4 4 ) a n d t e n d e n c y t o d r y o u t a n d p a r t i a l l y s o l i d i f y t h u s m a k i n g i t m o r e d i f f i c u l t t o e x t r a c t t h a n t h e l e s s v i s c o u s d e x t r o s e T h e r e l a t i v e l y h i g h c o n s u m p t i o n o f E n s u r e i n t h i s c h o i c e t e s t c o n t r a s t e d w i t h t h e l e s s e r c o n s u m p t i o n o f E n s u r e i n t h e n o c h o i c e t e s t ( C h a p t e r 3 ) P h a r a o h a n t s p r e f e r r e d f l u i d s w i t h h i g h c o n c e n t r a t i o n s o f s u g a r A l t h o u g h P h a r a o h a n t c o n s u m p t i o n o f E n s u r e

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5 1 w a s s e c o n d o n l y t o c o n s u m p t i o n o f d e x t r o s e i t w a s s i g n i f i c a n t l y l e s s t h a n t h a t o f d e x t r o s e E n s u r e h a s f e w e r s i m p l e s u g a r s a n d m o r e c o m p l e x s u g a r s t h a n d e x t r o s e m a k i n g i t a m o r e c o m p l e x f l u i d I n s e c t s r a p i d l y c o n v e r t i n g e s t e d g l u c o s e i n t r e h a l o s e f o r s t o r a g e i n h e m o l y m p h a n d m u s t t a k e a n a d d i t i o n a l h i g h e n e r g y s t e p o f c o n v e r t i n g c o m p l e x s u g a r s i n t o g l u c o s e f i r s t ( N a t i o n 2 0 0 3 ) E n s u r e d i e t a r y s u p p l e m e n t i s f u r t h e r c o m p l i c a t e d s i n c e i t c o n t a i n s h i g h c o n c e n t r a t i o n s o f p r o t e i n s a n d l i p i d s i n a d d i t i o n t o s u g a r s F i n a l l y E n s u r e h a d m o r e a n t s s t u c k i n i t t h a n a l l o t h e r f l u i d s ( T a b l e 4 1 F i g 4 2 ) S o m e a n t s r e m a i n i n g i n f l u i d w e r e d e a d d u r i n g f i n a l w e i g h i n g ; S u d d ( 1 9 5 7 a ) o b s e r v e d P h a r a o h a n t s r e s p o n d i n a n e g a t i v e m a n n e r t o d e a d P h a r a o h a n t s H e r e p o r t e d t h a t w h e n a P h a r a o h a n t e n c o u n t e r e d d e a d o f i t s o w n s p e c i e s i t b e c a m e a l a r m e d a n d r a n a w a y w h e n w i t h i n 1 c m o f d e a d a n d ; h o w e v e r t h o s e a n t s w e r e k i l l e d b y c r u s h i n g I n t h i s e x p e r i m e n t m o s t a n t s r e m a i n i n g i n t h e f l u i d w e r e s t i l l a l i v e ; a n t s w h i c h w e r e d e a d h a d n o t b e e n c r u s h e d A n y c h e m i c a l s e m i t t e d v i a c r u s h i n g t h a t m a y h a v e s e r v e d a s a n a l a r m p h e r o m o n e w e r e p r o b a b l y n o t p r e s e n t i n t h i s e x p e r i m e n t T h e r e f o r e t h e p r e s e n c e o f d e a d a n t s i n t h e t u b e s w a s n o t e x p e c t e d t o i n f l u e n c e f e e d i n g b e h a v i o r V i s u a l o b s e r v a t i o n o f a n t s i n m i c r o c a p i l l a r y t u b e s d i d n o t n o t e a n y f o r a g e r e l u c t a n c e t o e n t e r t u b e s c o n t a i n i n g a n t s U l t i m a t e l y t h e n u t r i t i o n a l c o m p l e x i t y a n d d e a d a n t a c c u m u l a t i o n i n E n s u r e d i d n o t p r e v e n t f o r a g e r s f r o m r e m o v i n g s i g n i f i c a n t q u a n t i t i e s o f f l u i d P l a s m a P l a s m a e v a p o r a t e d a t a s i g n i f i c a n t l y h i g h e r r a t e t h a n a l l o t h e r f l u i d s a n d w a s t h e o n l y f l u i d w i t h a h i g h e r e v a p o r a t i o n r a t e t h a n E n s u r e W h i l e p l a s m a e v a p o r a t e d a t t h e h i g h e s t r a t e c o n s u m p t i o n o f p l a s m a ( 9 3 1 2 + 3 1 7 ) w a s n o t s i g n i f i c a n t l y d i f f e r e n t t h a n t h a t o f s a l i n e ( 3 0 9 3 + 1 1 4 ) T h e h i g h e v a p o r a t i o n r a t e o f p l a s m a p r o b a b l y i n c r e a s e d t h e

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5 2 c o n c e n t r a t i o n o f i o n s w i t h i n t h e s o l u t i o n m o r e r a p i d l y t h a n o c c u r r e d i n s a l i n e T h e p r e s e n c e o f h e p a r i n a n t i c o a g u l a n t m a y h a v e i n f l u e n c e d a n t c o n s u m p t i o n o f p l a s m a a n d m a y h a v e i n f l u e n c e d t h e r a t e o f e v a p o r a t i o n a s w e l l B o t h h u m a n p l a s m a a n d s a l i n e s o l u t i o n a r e v e r y a q u e o u s f l u i d s w h i c h a r e h i g h i n s o d i u m a n d c h l o r i n e e l e c t r o l y t e s I n f a c t t h e e l e c t r o l y t e c o n t e n t o f 0 9 % N a C l s o l u t i o n a n d h u m a n p l a s m a a r e i d e n t i c a l ; s a l i n e i s o f t e n u s e d a s a f l u i d r e p l a c e m e n t t o i n c r e a s e b l o o d v o l u m e ( A A B B 2 0 0 2 ) S o i t w a s n o t s u r p r i s i n g t h a t P h a r a o h a n t p r e f e r e n c e f o r t h e s e t w o f l u i d s w a s n o t s t a t i s t i c a l l y d i f f e r e n t N a C l S a l i n e s o l u t i o n i s a v e r y a q u e o u s s o l u t i o n c o m p o s e d m o s t l y o f w a t e r w i t h 0 9 % s o d i u m c h l o r i d e i o n s E v a p o r a t i o n o f s a l i n e w a s t h e s e c o n d l o w e s t o f t e s t e d f l u i d s ; e v a p o r a t i o n o f s a l i n e w a s h i g h e r t h a n t h a t o f d e x t r o s e B o t h f l u i d s w e r e h i g h l y a q u e o u s s o l u t i o n s w i t h l o w d e n s i t y s o t h a t a n t i n g e s t i o n w a s e a s y C o n s u m p t i o n o f s a l i n e w a s s i g n i f i c a n t b u t w a s l e s s t h a n t h a t o f h i g h s u g a r f l u i d s M a n y e l e c t r o l y t e s o l u t i o n s a r e r e p e l l e n t t o r e d i m p o r t e d f i r e a n t s ( V i n s o n 1 9 7 0 ) H o w e v e r V i n s o n d e t e r m i n e d t h a t t h e a d d i t i o n o f s a l t i n c r e a s e d t h e a c c e p t a n c e o f s u g a r s b y r e d i m p o r t e d f i r e a n t s T h u s t h e a v a i l a b l e c h o i c e o f N a C l s o l u t i o n i n t e s t a r e n a s m a y h a v e r e s u l t e d i n i n c r e a s e d c o n s u m p t i o n o f h i g h s u g a r f l u i d s d e x t r o s e a n d E n s u r e I n t h i s e x p e r i m e n t P h a r a o h a n t s c o n s u m e d 0 9 % N a C l s o l u t i o n ( 3 0 9 3 + 1 1 4 ) d e s p i t e t h e a d l i b i t u m p r e s e n c e o f w a t e r T h i s c l e a r l y i n d i c a t e d t h a t t h e P h a r a o h a n t s w e r e i n t e r e s t e d i n s a l i n e s o l u t i o n f o r n u t r i t i o n a l r e a s o n s e s p e c i a l l y w h e n b a s e l i n e w a t e r c o n s u m p t i o n t e s t r e s u l t s ( 0 4 8 + 0 0 7 m g ) w e r e t a k e n i n t o a c c o u n t

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5 3 B l o o d B l o o d c e l l s w e r e t h e l e a s t c o n s u m e d o f a l l t e s t e d f l u i d s B l o o d w a s t h e o n l y f l u i d w h i c h h a d a h i g h e r r a t e o f e v a p o r a t i o n ( 3 3 2 + 0 2 5 ) t h a n o f c o n s u m p t i o n ( 2 0 4 6 + 0 4 8 ) a l t h o u g h b l o o d l o s s i n t h e t e s t a r e n a ( 5 2 0 + 0 5 8 ) w a s g r e a t e r t h a n e v a p o r a t i o n ( T a b l e 4 2 ) V e r y l i t t l e f o r a g i n g a c t i v i t y w a s o b s e r v e d o n m i c r o c a p i l l a r y t u b e s c o n t a i n i n g b l o o d c e l l s T h e a n t s d i d n o t r e c r u i t t o i t v e r y s t r o n g l y a s a f o o d s o u r c e a l t h o u g h t h e y d i d c o n s u m e i t i n s i g n i f i c a n t a m o u n t s S o l i d i f i c a t i o n o f t h e b l o o d c e l l s w a s o b s e r v e d T h e s m a l l i n n e r d i a m e t e r o f t h e m i c r o c a p i l l a r y t u b e a n d a h a r d e n e d s o l i d i f i e d s u r f a c e p r o b a b l y m a d e e x t r a c t i o n v e r y d i f f i c u l t f o r f o r a g e r s B l o o d w a s o n e o f t h e t w o m o s t v i s c o u s f l u i d s i n t h i s e x p e r i m e n t a n d c o n s u m p t i o n o f v e r y d e n s e a n d h i g h l y n u t r i t i o u s b l o o d c e l l s m a y h a v e r e s u l t e d i n s a t i a t i o n a t l o w e r l e v e l s o f c o n s u m p t i o n A l t e r n a t i v e l y b l o o d c e l l s a r e h i g h i n p r o t e i n b u t a r e n o t h i g h i n s u g a r s s o t h e l o w e r c o n s u m p t i o n o f b l o o d c e l l s a s c o m p a r e d t o t h a t o f h i g h s u g a r f l u i d s d e x t r o s e a n d E n s u r e m i g h t h a v e i n d i c a t e d t h a t t h e P h a r a o h a n t s ’ n u t r i t i o n a l n e e d f o r c a r b o h y d r a t e s w a s h i g h e r t h a n f o r p r o t e i n d u r i n g t h e t e s t p e r i o d I t i s c l e a r t h a t i n t h i s s t u d y d e x t r o s e w a s p r e f e r r e d o v e r a l l o t h e r f l u i d s I t i s p o s s i b l e t h a t h i g h s u g a r ( s o l e n u t r i e n t ) c o n c e n t r a t i o n i s f a v o r a b l e f o r P h a r a o h a n t f l u i d c o n s u m p t i o n S u m p t e r a n d B e e k m a n ( 2 0 0 3 ) r e p o r t e d P h a r a o h a n t s p r e f e r e n t i a l l y r e c r u i t t o f o o d s w i t h h i g h e s t s u c r o s e c o n c e n t r a t i o n W h e n a n t s a r e p l a c e d i n t e s t a r e n a t h e y c o n s u m e d a l l p a t i e n t c a r e f l u i d s K n o w i n g t h a t P h a r a o h a n t s a r e a t t r a c t e d t o a n d w i l l c o n s u m e f l u i d s u s e d i n p a t i e n t c a r e e s p e c i a l l y d e x t r o s e h o s p i t a l s t a f f s h o u l d n o t s p r e a d t h e s e f l u i d s i n h o s p i t a l ; i f t h e s e f l u i d s a r e p r e s e n t ( e s p e c i a l l y n e a r p a t i e n t ) t h e y s h o u l d b e c l e a n e d u p q u i c k l y t o d e c r e a s e p o t e n t i a l f o r P h a r a o h a n t a t t r a c t i o n t o p a t i e n t s w i t h i n h o s p i t a l a n d o t h e r h e a l t h c a r e e n v i r o n m e n t s

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5 4 T a b l e 4 1 M e a n n u m b e r o f a n t s r e m a i n i n g ( e i t h e r f e e d i n g s t u c k o r d e a d ) i n f l u i d s a t e n d o f 2 4 h c h o i c e t e s t f e e d i n g p e r i o d L i q u i d F o o d T e s t a r e n a s M e a n n u m b e r a n t s S E B l o o d 0 7 1 + 0 4 9 b 5 % D e x t r o s e 1 0 6 + 0 3 4 b E n s u r e 5 7 7 + 0 9 6 a 0 9 % N a C l 1 9 4 + 0 5 6 b P l a s m a 1 1 2 + 0 4 2 b W a t e r a 2 7 7 + 1 1 0 M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s

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T a b l e 4 2 M e a n ( c h o i c e t e s t ) i n i t i a l w e i g h t s o f f l u i d q u a n t i t y p r o v i d e d ( m g ) a n d m e a n f l u i d l o s s ( m g ) f o r e a c h o f f i v e d i f f e r e n t p a t i e n t c a r e f l u i d s u s e d i n h o s p i t a l e n v i r o n m e n t s ( 2 4 h ) f o r s t a r v e d ( 3 d ) p h a r a o h a n t c o l o n i e s M e a n ( m g ) S E M _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ C o n t r o l a r e n a s T e s t a r e n a s P a t i e n t c a r e _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ F l u i d I n i t i a l w t F l u i d L o s sa I n i t i a l w ta F l u i d L o s sb B l o o d 7 5 8 1 + 0 2 8 b 3 3 2 + 0 2 5 a b 7 6 0 3 + 0 4 2 b 5 2 0 + 0 5 8 c 5 % D e x t r o s e 7 0 4 7 + 0 1 7 c d 2 5 9 + 0 1 9 b 7 0 5 1 + 0 2 5 c d 5 0 8 0 + 3 4 6 a E n s u r e 7 6 9 2 + 0 1 8 a 4 0 0 + 0 2 4 a b 7 7 2 5 + 0 3 5 a 1 9 6 2 + 6 0 8 b 0 9 % N a C l 6 9 7 6 + 0 1 3 d 2 9 8 + 0 4 8 b 6 9 9 9 + 0 1 8 d 5 8 2 + 1 1 6 c P l a s m a 7 1 1 5 + 0 2 4 c 4 8 7 + 0 6 8 a 7 1 4 8 + 0 1 7 c 1 3 0 7 + 3 1 9 b c W a t e rc 6 9 4 8 + 0 1 2 3 2 1 + 0 1 9 6 9 5 4 + 0 1 6 5 6 4 + 0 5 9 M e a n s w i t h i n a c o l u m n f o l l o w e d b y t h e s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ) a F l u i d L o s s i n c o n t r o l a r e n a s i s t o e v a p o r a t i o n o n l y bF l u i d l o s s i n t e s t a r e n a s d u e t o e v a p o r a t i o n a n d c o n s u m p t i o n c o m b i n e d c W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s 5 5

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T a b l e 4 3 M e a n ( c h o i c e t e s t ) c o n s u m p t i o n ( m g ) f o r e a c h o f f i v e d i f f e r e n t p a t i e n t c a r e f l u i d s u s e d i n h o s p i t a l e n v i r o n m e n t s ( 2 4 h ) f o r s t a r v e d ( 3 d ) p h a r a o h a n t c o l o n i e s L i q u i d F o o d T e s t a r e n a s d f t P M e a n c o n s u m p t i o n S E M ( m g ) B l o o d 2 0 4 6 + 0 4 8 c 3 8 4 2 3 0 0 0 0 1 5 % D e x t r o s e 4 9 1 4 0 + 3 5 1 a 3 8 1 4 0 2 < 0 0 0 0 1 E n s u r e 1 6 0 5 4 + 6 2 3 b 3 8 2 5 8 0 0 0 5 6 0 9 % N a C l 3 0 9 3 + 1 1 4 b c 3 8 2 7 1 0 0 1 0 1 P l a s m a 9 3 1 2 + 3 1 7 b c 3 8 2 9 4 0 0 0 5 6 W a t e ra 0 4 8 + 0 0 7 1 6 7 2 5 < 0 0 0 0 1 M e a n c o n s u m p t i o n w i t h i n a r o w ( H0: = 0 ) a n a l y z e d v i a S t u d e n t ’ s t t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) M e a n c o n s u m p t i o n o f f l u i d s ( m g ) l i s t e d i n c o l u m n s a n a l y z e d v i a T u k e y ’ s m e a n s s e p a r a t i o n t e s t ( P = 0 0 5 ) M e a n s w i t h i n a c o l u m n f o l l o w e d b y s a m e l e t t e r s a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ) a W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s 5 6

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2 7 7 0 7 1 b 1 0 6 b 1 1 2 b 1 9 4 b 5 7 7 a 0 1 2 3 4 5 6 7 W a t e r B l o o d 5 % D e x t r o s e E n s u r e 0 9 % N a C l P l a s m aN o A n t s i n F l u i d F i g u r e 4 2 M e a n n u m b e r o f a n t s ( e i t h e r d e a d o r f e e d i n g ) r e m a i n i n g i n f l u i d a t e n d o f 2 4 h c h o i c e t e s t f e e d i n g p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) W a t e r a n a l y s i s d o n e s e p a r a t e l y a s b a s e l i n e s t u d y ; w a t e r d a t a n o t p a r t o f t e s t f l u i d a n a l y s i s 5 7

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2 5 9 b 2 9 8 b 3 3 2 a b 4 0 0 a b 4 8 7 a 0 0 0 1 0 0 2 0 0 3 0 0 4 0 0 5 0 0 6 0 0 B l o o d 5 % D e x t r o s e E n s u r e 0 9 % N a C l P l a s m a E v a p o r a t i o n ( m g ) F i g u r e 4 3 M e a n e v a p o r a t i o n ( c h o i c e t e s t ) o f f l u i d s u s e d i n h o s p i t a l e n v i r o n m e n t s f o r p a t i e n t c a r e (g ) ( 2 4 h o u r s ) D a t a w a s c o l l e c t e d f r o m t h e c o n t r o l a r e n a s : e v a p o r a t i o n w a s c a l c u l a t e d b y s u b t r a c t i n g f i n a l m a s s o f f l u i d r e m a i n i n g a t t h e e n d o f t h e 2 4 h t e s t p e r i o d f r o m t h e a m o u n t o f f l u i d i n i t i a l l y p r o v i d e d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) 5 8

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3 0 9 b c 9 3 1 b c 1 6 0 5 a b 4 9 1 4 a 2 0 5 c 0 1 0 2 0 3 0 4 0 5 0 6 0 B l o o d 5 % D e x t r o s e E n s u r e 0 9 % N a C l P l a s m aC o n s u m p t i o n ( m g ) F i g u r e 4 4 M e a n ( c h o i c e t e s t ) c o n s u m p t i o n ( m g ) f o r t e s t e d f l u i d s u s e d d u r i n g t e s t p e r i o d ( 2 4 h ) f o r s t a r v e d ( 3 d ) p h a r a o h a n t c o l o n i e s M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t b y T u k e y ’ s H S D t e s t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) 5 9

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6 0 C H A P T E R 5 N O C H O I C E D E H Y D R A T E D F L U I D T E S T I n t r o d u c t i o n H o s p i t a l s n u r s i n g h o m e s a n d o t h e r m e d i c a l t r e a t m e n t f a c i l i t i e s a r e o f t e n i n f e s t e d w i t h P h a r a o h a n t s ( G r e e n e t a l 1 9 5 4 E d w a r d s a n d B a k e r 1 9 8 1 a E i c h l e r 1 9 9 0 ) a t r o p i c a l p e s t s p e c i e s t h a t h a s w o r l d w i d e d i s t r i b u t i o n ( H l l d o b l e r a n d W i l s o n 1 9 9 0 F o w l e r e t a l 1 9 9 4 ) P h a r a o h a n t s h a v e b e e n d i s c o v e r e d i n s e n s i t i v e p a t i e n t c a r e a r e a s s u c h a s b u r n u n i t s w h e r e t h e y w e r e o b s e r v e d f e e d i n g o n o p e n w o u n d s ( W e i d n e r 1 9 8 2 ) i n n e o n a t a l u n i t s w h e r e t h e y w e r e o b s e r v e d f e e d i n g u p o n t h e i n t a c t s k i n o f n e w b o r n s ( S t e i n b r i n k 1 9 7 8 ) a s c i t e d b y E i c h l e r ( 1 9 9 0 ) B u r n v i c t i m s a n d n e w b o r n s t y p i c a l l y l i e o n b e d s c o n t a i n i n g s h e e t s a n d a r e p r o v i d e d w i t h v a r i o u s f l u i d s t o m e e t h y d r a t i o n a n d n u t r i t i o n a l n e e d s F l u i d s c a n s p i l l a n d d e s i c c a t e o n s h e e t s a n d m i g h t a t t r a c t P h a r a o h a n t s t o t h e p a t i e n t C o n s u m p t i o n a n d p r e f e r e n c e t e s t s w e r e p e r f o r m e d f o r 5 p a t i e n t c a r e f l u i d s u s e d i n h o s p i t a l s t o d e t e r m i n e i f P h a r a o h a n t s a c t u a l l y c o n s u m e d e a c h f l u i d ( C h a p t e r 3 ) a n d t o d e t e r m i n e w h i c h f l u i d s t h e y p r e f e r r e d ( C h a p t e r 4 ) i n t h e i r l i q u i d s t a t e w h e n g i v e n a c h o i c e W h i l e t h e y c o n s u m e d a l l f l u i d s c o n s u m p t i o n o f l i q u i d d e x t r o s e w a s s i g n i f i c a n t l y h i g h e r t h a n a l l o t h e r f l u i d s a n d w a s t h e p r e f e r r e d f l u i d i n b o t h t h e n o c h o i c e a n d t h e c h o i c e t e s t s T h e s e f i v e f l u i d s a r e t y p i c a l l y a d m i n i s t e r e d d i r e c t l y t o t h e p a t i e n t v i a i n s e r t e d t u b e s S a l i n e 5 % d e x t r o s e r e d b l o o d c e l l s a n d s e r u m a r e u s e d t o r e p l a c e l o s t f l u i d s ( A A B B 2 0 0 2 ) ; s a l i n e a n d d e x t r o s e c a n a l s o b e i n j e c t e d p e r i t o n e a l l y ( i n t o t h e

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6 1 a b d o m e n / s t o m a c h ) ( W e n z e l e t a l 2 0 0 2 ) E n s u r e d i e t a r y s u p p l e m e n t i s u n i q u e a m o n g t h e s e f i v e f l u i d s s i n c e i t i s a d m i n i s t e r e d s o l e l y t h r o u g h i n j e c t i o n i n t o t h e p a t i e n t ’ s s t o m a c h ( W e n z e l e t a l 2 0 0 2 ) R e g a r d l e s s o f t h e m e t h o d u s e d t o p r o v i d e t h e s e f l u i d s t o p a t i e n t s a n y o f t h e s e f l u i d s m i g h t e a s i l y s p i l l o n t o t h e p a t i e n t b e d a n d s o l i d i f y u p o n t h e b e d s h e e t s I t w a s d e t e r m i n e d t h a t t h e s e f i v e f l u i d s n e e d e d t o b e p r o v i d e d t o t h e P h a r a o h a n t s i n t h e i r d e s i c c a t e d s t a t e ( i e a s s o l i d s ) t o d e t e r m i n e i f t h e a n t s d e m o n s t r a t e d i n t e r e s t i n s p i l l e d p a t i e n t c a r e f l u i d s i n h o s p i t a l s T h e p o t e n t i a l a v a i l a b i l i t y o f t h e s e s u b s t a n c e s m i g h t b e s u b s t a n t i a l l y g r e a t e r i n t h e i r d r i e d f o r m t h a n i n t h e i r l i q u i d f o r m a n d c o u l d b e a f a c t o r i n a t t r a c t i n g t h e a n t s t o p a t i e n t b e d s a n d t o t h e p a t i e n t T h e o b j e c t i v e o f t h i s e x p e r i m e n t w a s t o d e t e r m i n e i f P h a r a o h a n t s w o u l d f o r a g e t o d e s i c c a t e d p a t i e n t c a r e f l u i d s u s e d i n h o s p i t a l u s i n g a n o c h o i c e t e s t M a t e r i a l s a n d M e t h o d s A n t s P h a r a o h a n t t e s t c o l o n i e s w e r e e x t r a c t e d f r o m l a r g e m a i n t e n a n c e / p a r e n t c o l o n i e s h o u s e d a n d r e a r e d a t t h e U r b a n E n t o m o l o g y L a b o r a t o r y U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F L S e e A p p e n d i x A f o r f u l l d e t a i l s o n m a i n t e n a n c e o f a n t s C o l o n i e s c o n s i s t e d o f 2 5 0 w o r k e r s 3 d e a l a t e q u e e n s a n d a p p r o x i m a t e l y 5 0 m g b r o o d O n e h u n d r e d t w e n t y f i v e w o r k e r s w e r e s e l e c t e d f r o m c e l l s t o e n s u r e p r e s e n c e o f n u r s e s a n d 1 2 5 w o r k e r s w e r e s e l e c t e d f r o m o u t s i d e c e l l s t o e n s u r e p r e s e n c e o f f o r a g e r s ( H a a c k 1 9 8 7 H a a c k a n d V i n s o n 1 9 9 0 H a a c k e t a l 1 9 9 5 ) B r o o d i n c l u d e d e g g s a l l l a r v a l s t a g e s a n d p u p a e S e e A p p e n d i x B f o r f u l l d e t a i l s o n s e l e c t i o n o f a n t s a n d p r e p a r a t i o n o f e x p e r i m e n t a l t e s t c o l o n i e s

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6 2 F o r a g i n g A r e n a a n d N e s t F o r a g i n g a r e n a s ( F i g 5 1 ) c o n s i s t e d o f c l e a r p l a s t i c b o x e s ( 2 7 c m b y 1 9 c m b y 1 0 c m d e e p ; C a t a l o g u e N o 1 9 5 C P i o n e e r P l a s t i c s D i x o n K Y ) w h i c h w e r e c o a t e d w i t h F l u o n ™ ( C a t a l o g u e N o A D 1 A s a h i G l a s s F l u o r o p o l y m e r s D o w n i n g t o n P A ) o n i n t e r i o r s i d e s t o p r e v e n t a n t e s c a p e O n e n e s t c e l l w a s p l a c e d w i t h i n o n e c o r n e r o f e a c h a r e n a C e l l s c o n s i s t e d o f s t e r i l e c o v e r e d p l a s t i c P e t r i d i s h e s ( 5 0 m m d i a m e t e r b y 1 5 m m d e p t h ; F a l c o n M o d e l N o 3 5 1 0 0 7 B e c t o n D i c k i n s o n a n d C o F r a n k l i n L a k e s N J ) f i l l e d h a l f w a y w i t h C a s t o n e d e n t a l p l a s t e r ( M o d e l N o 9 9 0 4 5 D e n t s p l y I n t e r n a t i o n a l Y o r k P A ) O n e e n t r a n c e h o l e ( ~ 5 m m d i a m e t e r ) w a s m e l t e d i n t o t h e u p p e r s i d e w a l l o f t h e P e t r i d i s h b a s e v i a i n s e r t i o n o f t h e m e t a l t i p o f a h o t g l u e g u n ( s a n s g l u e ) j u s t a b o v e t h e l e v e l o f t h e h a r d e n e d p l a s t e r T h e P e t r i d i s h l i d w a s p l a c e d o n t o p o f t h e b a s e a n d t h e e n t i r e n e s t i n g c e l l w a s p l a c e d i n t o t h e f o r a g i n g a r e n a w i t h t h e e n t r a n c e h o l e f a c i n g t h e o p p o s i t e d i a g o n a l c o r n e r o f t h e a r e n a W a t e r w a s p r o v i d e d a d l i b i t u m i n a 1 5 m l v i a l ( M o d e l N o 5 5 9 T h o r n t o n P l a s t i c s S a l t L a k e C i t y U T ) w i t h t h r e e l a r g e c o t t o n b a l l s i n s e r t e d i n t o t h e t o p t o p r e v e n t l e a k a g e a n d t o p r o v i d e e a s y a c c e s s f o r t h e a n t s T h e w a t e r v i a l w a s p l a c e d i n t h e o p p o s i t e d i a g o n a l c o r n e r o f t h e t e s t u n i t a r e n a a c r o s s f r o m t h e c e l l H o s p i t a l S h e e t s H o s p i t a l s h e e t s ( F o r e s t G e n e r a l H o s p i t a l N e w b e r r y R o a d G a i n e s v i l l e F L ) w e r e w a s h e d i n e x t r e m e l y h o t t e m p e r a t u r e s a n d u n d e r w e n t a r i g o r o u s s t e r i l i z a t i o n p r o c e s s ( J o h n B e n n e t p e r s o n a l c o m m u n i c a t i o n ) F r e s h l y l a u n d e r e d h o s p i t a l s h e e t s w e r e o b t a i n e d

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6 3 W a t e r V i a l ( 1 5 m l ) N e s t i n g c e l l ( 5 0 m m d i a m e t e r ; 1 5 m m d e e p ) 2 7 c m 1 9 c m h o s p i t a l s h e e t s q u a r e ( 2 5 b y 2 5 c m ) o n t o p o f a l u m i n u m f o i l s q u a r e ( a l s o 2 5 b y 2 5 c m ) d e h y d r a t e d f l u i d ; i n s i d e p e n c i l e d c i r c l e ( 2 0 c m d i a m ) F i g u r e 5 1 A r e n a s f o r d e h y d r a t e d l i q u i d f o o d n o c h o i c e e x p e r i m e n t A r e n a s c o n t a i n e d 1 2 5 i n s i d e w o r k e r s 1 2 5 o u t s i d e w o r k e r s 3 d e a l a t e q u e e n s a n d ~ 5 0 m g b r o o d ( a l l s t a g e s ) F l u i d s d r o p s p l a c e d o n h o s p i t a l s h e e t s d e s i c c a t e d f o r 4 d a n d p l a c e d i n c e n t e r o f a r e n a r e s t i n g o n a l u m i n u m f o i l F l u i d s p l a c e d w i t h i n p e n c i l e d c i r c l e ( 2 0 c m d i a m e t e r )

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6 4 f r o m a l o c a l h o s p i t a l a n d w e r e c u t i n t o s m a l l s q u a r e ( 2 5 b y 2 5 c m ) C i r c l e s ( 2 0 b y 2 0 c m ) w e r e p e n c i l e d o n t o h o s p i t a l s h e e t s a n d w e r e c e n t e r e d s o t h a t a 2 5 0 c m b o r d e r e x i s t e d b e t w e e n t h e p e n c i l e d c i r c l e a n d t h e e x t e r i o r b o r d e r o f t h e s h e e t P e n c i l n o t a t i o n s w e r e m a d e a l o n g t h e e d g e o f t h e h o s p i t a l s h e e t o u t s i d e o f t h e c i r c l e d b o r d e r t o i d e n t i f y t h e f l u i d a p p l i e d t o a s h e e t H e a v y d u t y a l u m i n u m f o i l w a s c u t i n t o s m a l l s q u a r e s ( 2 5 b y 2 5 c m ) a n d p l a c e d u n d e r t h e h o s p i t a l s h e e t s E a c h s q u a r e o f a l u m i n u m f o i l w i t h i t s r e s p e c t i v e h o s p i t a l s h e e t s q u a r e w a s p l a c e d i n t o a l a r g e ( 4 0 b y 5 2 b y 1 2 c m d e e p ) p l a s t i c t r a y ( M o d e l N o 4 0 0 5 N P a n e l C o n t r o l s G r e e n v i l l e S C ) a n d s u s p e n d e d ~ 0 3 m a b o v e a p o r t a b l e r o o m h e a t e r i n p r e p a r a t i o n f o r f l u i d a p p l i c a t i o n a n d d e h y d r a t i o n D e h y d r a t i o n o f F l u i d s F l u i d s u s e d i n t h i s e x p e r i m e n t c o n s i s t e d o f t a p w a t e r a n d 5 l i q u i d p a t i e n t c a r e f l u i d s : h u m a n b l o o d c e l l s h u m a n p l a s m a 5 % d e x t r o s e a n d 0 9 % N a C l E a c h f l u i d w a s p l a c e d i n a s t e r i l e t e s t t u b e C l e a n d r y p l a s t i c 1 5 m l d i s p o s a b l e p i p e t t e s w e r e u s e d t o w i t h d r a w f l u i d a n d p l a c e a d r o p o f ~ 1 m l v o l u m e i n t h e c e n t e r o f t h e 2 5 b y 2 5 c m h o s p i t a l s h e e t w i t h i n t h e p e n c i l e d c i r c l e ’ s 2 0 c m b o u n d a r i e s A s e p a r a t e p i p e t t e w a s u s e d f o r e a c h f l u i d a n d f r e s h p i p e t t e s w e r e u s e d e a c h t i m e N o m i x i n g o f p i p e t t e s o c c u r r e d D r o p s w e r e l e f t t o a i r d r y f o r 1 2 h u n t i l v i s u a l o b s e r v a t i o n i n d i c a t e d t h o r o u g h d e s i c c a t i o n D r o p s w e r e a d d e d i n t h i s m a n n e r u n t i l a t o t a l o f 5 d r o p s w e r e p l a c e d o n e a c h h o s p i t a l s h e e t A f t e r t h e f i n a l d r o p d r i e d t h e l a r g e t r a y w a s p l a c e d o n a l o w e r w a l l r a c k i n t h e e x p e r i m e n t a l l a b o r a t o r y i n a w a y f r o m t h e l i g h t a n d r e m a i n e d t h e r e f o r 4 d u n t i l u s e d i n t h e e x p e r i m e n t a l t e s t T h e t r a y w a s n o t c o v e r e d R o o m t e m p e r a t u r e w a s m a i n t a i n e d a t 2 6 7 C – 2 8 9 C a n d r e l a t i v e h u m i d i t y f l u c t u a t e d b e t w e e n 4 0 5 6 % F l u i d s w e r e p e r m i t t e d t o d e h y d r a t e d u r i n g t h e s a m e p e r i o d

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6 5 t h a t a n t s w e r e s t a r v e d S t a n d a r d i z e d P h a r a o h a n t c o l o n i e s w e r e s t a r v e d ( o n l y w a t e r p r o v i d e d ) f o r 7 2 h a f t e r b e i n g p l a c e d i n t h e a r e n a s T h e h o s p i t a l s h e e t s q u a r e c o n t a i n i n g t h e d r i e d s u b s t a n c e w a s p l a c e d i n t h e c e n t e r o f t h e t e s t u n i t h a l f w a y b e t w e e n t h e n e s t c e l l a n d t h e w a t e r E a c h a r e n a w a s c o v e r e d w i t h a c l e a r p l a s t i c l i d t o m a i n t a i n h u m i d i t y a n d t e m p e r a t u r e w i t h i n a r e n a E x p e r i m e n t a l D e s i g n T h e e x p e r i m e n t w a s d e s i g n e d a s a r a n d o m i z e d c o m p l e t e b l o c k b l o c k i n g o n c o l o n y E a c h b l o c k c o n s i s t e d o f P h a r a o h a n t c o l o n i e s ( 1 2 5 i n s i d e w o r k e r s 1 2 5 o u t s i d e w o r k e r s 3 d e a l a t e q u e e n s a n d ~ 5 0 m g b r o o d ) o b t a i n e d f r o m t h e s a m e p a r e n t c o l o n y a n d r a n d o m l y a s s i g n e d t o e a c h t r e a t m e n t ( d e h y d r a t e d f l u i d t y p e ) A n t s w e r e s e l e c t e d f r o m 6 d i f f e r e n t p a r e n t c o l o n i e s A s i n g l e f l u i d w a s o f f e r e d t o e a c h t e s t c o l o n y C o n t r o l a r e n a s l o c a t e d a d j a c e n t t o t e s t a r e n a s w e r e i d e n t i c a l e x c e p t t h a t t h e y l a c k e d a n t s T i m e d O b s e r v a t i o n s O b s e r v a t i o n s o f f o r a g i n g b e h a v i o r w e r e n o t e d u p o n i m m e d i a t e p l a c e m e n t o f f l u i d s i n t o a r e n a a n d t h e n u m b e r o f a n t s t h a t a p p e a r e d t o b e f e e d i n g o n t h e d r i e d f l u i d s b e g a n 1 5 m i n a f t e r i n i t i a l e x p o s u r e a n d c o n t i n u e d a t 1 5 m i n i n t e r v a l s f o r 2 h A n t s w e r e c o n s i d e r e d t o b e f e e d i n g i f t h e y w e r e s t a n d i n g s t a t i o n a r y a n d a s s u m e d a f e e d i n g p o s t u r e w i t h a n t e n n a e a n d / o r p a l p s t o u c h i n g d r i e d f l u i d s t h a t w e r e c o n t a i n e d w i t h i n t h e c i r c l e d p o r t i o n o f t h e h o s p i t a l s h e e t r e g a r d l e s s o f h o w m u c h o f t h e i r b o d y w a s w i t h i n t h e c i r c l e A n t s f e e d i n g o n f l u i d t h a t s p i l l e d b e y o n d t h e p e n c i l e d c i r c l e w e r e n o t c o u n t e d S i m i l a r l y a n t s e n g a g i n g i n t r o p h a l l a x i s w e r e n o t c o u n t e d e v e n i f t h i s a c t i v i t y o c c u r r e d i n t h e c e n t e r o f t h e c i r c l e A n t s t h a t w e r e i n a r e s t i n g p o s t u r e w i t h i n t h e c i r c l e w e r e a l s o n o t c o u n t e d A n t s t h a t w e r e w a l k i n g o r o t h e r w i s e a c t i v e l y m o v i n g d u r i n g t h e t i m e d

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6 6 o b s e r v a t i o n i n t e r v a l s w e r e n o t c o u n t e d e v e n i f a n t e n n a e w e r e i n c o n t a c t w i t h d e h y d r a t e d f l u i d s S t a t i s t i c a l A n a l y s i s F o u r a n a l y s e s w e r e d o n e u s i n g a n a l y s i s o f v a r i a n c e ( A N O V A ) w i t h m e a n s s e p a r a t e d u s i n g T u k e y ’ s H S D ( S A S I n s t i t u t e 2 0 0 0 ) F i r s t c o u n t s o f f e e d i n g a n t s o v e r a l l d r i e d f l u i d s a m o n g t h e 1 5 m i n i n t e r v a l s w e r e c o m p a r e d S e c o n d t h e t o t a l n u m b e r o f a n t s f e e d i n g o n e a c h d r i e d f l u i d f o r t h e e n t i r e 2 h t e s t p e r i o d w a s c o m p a r e d T h i r d f o r e a c h t i m e i n t e r v a l t h e m e a n n u m b e r s o f f e e d i n g a n t s w e r e c o m p a r e d a m o n g f l u i d t y p e s F i n a l l y f o r e a c h f l u i d t h e m e a n n u m b e r s o f a n t s f e e d i n g w e r e c o m p a r e d a m o n g e a c h o f t h e 1 5 m i n i n t e r v a l s R e s u l t s U s u a l l y 1 o r 2 a n t s w e r e f o r a g i n g i n t h e a r e n a s d u r i n g p l a c e m e n t o f d e h y d r a t e d f l u i d s i n t o a r e n a s F o r a g e r s i n a r e n a s c o n t a i n i n g E n s u r e a n d d e x t r o s e e x h i b i t e d a n i m m e d i a t e c h a n g e i n b e h a v i o r t h e y r a p i d l y i n c r e a s e d t h e i r w a l k i n g s p e e d a n d b e g a n w a v i n g a n t e n n a e i n f r o n t a n d a b o v e t h e i r h e a d s i n a n e x c i t e d m a n n e r T h e y t o o k f a s t r u n n i n g s t e p s i n s h o r t b u r s t s t o w a r d s t h e d e h y d r a t e d f l u i d a n d l o c a t e d t h e f l u i d w i t h i n a f e w s e c o n d s ; r e c r u i t m e n t o f a d d i t i o n a l a n t s i n a r e n a s c o n t a i n i n g E n s u r e a n d d e x t r o s e f r e q u e n t l y o c c u r r e d i n l e s s t h a n 5 m i n I n a r e n a s w i t h d e h y d r a t e d d e x t r o s e f o r a g e r s e x h i b i t e d a t e n d e n c y t o f e e d f o r s e v e r a l m i n u t e s b e f o r e r e c r u i t i n g o t h e r a n t s I n a r e n a s w i t h d e h y d r a t e d E n s u r e i n i t i a l f o r a g e r s t e n d e d t o e n g a g e i n e x t e n s i v e a n t e n n a l t a p p i n g o f d e h y d r a t e d f l u i d w h i l e r a p i d l y e n c i r c l i n g t h e e n t i r e h o s p i t a l s h e e t ; a f t e r c o m p l e t e l y c i r c l i n g t h e f l u i d t h e y r a p i d l y r a n t o t h e n e s t i n g c e l l S h o r t l y a f t e r w a r d s s e v e r a l a n t s w e r e o b s e r v e d p r o c e e d i n g i n a h u r r i e d m a n n e r t o w a r d s t h e E n s u r e w h e r e m o s t e n g a g e d i n f e e d i n g b e h a v i o r f o r s e v e r a l m i n u t e s A f e w r e t u r n e d t o t h e n e s t a g a i n r e s u l t i n g

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6 7 i m m e d i a t e l y i n a n i n c r e a s e d n u m b e r o f f o r a g e r s e m e r g i n g f r o m t h e n e s t A n t b e h a v i o r i n a r e n a s i n w h i c h h o s p i t a l s h e e t s s o a k e d w i t h b l o o d c e l l s p l a s m a s a l i n e o r w a t e r d i d n o t e l i c i t a n y i n c r e a s e d f o r a g i n g a c t i v i t y o r i m m e d i a t e r e c r u i t m e n t b e h a v i o r F o r a g e r s i n t h e s e a r e n a s c o n t i n u e d t o w a l k a r o u n d t h e a r e n a i n a s l o w u n h u r r i e d m a n n e r a n d d i d n o t s e e m t o r e s p o n d t o d e h y d r a t e d b l o o d c e l l s p l a s m a s a l i n e o r w a t e r a s a f o o d s o u r c e O c c a s i o n a l l y a f o r a g e r w a s o b s e r v e d f e e d i n g o n b l o o d c e l l s p l a s m a o r s a l i n e b u t n o f o r a g e r s w e r e o b s e r v e d o n s h e e t s t h a t h a d b e e n s o a k e d w i t h w a t e r F o r a g e r s d i d n o t e x h i b i t t h e s a m e t y p e o f e x c i t e d b e h a v i o r i n t h e s e a r e n a s a s w a s o b s e r v e d i n a r e n a s w i t h h i g h s u g a r d e h y d r a t e d f l u i d s E v e n a f t e r f e e d i n g f o r s e v e r a l m i n u t e s a n t s w e r e n o t o b s e r v e d r u n n i n g b a c k t o t h e n e s t a n d n o m a s s r e c r u i t m e n t w a s o b s e r v e d i n a r e n a s l a c k i n g s u g a r P e a k f e e d i n g o f a l l f l u i d s s i m u l t a n e o u s l y o c c u r r e d a t 4 5 m i n a n d s t e a d i l y d e c r e a s e d a f t e r w a r d s H o w e v e r a n t c o u n t s w e r e n o t s i g n i f i c a n t l y d i f f e r e n t a m o n g s t t i m e i n t e r v a l s ( F = 0 9 7 d f = 7 1 1 4 4 ; P = 0 4 4 9 9 ) ( F i g 5 2 ) T h e n u m b e r o f P h a r a o h a n t s f e e d i n g u p o n E n s u r e ( 1 3 8 9 + 0 5 3 ) w a s s i g n i f i c a n t l y h i g h e r t h a n o n a l l o t h e r d e h y d r a t e d f l u i d s F e e d i n g u p o n 5 % d e x t r o s e w a s s i g n i f i c a n t l y h i g h e r t h a n o n p l a s m a b l o o d c e l l s a n d s a l i n e ( F = 6 1 7 7 5 d f = 5 1 1 4 6 ; P < 0 0 0 0 1 ) ( F i g 5 3 ) N u m b e r s o f a n t s f e e d i n g u p o n E n s u r e w a s s i g n i f i c a n t l y h i g h e r a t e a c h t i m e i n t e r v a l t h a n o n a l l o t h e r f l u i d s ( F i g 5 4 t h r o u g h 5 1 1 ) T h e r e w a s n o s i g n i f i c a n t d i f f e r e n c e i n n u m b e r s o f a n t s f e e d i n g o n a l l o t h e r f l u i d s p e r t i m e i n t e r v a l O n l y E n s u r e a n d d e x t r o s e h a d s i g n i f i c a n t f e e d i n g a m o n g t i m e i n t e r v a l s d u r i n g t h e 2 h e x p e r i m e n t F e e d i n g o f E n s u r e p e a k e d a t 4 5 m i n ( 1 8 0 + 2 0 a n t s ) ( F i g 5 1 2 ) a n d f e e d i n g o f d e x t r o s e a l s o p e a k e d a t 4 5 m i n ( 2 4 + 0 4 a n t s ) ( F i g

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6 8 5 1 3 ) F e e d i n g o n t h e s a l i n e b l o o d c e l l s a n d p l a s m a w e r e v i r t u a l l y n o n e x i s t e n t a s m e a n c o u n t s a t a n y t i m e i n t e r v a l w a s n e v e r h i g h e r t h a n 0 2 a n t s ( F i g 5 1 4 5 1 5 5 1 6 ) D i s c u s s i o n P h a r a o h a n t s w e r e s t a r v e d f o r 7 2 h i n t h i s n o c h o i c e s o l i d f o o d t e s t – t h i s w a s 2 4 h l o n g e r t h a n s t a r v a t i o n p e r i o d o f 4 8 h u s e d i n m y l i q u i d c o n s u m p t i o n t e s t s ( C h a p t e r s 3 4 ) I n c o n t r a s t a 7 2 h s t a r v a t i o n p e r i o d r e s u l t e d i n o b s e r v a b l e f o r a g i n g b e h a v i o r f o r n e a r l y a l l t e s t a r e n a s S t a r v a t i o n p e r i o d s > 7 2 h w e r e n o t a t t e m p t e d b e c a u s e P h a r a o h a n t r e c r u i t m e n t t o c a r b o h y d r a t e s c a n b e v e r y h i g h a f t e r e x t e n s i v e s t a r v a t i o n d u e t o a m o r e r a p i d d e p l e t i o n o f c a r b o h y d r a t e s t o r e s ( N a t i o n 2 0 0 3 ) H a a c k ( 1 9 8 7 ) r e p o r t e d t h a t 7 d s t a r v a t i o n m a d e i t d i f f i c u l t t o c o u n t n u m b e r s o f f o r a g i n g P h a r a o h a n t s b e c a u s e t h e y o v e r w h e l m e d h i g h s u g a r f o o d s u p o n i n i t i a l e x p o s u r e H a a c k e t a l ( 1 9 9 5 ) r e p o r t e d t h a t 2 t o 3 d s t a r v a t i o n w a s s u f f i c i e n t t o d e p l e t e p r o t e i n s t o r e s e n o u g h t o e l i c i t s t r o n g f o r a g i n g b e h a v i o r f o r p r o t e i n s E n s u r e w a s t h e o n l y f l u i d w i t h s i g n i f i c a n t c o n s u m p t i o n t h r o u g h o u t t h e e x p e r i m e n t a n d i t w a s n o t e a b l e t h a t i t w a s t h e o n l y c o m p l e x f l u i d p r o v i d e d i n t h i s e x p e r i m e n t I t h a s h i g h c o n c e n t r a t i o n s o f a l l t h r e e m a j o r n u t r i e n t s w i t h 1 7 % p r o t e i n 1 7 % l i p i d a n d 2 6 % c a r b o h y d r a t e T h e n u m b e r o f a n t s r e c o r d e d f e e d i n g a t b o t h E n s u r e a n d d e x t r o s e p e a k e d a t t h e 4 5 m i n t i m e i n t e r v a l a l t h o u g h i s w a s n o t s i g n i f i c a n t l y d i f f e r e n t f r o m a n t c o u n t s o f m o s t o t h e r t i m e i n t e r v a l s T h e s e d a t a d i f f e r s l i g h t l y f r o m t h a t c o l l e c t e d b y H a a c k ( 1 9 8 7 ) w h o r e p o r t e d t h a t 9 3 % o f w o r k e r s f e d w i t h i n t h e f i r s t 3 0 m i n o f f o o d p l a c e m e n t i n t h e a r e n a T h e E n s u r e a n d d e x t r o s e w e r e t h e o n l y f l u i d s t o h a v e s i g n i f i c a n t n u m b e r s o f a n t s f e e d i n g u p o n t h e m a n d t h e y s h a r e d a s i m i l a r t r a i t : h i g h s u g a r c o n t e n t A l t h o u g h t h e t y p e s o f s u g a r s p r e s e n t e d d i f f e r e d a n d w e r e p r e s e n t i n t h e i r s o l i d f o r m t h e y e l i c i t e d f o r a g i n g a n d f e e d i n g b e h a v i o r o n a s i g n i f i c a n t n u m b e r o f f o r a g i n g a n t s F o r a g i n g t o

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6 9 s o l i d d e h y d r a t e d s u g a r h e r e d i f f e r s f r o m t h e r e s u l t s o f H a a c k e t a l ( 1 9 9 5 ) w h o o b s e r v e d t h a t P h a r a o h a n t s d i d n o t r e s p o n d t o s o l i d s u g a r c r y s t a l s a s a f o o d s o u r c e I n f a c t t h e y o b s e r v e d t h e a n t s f i l l i n g t i n y c r a c k s i n t h e n e s t i n g c e l l w a l l s w i t h t h e s u g a r c r y s t a l s S i n c e t h e s o l i d s u g a r s u s e d b y H a a c k e t a l ( 1 9 9 5 ) w e r e m a n u f a c t u r e d a n d s o l d a s t a b l e s u g a r b o t h e n a n t i o m e r s o f g l u c o s e w e r e p r o b a b l y p r e s e n t i n t h o s e c r y s t a l s I n c o n t r a s t t h e s o l i d s u g a r p r e s e n t i n m y e x p e r i m e n t w a s c r y s t a l l i n e s t r u c t u r e s o f D g l u c o s e t h e b i o l o g i c a l l y a c t i v e e n a n t i o m e r w h i c h d o e s n o t r e q u i r e e n e r g y f o r d i g e s t i o n s i n c e i t i s p a s s i v e l y a b s o r b e d t h r o u g h t h e m i d g u t w a l l a n d s t o r e d p r i m a r i l y a s g l y c o g e n ( N a t i o n 2 0 0 3 ) A l t h o u g h P h a r a o h a n t s a p p e a r e d t o f e e d u p o n d e x t r o s e t h e y a g g r e g a t e d a r o u n d a n d a p p e a r e d t o f e e d u p o n E n s u r e a t m u c h h i g h e r l e v e l s E n s u r e w a s t h e m o s t v i s c o u s f l u i d u s e d i n t h i s e x p e r i m e n t a n d w a s t h e o n l y f o o d t h a t c o n t a i n e d m u l t i p l e n u t r i e n t s I t t o o k l o n g e r t o d e s i c c a t e t h a n o t h e r f l u i d s a n d w h e n f i n a l l y d r y a p p e a r e d a s a b r o w n c o l o r e d t e x t u r e d g l o b t h a t h a d a t h r e e d i m e n s i o n a l t e x t u r e I n c o n t r a s t a l l c l e a r f l u i d s w e r e d i f f i c u l t t o s e e R e p e a t e d a p p l i c a t i o n s o f d e x t r o s e w e r e v i s i b l e b e c a u s e t h e y r e s u l t e d i n a n o t i c e a b l e s t i f f e n i n g o f t h e h o s p i t a l s h e e t a n d o f t e n a d h e r e d t o t h e s h e e t t o t h e u n d e r l y i n g a l u m i n u m f o i l P l a s m a w a s o b s e r v a b l e b e c a u s e i t l e f t a s t r a w c o l o r e d s t a i n o n t h e h o s p i t a l s h e e t B l o o d w a s e a s y t o s e e b u t i t w a s d i f f i c u l t t o a p p l y t o t h e s h e e t i n c o n t r o l l e d a m o u n t s s i n c e t h e b l o o d c e l l s w e r e v e r y s t i c k y B l o o d c e l l s d r i e d i n g l o b s t h a t w e r e r e l a t i v e l y l a r g e i n c o m p a r i s o n t o c l e a r f l u i d s a l t h o u g h b l o o d c e l l s a n d E n s u r e h a d a l m o s t t h e s a m e c o n s i s t e n c y S a l i n e a n d w a t e r w e r e n e a r l y i m p o s s i b l e t o s e e v i s u a l l y a l t h o u g h t h e a p p l i c a t i o n l e f t a w e t s p o t s o t h a t a p p l i c a t i o n c o u l d b e c o n f i r m e d

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7 0 i m m e d i a t e l y a f t e r a p p l i c a t i o n D e s i c c a t i o n o f s a l i n e r e s u l t e d i n a c r i s p y h o s p i t a l s h e e t w h i l e a p p l i c a t i o n o f w a t e r d i d n o t a p p e a r t o h a v e a n y e f f e c t S i g n i f i c a n t f e e d i n g u p o n h i g h p r o t e i n s u b s t a n c e s o r w a t e r w a s n o t e x p e c t e d H i g h e r f e e d i n g o f d e x t r o s e s o l i d s a n d l o w e r f e e d i n g u p o n E n s u r e w e r e a n t i c i p a t e d I n p r e v i o u s c o n s u m p t i o n s t u d i e s P h a r a o h a n t s p r e f e r e n t i a l l y c o n s u m e d 5 % d e x t r o s e a b o v e a l l o t h e r f l u i d s s o i t w a s i n t e r e s t i n g t h a t w h e n d r i e d P h a r a o h a n t s f e e d i n g w a s l e s s t h a n t h e E n s u r e d i e t a r y s u p p l e m e n t P h a r a o h a n t r e c r u i t m e n t t o E n s u r e o c c u r r e d a f t e r 4 d d e s i c c a t i o n s o t h i s f l u i d s t i l l e l i c i t e d f o r a g i n g a f t e r s e v e r a l d a y s T h u s i f s p i l l e d E n s u r e m a y a t t r a c t P h a r a o h a n t s a t l e a s t 4 d a f t e r d r y i n g o u t T h e c o m p l e t e n u t r i t i o n a l s t a t u s o f E n s u r e a v a i l a b l e t o t h e m a f t e r 3 d o f s t a r v a t i o n e l i m i n a t e d t h e n e e d f o r r e s o u r c e s w i t c h i n g s i n c e i t c o n t a i n e d a l l t h r e e m a j o r n u t r i e n t s A d d i t i o n a l l y t h e w o r k e r s m a y h a v e u s e d t h e i r 4 t o o t h e d ( O g a t a e t a l 1 9 9 8 ) m a n d i b l e s t o s c r a p e a n d e x t r a c t s o l i d p a r t i c l e s o f t h e l u m p y t e x t u r e d E n s u r e r e s i d u e A d u l t w o r k e r s d o n o t e a t s o l i d f o o d a l t h o u g h t h e y c a n d i g e s t s o l i d s a s l a r v a e ( L e M a s n e 1 9 5 3 ) a s c i t e d i n ( P e t t i 1 9 9 8 ) I n s t e a d w h e n c o n s u m i n g s o l u t i o n s o r s e m i s o l i d s c o n t a i n i n g l i q u i d s a d u l t s f i l t e r t h e l i q u i d p o r t i o n t h r o u g h t h e i r b u c c a l t u b e ( P e t t i 1 9 9 8 ) a n d s t o r e f i l t e r e d s o l i d s i n t h e i n f r a b u c c a l p o u c h T h e r e s u l t i n g c o m p a c t e d s o l i d s a r e l a t e r e j e c t e d a s a p e l l e t b y e v e r s i o n o f t h e i n f r a b u c c a l p o c k e t a f t e r t h e a n t ’ s r e t u r n t o t h e n e s t c o n t a i n i n g t h e d e v e l o p i n g l a r v a e ( P e t t i 1 9 9 8 ) A l s o w o r k e r a n t s c a n p i c k u p l a r g e r p a r t i c l e s i n t h e i r m a n d i b l e s t o t r a n s p o r t t h o s e b a c k t o t h e l a r v a e S o l e n o p s i s i n v i c t a ( B u r e n ) a n o t h e r m y r m i c i n e s p e c i e s h a s e v e n d e m o n s t r a t e d t h e u s e o f t o o l s t o a s s i s t i n f o o d t r a n s p o r t a t i o n b a c k t o t h e n e s t ( B a r b e r e t a l 1 9 8 9 ) H i g h f o r a g i n g n u m b e r s o f a n t s a t t h e d e s i c c a t e d E n s u r e f l u i d p r o b a b l y i n d i c a t e d t h a t t h i s d e s i c c a t e d f l u i d c o n t a i n e d

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7 1 n u t r i e n t s n e e d e d b y t h e l a r v a e o r w a s s u i t a b l e f o r l a r v a l c o n s u m p t i o n a n d s u b s e q u e n t l a r v a e t o w o r k e r t r o p h a l l a x i s D u e t o i t s h i g h s u g a r a n d l i p i d c o n t e n t E n s u r e f l u i d c o n t a i n e d l a r g e e n e r g y r e s e r v e s d e s p i t e i t s d e h y d r a t e d s t a t e T h u s i t m a y h a v e a t t r a c t e d t h e f o r a g e r s w i t h i t s h i g h n u t r i t i o n a l v a l u e e v e n t h o u g h t h e f o o d s t i l l h a d t o b e t r a n s p o r t e d b a c k t o t h e n e s t a n d b i o l o g i c a l l y m o d i f i e d b y t h e l a r v a l t r o p h a l l a x i s N u t r i t i o n a l r e q u i r e m e n t s o f a n t c o l o n i e s m a y v a r y w i t h c o l o n y s i z e ; n a t u r a l P h a r a o h a n t c o l o n i e s t e n d t o w a r d s 1 : 1 : 1 r a t i o s o f w o r k e r s : e g g s : l a r v a e ( P e a c o c k e t a l 1 9 5 5 b ) A l t h o u g h t h e e x p e r i m e n t a l c o l o n i e s u s e d i n t h i s e x p e r i m e n t w e r e s m a l l e r t h a n n a t u r a l c o l o n i e s d i s c o v e r e d i n h u m a n s t r u c t u r e s s u c h a s t h a t r e p o r t e d a n e c d o t a l l y b y B e l l e v o y e ( 1 8 8 9 ) t h e y w e r e r e p r e s e n t a t i v e o f t h e r a t i o s a n d w e r e s i m i l a r t o e x p e r i m e n t a l c o l o n y s i z e s u s e d b y H a a c k a n d V i n s o n ( 1 9 9 0 ) T h e a t t r a c t i v e n e s s o f d r i e d E n s u r e a f t e r a t l e a s t 4 d p o r t e n d s p o s s i b l e p r o b l e m s b y a t t r a c t i n g a n t s t o p a t i e n t s d u e t o s p i l l a g e o n h o s p i t a l l i n e n s H o w e v e r e x t e n d e d a t t r a c t i v e n e s s p o t e n t i a l l y i n d i c a t e s t h a t t h i s f l u i d m i g h t b e u s e f u l a s p a r t o f b a i t m a t r i x f o r P h a r a o h a n t c o n t r o l O f c o u r s e f u r t h e r t e s t i n g w o u l d b e r e q u i r e d t o a s s e s s t h i s p o t e n t i a l r e l a t i v e t o o t h e r b a i t a t t r a c t a n t s

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1 8 5 2 2 9 2 5 1 2 4 5 2 9 7 3 4 2 2 9 0 2 3 3 0 0 5 1 1 5 2 2 5 3 3 5 4 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0M i n u t e s a f t e r F l u i d I n t r o d u c t i o nM e a n N o A n t s F i g u r e 5 2 M e a n n u m b e r o f a n t s f e e d i n g o n a l l d e h y d r a t e d f l u i d s a m o n g 1 5 m i n i n t e r v a l s d u r i n g 2 h t e s t p e r i o d N o s i g n i f i c a n t d i f f e r e n c e s a m o n g t i m e i n t e r v a l s 7 2

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0 0 3 c 0 0 6 c 0 0 7 c 1 4 9 b 1 3 8 9 a 0 0 0 c 0 2 4 6 8 1 0 1 2 1 4 1 6 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 3 M e a n n u m b e r o f f o r a g i n g a n t s e x h i b i t i n g f e e d i n g b e h a v i o r a t d e h y d r a t e d f l u i d s o v e r e n t i r e 2 h t e s t p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t . 7 3

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0 0 b 0 0 b 0 0 b 1 7 b 1 2 2 a 0 0 b 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 4 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 1 5 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 7 4

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0 2 b 0 0 b 0 0 b 2 3 b 1 5 0 a 0 0 b 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 5 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 3 0 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 7 5

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0 0 b 0 1 b 0 0 b 2 3 b 1 8 0 a 0 0 b 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 6 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 4 5 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 7 6

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0 0 b 0 2 b 0 0 b 1 9 b 1 5 6 a 0 0 b 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 7 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 6 0 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 7 7

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0 0 b 0 0 b 0 2 b 0 6 b 0 0 b 1 3 9 a 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 8 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 7 5 m i n t i m e i n t e r v a l . M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 7 8

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0 0 b 0 0 b 0 1 b 1 0 b 1 4 0 a 0 0 b 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 9 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 9 0 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 7 9

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0 0 b 0 1 b 0 0 b 1 2 b 0 0 b 1 2 5 a 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 1 0 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 1 0 5 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P < 0 0 5 ) b y A N O V A a n d T u k e y ’ s H S D t e s t 8 0

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0 0 b 0 0 b 0 0 b 1 0 b 0 0 b 1 0 0 a 0 2 4 6 8 1 0 1 2 W a t e r E n s u r e 5 % D e x t r o s e P l a s m a B l o o d 0 9 % N a C lM e a n N o A n t s F i g u r e 5 1 1 M e a n n u m b e r o f a n t s f e e d i n g o n a t t h e 6 d r i e d f l u i d s a t 1 2 0 m i n t i m e i n t e r v a l M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 8 1

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1 2 2 a b 1 5 0 a b 1 5 6 a b 1 3 9 a b 1 4 0 a b 1 2 5 a b 1 0 0 b 1 8 0 a 0 0 2 0 4 0 6 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 0 2 0 0 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0M e a n N o A n t s F i g u r e 5 1 2 M e a n n u m b e r o f a n t s f e e d i n g o n E n s u r e a m o n g 1 5 m i n i n t e r v a l s o v e r t h e 2 h t e s t p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 8 2

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1 0 a b c 1 2 a b c 1 0 b c 0 6 c 1 9 a b c 2 4 a 2 3 a b 1 7 a b c 0 0 5 1 1 5 2 2 5 3 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0M e a n N o A n t s F i g u r e 5 1 3 M e a n n u m b e r o f a n t s f e e d i n g o n 5 % d e x t r o s e a m o n g 1 5 m i n i n t e r v a l s o v e r t h e 2 h t e s t p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 8 3

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0 0 0 c 0 0 0 c 0 0 0 c 0 0 0 c 0 0 0 c 0 0 4 b 0 0 4 b 0 1 7 a 0 0 0 0 0 2 0 0 4 0 0 6 0 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0M e a n N o A n t s F i g u r e 5 1 4 M e a n n u m b e r o f a n t s f e e d i n g o n s a l i n e a m o n g 1 5 m i n i n t e r v a l s o v e r t h e 2 h t e s t p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 8 4

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0 0 0 a 0 0 8 a 0 0 0 a 0 0 4 a 0 0 0 a 0 0 8 a 0 2 0 a 0 0 4 a 0 0 0 0 0 5 0 1 0 0 1 5 0 2 0 0 2 5 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0M e a n N o A n t s F i g u r e 5 1 5 M e a n n u m b e r o f a n t s f e e d i n g o n b l o o d c e l l s a m o n g 1 5 m i n i n t e r v a l s o v e r t h e 2 h t e s t p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P < 0 0 5 ) b y A N O V A a n d T u k e y ’ s H S D t e s t 8 5

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0 0 0 a 0 0 4 a 0 0 4 a 0 0 4 a 0 0 8 a 0 0 8 a 0 0 8 a 0 1 7 a 0 0 0 0 0 2 0 0 4 0 0 6 0 0 8 0 1 0 0 1 2 0 1 4 0 1 6 0 1 8 1 5 3 0 4 5 6 0 7 5 9 0 1 0 5 1 2 0M e a n N o A n t s F i g u r e 5 1 6 M e a n n u m b e r o f a n t s f e e d i n g o n h u m a n p l a s m a a m o n g 1 5 m i n i n t e r v a l s o v e r t h e 2 h t e s t p e r i o d M e a n s f o l l o w e d b y s a m e l e t t e r a r e n o t s i g n i f i c a n t l y d i f f e r e n t ( P = 0 0 5 ; S A S I n s t i t u t e 2 0 0 0 ) a n d T u k e y ’ s H S D t e s t 8 6

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8 7 C H A P T E R 6 S U M M A R Y A N D C O N C L U S I O N S T h e d i s e a s e v e c t o r i n g p o t e n t i a l o f P h a r a o h a n t s l i k e t h a t o f o t h e r a r t h r o p o d s m o r e t r a d i t i o n a l l y a s s o c i a t e d w i t h d i s e a s e t r a n s m i s s i o n s u c h a s c o c k r o a c h e s a n d f l i e s i s d i r e c t l y l i n k e d t o t h e i r f o o d p r e f e r e n c e s a n d f e e d i n g b e h a v i o r T h e r e f o r e P h a r a o h a n t c o n s u m p t i o n o f l i q u i d s u b s t a n c e s t h a t a r e s p e c i f i c a l l y u s e d i n p a t i e n t c a r e n e e d s t o b e e x a m i n e d m o r e c l o s e l y T h e g o a l o f t h i s r e s e a r c h w a s t o s t u d y P h a r a o h a n t c o n s u m p t i o n a n d p r e f e r e n c e s f o r p a t i e n t c a r e f l u i d s a n d t o d e t e r m i n e w h a t f l u i d ( s ) u s e d i n p a t i e n t c a r e a t t r a c t P h a r a o h a n t s I a c c o m p l i s h e d t h i s r e s e a r c h b y c o n d u c t i n g t h r e e d i s t i n c t e x p e r i m e n t s M o n o m o r i u m p h a r a o n i s ( L ) t h e P h a r a o h a n t i s a p e s t s p e c i e s w h i c h i n h a b i t s b u i l d i n g s t h r o u g h o u t t h e w o r l d ; i t s p o p u l a t i o n s a r e d i f f i c u l t t o c o n t r o l I t i s a h i g h l y s y n a n t h r o p i c ( s t r o n g l y a s s o c i a t e d w i t h h u m a n s ) ( E d w a r d s 1 9 9 1 ) p e s t t r a m p s p e c i e s a n d i s p r o c l a i m e d a s t h e “ m o s t w i d e l y d i s t r i b u t e d o f a l l a n t p e s t s ” b y E d w a r d s a n d S h o r t ( 1 9 9 0 ) w h o s u m m a r i z e d i t s d o c u m e n t e d p r e s e n c e i n t h e U n i t e d S t a t e s C a n a d a S o u t h A m e r i c a E g y p t N o r t h A f r i c a E u r o p e R u s s i a A u s t r a l i a a n d J a p a n W h e n i n h o s p i t a l s a n d o t h e r h e a l t h c a r e e n v i r o n m e n t s t h e s e a n t s a r e o f t e n f o u n d n e s t i n g t h r o u g h o u t t h e h o s p i t a l i n p l a c e s a s d i v e r s e a s o p e r a t i n g r o o m s l i n e n c l o s e t s b a b y i n c u b a t o r s a n d i n s i d e u n o p e n e d i n t r a v e n o u s t u b i n g p a c k a g e s P h a r a o h a n t s h a v e b e e n o b s e r v e d r e c r u i t i n g t o h u m a n s e c r e t i o n s i n t a c t e p i d e r m i s a n d p a t i e n t c a r e f l u i d s s u c h a s d e x t r o s e a n d s a l i n e s o l u t i o n s T h e o b j e c t i v e o f t h i s s t u d y w a s t o d e t e r m i n e t h e P h a r a o h a n t c o n s u m p t i o n o f p a t i e n t c a r e f l u i d s i n c l u d i n g h u m a n b l o o d h u m a n p l a s m a 5 % d e x t r o s e

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8 8 0 9 % N a C l a n d E n s u r e d i e t a r y s u p p l e m e n t i n b o t h n o c h o i c e a n d c h o i c e s i t u a t i o n s a n d t o d e t e r m i n e t h e i r r e c r u i t m e n t t o t h e s e f l u i d s i n t h e i r d e s i c c a t e d s t a t e P h a r a o h a n t s e n g a g e i n e x t e n s i v e f o r a g i n g a n d a r e m o s t a c t i v e a t n i g h t ( H e d g e s 1 9 9 7 ) T h e y p r o b a b l y u s e “ g u i d e l i n e o r i e n t a t i o n ” t o f o l l o w e d g e s a n d c r a c k s o f s t r u c t u r e s t o f i n d f o o d s o u r c e s s i m i l a r t o t h a t o b s e r v e d i n C a m p o n o t u s p e n n s y l v a n i c u s a n d T a p i n o m a s e s s i l e ( K l o t z a n d R e i d 1 9 9 2 ) T h e y w i l l a l s o f o r a g e v i a d u c t s a n d w a l l s p a c e s a l u m i n u m w i n d o w a n d d o o r f r a m e s a n d t h e y c a n f o r a g e u p t o d i s t a n c e s o f 4 5 m ( V a i l a n d W i l l i a m s 1 9 9 4 ) P h a r a o h a n t s a r e w e l l k n o w n f o r t h e i r c o n s u m p t i o n o f a w i d e v a r i e t y o f f o o d s ( E d w a r d s a n d B a k e r 1 9 8 1 a ) O v e r a l l t h e P h a r a o h a n t n u t r i t i o n a l n e e d s a r e c o m p l e x e n o u g h t o i n d i c a t e t h a t t h e y d o n e e d e a c h o f t h e m a j o r n u t r i t i o n a l g r o u p s ( l i p i d s c a r b o h y d r a t e s a n d p r o t e i n s ) b u t t h e P h a r a o h a n t i s a n o m n i v o r o u s c a r n i v o r e ( E b e l i n g 1 9 7 8 ) S o m e r e s e a r c h e r s b e l i e v e t h a t t h e P h a r a o h a n t i s p r i m a r i l y a t t r a c t e d t o s u g a r s ( G r a n o v s k y a n d H o w e l l 1 9 8 3 ) w h i l e o t h e r s t h i n k t h a t t h e y p r i m a r i l y p r e f e r t o c o n s u m e p r o t e i n s ( E d w a r d s a n d A b r a h a m 1 9 9 0 ) N o c h o i c e r e p l i c a t i o n s w e r e c o n d u c t e d b y p l a c i n g s t a r v e d ( 2 d ) P h a r a o h a n t c o l o n i e s i n a n a r e n a w i t h o n e f l u i d l o a d e d m i c r o c a p i l l a r y t u b e T e s t a r e n a s w e r e p a i r e d w i t h a d j a c e n t c o n t r o l a r e n a s t o c o m p r i s e a n e x p e r i m e n t a l u n i t C h o i c e r e p l i c a t i o n s w e r e p e r f o r m e d i n t h e s a m e m a n n e r e x c e p t t h a t 5 f l u i d s w e r e p r o v i d e d s i m u l t a n e o u s l y W o r k e r s f o r a g e d f r o m t h e n e s t i n g c e l l s t o t h e m i c r o c a p i l l a r y t u b e s i n o r d e r t o o b t a i n f o o d a n d w a t e r C o n s u m p t i o n w a s m e a s u r e d a t 2 4 h N o c h o i c e s t u d i e s d e m o n s t r a t e d t h a t P h a r a o h a n t s c o n s u m e d s i g n i f i c a n t a m o u n t s o f a l l p a t i e n t c a r e a n d h u m a n f l u i d s p r o v i d e d C o n s u m p t i o n o f 5 % d e x t r o s e ( 3 7 9 5 m g )

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8 9 w a s s i g n i f i c a n t l y h i g h e r t h a n t h a t o f a l l o t h e r f l u i d s C o n s u m p t i o n o f b a s e l i n e w a t e r c o n t r o l s w a s 2 5 4 m g C h o i c e s t u d i e s d e m o n s t r a t e d t h a t P h a r a o h a n t s c o n s u m e d s i g n i f i c a n t a m o u n t s o f a l l p a t i e n t c a r e a n d h u m a n f l u i d s p r o v i d e d C o n s u m p t i o n o f 5 % d e x t r o s e ( 4 9 1 4 m g ) w a s s i g n i f i c a n t l y h i g h e r t h a n t h a t o f a l l o t h e r f l u i d s B l o o d w a s l e a s t c o n s u m e d a t 2 0 5 m g S t a r v e d ( 3 d ) P h a r a o h a n t s r e c r u i t e d t o d r i e d r e s i d u e s o f p a t i e n t c a r e f l u i d s F o r a g i n g p e a k e d a t 4 5 m i n a f t e r p l a c e m e n t w i t h a m e a n n u m b e r o f 3 4 2 a n t s p e r s u b s t a n c e a n d d e c l i n e d t o a n a v e r a g e o f 1 8 5 a n t s p e r s u b s t a n c e a t 1 2 0 m i n a f t e r p l a c e m e n t F o r a g i n g t o E n s u r e d i e t a r y s u p p l e m e n t ( 1 3 8 9 a n t s ) w a s s i g n i f i c a n t l y h i g h e s t f o l l o w e d b y r e c r u i t m e n t t o 5 % d e x t r o s e ( 1 4 9 a n t s ) F e w a n t s ( 0 0 3 0 0 7 a n t s p e r s u b s t a n c e ) r e c r u i t e d t o p l a s m a b l o o d c e l l s o r s a l i n e T h i s s t u d y d e m o n s t r a t e d t h a t P h a r a o h a n t s o c c u r r i n g i n h o s p i t a l s a r e c a p a b l e o f c o n s u m i n g p a t i e n t c a r e f l u i d s e s p e c i a l l y 5 % d e x t r o s e i n t h e l i q u i d s t a t e a n d E n s u r e i n t h e d e s i c c a t e d s t a t e R e c r u i t m e n t a n d c o n s u m p t i o n o c c u r r e d d e s p i t e t h e a d l i b i t u m p r o v i s i o n o f w a t e r i n t e s t a r e n a s P h a r a o h a n t s l i v e i n h o s p i t a l s a n d h a v e b e e n o b s e r v e d f e e d i n g u p o n p a t i e n t s a s w e l l a s u p o n p a t i e n t c a r e f l u i d s T h e i r s t r o n g s y n a n t h r o p i c b e h a v i o r i n c r e a s e s t h e i r p o t e n t i a l t o t r a n s m i t n o s o c o m i a l a n d o t h e r i n f e c t i o u s d i s e a s e s t o h u m a n s i n h o s p i t a l e n v i r o n m e n t s C o n t r o l i s d i f f i c u l t d u e t o t h e r e p r o d u c t i v e s t r a t e g i e s o f t h i s i n s e c t ( E d w a r d s 1 9 8 6 ) T h e h e a l t h t h r e a t w h i c h t h e y p o s e c a n b e e f f e c t i v e l y d e c r e a s e d b y r e d u c i n g s p i l l a g e o f p a t i e n t c a r e f l u i d s e s p e c i a l l y h i g h s u g a r f l u i d s s u c h a s 5 % d e x t r o s e a n d E n s u r e d i e t a r y s u p p l e m e n t

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9 0 A P P E N D I X A A N T M A I N T E N A N C E A l l p a r e n t a n t c o l o n i e s ( > 2 0 0 0 0 0 w o r k e r s ) w e r e h o u s e d a n d r e a r e d a t t h e U r b a n E n t o m o l o g y L a b o r a t o r y U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F L R e a r i n g t e m p e r a t u r e s r a n g e d f r o m 2 1 9 C – 2 9 4 C ( 7 1 5 F 8 5 F ) a n d r e l a t i v e h u m i d i t y f l u c t u a t e d b e t w e e n 2 0 5 3 % A t t e m p t s w e r e m a d e t o c o n t r o l r o o m t e m p e r a t u r e a n d h u m i d i t y b y r u n n i n g a s p a c e h e a t e r 2 4 h o u r s p e r d a y a n d f i l l i n g a h u m i d i f i e r o n c e o r t w i c e p e r d a y a s n e e d e d t o c o m p e n s a t e f o r t h e r o o m ’ s e x t r e m e a i r f l o w ( r e a r i n g r o o m c o n n e c t e d t o o f f i c e s w i t h i n t e g r a t e d a i r f l o w a n d u s e d o u t s i d e c o l d w i n t e r a i r f o r a i r f l o w r e p l e n i s h m e n t ) E a c h c o l o n y w a s h o u s e d w i t h i n a l a r g e ( 4 0 b y 5 2 b y 1 2 c m d e e p ) p l a s t i c t r a y ( M o d e l N o 4 0 0 5 N P a n e l C o n t r o l s G r e e n v i l l e S C ) T r a y s i d e s w e r e c o a t e d w i t h F l u o n ™ ( A s a h i G l a s s F l u o r o p o l y m e r s D o w n i n g t o n P A ) t o p r e v e n t a n t e s c a p e F o u r t o e i g h t c e l l s ( d e p e n d i n g o n t h e s i z e o f t h e c o l o n y ) w e r e p r o v i d e d a s h a r b o r a g e C e l l s c o n s i s t e d o f s q u a r e p l a s t i c P e t r i d i s h e s ( 1 0 0 b y 1 0 0 b y 1 5 m m ) ( W i l l i a m s 1 9 9 0 ) w i t h a c i r c u l a r ( 5 0 b y 2 m m d e e p ) d e p r e s s i o n i n C a s t o n e d e n t a l p l a s t e r ( M o d e l N o 9 9 0 4 5 D e n t s p l y I n t e r n a t i o n a l Y o r k P A ) C e l l l i d s c o n t a i n e d 4 m m h o l e s i n e a c h c o r n e r t o p r o v i d e a n t s w i t h m u l t i p l e i n g r e s s a n d e g r e s s p o i n t s Y e l l o w o r r e d a c e t a t e s h e e t s w e r e t a p e d o n t o p o f c e l l l i d s o s t e n s i b l y t o i n c r e a s e d e s i r a b i l i t y o f h a r b o r a g e t o t h e a n t s A c e t a t e s h e e t s t h a t f e l l o f f o v e r t i m e ( a n t s c o n s u m e d t a p e a n d b u i l t n e s t s b e t w e e n a c e t a t e s h e e t s a n d l i d ) w e r e n o t r e p l a c e d I n a l l c a s e s l i d s o f c e l l s b e c a m e c o m p l e t e l y c o a t e d w i t h h a r d b r o w n f i n e l y g r a i n e d a n t r e g u r g i t a t i o n a n d p a r t i a l l y d i g e s t e d f o o d ( P e a c o c k a n d B a x t e r 1 9 4 9 )

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9 1 W a t e r a n d 1 0 % s u c r o s e w e r e p r o v i d e d a d l i b i t u m i n 1 5 m l v i a l s ( M o d e l N o 5 5 9 T h o r n t o n P l a s t i c s S a l t L a k e C i t y U T w i t h t h r e e l a r g e c o t t o n b a l l s i n s e r t e d i n t o t h e t o p t o p r e v e n t l e a k a g e a n d t o p r o v i d e e a s y a c c e s s f o r t h e a n t s N u m b e r o f v i a l s v a r i e d b e t w e e n c o l o n i e s b a s e d o n n u m b e r o f c e l l s w i t h i n e a c h c o l o n y : o n e v i a l o f w a t e r a n d o n e v i a l o f s u c r o s e w a s p r o v i d e d f o r e a c h n e s t i n g c e l l w i t h i n p a r e n t c o l o n y W a t e r a n d s u c r o s e v i a l s w e r e r e p l a c e d o n e t w o t i m e s p e r w e e k ( a s n e e d e d b a s e d o n v i s u a l d e t e r m i n a t i o n o f f l u i d q u a n t i t y r e m a i n i n g p r e s e n c e o f m o l d o r t u n n e l i n g b y a n t s ) F r e s h l y k i l l e d ( f r o z e n ) A m e r i c a n c o c k r o a c h e s P e r i p l a n e t a a m e r i c a n a ( U r b a n E n t o m o l o g y L a b o r a t o r y U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F l o r i d a ) w e r e p r o v i d e d t w o t h r e e t i m e s p e r w e e k f o r p r o t e i n T h a w e d a d u l t a n d i m m a t u r e P a m e r i c a n a w e r e p l a c e d i n a s m a l l s o u f f l c u p ( 2 9 5 m l M o d e l P 1 0 0 S o l o C o m p a n y U r b a n a I L ) w i t h s i d e s c o a t e d w i t h F l u o n ™ A l l o l d f o o d w a s r e m o v e d f r o z e n a n d d i s p o s e d o f w h e n n e w f o o d w a s p r o v i d e d t o p r e v e n t m o l d g r o w t h p h o r i d f l y d e v e l o p m e n t a n d a n t n e s t i n g i n i n s e c t c o r p s e s A n t c o l o n i e s w e r e s u p e r f i c i a l l y c l e a n e d o f d e a d a n t p i l e s a n d f o o d d e b r i s e v e r y w e e k M o r e t h o r o u g h c l e a n i n g s o f a n t r e g u r g i t a t i o n a n d p a r t i a l l y d i g e s t e d f o o d p e l l e t s w e r e p e r f o r m e d o n c e p e r m o n t h a n d w e r e p e r f o r m e d b y r e m o v i n g a l l v i a l s a n d f o o d f r o m t h e l a r g e p l a s t i c t r a y s t a c k i n g c e l l s a t o p o n e a n o t h e r a t a n a n g l e ( k e e p i n g e n t r a n c e / e x i t h o l e s a l w a y s o p e n ) a t o n e s i d e o f t h e t r a y a n d q u i c k l y w i p i n g t r a y b o t t o m w i t h a l a r g e d a m p c o t t o n b a l l t o p i c k u p r e g u r g i t a t e d m a t t e r a n d i n s e c t d e b r i s O n c e c l e a n a l a r g e d r y c o t t o n b a l l w a s w i p e d o v e r t h e s a m e a r e a t o r e m o v e a n y r e m a i n i n g w a t e r t h a t m i g h t d r o w n t h e a n t s T h e s e c o n d s i d e o f t h e l a r g e p l a s t i c t r a y w a s c l e a n e d i n

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9 2 a s i m i l a r m a n n e r A f t e r c l e a n i n g a l l c e l l s w e r e p l a c e d g e n t l y b a c k i n t o o r i g i n a l p o s i t i o n a n d f o o d w a t e r a n d s u c r o s e w e r e a l s o r e t u r n e d t o c o l o n y

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9 3 A P P E N D I X B S E L E C T I O N O F A N T S E a c h t e s t u n i t w a s p o p u l a t e d w i t h 1 2 5 o u t s i d e w o r k e r s t o e n s u r e p r e s e n c e o f f o r a g e r s 1 2 5 i n s i d e w o r k e r s t o e n s u r e p r e s e n c e o f n u r s e s ( E r p e n b e c k 1 9 7 6 M i r e n d a a n d V i n s o n 1 9 8 1 H a a c k 1 9 8 7 ) 3 d e a l a t e q u e e n s a n d 5 0 m g b r o o d ( p u p a e l a r v a e a n d e g g s ) N o a t t e m p t w a s m a d e t o i n c l u d e s p e c i f i c p r o p o r t i o n s o f t h i r d i n s t a r s F o l l o w i n g t h e m e t h o d o f O i e t a l ( 2 0 0 0 ) I r a n d o m l y s e l e c t e d l a r v a e t o c o m p r i s e ~ 7 5 % o f b r o o d w i t h a l l l a r v a l s t a g e s p r e s e n t E g g s a c c o u n t e d f o r ~ 2 5 % o f b r o o d F o r a g i n g a n t s w e r e s e l e c t e d f i r s t b y p l a c i n g a 1 0 c m b y 1 0 c m p i e c e o f w h i t e p h o t o c o p y p a p e r o n t o p o f w a t e r v i a l s i n l a r g e ( > 2 0 0 0 0 0 w o r k e r s ) p a r e n t c o l o n y t o c o l l e c t f o r a g i n g a n t s A n t s w e r e t h e n s h a k e n o f f p a p e r i n t o a n e m p t y o p a q u e p l a s t i c s t o r a g e b o x ( 1 8 c m b y 3 2 5 c m b y 1 0 6 c m d e e p ; M o d e l N o 1 6 5 2 S t e r l i t e C o r p T o w n s e n d M A ) S t o r a g e b o x i n t e r i o r s i d e s w e r e c o a t e d w i t h F l u o n ™ ( A s a h i G l a s s F l u o r o p o l y m e r s D o w n i n g t o n P A ) t o p r e v e n t e s c a p e A n t s w e r e a s p i r a t e d o u t o f s t o r a g e b o x a n d t r a n s f e r r e d t o a s m a l l s o u f f l c u p ( 2 9 5 m l ) w i t h F l u o n ™ c o a t e d s i d e F i v e d i f f e r e n t b a t c h e s ( c o l o n i e s ) o f 1 2 5 a n t s e a c h w e r e c o l l e c t e d ( o n e f o r e a c h t e s t a r e n a ) a n d p l a c e d i n t o i n d i v i d u a l c u p s W h e n e a c h o f t h e f i v e c u p s c o n t a i n e d 1 2 5 f o r a g e r s a l l c u p s c o n t a i n i n g a n t s w e r e p l a c e d i n t o a s t o r a g e b o x a n d s e t a s i d e u n t i l n u r s e s a n d b r o o d w e r e c o l l e c t e d N u r s e s a n d d e a l a t e q u e e n s w e r e s e l e c t e d b y e m p t y i n g i n d i v i d u a l p a r e n t c o l o n y r e a r i n g c e l l s i n t o a n e m p t y s t o r a g e b o x w i t h s i d e s F l u o n ™ c o a t e d t o p r e v e n t e s c a p e A n t s n o t c a r r y i n g b r o o d ( H a a c k 1 9 8 7 ) w e r e a s p i r a t e d o u t o f t h i s a r e n a A s w i t h f o r a g e r s

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9 4 1 2 5 n u r s e s w e r e c o l l e c t e d a n d p l a c e d i n t o e a c h o f f i v e s m a l l s o u f f l c u p s T h r e e d e a l a t e q u e e n s w e r e t h e n a s p i r a t e d a n d p l a c e d i n t o e a c h c u p a l o n g w i t h n u r s e s T h e s e c u p s w e r e t h e n p l a c e d i n t o a s t o r a g e b o x a n d s e t a s i d e d u r i n g c o l l e c t i o n o f b r o o d B r o o d o f v a r i o u s a g e s w e r e c o l l e c t e d ; a n y a n t s r e m a i n i n g w i t h b r o o d w e r e m a n u a l l y s e p a r a t e d f r o m b r o o d a n d r e t u r n e d t o p a r e n t c o l o n y B r o o d ( 5 0 m g a l l s t a g e s ) w e r e w e i g h e d o n a d i g i t a l s c a l e a n d p l a c e d i n t o e a c h s m a l l c u p c o n t a i n i n g n u r s e s N u r s e s a n d q u e e n s w i t h i n e a c h c u p w e r e p e r m i t t e d 1 h t o o r g a n i z e b r o o d A f t e r 1 h a d j u s t m e n t t i m e o n e e n t i r e c u p w a s e m p t i e d i n t o e a c h a r e n a b y t u r n i n g t h e c u p u p s i d e d o w n o v e r c o v e r e d P e t r i d i s h ( n e s t i n g c e l l ) a n d t a p p i n g l i g h t l y s o t h a t n u r s e s q u e e n s a n d b r o o d l a n d e d o n t o p o f h a r b o r a g e c e l l I m m e d i a t e l y a f t e r w a r d s f o r a g e r s w e r e e m p t i e d i n t o t e s t a r e n a i n s a m e m a n n e r A r e n a w a s c o v e r e d w i t h p l a s t i c l i d P l a c e m e n t i n t o a r e n a b e g a n s t a r v a t i o n p e r i o d A n t s w e r e s t a r v e d i n t e s t a r e n a 4 8 h p r i o r t o t e s t W a t e r w a s a v a i l a b l e a d l i b i t u m d u r i n g t h e e n t i r e s t a r v a t i o n p e r i o d

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9 5 A P P E N D I X C C O N S U M P T I O N F O R M U L A S F l u i d l o s s i n t e s t a r e n a s w a s c o m p r i s e d o f c o n s u m p t i o n a n d e v a p o r a t i o n ; t h e r e f o r e t o d e t e r m i n e c o n s u m e d q u a n t i t i e s t h e a m o u n t o f f l u i d l o s t t o e v a p o r a t i o n m u s t b e s u b t r a c t e d M a n y r e s e a r c h e r s h a v e d e s i g n e d f o r m u l a s t o a d j u s t c o n s u m p t i o n f o r f l u i d l o s s a n d I e x a m i n e d e a c h o f t h e f o l l o w i n g b e f o r e s e l e c t i n g a n d m o d i f y i n g W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a I n i t i a l l y I m e a s u r e d c o n s u m p t i o n b y p l a c i n g a m e t r i c r u l e r ( m m s c a l e ) a l o n g s i d e a 3 0 l ( 1 m m I D ) m i c r o c a p i l l a r y t u b e i n a m e t h o d s i m i l a r t o t h a t o f ( M a y 1 9 8 5 a 1 9 8 5 b ) T h i s m e t h o d w a s q u i c k l y a b a n d o n e d w h e n I d e t e r m i n e d t h a t g r a v i m e t r i c m e a s u r e m e n t o f f l u i d s w a s m o r e p r e c i s e f o r t h e s m a l l v o l u m e s o f f l u i d I w a s u s i n g T h e r e f o r e I c o n s i d e r e d c a l c u l a t i n g c o n s u m p t i o n b y g r a v i m e t r i c m e a s u r e m e n t ( E v a n s 1 9 3 9 W e i a n d J o h n s o n 1 9 9 6 ) E v a n s ’ ( 1 9 3 9 ) f o r m u l a w a s b a s e d o n t h e a s s u m p t i o n t h a t t h e n a t u r a l m o i s t u r e l o s s r a t e s o f l e a v e s k e p t i n c o n t a i n e r s w i t h o u t i n s e c t s w a s s i m i l a r t o t h a t o f l e a v e s p r o v i d e d a s f o o d t o i n s e c t s a n d h e c o n t r o l l e d c o n s u m p t i o n f o r n a t u r a l m o i s t u r e l o s s b y t a k i n g t h e d i f f e r e n c e o f u n e a t e n a n d i n t r o d u c e d f o o d w e i g h t s a n d m u l t i p l y i n g b y m o i s t u r e l o s s r a t e : ( % w t l o s s o f a l i q u o t ) x ( w t f o o d i n t r o d u c e d + w t u n e a t e n f o o d ) / 2 A n y a s s u m p t i o n s W e i a n d J o h n s o n ( 1 9 9 6 ) m a y h a v e u s e d i n t h e d e v e l o p m e n t o f t h e i r g r a v i m e t r i c m e t h o d t o d e t e r m i n e a d u l t L e p i d o p t e r a c o n s u m p t i o n o f s u c r o s e s o l u t i o n s v i a a m i c r o c a p i l l a r y t u b w e r e n o t r e p o r t e d

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9 6 M i c r o c a p i l l a r y t u b e s p r o v i d e o n e o f t h e m o s t c o n t r o l l e d m e t h o d s f o r l i q u i d f o o d c o n s u m p t i o n s t u d i e s I n a t u b e w a t e r l o s s o c c u r s o n l y a s e v a p o r a t i o n f r o m t h e o p e n e n d s i n c o n t r a s t t o a l e a f w h e r e s u r f a c e a r e a d e c r e a s e s w i t h c o n s u m p t i o n s o t h a t w a t e r l o s s i s a f f e c t e d b y c h a n g i n g t r a n s p i r a t i o n a n d m e t a b o l i s m i n a d d i t i o n t o e v a p o r a t i o n W h i l e t h e s u b t r a c t i o n m e t h o d i s s i m p l e a n d i n t u i t i v e i t d o e s n o t t a k e i n t o a c c o u n t t h e f a c t t h a t e v a p o r a t i o n r a t e s o f f l u i d s c a n c h a n g e o v e r t i m e a s f l u i d l o s s o c c u r s ( E v a n s 1 9 3 9 C a n d y a n d B a k e r 2 0 0 2 ) I t a l s o d o e s n o t a c c o u n t f o r p o t e n t i a l l y i n c r e a s e d s u r f a c e a r e a o f d e s i c c a t i n g f l u i d s E v a n s ( 1 9 3 9 ) d e t e r m i n e d a w e i g h t l o s s r a t i o b y m u l t i p l y i n g t h e p e r c e n t w e i g h t l o s s i n c o n t r o l a r e n a s b y t h e m e a n w e i g h t o f p r o v i d e d a n d r e m a i n i n g f o o d i n t e s t a r e n a s A s m e n t i o n e d i n C a n d y a n d B a k e r ( 2 0 0 2 ) t h i s a d j u s t s c o n s u m p t i o n o n l y f o r e v a p o r a t i o n r a t e s o f i n i t i a l w e i g h t o f f o o d p r o v i d e d i n c o n t r o l a r e n a s W a l d b a u e r ( 1 9 6 2 1 9 6 8 ) e x p a n d e d t h i s f o r m u l a t o i n c l u d e a d j u s t m e n t r a t i o s f o r e v a p o r a t i o n r a t e s o f b o t h p r o v i d e d a n d r e m a i n i n g f o o d A l t h o u g h h e a s s u m e d t h a t m o i s t u r e l o s s d e c r e a s e d b y h a l f W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a i s a s t a n d a r d m e t h o d o f m e a s u r i n g c o n s u m p t i o n i n n u t r i t i o n a l e c o l o g y l i t e r a t u r e ( C a n d y a n d B a k e r 2 0 0 2 ) T h e r e f o r e h i s f o r m u l a ( s h o w n b e l o w ) c a l c u l a t e s c o n s u m p t i o n c o r r e c t e d f o r f l u i d l o s s o f v a r i a b l e r a t e s b a s e d u p o n a n a s s u m p t i o n o f m o i s t u r e l o s s r a t e s o v e r t i m e : C = ( 1 – a / 2 ) ( W [ L + b L ] ) w h e r e C = t r u e q u a n t i t y o f f l u i d c o n s u m e d a f t e r a d j u s t e d f o r e v a p o r a t i o n W = m a s s o f f l u i d p r o v i d e d i n t e s t a r e n a L = m a s s o f u n e a t e n f l u i d a n d a n t s r e m a i n i n g i n f l u i d i n t e s t a r e n a

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9 7 a = r a t i o o f c o n t r o l f l u i d w e i g h t l o s s a n d c o n t r o l f l u i d i n i t i a l m a s s b = r a t i o o f c o n t r o l f l u i d e v a p o r a t i o n a n d f i n a l m a s s W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a w a s b a s e d o n s e v e r a l a s s u m p t i o n s : ( 1 ) m o i s t u r e l o s s w o u l d d e c r e a s e a t a c o n s t a n t r a t e o v e r t h e t e s t p e r i o d s o t h a t f i n a l m o i s t u r e l o s s r a t e s w e r e h a l f t h e i n i t i a l m o i s t u r e l o s s r a t e s ( 2 ) p e r c e n t w e i g h t l o s s w a s i n d e p e n d e n t o f f o o d w e i g h t a n d ( 3 ) f e e d i n g d o e s n o t i n c r e a s e w e i g h t l o s s ( C a n d y a n d B a k e r 2 0 0 2 ) d e r i v e d d i f f e r e n t i a l e q u a t i o n s f r o m W a l d b a u e r ’ s ( 1 9 6 8 ) w o r k f o r ( 1 ) w e i g h t l o s s d u e t o f e e d i n g a n d m o i s t u r e l o s s a n d ( 2 ) w e i g h t l o s s o f c o n t r o l s d u e t o m o i s t u r e l o s s o v e r s e v e r a l d a y s a n d r e p o r t e d t h a t h i s f o r m u l a r e s u l t e d i n n e g l i g i b l e c o n s u m p t i o n o v e r e s t i m a t e s ( < 0 2 % ) w h e n p e r c e n t a g e o f m o i s t u r e l o s s o f c o n t r o l f o o d s w a s < 1 0 % b u t t h a t c o n s u m p t i o n o v e r e s t i m a t e s w e r e s t a t i s t i c a l l y s i g n i f i c a n t w h e n m o i s t u r e l o s s o f c o n t r o l f o o d s w a s l a r g e M y e x p e r i m e n t l a s t e d f o r o n l y o n e d a y s o t h e d i f f e r e n t i a l e q u a t i o n m o d e l o f C a n d y a n d B a k e r ( 2 0 0 2 ) w a s n o t a p p l i c a b l e W a l d b a u e r ’ s ( 1 9 6 8 ) o v e r e s t i m a t i o n o f c o n s u m p t i o n w a s n o t a l i m i t a t i o n f o r m y e x p e r i m e n t s i n c e p e r c e n t a g e o f m o i s t u r e l o s s i n c o n t r o l a r e n a s w a s < 1 0 % T h u s W a l d b a u e r ’ s ( 1 9 6 8 ) f o r m u l a a c c o m m o d a t e d m y d a t a s e t W a l d b a u e r ( 1 9 6 8 ) s t u d i e d c a t e r p i l l a r c o n s u m p t i o n o f f r e s h l e a v e s C a n d y a n d B a k e r ’ s ( 2 0 0 2 ) f o r m u l a w a s b a s e d o n t h e f o l l o w i n g a s s u m p t i o n s : ( 1 ) p e r c e n t a g e w e i g h t l o s s w a s i n d e p e n d e n t o f f o o d w e i g h t ( 2 ) f e e d i n g d o e s n o t i n c r e a s e w e i g h t l o s s ( 3 ) f e e d i n g r a t e p e r c a t e r p i l l a r w a s c o n s t a n t a n d ( 4 ) m o i s t u r e l o s s r a t e w a s c o n s t a n t p r o p o r t i o n t o l e a f w e i g h t w i t h a n e x p o n e n t i a l d e c r e a s e i n w e i g h t l o s s B e c a u s e E v a n s ( 1 9 3 9 ) W a l d b a u e r ( 1 9 6 8 ) a n d C a n d y a n d B a k e r ( 2 0 0 2 ) a l l s t u d i e d c o n s u m p t i o n o f l e a v e s b y c a t e r p i l l a r s t h e i r a s s u m p t i o n t h a t f e e d i n g d o e s n o t i n c r e a s e n a t u r a l w e i g h t l o s s b y c r e a t i n g a f r e s h l y c u t s u r f a c e A l t h o u g h I u s e d l i q u i d s a n d w a s

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9 8 n o t c o n c e r n e d a b o u t f r e s h l y c u t s u r f a c e s I w a s c o n c e r n e d a b o u t c h a n g i n g r a t i o s o f e x p o s e d s u r f a c e a r e a i n r e g a r d t o l i q u i d v o l u m e s i n c e e i t h e r a n i n c r e a s e d s u r f a c e a r e a o r a d e c r e a s e d v o l u m e w o u l d r e s u l t i n i n c r e a s e d m o i s t u r e l o s s r a t e s o v e r t i m e S u r f a c e a r e a i n c r e a s e d d u e t o d e s i c c a t i o n i n b l o o d c e l l s a n d E n s u r e a n d r e m a i n e d c o n s t a n t i n a l l o t h e r f l u i d s F e e d i n g r e s u l t e d i n d e c r e a s e d v o l u m e i n a l l f l u i d s T h u s t h e m o i s t u r e l o s s r a t e s o f a l l f l u i d s c h a n g e d o v e r t i m e a n d W a l d b a u e r ’ s ( 1 9 6 8 ) a c c o m m o d a t e s m y d a t a b y p e r m i t t i n g a d j u s t m e n t s f o r c h a n g i n g m o i s t u r e l o s s r a t e s e v e n t h o u g h I u s e d l i q u i d s r a t h e r t h a n f r e s h l y c u t l e a v e s

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9 9 L I S T O F R E F E R E N C E S A m e r i c a n A s s o c i a t i o n o f B l o o d B a n k s ( A A B B ) 2 0 0 2 C i r c u l a r o f i n f o r m a t i o n f o r t h e u s e o f h u m a n b l o o d a n d b l o o d c o m p o n e n t s A m e r i c a n A s s o c i a t i o n o f B l o o d B a n k s B e t h e s d a M D 3 7 p p A n o n 1 9 7 4 P h a r a o h s a n t s i n h o s p i t a l s B r i t M e d J 2 : 6 6 A n o n 1 9 8 6 P h a r a o h a n t s M o n o m o r i u m p h a r a o n i s ( L ) N a t i o n a l P e s t C o n t r o l A s s o c i a t i o n E S P C 0 3 2 1 5 5 D u n n L o r i n g V A 6 p p A r n o l d G 1 9 1 6 A m o n o g r a p h o f t h e F o r m i c i d a e o f S o u t h A f r i c a P a r t I I P o n e r i n a e D o r y l i n a e A n n S A f r M u s 1 4 : 1 5 9 2 7 0 B a r b e r J T E G E l l g a a r d L B T h i e n a n d A E S t a c k 1 9 8 9 T h e u s e o f t o o l s f o r f o o d t r a n s p o r t a t i o n b y t h e i m p o r t e d f i r e a n t S o l e n o p s i s i n v i c t a A n i m B e h a v 3 8 : 5 5 0 5 5 2 B e a t s o n S H 1 9 7 2 P h a r a o h s a n t s a s p a t h o g e n v e c t o r s i n h o s p i t a l s L a n c e t 1 : 4 2 5 4 2 7 B e a t s o n S H 1 9 7 3 P h a r a o h s a n t s e n t e r g i v i n g s e t s L a n c e t 1 : 6 0 6 B e l l e v o y e M A 1 8 8 9 O b s e r v a t i o n s o n M o n o m o r i u m p h a r a o n i s L a t r I n s e c t L i f e 2 : 2 3 0 2 3 3 B e r n d t K P 1 9 7 7 P h y s i o l o g y o f r e p r o d u c t i o n i n t h e P h a r a o h s a n t ( M o n o m o r i u m p h a r a o n i s L ) 1 P h e r o m o n e m e d i a t e d c y c l i c p r o d u c t i o n o f s e x u a l s W i a d o m P a r a z y t 2 3 : 1 6 3 1 6 6 B o l t o n B 1 9 8 7 A r e v i e w o f t h e S o l e n o p s i s g e n u s g r o u p a n d r e v i s i o n o f A f r o t r o p i c a l M o n o m o r i u m M a y r ( H y m e n o p t e r a : F o r m i c i d a e ) B u l l B r M u s E n t o m o l 5 4 : 2 6 3 4 5 2 B o l t o n B 1 9 9 4 I d e n t i f i c a t i o n g u i d e t o t h e a n t g e n e r a o f t h e w o r l d H a r v a r d U n i v e r s i t y P r e s s C a m b r i d g e M A 5 1 2 p p B o l t o n B 1 9 9 5 A n e w g e n e r a l c a t a l o g u e o f t h e a n t s o f t h e w o r l d H a r v a r d U n i v e r s i t y P r e s s C a m b r i d g e M A 5 0 4 p p

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1 0 0 B r g e s e n L W a n d P V J e n s e n 1 9 9 5 I n f l u e n c e o f l a r v a e a n d w o r k e r s o n e g g p r o d u c t i o n o f q u e e n s o f t h e p h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) I n s e c t S o c 4 2 : 1 0 3 1 1 2 B o y d A M a n d B H K a y 2 0 0 0 V e c t o r c o m p e t e n c e o f A e d e s a e g y p t i C u l e x s i t i e n s C u l e x a n n u l i r o s t r i s a n d C u l e x q u i n q u e f a s c i a t u s ( D i p t e r a : C u l i c i d a e ) f o r B a r m a h F o r e s t V i r u s J M e d E n t o m o l 3 7 : 6 6 0 6 6 3 B u e n o O C a n d H G F o w l e r 1 9 9 4 E x o t i c a n t s a n d n a t i v e a n t f a u n a o f B r a z i l i a n h o s p i t a l s p p 1 9 1 1 9 8 I n D F W i l l i a m s [ e d ] E x o t i c a n t s W e s t v i e w P r e s s B o u l d e r C O 3 3 2 p p B u t l e r C G 1 9 4 0 T h e c h o i c e o f d r i n k i n g w a t e r o f t h e h o n e y b e e J E x p B i o l 1 7 : 2 5 3 2 6 1 C a e t a n o F H 1 9 9 0 M o r p h o l o g y o f t h e d i g e s t i v e t r a c t a n d a s s o c i a t e d e x c r e t o r y o r g a n s o f a n t s p p 1 1 9 1 2 9 I n R K V a n d e r M e e r K J a f f e a n d A C e d e n o [ e d s ] A p p l i e d m y r m e c o l o g y : a w o r l d p e r s p e c t i v e W e s t v i e w P r e s s B o u l d e r C O 7 4 1 p p C a n d y S G a n d S C B a k e r 2 0 0 2 C a l c u l a t i n g f o o d c o n s u m p t i o n i n t h e l a b o r a t o r y : a f o r m u l a t o a d j u s t f o r n a t u r a l f o o d w e i g h t l o s s A u s t r J E n t o m o l 4 1 : 1 7 0 1 7 3 C a r t w r i g h t R Y a n d C M C l i f f o r d 1 9 7 3 P h a r a o h s a n t s T h e L a n c e t : 1 4 5 5 1 4 5 6 C a s s i l l D L a n d W R T s c h i n k e l 1 9 9 9 T a s k s e l e c t i o n b y w o r k e r s o f t h e f i r e a n t S o l e n o p s i s i n v i c t a B e h a v E c o l S o c i o b i o l 4 5 : 3 0 1 3 1 0 C h a p m a n R F 1 9 9 8 T h e i n s e c t s C a m b r i d g e U n i v e r s i t y P r e s s C a m b r i d g e M A 4 9 0 p p E b e l i n g W 1 9 7 8 U r b a n E n t o m o l o g y U n i v e r s i t y o f C a l i f o r n i a B e r k e l e y C A 6 9 5 p p E d w a r d s J P 1 9 7 5 T h e e f f e c t s o f a j u v e n i l e h o r m o n e a n a l o g u e o n l a b o r a t o r y c o l o n i e s o f P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) ( H y m e n o p t e r a F o r m i c i d a e ) B u l l E n t o m o l R e s 6 5 : 7 5 8 0 E d w a r d s J P 1 9 8 6 T h e b i o l o g y e c o n o m i c i m p o r t a n c e a n d c o n t r o l o f t h e P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) p p 2 5 7 2 7 1 I n S B V i n s o n [ e d ] E c o n o m i c i m p a c t a n d c o n t r o l o f s o c i a l i n s e c t s P r a e g e r P u b l i s h e r s N e w Y o r k N Y 4 3 2 p p E d w a r d s J P 1 9 8 7 C a s t e r e g u l a t i o n i n t h e P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s : t h e i n f l u e n c e o f q u e e n s o n t h e p r o d u c t i o n o f n e w s e x u a l f o r m s P h y s i o l E n t o m o l 1 2 : 3 1 3 9

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1 0 1 E d w a r d s J P 1 9 9 1 C a s t e r e g u l a t i o n i n t h e P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s : r e c o g n i t i o n a n d c a n n i b a l i s m o f s e x u a l b r o o d b y w o r k e r s P h y s i o l E n t o m o l 1 6 : 2 6 3 2 7 1 E d w a r d s J P a n d L A b r a h a m 1 9 9 0 C h a n g e s i n f o o d s e l e c t i o n b y w o r k e r s o f t h e P h a r a o h a n t M o n o m o r i u m p h a r a o n i s M e d V e t E n t o m o l 4 : 2 0 5 2 1 1 E d w a r d s J P a n d L F B a k e r 1 9 8 1 a D i s t r i b u t i o n a n d i m p o r t a n c e o f t h e P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) i n N a t i o n a l H e a l t h S e r v i c e H o s p i t a l s i n E n g l a n d J H o s p I n f e c t 2 : 2 4 9 2 5 4 E d w a r d s J P a n d L F B a k e r 1 9 8 1 b P h a r a o h s a n t s I n t P e s t C o n t r 2 3 : 3 5 3 6 E d w a r d s J P a n d J E S h o r t 1 9 9 0 P h a r a o h s a n t a n o v e r v i e w p p 1 1 1 6 I n P r o c e e d i n g s N a t i o n a l C o n f e r e n c e o f U r b a n E n t o m o l o g y S l o u g h B e r k s h i r e s U K E i c h l e r W 1 9 9 0 H e a l t h a s p e c t s a n d c o n t r o l o f M o n o m o r i u m p h a r a o n i s p p 6 7 1 6 7 5 I n R K V a n d e r M e e r K J a f f e a n d A C e d e n o [ e d s ] A p p l i e d m y r m e c o l o g y : a w o r l d p e r s p e c t i v e W e s t v i e w P r e s s B o u l d e r C O 7 4 1 p p E m e r y C 1 9 2 2 H y m e n o p t e r a F a m F o r m i c i d a e S u b f a m M y r m i c i n a e G e n e r a I n s e c t o r u m 1 7 4 B : 9 5 2 0 6 E r p e n b e c k A 1 9 7 6 T r a c e r u n t e r s u c h u n g e n z u r N a h r u n g s v e r t e i l u n g i n S o z i e t t e n d e r P h a r a o a m e i s e M o n o m o r i u m p h a r a o n i s ( L ) ( H y m e n o p t e r a : F o r m i c i d a e ) P h D d i s s e r t a t i o n B o n n U n i v e r s i t y B o n n G e r m a n y 7 0 p p E v a n s A C 1 9 3 9 T h e u t i l i s a t i o n o f f o o d b y t h e l a r v a e o f t h e b u f f t i p P h a l e r a b u c e r h a l a ( L i n n ) ( L e p i d o p t e r a ) p p 2 5 3 0 I n P r o c e e d i n g s R o y a l E n t o m o l o g i c a l S o c i e t y L o n d o n U K F o w l e r H G 1 9 9 0 T h e p e s t a n t s o f S o u t h A m e r i c a p p 2 6 9 2 7 0 I n G K V e e r e s h B M a l l i k a n d C A V i r a k t a m a t h [ e d s ] S o c i a l i n s e c t s a n d t h e e n v i r o n m e n t O x f o r d & I B H P u b l C o P v t L t d N e w D e l h i I n d i a 7 6 5 p p F o w l e r H G M N S c h l i n d w e i n a n d M A d e M e d e i r o s 1 9 9 4 E x o t i c a n t s a n d c o m m u n I t y s i m p l i f i c a t i o n i n B r a z i l : a r e v i e w o f t h e i m p a c t o f e x o t i c a n t s o n n a t i v e a n t a s s e m b l a g e s p p 1 5 1 1 6 2 I n D W h e e l e r [ e d ] E x o t i c a n t s W e s t v i e w P r e s s B o u l d e r C O 3 3 2 p p G r a c e J K D L W o o d a n d B W G r u n b a u m 1 9 8 6 E f f e c t o f A r g e n t i n e a n t c o n t a m i n a t i o n o n A B O b l o o d t y p i n g o f h u m a n s a l i v a s a m p l e s B u l l E n t o m o l S o c A m 3 2 : 1 4 7 1 4 9

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1 0 2 G r a n o v s k y T A a n d H H H o w e l l 1 9 8 3 T e x a s A & M r e s e a r c h t e a m d e v e l o p s n e w P h a r a o h s a n t c o n t r o l t e c h n i q u e P e s t C o n t r T e c h 1 1 : 3 0 3 4 G r e e n A A M J K a n e P S T y l e r a n d D G H H a l s t e a d 1 9 5 4 T h e c o n t r o l o f P h a r a o h s a n t i n h o s p i t a l s P e s t I n f e s t R e s 1 : 2 4 H a a c k K D 1 9 8 7 A s p e c t s o f t h e f o o d h a n d l i n g b e h a v i o r o f t h e P h a r a o h a n t M o n o m o r i u m p h a r a o n i s ( L ) ( H y m e n o p t e r a : F o r m i c i d a e ) M S t h e s i s T e x a s A & M U n i v e r s i t y C o l l e g e S t a t i o n T X 1 2 5 p p H a a c k K D 1 9 9 1 E l i m i n a t i o n o f P h a r a o h a n t s : a n a n a l y s i s o f f i e l d t r i a l s w i t h P r o c o n t r o l a n d M a x f o r c e a n t b a i t s P e s t C o n t r T e c h 1 9 : 3 2 4 2 H a a c k K D a n d S B V i n s o n 1 9 9 0 F o r a g i n g o f P h a r a o h a n t s M o n o m o r i u m p h a r a o n i s ( L ) ( H y m e n o p t e r a : F o r m i c i d a e ) i n t h e l a b o r a t o r y p p 4 5 2 4 6 0 I n R K V a n d e r M e e r K J a f f e a n d A C e d e n o [ e d s ] A p p l i e d m y r m e c o l o g y : a w o r l d p e r s p e c t i v e W e s t v i e w P r e s s B o u l d e r C O 7 4 1 p p H a a c k K D S B V i n s o n a n d J S O l s o n 1 9 9 5 F o o d d i s t r i b u t i o n a n d s t o r a g e i n c o l o n i e s o f M o n o m o r i u m p h a r a o n i s ( L ) ( H y m e n o p t e r a : F o r m i c i d a e ) J E n t o m o l S c i 3 0 : 7 0 8 1 H a r r i s J R 1 9 9 1 E n v i r o n m e n t a l f a c t o r s a f f e c t i n g t h e d i s t r i b u t i o n o f e s t a b l i s h e d a n d t h e f o r m a t i o n o f s a t e l l i t e c o l o n i e s o f t h e P h a r a o h a n t M o n o m o r i u m p h a r a o n i s ( L ) ( H y m e n o p t e r a : F o r m i c i d a e ) P h D d i s s e r t a t i o n T e x a s A & M U n i v e r s i t y C o l l e g e S t a t i o n T X 2 0 3 p p H e d g e s S A 1 9 9 7 A n t s p p 5 0 3 5 8 9 I n D M o r e l a n d [ e d ] H a n d b o o k o f P e s t C o n t r o l 8 t h e d M a l l i s H a n d b o o k a n d T e c h n i c a l T r a i n i n g C o m p a n y N e w Y o r k N Y 1 4 5 6 p p H l l d o b l e r B a n d E O W i l s o n 1 9 9 0 T h e A n t s B e l k n a p P r e s s o f H a r v a r d U n i v e r s i t y P r e s s C a m b r i d g e M A 7 3 2 p p J e a n s o n R F L W R a t n i e k s a n d J L D e n e u b o u r g 2 0 0 3 P h e r o m o n e t r a i l d e c a y r a t e s o n d i f f e r e n t s u b s t r a t e s i n t h e P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s P h y s i o l E n t o m o l 2 8 : 1 9 2 1 9 8 K l o t z J H D H O i K M V a i l a n d D F W i l l i a m s 1 9 9 6 L a b o r a t o r y e v a l u a t i o n o f a b o r i c a c i d l i q u i d b a i t o n c o l o n i e s o f T a p i n o m a m e l a n o c e p h a l u m A r g e n t i n e a n t s a n d P h a r a o h a n t s ( H y m e n o p t e r a : F o r m i c i d a e ) J E c o n E n t o m o l 8 9 : 6 7 3 6 7 7 K l o t z J H a n d B L R e i d 1 9 9 2 T h e u s e o f s p a t i a l c u e s f o r s t r u c t u r a l g u i d e l i n e o r i e n t a t i o n i n T a p i n o m a s e s s i l e a n d C a m p o n o t u s p e n n s y l v a n i c u s ( H y m e n o p t e r a : F o r m i c i d a e ) J I n s e c t B e h a v 5 : 7 1 8 2

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1 0 3 L e M a s n e G 1 9 5 3 O b s e r v a t i o n s s u r l e s r e l a t i o n s e n t r e l e c o u v a i n e t l e s a d u l t e s c h e z l e s f o u r m i s A n n S c i N a t Z o o l 1 5 : 1 5 6 M a i l l e u x A C J L D e n e u b o u r g a n d C D e t r a i n 2 0 0 0 H o w d o a n t s a s s e s s f o o d v o l u m e ? A n i m B e h a v 5 9 : 1 0 1 1 0 6 9 M a y P G 1 9 8 5 a A s i m p l e m e t h o d f o r m e a s u r i n g n e c t a r e x t r a c t i o n r a t e s i n b u t t e r f l i e s J L e p i d o p S o c 3 9 : 5 3 5 5 M a y P G 1 9 8 5 b N e c t a r u p t a k e r a t e s a n d o p t i m a l n e c t a r c o n c e n t r a t i o n s o f t w o b u t t e r f l y s p e c i e s O e c o l o g i a 6 6 : 3 8 1 3 8 6 M e m m l e r R L B J C o h e n a n d D L W o o d 1 9 9 6 T h e h u m a n b o d y i n h e a l t h a n d d i s e a s e L i p p i n c o t t P h i l a d e l p h i a P A 5 1 9 p p M i r e n d a J T a n d S B V i n s o n 1 9 8 1 D i v i s i o n o f l a b o r a n d s p e c i f i c a t i o n o f c a s t e s i n t h e r e d i m p o r t e d f i r e a n t S o l e n o p s i s i n v i c t a B u r e n J A n i m B e h a v 2 9 : 4 1 0 4 2 0 M u l l e n G a n d L A D u r d e n 2 0 0 2 M e d i c a l a n d V e t e r i n a r y E n t o m o l o g y E l s e v i e r S c i e n c e S a n D i e g o C A 5 9 7 p p N a t i o n J L 2 0 0 3 I n s e c t P h y s i o l o g y C R C P r e s s B o c a R a t o n F L 4 8 5 p p O g a t a K M T e r a y a m a a n d R W T a y l o r 1 9 9 8 M o n o m o r i u m p h a r a o n i s ( L i n n a e u s ) J a p a n e s e A n t D a t a b a s e G r o u p ( h t t p : / / a n t e d b / m i y a k y o u a c j p / T a x o E / F 4 1 1 0 7 h t m l ) O c t o b e r 2 2 0 0 3 O i D H K M V a i l a n d D F W i l l i a m s 1 9 9 6 F i e l d e v a l u a t i o n o f p e r i m e t e r t r e a t m e n t s f o r P h a r a o h a n t ( H y m e n o p t e r a : F o r m i c i d a e ) c o n t r o l F l E n t o m o l 7 9 : 2 5 2 2 6 3 O i D H K M V a i l a n d D F W i l l i a m s 2 0 0 0 B a i t d i s t r i b u t i o n a m o n g m u l t i p l e c o l o n i e s o f P h a r a o h a n t s ( H y m e n o p t e r a : F o r m i c i d a e ) J E c o n E n t o m o l 9 3 : 1 2 4 7 1 2 5 5 O i D H K M V a i l D F W i l l i a m s a n d D N B i e m a n 1 9 9 4 I n d o o r a n d o u t d o o r f o r a g i n g l o c a t i o n s o f P h a r a o h a n t s ( H y m e n o p t e r a : F o r m i c i d a e ) a n d c o n t r o l s t r a t e g i e s u s i n g b a i t s t a t i o n s F l E n t o m o l 7 7 : 8 5 9 1 O s t e r G F a n d E O W i l s o n 1 9 7 8 C a s t e a n d e c o l o g y i n t h e s o c i a l i n s e c t s P r i n c e t o n U n i v e r s i t y P r e s s P r i n c e t o n N J 3 7 2 p p P a s s e r a L 1 9 9 4 C h a r a c t e r i s t i c s o f t r a m p s p e c i e s p p 2 3 4 3 I n D F W i l l i a m s [ e d ] E x o t i c a n t s W e s t v i e w P r e s s B o u l d e r C O 3 3 2 p p

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1 0 4 P e a c o c k A D 1 9 5 0 S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 4 E g g p r o d u c t i o n E n t o m o l M o n M a g 8 6 : 2 9 4 2 9 8 P e a c o c k A D a n d A T B a x t e r 1 9 4 9 S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 1 T h e r e a r i n g o f a r t i f i c i a l c o l o n i e s E n t o m o l M o n M a g 8 5 : 2 5 6 2 6 0 P e a c o c k A D a n d A T B a x t e r 1 9 5 0 S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 3 L i f e h i s t o r y E n t o m o l M o n M a g 8 6 : 1 7 1 1 7 8 P e a c o c k A D D W H a l l I C S m i t h a n d A G o o d f e l l o w 1 9 5 0 T h e b i o l o g y a n d c o n t r o l o f t h e a n t p e s t M o n o m o r i u m p h a r a o n i s ( L ) D e p t A g r S c o t l a n d M i s c P u b N o 1 7 : 1 5 1 P e a c o c k A D I C S m i t h D W H a l l a n d A T B a x t e r 1 9 5 4 S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 8 M a l e p r o d u c t i o n b y p a r t h e n o g e n e s i s E n t o m o l M o n M a g 9 0 : 1 5 4 1 5 8 P e a c o c k A D J H S u d d a n d A T B a x t e r 1 9 5 5 a S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 1 1 C o l o n y f o u n d a t i o n E n t o m o l M o n M a g 9 1 : 1 2 5 1 2 9 P e a c o c k A D J H S u d d a n d A T B a x t e r 1 9 5 5 b S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 1 2 D i s s e m i n a t i o n E n t o m o l M o n M a g 9 1 : 1 3 0 1 3 3 P e a c o c k A D F L W a t e r h o u s e a n d A T B a x t e r 1 9 5 5 c S t u d i e s i n P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) 1 0 V i a b i l i t y i n r e g a r d t o t e m p e r a t u r e a n d h u m i d i t y E n t o m o l M o n M a g 9 1 : 3 7 4 2 P e t t i J M 1 9 9 8 T h e s t r u c t u r e a n d f u n c t i o n o f t h e b u c c a l t u b e f i l t e r i n w o r k e r s o f t h e a n t s p e c i e s S o l e n o p s i s i n v i c t a C a m p o n o t u s f l o r i d a n u s a n d M o n o m o r i u m p h a r a o n i s P h D d i s s e r t a t i o n U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F L 1 3 2 p p P o r t e r S D a n d W R T s c h i n k e l 1 9 8 7 F o r a g i n g i n S o l e n o p s i s i n v i c t a ( H y m e n o p t e r a : F o r m i c i d a e ) : e f f e c t s o f w e a t h e r a n d s e a s o n E n v i r o n E n t o m o l 1 6 : 8 0 2 8 0 8 R e i m e r N J W B e a r d s l e y a n d G J a h n 1 9 9 0 P e s t a n t s i n t h e H a w a i i a n I s l a n d s p p 4 0 5 0 I n R K V a n d e r M e e r K J a f f e a n d A C e d e n o [ e d s ] A p p l i e d m y r m e c o l o g y : a w o r l d p e r s p e c t i v e W e s t v i e w P r e s s B o u l d e r C O 7 4 1 p p R i c h a r d s M J J R E d w a r d s D H C u l v e r a n d R P G a y n e s 1 9 9 9 N o s o c o m i a l i n f e c t i o n s i n p e d i a t r i c i n t e n s i v e c a r e u n i t s i n t h e U n i t e d S t a t e s P e d i a t r i c s 1 0 3 : 7 R i l e y C V 1 8 8 9 S o m e i n s e c t p e s t s o f t h e h o u s e h o l d B e d b u g s a n d r e d a n t s I n s e c t L i f e 2 : 1 0 4 1 0 8

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1 0 5 S t a t i s t i c a l A n a l y s i s S o f t w a r e I n s t i t u t e ( S A S ) 2 0 0 0 S t a t i s t i c a l a n a l y s i s s o f t w a r e c o m p u t e r p r o g r a m v e r s i o n 8 0 1 S A S I n s t i t u t e C a r y N C S e r v i c e M W 2 0 0 0 M e d i c a l E n t o m o l o g y f o r S t u d e n t s C a m b r i d g e U n i v e r s i t y P r e s s C a m b r i d g e U K 2 8 3 p p S m i t h M R 1 9 6 5 H o u s e i n f e s t i n g a n t s o f t h e e a s t e r n U n i t e d S t a t e s : t h e i r r e c o g n i t i o n b i o l o g y a n d e c o n o m i c i m p o r t a n c e U S D A T e c h B u l l 1 3 2 6 : 1 1 0 5 S t e i n b r i n k H 1 9 7 8 P h a r a o a m e i s e n a u f e i n e r F r u e h g e b u r t e n s t a t i o n A n g e w P a r a s i t o l 1 9 : 2 8 3 1 S u d d J H 1 9 5 7 a A r e s p o n s e o f w o r k e r a n t s t o d e a d a n t s o f t h e i r o w n s p e c i e s N a t u r e 1 7 9 : 4 3 1 4 3 2 S u d d J H 1 9 5 7 b C o m m u n i c a t i o n a n d r e c r u i t m e n t i n P h a r a o h s A n t M o n o m o r i u m p h a r a o n i s ( L ) B r i t J A n i m B e h a v 3 : 1 0 4 1 0 9 S u d d J H 1 9 6 0 T h e f o r a g i n g m e t h o d o f P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s ( L ) A n i m B e h a v 8 : 6 7 7 5 S u m p t e r D J T a n d M B e e k m a n 2 0 0 3 F r o m n o n l i n e a r i t y t o o p t i m a l i t y : p h e r o m o n e t r a i l f o r a g i n g b y a n t s A n i m B e h a v 6 6 : 2 7 3 2 8 0 U n i t e d S t a t e s D e p a r t m e n t o f A g r i c u l t u r e ( U S D A ) 2 0 0 2 U S D A N a t i o n a l N u t r i e n t D a t a b a s e f o r S t a n d a r d R e f e r e n c e R e l e a s e 1 5 ( w w w n a l u s d a g o v / f n i c / f o o d c o m p ) M a r c h 3 2 0 0 4 V a i l K M 1 9 9 6 F o r a g i n g s p a t i a l d i s t r i b u t i o n a n d c o n t r o l o f t h e P h a r a o h a n t M o n o m o r i u m p h a r a o n i s ( L ) P h D d i s s e r t a t i o n U n i v e r s i t y o f F l o r i d a G a i n e s v i l l e F L 1 0 8 p p V a i l K M a n d D F W i l l i a m s 1 9 9 4 F o r a g i n g o f t h e p h a r a o h a n t M o n o m o r i u m p h a r a o n i s : a n e x o t i c i n t h e u r b a n e n v i r o n m e n t p p 2 2 8 2 3 9 I n D F W i l l i a m s [ e d ] E x o t i c a n t s W e s t v i e w P r e s s B o u l d e r C O 3 3 2 p p V a i l K M a n d D F W i l l i a m s 1 9 9 5 P h a r a o h a n t ( H y m e n o p t e r a : F o r m i c i d a e ) c o l o n y d e v e l o p m e n t a f t e r c o n s u m p t i o n o f p y r i p r o x y f e n b a i t s J E c o n E n t o m o l 8 8 : 1 6 9 4 1 7 0 2 V a i l K M D F W i l l i a m s a n d D H O i 1 9 9 6 P e r i m e t e r t r e a t m e n t s w i t h t w o b a i t f o r m u l a t i o n s o f p y r i p r o x y f e n f o r c o n t r o l o f P h a r a o h a n t ( H y m e n o p t e r a : F o r m i c i d a e ) J E c o n E n t o m o l 8 9 : 1 5 0 1 1 5 0 7

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1 0 6 V i n s o n S B 1 9 7 0 G u s t a t o r y r e s p o n s e b y t h e i m p o r t e d f i r e a n t t o v a r i o u s e l e c t r o l y t e s A n n E n t o m o l S o c A m 6 3 : 9 3 2 9 3 5 W a l d b a u e r G P 1 9 6 2 T h e g r o w t h a n d r e p r o d u c t i o n o f m a x i l l e c t o m i z e d t o b a c c o h o r n w o r m s f e e d i n g o n n o r m a l l y r e j e c t e d n o n s o l a n a c e o u s p l a n t s E n t o m o l E x p A p p 5 : 1 4 7 1 5 8 W a l d b a u e r G P 1 9 6 8 T h e c o n s u m p t i o n a n d u t i l i z a t i o n o f f o o d b y i n s e c t s A d v I n s e c t P h y s 5 : 2 2 9 2 8 8 W e i X a n d S J J o h n s o n 1 9 9 6 G r a v i m e t r i c m e t h o d f o r t h e m e a s u r e m e n t o f s u g a r c o n s u m p t i o n b y a d u l t v e l v e t b e a n c a t e r p i l l a r ( L e p i d o p t e r a : N o c t u i d a e ) F l E n t o m o l 7 9 : 3 8 4 3 9 2 W e i d n e r T 1 9 8 2 P h a r a o h a n t s : a g r o w i n g p r o b l e m i n A m e r i c a s h o s p i t a l s ? P e s t C o n t r T e c h 1 0 : 3 7 6 2 W e n z e l R P T B r e w e r a n d J P B u t z l e r 2 0 0 2 A g u i d e t o i n f e c t i o n c o n t r o l i n t h e h o s p i t a l D e c k e r I n c R i c h m o n d V A 1 8 4 p p W h e e l e r W M 1 9 1 0 A n t s : t h e i r s t r u c t u r e d e v e l o p m e n t a n d b e h a v i o r C o l u m b i a U n i v e r s i t y P r e s s N e w Y o r k N Y 6 6 3 p p W i l l i a m s D F 1 9 9 0 E f f e c t s o f f e n o x y c a r b b a i t s o n l a b o r a t o r y c o l o n i e s o f t h e P h a r a o h s a n t M o n o m o r i u m p h a r a o n i s p p 6 7 6 6 8 3 I n R K V a n d e r M e e r K J a f f e a n d A C e d e n o [ e d s ] A p p l i e d m y r m e c o l o g y : a w o r l d p e r s p e c t i v e W e s t v i e w P r e s s B o u l d e r C O 7 4 1 p p W i l l i a m s D F a n d K M V a i l 1 9 9 4 C o n t r o l o f a n a t u r a l i n f e s t a t i o n o f t h e P h a r a o h a n t ( H y m e n o p t e r a : F o r m i c i d a e ) w i t h a c o r n g r i t b a i t o f f e n o x y c a r b J E c o n E n t o m o l 8 7 : 1 0 8 1 1 5 W i l s o n E O a n d R W T a y l o r 1 9 6 7 T h e a n t s o f P o l y n e s i a ( H y m e n o p t e r a : F o r m i c i d a e ) P a c I n s e c t s M o n o g r 1 4 : 1 1 0 9 W r i g h t C G a n d D S t o u t 1 9 7 8 F o r b e t t e r P h a r a o h a n t c o n t r o l P e s t C o n t 4 6 : 2 6 3 2

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1 0 7 B I O G R A P H I C A L S K E T C H R o x a n n e G r a c e B u r r u s w a s b o r n o n M a y 7 1 9 6 5 a t t h e B e t h e s d a N a v a l H o s p i t a l i n M a r y l a n d t o J a m e s D a n d D a w n E B u r r u s w h i l e t h e y w e r e l i v i n g i n n e a r b y V i r g i n i a H e r p a r e n t s m e t i n W a s h i n g t o n D C a s b o t h w e r e a c t i v e d u t y N a v y s t a t i o n e d i n t h a t c i t y H e r y o u t h w a s s p e n t i n a v a r i e t y o f c o u n t r i e s i n c l u d i n g T a i w a n C y p r u s a n d S p a i n S h e h a s o n e y o u n g e r s i s t e r K a r e n w h o w a s b o r n i n T a i w a n i n 1 9 6 7 R o x a n n e a t t e n d e d P i n e F o r e s t H i g h S c h o o l i n P e n s a c o l a F l o r i d a w h e r e h e r f a t h e r c o m p l e t e d h i s 2 0 y e a r s o f s e r v i c e i n t h e U S N a v y A f t e r h i g h s c h o o l R o x a n n e a t t e n d e d t h e U n i v e r s i t y o f S o u t h e r n M i s s i s s i p p i w h e r e s h e o b t a i n e d a B a c h e l o r o f S c i e n c e i n m a t h e m a t i c s w i t h a m i n o r i n c o m p u t e r s c i e n c e I m m e d i a t e l y a f t e r g r a d u a t i o n f r o m c o l l e g e s h e j o i n e d t h e U S A i r F o r c e a n d w a s s t a t i o n e d a t H a n s c o m A F B i n B e d f o r d M a s s a c h u s e t t s w h e r e h e r w o r k a t t h e A i r F o r c e C o m p u t e r A c q u i s i t i o n C e n t e r ( A F C A C ) h e l p e d t o e s t a b l i s h L o c a l A r e a N e t w o r k s ( L A N s ) a n d W i d e A r e a N e t w o r k s ( W A N s ) i n v a r i o u s D e p a r t m e n t o f D e f e n s e ( D O D ) a g e n c i e s T h e L A N a n d W A N t e c h n o l o g y w a s c u t t i n g e d g e t e c h n o l o g y a t t h e t i m e ; W A N s h e l p e d p a v e t h e w a y f o r t h e c u r r e n t i n t e r n e t t h a t i s s o f a m i l i a r n o w b u t w a s r e l a t i v e l y u n k n o w n t h e n A f t e r c o m p l e t i n g h e r m i l i t a r y s e r v i c e R o x a n n e r e m a i n e d i n N e w E n g l a n d a n d o b t a i n e d a p o s i t i o n a t E d w a r d s H i g h V a c u u m a d i v i s i o n o f B O C G r o u p L T D b e g i n n i n g a s a n I n s i d e S a l e s C o o r d i n a t o r a n d e n d i n g a s a D a t a b a s e P r o j e c t M a n a g e r D u r i n g h e r y e a r s a t E d w a r d s s o m e f r i e n d s i n t r o d u c e d h e r t o t h e s p o r t o f b a c k p a c k i n g i n

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1 0 8 t h e W h i t e M o u n t a i n s o f N e w H a m p s h i r e W h i l e e x p l o r i n g t h e v a s t o u t d o o r s w i t h t h e m s h e b e c a m e a v i d l y i n t e r e s t e d i n t h e c r e a t u r e s a n d p l a n t s s h e o b s e r v e d a n d d e s i r e d t o l e a r n m o r e a b o u t t h e i n t e r a c t i o n s a n d e c o l o g y o f t h e s e b i o l o g i c a l o r g a n i s m s T h i s d e s i r e t o l e a r n l e d h e r t o t a k e a n e d u c a t i o n a l p a t h t h a t r e s u l t e d i n l e a v i n g E d w a r d s t o a c c e p t a s u m m e r i n t e r n s h i p p o s i t i o n h e l p i n g a P h D s t u d e n t a t t h e U n i v e r s i t y o f M a i n e O r o n o c o m p l e t e a f i n a l f i e l d s e a s o n ’ s d o c u m e n t a t i o n o f i n s e c t d i v e r s i t y i n b o t h s t a n d i n g a n d f a l l e n t i m b e r A f t e r a s u m m e r c o u n t i n g w h a t s e e m e d l i k e m i l l i o n s o f c o l l e m b o l a a n d o t h e r m i n u t e a r t h r o p o d s i n s o i l s a m p l e s R o x a n n e w a s e s p e c i a l l y f a s c i n a t e d w i t h i n s e c t s a n d t h e e v e n t s t h a t u l t i m a t e l y l e d h e r t o t h e U n i v e r s i t y o f F l o r i d a s e e m e d t o f a l l i n t o p l a c e o f t h e i r o w n v o l i t i o n f o l l o w i n g t h a t s u m m e r S h e e n j o y e d l e a r n i n g b i o l o g y s o m u c h t h a t s h e a t t e n d e d M i d d l e s e x C o m m u n i t y C o l l e g e t o c o m p l e t e a n a s s o c i a t e ’ s d e g r e e f o c u s i n g o n l i f e s c i e n c e s S h e t h e n i m m e d i a t e l y t r a n s f e r r e d t o t h e U n i v e r s i t y o f M a s s a c h u s e t t s A m h e r s t w h e r e s h e o b t a i n e d a b a c h e l o r ’ s d e g r e e i n b i o l o g y W h i l e a t U M A S S s h e b e c a m e i n t e r e s t e d i n p r e v e n t i v e m e d i c i n e a n d h e a l t h a n d w a n t e d t o p u r s u e b o t h e n t o m o l o g y a n d m e d i c a l i n t e r e s t s A n i n t e r n e t s e a r c h o f m e d i c a l s c h o o l s u n e x p e c t e d l y t u r n e d u p t h e U S N a v y ’ s w e b s i t e d e s c r i b i n g a f i e l d c a l l e d m e d i c a l e n t o m o l o g y B y b e c o m i n g a M e d i c a l E n t o m o l o g i s t i n t h e U S N a v y s h e c o u l d b e a n e n t o m o l o g i s t a n d h e l p p r e v e n t a r t h r o p o d t r a n s m i s s i o n o f h u m a n d i s e a s e s T h e p i v o t a l d e c i s i o n t o b e c o m e a M e d i c a l E n t o m o l o g i s t l e d R o x a n n e t o t h e U n i v e r s i t y o f F l o r i d a w h e r e s h e c o m p l e t e d a m a s t e r ’ s d e g r e e i n m e d i c a l a n d u r b a n e n t o m o l o g y i n p r e p a r a t i o n f o r h e r c a r e e r i n t h e N a v a l M e d i c a l S e r v i c e s C o r p ( M S C )


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Title: Pharaoh Ant Consumption of Fluids Used in Hospital Environments
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Copyright Date: 2008

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Permanent Link: http://ufdc.ufl.edu/UFE0003202/00001

Material Information

Title: Pharaoh Ant Consumption of Fluids Used in Hospital Environments
Physical Description: Mixed Material
Copyright Date: 2008

Record Information

Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: All rights reserved by the source institution and holding location.
System ID: UFE0003202:00001


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PHARAOH ANT CONSUMPTION OF FLUIDS USED IN HOSPITAL
ENVIRONMENTS














By

ROXANNE GRACE BURRUS


A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF SCIENCE

UNIVERSITY OF FLORIDA


2004







































Copyright 2004

by

Roxanne Grace Burrus


































I dedicate this thesis to God, The Risen Lord and Creator of all things.















ACKNOWLEDGMENTS

I am deeply indebted to family and friends whose advice, moral support, and

assistance in a multitude of ways have proven so vitally important and uplifting. First, I

am grateful to God for an inquisitive mind, a delight in His creation, and an unending

curiosity about how things came to be the way they are. Even as a child, I enjoyed

exploring outdoors.

I thank my parents for their guidance, support and encouragement over the past

several years in so many ways. In addition to being great parents, they are good friends,

whose company I enjoy and whose counsel I appreciate. They are really strong in the

common sense department, which has been good ballast against the idealistic attitude I

tackle life's challenges with. Two years ago, when circumstances prevented me from

collecting my household goods during the university's spring break, my parents drove

from Florida to Massachusetts, packed my belongings and cats into a rental truck, and

delivered it to me in Gainesville, FL all within one week. That kind of generosity and

kindness will always be remembered and greatly appreciated.

I thank my sister, Karen, whose unwavering support and selfless attitude has

given me the energy to persevere more than she knows. She has been a role model of

encouragement at every step of the way. I am fortunate to count her as a friend as well as

a sibling.









Sarah Smith has been both friend and mentor in many ways. As a biologist and

naturalist, her knowledge and appreciation for life and living organisms have been

inspiring. She put useful tools in my path so that I experienced the joy of discovery.

Other friends have firmly entrenched themselves in my life. Marsha Asquith,

Mike Magee, Dave Cotton Doug Nichols, Aline Rober, John Crew, and Mark Alarie

spoke words of wisdom at just the right time on so many occasions, and helped me stay

the course during difficult times. They provided ideas and insights that proved invaluable

in both my personal life and in my research.

While at the University of Massachusetts, Amherst, I had the good fortune to

meet Reg Coler. His enthusiasm and vivacity are topped only by his keen interest and

knowledge of entomology. As my first graduate advisor, he taught me research skills that

will prove invaluable for the rest of my life. The knowledge that he imparted is

especially remarkable when one considers the short amount of time we were able to work

together.

My tenure at the University of Florida brought new friends and mentors into my

life. Gene Gerberg has mentored me from his strong background of a lifetime of work in

the field of medical entomology. His advice has already proven valuable in many

respects, and I am sure it will continue to do so. He took this fledgling entomologist

under his wings and ensured that I stayed on track during my time here in Florida.

Knowing him has been a real pleasure; I have enjoyed and benefited from his anecdotes

of different entomologists he worked with. His autobiography will be interesting to read.

Faith Oi has spent a great deal of time helping me with statistics and fine-tuning

the manuscript. In addition, she taught me useful research skills that have been time-









saving tools. I appreciate her help and advice. I am a better researcher due to her

influence.

David Oi has also spent a great deal of time helping with the thesis and with

statistics. His contributions improved the experimental design, the analysis of data, and

the manuscript. His knowledge of ant behavior and of the literature has been inspiring.

The knowledge and research skills he imparted to me have already made me a better

scientist, and will continue to prove helpful.

Phil Koehler has helped me put together nice presentations. Much of what I have

learned from working with him and in his lab extends beyond textbook knowledge, and

will prove useful in many situations.

I thank the staff in the entomology department, especially Debbie Hall and Myma

Litchfield. They have been wonderful in every respect.

Finally, I thank my fellow Tri-Gator athletes and coaches. Training for and

participating in triathlons have become an integral part of my life. Completion of long

events has helped clear my focus and steel my mind. This has improved my research,

and helped me enjoy the journey.
















TABLE OF CONTENTS



ACKNOWLEDGMENTS .................................................... iv

L IST O F T A B L E S ....................................................... ............................ ..................... x

L IST O F F IG U R E S ..................................................... ................................................ xi

A B ST R A C T ...................................................................................................... ............ xiii

CHAPTER

1 IN T R O D U C T IO N ............................................................................. .....................1......

2 LITERATURE REVIEW ............. ...........................................................3......

G general A nts .................................................. ......................................................3...
T ram p A n ts ................................................................................. ............................ 3
Pharaoh A nt .................................................. .......................................................4...
Origin and Distribution................. .........................................................5......
Biology ........... ........... .. ........................................ .5
F oraging and F ood ....................................................................... ........................ 7
Medical Importance.....................................................................................7......
Control ..................... .. ... ............................................. 8

3 NO-CHOICE LIQUID TEST..................................................................................13

In tro du ctio n ......................................................... .................................................. 13
M materials and M ethods ........................................................................ ..................... 15
Ants ...................... ........... ..................... 15
F raging A rena .................................................. ............................................ 15
E xperim ental F luids.................................................................... ...................... 17
E xperim ental D esign .................................................................. ...................... 19
Fluid Measurements ...................................................................................21
Statistical A nalysis................................................... ............................................. 22
Results.. ........... ........... ............. ..................... 23
D iscu ssio n ........................................................... .................................................. 2 5
Fluid Measurements ...................................................................................26
Water Consumption.....................................................................................26
Dextrose Consumption ..............................................................................28









Plasma and Blood Consumption ...............................................................29
Saline (=0.9% NaCl) Consumption...................................................................31
E nsure C onsum ption .............. .......................................... ....................... 31

4 C H O IC E L IQ U ID TE ST .................................................................. ....................... 38

In tro du ctio n .......................................................... ................................................. 3 8
Materials and Methods ................................. ..............................40
Ants ....... ............... ........... ..................... 40
F raging A rena .................................................. ............................................ 40
Experim mental Fluids............................... ...................................................... 42
Experimental Design ............. ................................... 42
Fluid M easurem ents ............................. ...................................................... 43
Statistical A nalysis................................................... ............................................. 45
Results.. ........... ........... ............. ..................... 45
Number of Ants .....................................................45
Fluid M easurem ents ............................. ...................................................... 46
C o n su m p tio n ................................................................................ ...................... 4 7
D iscu ssio n ........................................................... .................................................. 4 7
Number of Ants .....................................................47
Fluid M easurem ents ............................. ...................................................... 48
C o n su m p tio n ................................................................................ ...................... 4 9
D ex tro se ........................................................ ................................................. 5 0
E n su re ........................................................ ................................................. 5 0
Plasma .... ................ ......... .. ..................... 51
NaC1.......... ............... ............. ..................... 52
B lo o d ........................................................... ................................................. 5 3

5 NO-CHOICE DEHYDRATED FLUID TEST.......................................................60

In tro du ctio n .......................................................... ................................................. 60
Materials and Methods ................................. .............................. 61
A nts .................................. ...................... 61
Foraging Arena and Nest.....................................................................62
H hospital Sheets .................................................. ............................................ 62
Dehydration of Fluids............................ ..................................................... 64
Experim mental D design ............................. ..................................................... 65
Tim ed Observations..................................................................................... 65
Statistical A nalysis................................................... ............................................. 66
Results.. ........... ........... ............. ..................... 66
D iscu ssio n ........................................................... .................................................. 6 8

6 SUMMARY AND CONCLUSIONS ..........................................................................87

APPENDIX

A ANT MAINTENANCE......................... .................................... 90


viii









B SELECTION OF ANTS ....................................................93

C CON SUM PTION FORM ULA S....................................................... .......................95

L IST O F R EFER EN C E S ................................................. ............................................ 99

BIOGRAPHICAL SKETCH ............. ...................................107















LIST OF TABLES

Table page

3-1. Mean (liquid no-choice) initial weights................................................................ 34

3-2. Mean (liquid no-choice) consumption...................................................................35

4-1. Mean number of ants remaining.................................................................54

4-2. Mean (choice test) initial weights................ .................................................55

4-3. Mean (choice test) consumption........................................................................... 56















LIST OF FIGURES

Figure page

3-1. N o-choice liquid arena. .............................................................................................16

3-2. Mean (liquid no-choice) evaporation (24 h) in control arenas. ................................36

3-3. Mean (liquid no-choice) consumption (24 h) in test arenas ......................................37

4-1. C choice liquid arena .................................................................. ............................ 4 1

4-2. Mean number of ants remaining in fluid after 24 h ................................................. 57

4-3. Mean (choice test) evaporation. ................................................................................58

4-4. Mean (choice test) consumption. ..............................................................................59

5-1. D dehydrated fluid arena. ............................................................................................. 63

5-2. Mean number of ants per 15 min. time intervals over entire 2 h. ............................72

5-3. Mean number of ants per dehydrated fluid over entire 2 h. ....................................73

5-4. Mean number of ants per fluid at 15 min .. ................................................................74

5-5. Mean number of ants per fluid at 30 min. ...............................................................75

5-6. Mean number of ants per fluid at 45 min. .................................................................76

5-7. Mean number of ants per fluid at 60 min. .................................................................77

5-8. Mean number of ants per fluid at 75 min. .................................................................78

5-9. Mean number of ants per fluid at 90 min. .................................................................79

5-10. Mean number of ants per fluid at 105 min. ........................... ............................ 80

5-11. Mean number of ants per fluid at 120 min. .............................................................81

5-12. Mean number of ants feeding on Ensure .................................. ...................... 82

5-13. Mean number of ants feeding on 5% dextrose ............................ ...................... 83









5-14 M ean number of ants feeding on saline......................................... ....................... 84

5-15. Mean number of ants feeding on human blood cells.............................................85

5-16. Mean number of ants feeding on human plasma................................................86















Abstract of Thesis Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Master of Science

PHARAOH ANT CONSUMPTION OF FLUIDS USED IN HOSPITAL
ENVIRONMENTS


By

Roxanne Grace Burrus

May 2004

Chair: Philip G. Koehler
Cochair: David H. Oi
Major Department: Entomology and Nematology

Monomorium pharaonis (L.), the Pharaoh ant, is a pest species which inhabits

buildings throughout the world. Its populations are difficult to control. When in

hospitals and other health-care environments, these ants are often found nesting in places

as diverse as operating rooms, linen closets, baby incubators, and inside unopened

packages of intravenous tubing. Pharaoh ants have been observed feeding upon human

secretions and on intact epidermis. In addition they have been reported to consume

patient-care fluids, such as dextrose and saline solutions.

Small Pharaoh ant colonies that were starved for 2 d consumed significant

amounts of either blood, 5% dextrose, Ensure dietary supplement, 0.9% NaCl solution

(saline), or plasma when given 24 h access to each fluid separately. Comparison of

consumption based on these no-choice exposures indicated that significantly more 5%

dextrose was consumed among all the other fluids. Simultaneous, or choice, exposures of









the same fluids again showed the most significant consumption occurred on 5% dextrose.

Counts of feeding Pharaoh ants, which were starved for 3 d and given individual access

to the fluids dried on hospital linens, were highest on the Ensure dietary supplement,

followed by the 5% dextrose.














CHAPTER 1
INTRODUCTION

Monomorium pharaonis (L.), commonly known as the Pharaoh ant, has a close

association with human habitats (Wheeler 1910), and is probably one of the world's most

studied ants. It is a tropical ant that needs both high humidity and high temperatures

(Hblldobler and Wilson 1990). In temperate zones, it lives in buildings, such as

hospitals, which are heated and have indoor plumbing throughout the structure (Edwards

and Short 1990). Beatson (1972) reported that Pharaoh ants can carry several pathogens,

including Streptococcus, Staphylococcus, Escherichia coli, Bacillus, Pseudomonas, and

others. This discovery sparked a brief interest in the pathogenic vector capacity of

Pharaoh ants that lasted into the 1980's. More recently, independent observations by

Edwards and Short (1990) and (Edwards and Short 1990, Eichler 1990) of Pharaoh ants

feeding on human patients rekindled interest in this species. Additionally, a limited

number of tramp ant species, including Pharaoh ants, have been implicated as vectors of

hospital-acquired diseases (Fowler 1990), and were associated with a higher incidence of

nosocomial infections (Richards et al. 1999). The ability of Pharaoh ants to move

between patients receiving medical treatment and hospital areas that may harbor

pathogens, such as sewage, plumbing, food-preparation areas, and even diseased patients

suggests that Pharaoh ants have the potential of spreading pathogenic diseases throughout

a hospital. Eichler (1990) considered Pharaoh ants to be one of the most dangerous

insects in health-care environments.







2

The disease vectoring potential of Pharaoh ants, like that of other arthropods more

traditionally associated with disease transmission, such as cockroaches and flies, may be

directly linked to their food preferences and foraging behavior. Therefore, Pharaoh ant

consumption of liquid substances that are specifically used in patient care needed to be

examined more closely. The objective of this research was to determine which fluid(s)

used in patient care attract Pharaoh ants. Two experiments were conducted to determine

if Pharaoh ants consume (1) liquid fluids used in hospitals (Chapter 3), and (2)

dehydrated hospital fluids (Chapter 5). A third experiment (Chapter 4) compared the

attraction among the liquid hospital fluids when presented simultaneously to Pharaoh

ants.














CHAPTER 2
LITERATURE REVIEW

General Ants

There are over 9,500 known species of ants in the world (Bolton 1995). Ants are

eusocial insects which live in colonies comprised of multiple castes, usually

morphological, but sometimes temporal (Oster and Wilson 1978), and multiple

developmental stages. Eusocial behavior in ants is seen in the division of labor/function

amongst different castes, in the presence of overlapping generations, and in the tending of

immature brood by non-parental adults (Hblldobler and Wilson 1990)

Tramp Ants

Ants which are harmful to human activity are classified as pests, and are often

considered to be tramp species. Tramp species have many traits that enabled them to be

synanthropic. They are attracted to changing environments that are linked strongly to

human activity (Passera 1994), and often nest inside buildings (Ebeling 1978). Many

have been transported to new countries around the world via human commerce (Riley

1889). Tramp species tend to migrate quickly in response to changes in environmental

conditions or food availability, or to overpopulated colony size (Passera 1994). Many are

polydomous colonies that lack aggression between neighboring nests, so that workers,

queens and brood are freely exchanged between nests (Passera 1994). Within each nest,

tramp species are often polygyne, that is, they have many fertilized queens (Ebeling

1978, Passera 1994). Sexual castes often engage in intranidal mating rather than having

nuptial flights, even when wings are present (Hblldobler and Wilson 1990). Colony







4

reproduction takes place by sociotomy, or budding, when workers and newly mated

queens leave to start a new nest nearby (Passera 1994). Tramp ants are usually

monomorphic, with only one morphological worker caste, and are always very small

(Passera 1994).

Pharaoh Ant

Monomorium pharaonis (L.) (Formicidae: Myrmicinae), the Pharaoh ant, is a

worldwide pest (Wheeler 1910, HOlldobler and Wilson 1990) that is closely associated

with humans and is considered a tramp ant (Passera 1994). Pharaoh ant colonies exhibit

a temporal division of labor (Sudd 1957b) in which younger workers spend most of their

time tending the brood and queens while older workers primarily forage. In the absence

of one caste, members of another caste will perform the additional tasks. Pharaoh ants

feed on a wide variety of foods (Bellevoye 1889, Peacock et al. 1950, Sudd 1960,

Granovsky and Howell 1983, Edwards 1986, Edwards and Abraham 1990). They also

feed on humans, where they have been observed recruiting to body and wound secretions,

(Beatson 1972, Edwards and Baker 1981a, Granovsky and Howell 1983, Edwards and

Short 1990, Eichler 1990). In addition, they have been observed recruiting to or feeding

on the unbroken skin of babies' cheeks, lips, neck folds and backs of ears has been

reported (Steinbrink 1978, Eichler 1990). Pharaoh ants are especially well-adapted to

taking advantage of any available space for nesting sites, and are primarily found nesting

inside walls and crevices (Wheeler 1910). However, colonies have been discovered in

even more unusual locations, such as in linens, between sheets of paper (Ebeling 1978),

in intravenous tubing packages, in surgical and resuscitation equipment (Beatson 1973),

and in baby incubators (Steinbrink 1978, Eichler 1990).









Origin and Distribution

The Pharaoh ant was originally designated as Formica pharaonis Linneaus 1758,

and had six different names until it was eventually named Monomorium pharaonis (L.)

(Bolton 1994). The Pharaoh ant's origin is uncertain, with many researchers contending

that the ant originated in the tropical regions of Africa (Ebeling 1978). However, Emery

(1922), Wilson and Taylor (1967) and Bolton (1987) agreed that India is the probable

origin of the Pharaoh ant. (Arnold 1916) stated South America was the original location

of this ant. The Pharaoh ant is common and widely prevalent in all these areas, and is

easily spread by human commerce. Hence, its true origin may never be determined.

The Pharaoh ant is a tramp species that is strongly associated with humans

(Edwards 1991), and was proclaimed as the "most widely distributed of all ant pests" by

Edwards and Short (1990). They listed its documented presence to the United States,

Canada, South America, Egypt, North Africa, Europe, Russia, Australia, and Japan. The

Pharaoh ant is also present in the Hawaiian Islands (Reimer et al. 1990).

Biology

Pharaoh ant workers are monomorphic, and are very small, with a length of 1.5-2.0

mm. They are reddish to orange-brown in color, have a two-segmented petiole with each

segment having a node, and their gasters have a white stripe on the anterior portion of its

dorsal surface (Haack 1987, Harris 1991) Each antenna has 12 segments, with the final 3

segments forming a visible club. Adult males are winged, black in color, and slightly

larger than the workers, but smaller than queens. In natural colonies, males appear

infrequently, although male eggs are laid continuously (Peacock et al. 1954). At 4 mm in

length, queens are the largest caste of the Pharaoh ant. Virgin queens can be







6

distinguished from egg-laying queens by the presence of their wings, which are bitten off

after mating.

Reproductive males and females emerge in the nest with wings, but they do not

have nuptial flights. Instead, they mate within the nest (Peacock and Baxter 1950).

Wing muscles in the thorax of mated queens hystolize so that the thorax is modified to

contain a thoracic crop that is supplemental to the gaster's crop (Caetano 1990).

Within the nest, brood is normally separated into distinct piles that are sorted by

age (Peacock and Baxter 1949). Pharaoh ant eggs are round and stick together (Harris

1991). Larvae are immobile, and must be transported and cared for by adult workers.

Sexual brood develops faster than worker brood, but differentiation between the two

brood castes is difficult until the gut color of worker larvae changes to black, or the

sexual larvae becomes noticeably larger than the worker larvae (Peacock and Baxter

1950).

Lifespans of Pharaoh ants differ among the castes. Workers live -9-10 weeks,

males normally live -3 weeks, and queens live -39 weeks, (Peacock and Baxter 1950),

although they can live as long as 52 weeks (Edwards 1987).Pharaoh ant colonies can vary

tremendously in size (Peacock et al. 1955a) and can contain thousands of individuals,

including many fertile queens. Queen fecundity is directly related to the number of last

instar larvae present in the colony (BOrgesen and Jensen 1995). Natural Pharaoh ant

colonies tend to have a 1:1:1 ratio of workers:eggs:larvae (Peacock et al. 1955b).

Pharaoh ant colonies are polydomous, and exchange queens, workers and brood among

nests (Edwards 1991). Migrating groups of ants often form a temporary nest while

searching for a more permanent nest site (Edwards and Baker 1981a). New nests are







7

formed via budding (Hedges 1997), since Pharaoh ants do not have nuptial flights

(Ebeling 1978).

Peacock and Baxter (1949) originally stated that Pharaoh ants preferred

temperatures -27C and 80% RH. Harris (1991) demonstrated that Pharaoh ants

preferred a lower relative humidity of -65%. In temperate regions, Pharaoh ants nest

primarily indoors (Ebeling 1978).

Foraging and Food

Pharaoh ants engage in extensive foraging, and are most active at night { Hedges,

1997 #721}. They likely use "guideline orientation" (Klotz and Reid 1992) to follow

edges and cracks of structures to find food sources. They will also forage via ducts and

wall spaces aluminum window and door frames, and they can forage up to distances of 45

m (Vail and Williams 1994).

Pharaoh ants are well-known for their consumption of a wide variety of foods

(Smith 1965, Edwards and Baker 1981a). Some researchers contend that the Pharaoh ant

is primarily attracted to sugars (Granovsky and Howell 1983), while others think that

they primarily consume proteins (Edwards and Abraham 1990). Other research indicated

that Pharaoh ant colonies compensated for what was lacking nutritionally, whereby

laboratory colonies that were reared on carbohydrates exhibited more interest in protein

baits, and vice-versa (Edwards and Abraham 1990). Thus, the food preference of

Pharaoh ants is variable and dynamic, changing with nutritional needs (Holldobler and

Wilson 1990), and time (Edwards 1986).

Medical Importance

The Pharaoh ant is a tropical ant and needs high humidity and high temperatures to

survive (Peacock and Baxter 1950, Peacock et al. 1955c). In temperate zones, it is







8

dependent upon heated buildings, and is a common pest species found in hospitals and

other health care facilities world-wide. Although hospital infestations of Pharaoh ants

were already recognized as an inconvenience, the discovery of several pathogenic

organisms, including Streptococcus spp., Staphylococcus spp., Escherichia coli, Bacillus

spp., Psuedomonas spp., Klebsiella spp., and Clostridium spp., on hospital populations of

Pharaoh ants (Beatson 1972), created interest in the potential for Pharaoh ants to transmit

diseases to humans.

In Brazil, the potential for Pharaoh ants to vector hospital-acquired (nosocomial)

infections may have been especially high, as this country had some of the highest rates of

intra-hospital infections. In these hospitals, it was determined that only tramp ant

species, including Pharaoh ants, were near patients (Bueno and Fowler 1994).

Besides being potential vectors, Pharaoh ants can cause skin irritations and lesions

(Eichler 1990). Their small size enables them to crawl through gauze covering bandaged

wounds and ants as small as 2.0 mm in length can cause intense pain when foragers walk

on open wounds (Eichler 1990). Pharaoh ants also have been found feeding directly on

open wounds of burn-unit patients (Anon. 1986), on or under bandages covering wounds

(Cartwright and Clifford 1973), on premature newborns in incubators (Edwards and Short

1990), on bedridden elderly or postoperative patients (especially patients who were

leaking body fluids)(Anon. 1974), and in intravenous tubes that supply fluids to patients

(Beatson 1973, Edwards and Baker 1981b).

Control

Once ensconced in a structure, Pharaoh ant populations are extremely difficult to

eradicate partially due to cryptic habits, including a tendency to live inside walls and

under foundations. It is often difficult to locate Pharaoh ant nests without damaging the







9

building's structure (Vail 1996). Many different methods of Pharaoh ant elimination

have been attempted, with varying degrees of success. The efficacy of pesticide sprays

and dusts, stomach poisons, baits containing toxicants, and, baits containing insect

growth regulators (IGRs) have all been investigated.

In the 1990s, sprays and dusts were usually ineffective against Pharaoh ants,

because they affected only a small percentage of workers (Williams 1990) due to the fact

that less than 25% of workers forage and many of the chemicals were in the pyrethroid

class and highly repellent. Their use was even hypothesized to result in budding

(Williams and Vail 1994), since Pharaoh ant colonies respond rapidly to disturbance

(Peacock et al. 1955b, Passera 1994). Eichler (1990) believed that insecticide spraying of

Pharaoh ant infestations to be a form of malpractice. In addition, the effectiveness of

sprays was believed to have been reduced if multiple nests are present in the general area

(Williams 1990).

Stomach poisons work too quickly and reduce worker numbers so much that

poisons are not distributed to queens and brood (Edwards 1975). Colonies can be

perpetuated by as few as 100 workers and 50 brood (Peacock et al. 1955a) or even with

as few as 5 workers, 30 eggs, 19 larvae, and 3 pupae (Vail 1996). Therefore, to eliminate

Pharaoh ant populations, it is essential for toxicants to reach the queens and brood.. Baits

should be comprised of a slow acting, non-repellent toxicant incorporated into an

attractive bait matrix so that the bait is consumed and spread through the colony via

trophallaxis. Eichler (1990) recommended meat-based bait matrices, and sodium

arsenate or chlordecone. Finally, he recommended pre-baiting to determine what food

attracts the Pharaoh ants at the time, since they are known to change food preferences, he







10

suggested placing baits near locations that they would likely forage, such as sinks. He

also suggested denying ants access to alternative food sources, so that the bait would be

their only food source.

Klotz et al. (1996) noted that boric acids were used against ants as early as the late

1800s (Riley 1889), and are still important in ant control. They cited a recommendation

by Wright and Stout (1978) for 2% boric acid in either a liquid or solid bait. Klotz et al.

(1996) tested the efficacy of 1% boric acid in sucrose solution and 0.9% hydramethylnon

with both limited and continuous availability in the laboratory. They determined that

colony growth was reduced, but not eliminated, after 3 d exposure to boric acid bait. In

contrast, continuous feeding of boric acid bait resulted in elimination of all colonies.

Colonies exposed to hydramethylnon had 90% reduction of workers by 3 wk and 90%

reduction of brood by 4 wk; queens were eliminated in 2 wk. They concluded that boric

acid-sucrose solutions might provide an alternative to other commercially available

Pharaoh ant baits.

Many toxicants such as Mirex are no longer used due to carcinogenicity (Eichler

1990). Insect growth regulators (IGRs) which mimic juvenile hormones, have been

developed (Nation 2003). IGRs often contain either the active ingredient methoprene or

pyriproxyfen. Insect growth regulators are slower-acting than stomach poisons because

they affect only queen and brood: they induce sterility in queen and disrupt brood

development (Edwards 1975). The workers remain active for several weeks, but are not

able to start new colonies due to death of queens and brood.

Oi et al. (1996) compared the efficacy of several different applications to

determine if indoor Pharaoh ant infestations could be controlled by exterior treatments,







11

since Pharaoh ants have been observed foraging on exterior wall surfaces and outside

buildings (Haack 1991, Oi et al. 1994, Vail and Williams 1994). They tested several

methods registered for ant control: a contact insecticide (cyfluthrin), a granular contact

insecticide (diazinon), a granular bait containing a contact insecticide (propoxur), and a

granular bait containing a delayed-action toxicant (hydramethylnon). They determined

that exterior application of the delayed-action toxicant hydramethylnon resulted in

successful control indoor Pharaoh ant infestations, while all other tested treatments

resulted in increased foraging. They concluded that exterior baiting could reduce the

need for indoor pesticide applications. In 2003, a new compound, fipronil (Termidor,

BASF, Research Triangle, NC) received registration for ant applications. It was an old

idea of spraying the perimeter of a building for ant control with a new compound that had

important bait-like characteristics. Fipronil is a non-repellent, slow acting toxicant.

Successful control of Pharaoh ants foraging outside and inside homes treated on the

perimeter only with fipronil, has been demonstrated (F. Oi and J. Jonovich, personal

communication) according to label directions (25 cm up and 25 cm out).

Vail et al. (1996) evaluated the efficacy of different bait formulations of the IGR

pyriproxyfen as an exterior perimeter treatment against Pharaoh ant infestations. They

noted that while pyriproxyfen had already been evaluated as an effective control agent

against most urban insects tested previously, and despite the demonstration of its

effectiveness against Pharaoh ants in the laboratory (Vail and Williams 1995), no IGRs

were commercially available for use against Pharaoh ants. They determined that the baits

formulated with various amounts of pyriproxyfen in two different bait matrices were all

successful in controlling the ant populations, although complete elimination did not occur







12

until 16 wk after treatment. They observed that brood rapidly declined in treated

colonies. Dyes used in bait formulations was visible in larvae, queens and workers,

indicating thorough distribution of baits throughout the colonies. Finally, they concluded

that application of an IGR bait 1 wk before application of a metabolic inhibitor might

provide both quick mortality and long-term control, and they stated that vertical

placement of baits aimed at Pharaoh ants might prevent other ant species from consuming

baits.

Oi et al. (2000) investigated the distribution of baits containing the IGR,

pyriproxyfen, and the metabolic inhibitor, hydramethylnon, among multiple Pharaoh ant

colonies. They determined that the IGR did not kill adult worker ants quickly so that

more worker ants lived long enough to distribute the bait to all nest sites. In contrast,

hydramethylnon caused rapid mortality in adult ants, and ants from nearby nests migrated

into sites whose populations declined due to metabolic inhibitor mortality. They further

determined that the IGR pyriproxyfen resulted in a slow, gradual decline of both workers

and queens in all nest sites, and lasted a long time, in contrast to the metabolic inhibitor

hydramethylnon, which worked more quickly, but did not last as long. Finally, they

postulated that longer survivorship of workers increased the amount of trophallaxis

between ants of different nest sites and increased the distribution of the IGR,

pyriproxyfen, among multiple nests, thereby reducing the possibility of re-infestation.

They concluded that the mode of action and speed of action of bait toxicants should be

evaluated when selecting ant baits for polydomous colonies such as the Pharaoh ant.














CHAPTER 3
NO-CHOICE LIQUID TEST

Introduction

Pharaoh ants' consumption of a wide variety of substances (Bellevoye 1889,

Granovsky and Howell 1983, Edwards and Abraham 1990) and their abundant presence

in hospitals is well-documented (Green et al. 1954, Beatson 1972, Edwards and Baker

1981b). Although Pharaoh ants are primarily sampled in kitchens and bathrooms because

it is assumed that there will be an abundance of water and food available to sustain ant

colonies, Vail and Williams (1994) found that they are capable of foraging up to 45 m.

Pharaoh ants have been found in hospitals and health-care facilities (Eichler 1990,

Fowler 1990). They have been discovered inside a permanently installed tracheotomy

tube and intravenous tubes (Weidner 1982). Pharaoh ants have been observed feeding

upon patients within different hospital areas. In neonatal units where newborns are

placed inside special incubating units, they have been observed feeding on intact skin of

newborn infants such as cheeks, lips, neck folds and backs of ears (Steinbrink 1978) as

cited in (Eichler 1990). They have been observed feeding on bum patient open wounds

(Weidner 1982). Their small size enables them to crawl through gauze; thus, they have

also been found under bandages of covered wounds (Beatson 1972). Finally, Pharaoh ant

infestations have been located in beds of bedridden elderly or postoperative patients

(especially patients who were leaking body fluids) (Anon. 1974).

Pharaoh ants can also be associated with other fluids present in a hospital

environment. Suppurating wounds exude plasma, which is the non-cellular portion of







14

whole blood. Additional fluids can also be present, such as saline or dextrose solutions

that are administered either intravenously or topically (AABB 2002, Wenzel et al. 2002).

Both types of administration of saline and dextrose decrease moisture loss in the absence

of epithelial tissue and in situations of limited fluid consumption (AABB 2002); topical

use also will sterilize wounds and decrease the potential for infection at injury site.

Pharaoh ants might be attracted to these fluids or to substances present in patient blood or

plasma. In addition to saline and dextrose intravenous and topical administrations, many

patients also receive liquid dietary supplements after surgery, via tubing that is inserted

into patient's stomach (Wenzel et al. 2002). These fluids provide another potentially

attractive food source to Pharaoh ants. Dextrose, saline and liquid dietary supplements

are administered directly into patient via tubing. Anecdotal reports from nurses (S. Cobb,

personal communication) indicate that medical practices such as tube clearing (i.e.,

procedure of squirting liquid out of tube prior to insertion of tube into patient) often result

in placement of liquids on or near the patient bed. Thus, each of these liquids is available

not only at insertion point in patient's body, but might also be available in abundance

throughout a hospital room, particularly close to the patient's bed.

Pharaoh ants occur in hospital rooms and have been documented to feed upon

patients, but no previous study has been undertaken to determine consumption of patient-

care fluids and human fluids. Even though ants were documented as feeding on wounds,

no examination has investigated consumption of human body fluids. Thus, my objective

was to determine if Pharaoh ants consume fluids used specifically for patient-care in

hospitals.







15

Materials and Methods

Ants

Pharaoh ant test colonies were extracted from large colonies housed and reared at

the Urban Entomology Laboratory, University of Florida, Gainesville, FL. Rearing

temperatures ranged from 21.90C 29.40C with relative humidity between 20-53%.

(Appendix A).

Colonies consisted of 250 workers, 3 dealate queens and approximately 50 mg

brood. Half of the workers were selected from the inside of nesting cells to ensure the

presence of nurses and 125 workers were selected from outside of the nesting cells, at

foraging locations such as exterior surface of water vials, to ensure the presence of

foragers (Haack 1987, Haack and Vinson 1990, Haack et al. 1995). Brood included eggs,

all larval stages, and pupae. See Appendix B for full details on selection of ants and

preparation of experimental test colonies.

Foraging Arena

Foraging arenas contained one nesting cell, one water vial, and one microcapillary

tube (Fig. 3-1). Foraging arenas consisted of clear plastic boxes (27 by 19 by 10 cm

deep; Pioneer Plastics, Dixon, KY) which were coated with Fluon (Asahi Glass

Fluoropolymers, Downington, PA) on interior sides to prevent ant escape. Nesting cells

consisted of sterile, covered plastic Petri dishes (50 mm diameter by 15 mm deep;

Falcons Model No. 351007, Becton Dickinson and Co., Franklin Lakes, NJ) filled

halfway with Castone dental plaster (Model No. 99045, Dentsply International, York

PA). A circular indentation was made in plaster by inserting the cap of a 15 ml vial

(Model No. 55-9, Thornton Plastics, Salt Lake City, UT) into freshly mixed plaster in the



























30Rl capillary tube,
with fluid


27 cm


4 19cm ,
Figure 3-1. No-choice liquid arena with water vial and nesting cell placed in diagonally
opposite comers of test arena. A microcapillary tube containing one fluid was
placed in the center of arena. Both test arenas (with ants) and control arenas
(no ants) were placed adjacent to each other to comprise test pair. Test arenas
contained 125 inside ants, 125 outside ants, 3 queens and -50 mg brood
(including pupae, eggs and all stages of larvae). Control arenas lacked ants.







17

base of a Petri dish. The vial cap was removed after plaster was thoroughly dried (24 h)

and hardened, leaving an indentation (33 mm diameter by 2 mm deep) and rough sloping

edge (Peacock and Baxter 1949) for ants to rear brood. One entrance hole (-5 mm

diameter) was melted into the upper edge of the Petri dish base via insertion of the metal

tip of a hot glue gun (sans glue) just above level of hardened plaster. Immediately prior

to placing ants in arenas, Petri dish bases were immersed in a container of tap water for

30 min to saturate the plaster. Saturated bases were removed from water and excess

water was blotted from all surfaces. A Petri dish lid was placed on top of the base and

the entire nesting cell was placed into the foraging arena with the entrance hole facing the

opposite diagonal comer of the arena.

Water was provided ad libitum in a 15 ml vial (Model No. 55-9, Thornton

Plastics, Salt Lake City, UT) with three large cotton balls inserted into the top to prevent

leakage and to provide easy access for ants. A water vial was placed in the opposite

diagonal comer of the arena across from the nesting cell (Fig. 3-1). One test arena

containing ants and one control arena without ants were placed adjacent to each other and

comprised a paired experimental unit for each replication. Control arena data were used

to quantify and adjust for evaporation of fluids. All arenas were covered with clear

plastic lids to maintain humidity and temperature. Airflow was maintained in the room

by the building's ventilation system during test period.

Experimental Fluids

Five fluids commonly used in hospital environments for patient-care were used in

this no-choice test. Test fluids can be provided to patients via insertion of tubing, either

intravenously (i.e., injected directly into bloodstream) or into intraperitoneally (i.e., into







18

stomach). Fluids consisted of human blood cells, human plasma, saline (0.9% sodium

chloride); 5% dextrose (D-glucose), and Ensure@ (Vanilla Plus, Ross Products Division,

Abbott Laboratories, Columbus, OH).

Blood cells and plasma (all other blood components except blood cells) were

obtained by centrifugal separation of donated whole blood, and were tested as individual

fluids since these two major constituents of whole blood differ greatly in composition and

structure. Blood cells can break down within 5 to 7 d (AABB 2002) without addition of

anti-coagulants due to enzymatic activity of intracellular blood proteins and cell death.

Whole blood samples contained heparin anticoagulant, which prevents clotting and

increases shelf-life of blood cells to -42 d. Plasma and blood cells were stored together

and shared a shelf-life of -42 d, which is typical for whole blood used in trauma and

emergency situations (Memmler et al. 1996, AABB 2002). Whole blood was obtained on

its expiration date after 30 d in refrigerated storage at LifeSouth Blood Bank (Gainesville,

FL). Therefore, all whole blood was aged 30 d and contained heparin anti-coagulant.

Whole blood was centrifuged to separate blood cells (bottom layer) from plasma

(top layer). Plasma was pipetted out and placed in separate test tubes. Plastic caps were

inserted into test tube openings to prevent leakage. Blood and plasma samples were

placed into a resealable plastic bag (0.95 L, Ziploc, S.C. Johnson & Son, Inc., Racine,

WI) with two paper towels folded inside to absorb fluid in case of spillage. Resealable

plastic bags were placed inside a thick-walled Styrofoam cooler. Ice (in separate

resealable plastic bag) was present in the cooler to keep blood cells and plasma samples

cool during transport. Blood cells and plasma were refrigerated and used within one

week of acquisition. Blood cells and plasma were both used with appropriate biohazard







19

procedures. Shands Hospital Burn Unit provided sealed, unopened bags of 5% dextrose

and 0.9% NaCl. Dextrose and saline were placed and maintained in a refrigerator (along

with blood cells and plasma) at -4.1 C before use. Dextrose and saline portions were

squirted into dry, clean test tubes (150 mm by 10 mm inner diameter, No. 9800, Pyrex

Labware, Corning Inc., Corning, NY) for use in experiments. Bags were sealed

immediately after use and remained out of the refrigerator for no longer than 5 min while

removing fluid for testing.

Ensure@ (Ensure Plus@ Vanilla), a commonly used flavor in hospital

environments (S. Cobb, personal communication), was used in experiments because

Pharaoh ants have been observed feeding upon dietary supplements in hospitals and other

health-care environments by pest management personnel (W. Wright, Jacksonville PMC

Seminar, personal communication). Ensure@ is a dietary supplement that contains

substantial proportions of proteins (26%), sugars (17%), and lipids (17%).

Experimental Design

The experiment was designed as a randomized complete block, blocking on

colony. Each block consisted of Pharaoh ant colonies (125 inside workers, 125 outside

workers, 3 dealate queens and -50 mg brood) obtained from the same parent colony and

randomly assigned to each treatment (fluid type). Ants were selected from nine different

parent colonies. A single fluid was offered to each test colony to determine the

consumption and preference of fluids under no-choice exposure. One test arena and one

control arena were located adjacent to each other. There were 17 replications for a total

of 85 test arenas and 85 control arenas. Fluids were tested over a six week period from







20

January 24, 2003 to March 10, 2003; room temperature and humidity were between 25.7

+ 3.70C and 20-53%, respectively.

The temperature and relative humidity of test arenas and control arenas, in

contrast to room temperature and humidity, was determined in a separate study on one

day (June 2003) when room temperature was 26.40C and room humidity was 38%. This

was accomplished by placing a digital thermometer and hygrometer inside each of 13

paired test and control arenas for 2 min, a length of time sufficient for readings to

stabilize but not long enough for ants to invade electronic equipment. Colonies inside

test arenas were identical in composition to those used in this no-choice consumption test.

Water consumption was determined in a separate study over a three week period

in November, 2003 during which room temperature and humidity were maintained

between 28.1 + 1.40C and 43-48%. Ants were provided with water in microcapillary

tubes (30 pl volume, 95 mm by 10 gm inner diameter; Model No. 21-380-9C, Fisher

Scientific, Atlanta, GA) in the same manner that test fluids were provided. Water control

tests were replicated using the same 9 colonies in the same sequence used for test fluids

with a total of 17 test arenas and 17 control arenas.

Pharaoh ant colonies were starved for 48 h before placement of fluids into arenas;

water was available ad libitum in all arenas, including water controls, via a 15 ml cotton-

stoppered vial during starvation period (Haack and Vinson 1990, Haack et al. 1995).

After 48 h of starvation, a microcapillary tube containing one test fluid was added to each

arena. Ants were allowed to feed for 24 h. Then, fluid loss was measured

gravimetrically in each arena.









Fluid Measurements

Each empty 30 pL (95 mm long) microcapillary tube was weighed via a digital

scale to +0.1 mg, and the mass of the empty microcapillary tube was recorded. Each tube

came pre-marked with a green fill line located 67 mm from tip of tube. Each

microcapillary tube was filled to the 67 mm mark and held -21.2 pL of test fluid. Mass

of the microcapillary tube with fluid was weighed and recorded. The amount of fluid

provided to all control and test arenas was calculated for each arena by subtracting the

mass of the empty microcapillary tube from the mass of the tube containing initial mass

of provided fluid (Wei and Johnson 1996). Thus, initial provided amounts of each fluid

were obtained for control arenas and for test arenas.

In addition to initial weights of food provided to arenas, data collected during the

experiment included post-evaporation weights of fluid-loaded tubes for control arenas.

Final, post-evaporation weights of control arena fluids were obtained by subtraction of

the appropriate empty microcapillary tube mass (Wei and Johnson 1996) from the initial

weight of the microcapillary tube loaded with food.

Finally, data collected during the experiment included post-consumption weights

of fluid-loaded tubes for test arenas. Final, post-consumption weights of test arena fluids

were obtained by subtraction of the appropriate empty microcapillary tube mass (Wei and

Johnson 1996).

Fluid loss in control arenas was calculated by subtracting the amount of fluid

initially provided from the amount of fluid remaining; in control arenas, fluid loss was

due entirely to evaporation. Fluid loss in test arenas was comprised of both consumption

and evaporation; therefore, consumed quantities were determined by adjusting for







22

evaporation. Provision of fluids via microcapillary tubes enabled fluids to retain constant

surface areas, and greatly minimize evaporation. Although several consumption formulas

were considered (Appendix C), because it is a standard method of measuring

consumption in nutritional ecology literature, a slightly modified version of Waldbauer's

(1968) formula was used to correct for evaporation rates of both initial and final fluid

quantities in test and control arenas as follows:

C = (1 2[Y/G]) (W [L + (Y/H)L])

where

C = corrected consumption,

W = mass of fluid provided (g) in test arena,

L = mass of uneaten fluid and ants remaining in fluid (g) in test arena,

G = mass of fluid provided (g) in control arena,

H = mass of fluid remaining (g) in control arena,

Y = G H = weight loss of control fluid,

Y/G = weight loss of control fluid / initial mass of control fluid (= a, in Waldbauer 1968),

Y/H = weight loss of control fluid / final mass of control fluid (= b, in Waldbauer 1968).

Statistical Analysis

Evaporation of patient-care fluids in control arenas was analyzed via a one-tailed

Student's t-test (P = 0.05; SAS Institute 2000) to determine if the amount of evaporation

was significantly different than zero (Ho: t = 0) (Fig. 3-2). Evaporation of each fluid was

also compared via analysis of variance, and means were separated using Tukey's HSD (P

= 0.05). In a separate study, means of humidity and temperature inside control arenas







23

were compared via analysis of variance; means were separated using Tukey's HSD (P =

0.05; SAS Institute 2000).

Negative consumption values (9 of 101 values) were truncated to zero.

Consumption of patient-care fluids and water was analyzed via a one-tailed Student's t-

test (P = 0.05; SAS Institute 2000) to determine whether consumption was significantly

different than zero (Ho: t = 0) (Table 3-2). Homogeneity of variances for consumption

among the different fluids was tested with Levene's Test. Variances were not

homogeneous, so data were logarithmically transformed logoo (x+1)). Transformed fluid

consumption was compared via analysis of variance, and means were separated using

Tukey's HSD (P = 0.05; SAS Institute 2000).

Results

Pharaoh ant workers foraged from the nesting cell to the microcapillary tubes in

order to obtain food and water. No queens, larvae, or pupae were observed in the

microcapillary tubes containing patient-care fluids or water. Workers were observed in

microcapillary tubes within 5 min of placing fluids in test arenas. They generally entered

and fed one at a time, due to the narrow tube diameter (1 mm ID), and exited by backing

out. Occasionally, an extremely small worker was able to turn around within a

microcapillary tube. In general, ants waited around the opening of the microcapillary

tube, sometimes entering immediately behind another ant. When this occurred, all ants

had to exit in reverse order to permit first ant to exit the tube. In relatively few instances,

ants became stuck in the fluid and were unable to extricate themselves. Because stuck

ants became embedded, the fluid spread out and extended past the stuck ant so that other

ants were still able to feed upon the fluid. Numbers of ants remaining in fluids were not







24

recorded. Although some ants died during the starvation and the 24 h test period,

mortality was not counted and was considered a natural event that did not significantly

affect mean consumption per arena of any specific fluid.

A volume of -21.2 pl of each patient-care fluid was provided. Because the tested

fluids have differing specific gravities and because of minor variations in the volumes

provided, initial weights were significantly different (Table 3-1). Ensure, the most

viscous fluid, had significantly more mass (-76 mg) placed in microcapillary tubes than

other fluids. Saline had the least mass of fluid provided (-68 mg). Mean data for initial

amounts of water provided in a separate study was measured (Table 3-1) but was not

analyzed with other fluids.

Evaporation of the tested patient-care fluids from the microcapillary tubes was

distinctly measurable despite the higher than room humidity in covered arenas containing

cotton-stoppered water vials. Fluid loss in control arenas, due to evaporation only,

ranged from 7-13 mg and did not differ significantly among the patient-care fluids (F =

2.10, df = 1, 80; P = 0.0879) (Table 3-1, Fig. 3-2). In contrast, there were significant

differences in fluid loss of test arenas (Table 3-1), which was due to combined

evaporation and consumption. Total fluid loss of dextrose (42.88 2.36 mg) in test

arenas was significantly higher than that of all other fluids; saline solution lost

significantly less fluid (16.37 3.60 mg) than all other fluids.

Temperature observations of 13 paired test units in May 2003, indicated that

temperature inside covered test arenas (27.36 + 0.3 10C) was statistically significantly

higher than inside covered control arenas (26.38 + 0.340C) and room temperature

(26.400C) by about oC, (F = 4.58; df = 1,25; P = 0.0428). The mean temperature of







25

control arenas was not significantly different than room temperature. Humidity was not

significantly different between the insides of covered test (51.69 + 0.88% RH) and

control arenas (51.77 + 1.34% RH) (F = 0.00; df = 1, 25; P = 0.9621), although they were

both significantly higher than room humidity (38% RH).

Pharaoh ants consumed significant quantities of all patient-care fluids (Table 3-2).

Consumption of 5% dextrose (37.95 + 5.21 mg) was significantly higher than the

consumption of all other fluids (F = 14.86; df = 4, 80; P < 0.0001; SAS Institute 2000).

Consumption of human plasma (17.63 + 3.26 mg), human blood cells (10.33 + 1.67 mg),

saline solution (9.94 + 3.60 mg) and Ensure dietary supplement (7.39 + 1.53 mg) (i.e.,

all fluids except dextrose) were not consumed significantly differently. Consumption of

baseline water controls (2.54 + 0.54 mg) was significantly greater than zero (Ho: t = 0).

Discussion

Pharaoh ants must forage to obtain energy and protein to feed queen and larvae.

Adult workers consume larval postpharyngeal regurgitations and anal secretions to help

meet their own metabolic and nutritional needs (Holldobler and Wilson 1990). Since

pupae do not feed, development of Pharaoh ants must be nutritionally supported entirely

during larval stages. Adult Pharaoh ants ingest to crop only particles smaller than 1.8 [tm

diameter (Petti 1998). Larger particles are filtered into the infrabuccal pocket and

transported back to the immobile larvae in the nest (Petti 1998), where the larvae can

digest these solid particles and later regurgitate fluids back to the workers and queens,

primarily through the labial glands (Holldobler and Wilson 1990).Since larvae provide

digested food to workers and provide up to 93% of substances that queens depend upon

(BOrgesen and Jensen 1995), colony vigor depends on healthy growing of larvae. Larvae







26

are especially important in Pharaoh ant colonies, because the ability of queens to found

new colonies is positively linked to larval presence (Borgesen and Jensen 1995). Protein

is a necessary constituent of cuticle and organ development; if protein availability is

limited, many species of ants will consume brood (Holldobler and Wilson 1990).

Peacock (1950) attributed brood mortality to various causes, including cannibalism of

sexual brood. In general, the priority for protein feeding is queens, larvae and workers

(Berndt 1977, Borgesen and Jensen 1995). Workers are last in the protein nutritional

hierarchy, since queens and brood need large amounts for egg-laying and developing,

respectively (Holldobler and Wilson 1990).

Fluid Measurements

The amount of fluid provided can impact the rate of evaporation of any given

fluid, since evaporation rates are dependent upon the volume of fluid present as well as

upon the surface area exposed to air. Despite some fluctuation in temperature and

humidity, the surface area from which evaporation could have occurred was limited to

two 1 mm diameter openings, and as a result there was no significant difference between

evaporation of the various liquids tested.

Finally, ants did not eat until depletion of provided fluids during the 24 h study.

Thus, while Waldbauer's (1968) formula is the standard for measuring consumption in

nutritional ecology studies, it would probably have been as acceptable to use a straight

subtraction of evaporation as an adjustment for consumption in this experimental set up.

Water Consumption

Water is an essential component of ant survival and reproduction (Chapman

1998). Foraging is a multi-step process for Pharaoh ant colonies (Sudd 1957b, 1960).







27

After discovery of water or food, foragers lay faranal pheromone trails as they quickly

return to the nest (Jeanson et al. 2003) and recruit additional workers. In this experiment,

workers discovered the water vials within 15 min of placement into arena at the

beginning of the 24 h starvation period and established a trail from the water vial to the

nesting cell. They maintained these trails during the entire starvation period so that

recruitment to water vials was established prior to placement of fluid-loaded

microcapillary tubes. Because faranal is a short-lived pheromone with a half life of -9

min on clean plastic surfaces (Jeanson et al. 2003), Pharaoh ants must continuously

deposit faranal on trails to maintain recruitment to desirable resources. At the end of the

starvation period, a microcapillary tube containing fluid was placed in arena; workers

discovered the new fluid source almost immediately. In the water baseline study, which

consisted of placement of water-loaded microcapillary tubes into the arena along with ad

libitum water in 15 ml vial, the choice available to ants was consumption of water via

different mechanisms, either via the microcapillary tube centered in the arena, or via the

vial placed in the opposing corner of the arena, rather than between different fluids.

Water consumption in the baseline study was significant, but appeared to be much

less than consumption of all other fluids, indicating that while Pharaoh ants consumed

water in microcapillary tubes, they probably obtained water primarily from water vials.

Observations in all test arenas of multiple ants feeding at water vials, of only 1 or 2 ants

feeding at microcapillary tubes, and maintenance of recruitment trails to water vials

support this possibility.









Dextrose Consumption

Simple sugars such as dextrose are immediately digested without need to break

dextrose down into simpler components, as is needed for more complex sugars such as

sucrose, fructose and other sugars. In fact, it is possible that dextrose is digested more

rapidly than glucose that contains both L-glucose and D-glucose, because L-glucose must

be converted to D-glucose first for digestion. Dextrose is actually composed entirely of

D-glucose molecules, the most easily digested enantiomer of glucose.

Consumption of dextrose in each arena was significantly higher than consumption

of Ensure@, the only other high-sugar fluid provided in this experiment. Foraging

studies revealed that Pharaoh ants are attracted to sugar food sources (Sudd 1960, Haack

and Vinson 1990, Vail and Williams 1994, Vail 1996). It is possible that foraging needs

of Pharaoh ants change with seasonal requirements in a manner similar to that of

Solenopsis invicta (L.) (Porter and Tschinkel 1987). Dextrose is probably more attractive

to Pharaoh ants than Ensure@ for several reasons: (1) sugar is the only nutrient present,

(2) sugar is present as D-glucose, the simplest and most easily digested and metabolized

form possible, and (3) concentration of sugar in 5% dextrose solution has a lower

viscosity to allow for easier extraction by adult workers, and may help satisfy moisture

requirements as well as energy requirements.

Significant consumption (Ho: [t = 0) of dextrose agrees with research of other

scientists who previously reported that ants feed readily and frequently on sugars. Many

researchers used sugar or honey in either liquid solutions or as solids in bait and foraging

studies for Pharaoh ants. Riley (1889) suggested coating pieces of crockery with sugar

solution to attract Pharaoh ants, and Bellevoye (1889) reported Pharaoh ants consuming







29

sweets in his apartment and neighboring bakery. Therefore, it was not unexpected that

consumption of 5% dextrose was significantly higher than consumption of other fluids.

Plasma and Blood Consumption

Pharaoh ants must forage for protein to meet metabolic needs of the queen for

egg-laying and to enable larvae to complete their development. Human whole blood

contains a significant amount of protein (-8%). Blood cells contain protein that is bound

in bi-lipid cellular membranes and inter-cellular structures. In contrast, plasma contains

most of whole blood's protein as free-floating particles in a solution that is 90% water.

Blood cells solidified in the microcapillary tubes during test, resulting in loss of

capillary action and greatly increased surface area of exposed fluid. Although heparin

slows coagulation of blood cells, blood cells solidified over time. Pipetting blood cells

out of test tubes became increasingly difficult as samples aged. All samples were used

within 7 d to reduce solidification. While heparin anticoagulant was present in whole

blood samples, initial removal of plasma via pipette probably included removal of

heparin, since plasma contains everything in whole blood except blood cells (AABB

2002). Therefore, blood cells as tested probably contained little anticoagulant during test

period; thus, coagulation and solidification was more likely in blood cells than in plasma.

Additionally, there was variability in coagulation levels between donated samples.

Whole blood samples were not analyzed for hematocrit percentages (percentage of blood

cells present in whole blood varies from person to person and even from day to day: i.e.,

hematocrit can increase when donor is dehydrated or decrease if donor has consumed

large quantities of salt in diet) prior to use in this experiment.







30

The effect of heparin upon Pharaoh ant consumption was not studied in this

experiment. There is evidence that heparin may negatively affect feeding ability of

mosquitoes (Boyd and Kay 2000), but mosquitoes are capillary feeders specialized to

feed directly from vertebrate host bloodstream (Service 2000, Mullen and Durden 2002),

and heparin mostly affects the successful bloodmeal acquisition in mosquito (Boyd and

Kay 2000). In contrast, Pharaoh ants are omnivorous feeders that feed upon exterior,

open wounds rather than penetrating below vertebrate epidermis into sealed veins and

arteries. Finally, ant colonies used in this experiment were small, thus, protein

requirements may have been lower, resulting in less feeding of high-protein fluids.

In regards to both plasma and blood cells, this experiment represents the first time

that blood, either as whole blood, or as its constituents, has been fed to Pharaoh ants,

although Grace et al. (1986) fed human sputum to Iridomyrmex (Linepithema) humilis

(Mayr) during a forensic murder investigation. While nobody has ever fed human plasma

or human blood (or any animal plasma or blood) to Pharaoh ants, many researchers have

determined that Pharaoh ants will feed on high-protein foods such as raw liver, liver

powder, flies, and cockroaches (Bellevoye 1889, Sudd 1960, Edwards 1986, Haack and

Vinson 1990). Therefore, significant consumption of human plasma, a high-protein fluid,

comes as no surprise, and I suspect that presence of plasma in open wounds and at burn

sites might be an important factor in Pharaoh ant attraction to patients in hospitals. In

contrast, consumption of blood cells was not expected to be large, since blood cells are

not as prevalent at open wound/burn sites as plasma. Bums tend to ooze plasma rather

than to bleed, due to body's immune system response, which facilitates delivery of white







31

blood cells and enzymes (proteins) to damaged epithelium to fight infection and close

open wounds by development of scar tissue.

Saline (=0.9% NaCI) Consumption

Electrolytes such as sodium (Na+) and chlorine (Cl-) ions (both present in saline

solution) are nutrients that Pharaoh ants need in their diet (Chapman 1998). Vinson

(1970) cited Butler's (1940) report that some dilute solutions of NaCl were preferred over

water. However, significant consumption of saline solution in each test arena over a 24 h

period after only 3 d of starvation was not expected. Saline was included because saline

solution is often used in patient-care in hospital environments, both to replace lost fluids

and to sterilize wounds, especially in burn units.

Saline consumption in this experiment agreed with Williams (1990) when he

compared acceptability of saline to acceptability of several sugars (he used raw honey as

test standard). It is interesting to note that the acceptance ratio he obtained for saline

consumption was nearly identical to those for consumption of 10% dextrose and 10%

glucose, both simple sugars. He contrasted saline's acceptance ratio (0.069 + 0.015)

against raw honey's acceptance ration (1.00 + 0.000) to determine that while saline was

accepted as a food source, it was much less preferable than most sugars.

Ensure Consumption

Ensure was the only test fluid that could be considered "complete," since it

contains all three major nutrients: 26% proteins, 17% sugars, and 17% lipids. This fluid

was expected to be highly attractive to Pharaoh ants and consumption of this complete

food was anticipated to be higher than that of other fluids. Consumption of Ensure may

have been affected by its nutritional completeness. Vail reported that a bait containing







32

all three nutrients (carbohydrates, lipids and proteins) would attract any foraging Pharaoh

ants. Although consumption of Ensure@ was less than that of other fluids, that may

reflect its completeness, since Pharaoh ants would not need to eat as much to meet their

nutritional requirements. In contrast, they would need to consume large quantities of a

low-density sole nutrient food to obtain the same energy and nutrition.

This experiment represents the first time that Ensure@ (or any dietary

supplement) has been provided to Pharaoh ants for consumption. These results provide

some interesting information about consumption of complete foods vs. simpler foods.

It is possible that ant preference for Ensure@ might have been limited for one or

more reasons: I observed that Ensure@ dried and partially solidified inside microcapillary

tubes while other fluids remained in original liquid form. Solidification of Ensure@ was

most closely matched by solidification of blood cells, but the manner of drying was

observed to be different. Specifically, blood cells lost microcapillary function and spread

out in a thin layer along microcapillary tube bottoms after solidification and desiccation.

In contrast, Ensure@ dried in microcapillary tubes in such a manner as to partially block

the tube, making ant penetration and extraction of fluid potentially more difficult.

Low consumption of plasma, blood cells and Ensure@ might have been due to the

presence of heparin anticoagulant or to high lipid content in high-protein fluids. Heparin

was present in whole blood samples prior to centrifugation, so proportions of heparin in

separated blood cells and plasma were unknown, and the effect of heparin on feeding was

not studied. Protein comprises approximately 8% of plasma, 8% of blood cells (AABB

2002), and 17% of Ensure@ (nutrition label). Previous research has shown that Pharaoh

ants deplete protein stores as rapidly as carbohydrate stores (Haack and Vinson 1990,







33

Haack et al. 1995), but proteins used in those experiments consisted of a powdered egg

yolk paste, and egg yolks have a high fat content of -74% in contrast to their protein

content of -20% (USDA 2002). Ensure@ has a much lower fat content (-17%) and a

slightly higher protein content (-26%) than egg yolks. Pharaoh ants have fed upon lipid

foods with no starvation (Williams 1990), so the effect of lipids upon consumption of

Ensure@ is unknown.

It is clear that in this study dextrose consumption exceeded that of all other fluids.

Sumpter and Beekman (2003) reported Pharaoh ants preferentially recruit to foods with

highest sucrose concentration. Knowing that Pharaoh ants are attracted to and will

consume fluids used in patient-care, especially dextrose, hospital staff should not spread

these fluids in hospital; if these fluids are present (especially near patient), they should be

cleaned up quickly to decrease potential for Pharaoh ant attraction to patients within

hospital and other health-care environments.

















-C
0


Ca







It 05


+1 +1 +1 +1 +1







to 05 -"c


00 0





too
::S 1 7o N C








. a +1 +1 +1 +1 + o +
0 0 -U O O o-






" a ,- 0 00 r 0 -o o


- (:5+ +1 +1 +1 +1 +1





Co C l o

C C) ON z (N 6











So 65












Table 3-2. Mean (liquid no-choice) consumption (24 h) for each of five different patient-care fluids used in hospital
environments by starved (3d) Pharaoh ant colonies (n=250).


Patient-care
Fluid


Test arenas
Mean consumption SEM (mg)


Blood


5% Dextrose

Ensure@

0.9% NaCl


10.33 + 1.67b

37.95 + 5.21a

7.39 + 1.53b

9.94 + 3.60b

17.63 + 3.26b


Plasma


Water 0.48 + 0.07 16 7.25 < 0.0001


Mean consumption within a row (Ho: t = 0) analyzed via Student's t-test (SAS Institute 2000). Mean consumption of fluids
(mg) and rank (number) listed in columns analyzed via Tukey's HSD test (P = 0.05). Means within a column followed by
same letters are not significantly different (F = 14.86; df = 4, 80; P < 0.0001); data were logarithmic-transformed before
analysis.
a Water analysis done separately as baseline study; water data not part of test fluid analysis.


6.17

7.28

4.83

2.76

5.41


< 0.0001

< 0.0001

< 0.0001

0.0095

< 0.0001



















14 13.53a
12.71a

12 -


10 9.25a
E 8.59a
0 o 7.54a
8 6-
0.



4 3.21


2


0 --
Water Blood 5% Dextrose Ensure@ 0.9% NaCI Plasma



Figure 3-2. Mean (liquid no-choice) evaporation (24 h) in control arenas for each tested patient-care fluid used in hospital
environments. Evaporation means followed by same letter are not significantly different (Tukey's t-test; F = 2.10; df = 4,
80; P < 0.0001). Water analysis done separately as baseline study; water data not part of test fluid analysis

















401 37.95a


35 -


30


E 25

0
C. 20 17.63b
E

C 15
0
S10.33b 9.94b
T 10 739b


5 -

0.48
0 ---- i i3----- ----- -----------i---------- ------ -----------------

Water 5% Dextrose Plasma Blood 0.9% NaCI Ensure



Figure 3-3. Mean (liquid no-choice) consumption (24 h) in test arenas for each tested patient-care fluid used in hospital by starved
(3d) Pharaoh ant colonies (n=250). Means followed by same letter are not significantly different (Tukey's t-test; F =13.97;
df = 4, 80; P < 0.0001); data were logarithmic-transformed before analysis. Water analysis done separately as baseline
study; water data not part of test fluid analysis..













CHAPTER 4
CHOICE LIQUID TEST

Introduction

Many social insects are omnivores, consuming a wide variety of foods from

multiple nutritional classifications. This is especially true of hymenopteran colonies.

Within the hymenoptera, omnivorous diets are frequently seen in many species of ants

(Hblldobler and Wilson 1990). This omnivorous diet differentiates them from other

social insects such as termites which are specialized to feed upon foods that are limited in

their availability, either nutritionally (dead wood in the case of termites) or seasonally

(pollen and nectar in the case of bees) (Wheeler 1910).

Like most other tramp ant species, Pharaoh ants feed on a wide variety of energy-

rich foods (Wheeler 1910) that include insects, nectar, fruit, dead animals, sweets,

pastries, meat, jelly, (Peacock et al. 1950, Sudd 1960). Disturbingly, Pharaoh ants also

feed on humans (Beatson 1972, Edwards and Short 1990, Eichler 1990)

Nutritional needs of social insects are complex and varied due to the structure of

their colonies. Ant colonies contain different castes and development stages: workers

and reproductive, eggs, larvae, and pupae; all stages except eggs and pupae have

nutritional needs and consume food. Nutritional needs differ among castes and stages:

for example, queens need more protein for egg-laying, larvae need protein to complete

development, and workers require more carbohydrates to supply energy for foraging and

tending of brood and queens (Hblldobler and Wilson 1990). The nutritional needs of the

colony reflect the combination of all individual members' nutritional needs within the







39

colony. Cassill and Tschinkel (1999) reported that the overlapping nutritional needs of

different caste members determines foraging and nursing behavior by worker ants within

the colony.

An interesting aspect of Pharaoh ants is their ability to change their consumption

to adjust for the nutritional need of the colony. Edwards and Abraham (1990)

demonstrated that Pharaoh ants alternate between high-protein and high-carbohydrate

foods. They concluded that foods containing both of these nutrients would prove more

attractive as Pharaoh ant bait matrices, since inclusion of both simultaneously would

provide the ants with both highly-recruited nutrients. They also noted that Pharaoh ant

colonies which have satiated upon one nutrient, (e.g. carbohydrates) might recruit more

to the other major nutrient, such as protein. Because Pharaoh ants vary their diet, their

foraging and feeding behavior is an especially important consideration when alternative

foods are available for consumption.

In hospitals, a wide variety of food is available throughout the hospital, in

kitchens and cafeterias, in food-carts wheeled through the hospital, and in snack vending

machines. In addition, Pharaoh ants have access to unique nutritional fluids, such as

human body fluids and fluids used in patient-care, available to them in health-care

environments. Pharaoh ants have been observed feeding around wounds and body

openings, which might have been leaking body fluid (Eichler 1990). Pharaoh ant

consumption of fluids associated with hospital patients was confirmed in a no-choice

bioassay (Chapter 3). Because patient-care fluids are used throughout the hospital,

further examination was deemed necessary to determine how the simultaneous

availability of these patient-care fluids might affect Pharaoh ant consumption.







40

Specifically, the objective was to determine the preference of small colonies of Pharaoh

ants for five liquid patient-care fluids using a choice test.

Materials and Methods

Ants

Pharaoh ant test colonies were extracted from large parent colonies housed and

reared at the Urban Entomology Laboratory, University of Florida, Gainesville, FL.

Rearing temperatures ranged from 21.90C 29.40C with relative humidity between 20-

53%. (Appendix A) Colonies consisted of 250 workers, 3 dealate queens and

approximately 50 mg brood. One hundred twenty five workers were selected from cells

to ensure the presence of nurses and 125 workers were selected from outside cells to

ensure the presence of foragers (Haack 1987, Haack and Vinson 1990, Haack et al.

1995). Brood included eggs, all larval stages and pupae. See Appendix B for full details

on selection of ants and preparation of experimental test colonies.

Foraging Arena

Foraging arenas contained one nesting cell, one water vial and five microcapillary

tubes with fluids (Figure 4-1). These arenas were identical to those used in the previous

no-choice liquid consumption test (Chapter 3) except for the addition of four

microcapillary tubes to permit fluid availability. A detailed description of the foraging

arena was provided in Chapter 3.

























Five 30 l capillary tubes,
with fluids


27 cm


4 19cm -



Figure 4-1. Arenas with water vial and nesting cells placed in diagonally opposite
comers. Capillary tubes placed in center. Test arenas (with ants) and control
arenas (no ants) were placed adjacent to each other and comprised a test pair.
Test arenas contained 125 inside ants, 125 outside ants, 3 queens and -50 mg
brood (including all stages of brood).







42

Water was provided ad libitum in a 15 ml vial (Model No. 55-9, Thornton Plastics, Salt

Lake City, UT) with three large cotton balls inserted into top to prevent leakage and to

provide easy access for ants. A water vial was placed in the opposite diagonal comer of

arena across from the nesting cell. Each replicate consisted of one test arena placed

adjacent to one control arena. Test arenas contained ants, while control arenas lacked

ants. Control arena data was used to quantify and adjust for evaporation of fluids. Each

arena was covered with a clear plastic lid to maintain humidity and temperature.

Experimental Fluids

The same five fluids used in the no-choice test in Chapter 3 were used in this

choice test. Fluids consisted of human blood cells, human plasma, saline (0.9% sodium

chloride); 5% dextrose (D-glucose), and Ensure (Vanilla Plus, Ross Products Division,

Abbott Laboratories, Columbus, OH). Fluids were chosen to replicate hospital conditions

and potential fluid sources as closely as possible. The detailed descriptions of each fluid

and handling procedures used in this study are identical to that provided in Chapter 3.

Experimental Design

The experiment was designed as a randomized complete block, blocking on

colony. Each block consisted of one Pharaoh ant colony (125 inside workers, 125 outside

workers, 3 dealate queens and -50 mg brood) obtained from a single parent colony. Five

fluids were simultaneously offered to each colony to determine the consumption among

the test fluids under a choice exposure. As in the previous no-choice experiment

(Chapter 3), an experimental unit consisted of a paired test and control arena located

adjacent to one another. The choice test was replicated 21 times. Ants were selected

from 6 different parent colonies. Fluids were tested over a two week period from May







43

21, 2003 to June 3, 2003; room temperature and relative humidity were maintained

between 24.9-28.60C and 35-57% RH.

Pharaoh ant colonies were starved for 48 h; water was available ad libitum in all

arenas, including water controls, via a 15 ml cotton-stoppered vial during starvation

period (Haack and Vinson 1990, Haack et al. 1995). After 48 h of starvation, five

microcapillary tubes, each containing one test fluid, were added to each arena, and the

water vial was not removed. Thus, water was always available in all arenas. Fluids were

offered simultaneously to each colony to determine consumption preference among

provided fluids.

Mortality after starvation period and prior to 24 h feeding period in test arenas

was observed, but not recorded. Dead ants were neither counted nor removed. Mortality

rates during starvation periods were not taken into consideration when recording

consumption from test arenas.

Fluid Measurements

Fluid placement into arenas was identical to the previous no-choice test (Chapter

3) except that five different fluids were simultaneously provided, each in its own

microcapillary tube. In brief, each microcapillary tube (Model No. 21-380-9C, Fisher

Scientific, Pittsburgh, PA) was weighed before and after the liquid was loaded. The

amount of liquid provided to all control and test arenas was calculated for each arena by

subtracting the mass of the empty microcapillary tube from the mass of the tube

containing initial mass of provided fluid (Wei and Johnson 1996).

Ants were allowed to feed for 24 h. Microcapillary tubes were then reweighed in

both control and test arenas. Due to frequent presence of ants still in microcapillary tubes

during final gravimetric measurement on a digital scale, weights of post-consumption test







44

fluid after 24 hour test period included the mass of ants remaining in microcapillary

tubes. Final ant biomass measurements were included in final mass weights recorded and

were subtracted later using an average mass value of 0.0002 g per Pharaoh ant worker.

Worker ant mass was determined by weighing several individual worker ants to

determine average mass per ant.

Fluid loss in control arenas was calculated by subtracting amount of fluid initially

provided from amount of fluid remaining, resulting in the amount of evaporation that

occurred in the 24 h test period. Fluid loss in test arenas was comprised of consumption

and evaporation; therefore, to determine consumed quantities, the amount of fluid lost to

evaporation was subtracted. Many researchers have designed formulas to adjust

consumption for fluid loss, and Waldbauer's (1968) formula was selected (Appendix C).

Because multiple ants remained in test arena fluids, Waldbauer's (1968) formula was

modified to account for mass of ants remaining in fluid. Thus, corrected consumption

was determined as follows:

C = (1 2[Y/G]) (W [(L A) + (Y/H)(L A)])

where

C= corrected consumption,

W = mass of fluid provided (g) in test arena,

L = mass of uneaten fluid and ants (g) in test arena,

A = n*(0.0002 g/ant) mass of ants stuck in tube, n = number of ants stuck in tube,

thus, (L-A) = mass of uneaten fluid,

G = mass of fluid provided (g) in control arena,

H = mass of fluid remaining (g) in control arena.







45

Y = G H = weight loss (g) of control fluid,

Y/G = weight loss of control fluid / initial mass of control fluid (= a, in Waldbauer 1968),

Y/H = weight loss of control fluid / final mass of control fluid (= b, in Waldbauer 1968).

Statistical Analysis

Dead or alive ants remaining in microcapillary tubes at the conclusion of the 24 h

test period were compared among test fluids. Mean number (n) of ants remaining in test

fluids was compared via analysis of variance and Tukey's HSD test (P = 0.05; SAS

Institute 2000)

Fluid loss from evaporation in the control arenas and fluid loss from evaporation

and consumption in the test arenas among types of hospital fluids were compared by

analysis of variance and Tukey's HSD (P = 0.05; SAS Institute 2000). Variances were

tested using Levene's Homogeneity of Variances test (P = 0.05; SAS Institute 2000).

Negative consumption values (15 of 100 values) were truncated to zero.

Consumption of patient-care fluids and water was analyzed via a one-tailed Student's t-

test to determine if there was significant consumption (Ho: [t = 0).

Consumption data were compared via analysis of variance, and means were

separated using Tukey's HSD test (P = 0.05; SAS Institute 2000). Homogeneity of

variances for consumption among the different fluids was tested with Levene's Test (P =

0.05; SAS Institute 2000).

Results

Number of Ants

Ants were observed entering microcapillary tubes within 5 min of placing fluids

in test arenas. Observations of ant behavior in this experiment were similar to those seen

in Chapter 3.







46

Mean number of ants remaining in Ensure (5.77 + 0.96) at the conclusion of the

test period was significantly higher (F =12.40; df = 4, 95; P < 0.0001) than in the other

fluids (Table 4-1). Numbers of ants remaining in all other fluids were low and did not

significantly differ, ranging from 0.71 to 1.94 (Fig. 4-2).

Fluid Measurements

Initial quantities of fluids provided to test and control arenas differed significantly

(F = 256.9, df=4, 95; P < 0.0001) among the tested patient-care fluids. The mass of

Ensure (-77 mg) placed into control arenas was higher than that of all other fluids. The

mass of blood (-76 mg) was significantly higher than that of plasma, dextrose, and

saline. The mass of provided saline (-69 mg) was less than that of all other fluids in

control arenas, but not in test arenas (Table 4-2).

In test arenas, initial amounts of fluids provided were also significantly different

(F = 133.39; df=4, 95; P <0.0001). As in control arenas, significantly more Ensure by

mass was placed into arenas than all other fluids, the mass of blood provided was

significantly higher than that of plasma, dextrose, and saline, and the mass of provided

saline was significantly less than that of all other fluids (Table 4-2).

Evaporation in control arenas was significantly different between fluids (F = 4.85;

df = 4, 95; P= 0.0013). Plasma had the highest fluid loss due to evaporation, while saline

and dextrose had the least amount of fluid loss due to evaporation (Table 4-2).

Fluid loss of tested patient-care fluids in test arenas was significantly different due

to combined evaporation and consumption (F = 29.01; df=4, 95; P < 0.0001). When

ants were present, significantly more dextrose (-77 mg) was lost than any other fluid.

Ensure lost more fluid than plasma, saline and blood. Saline and blood had the smallest

amount of fluid loss (Table 4-2). Fluid loss in control arenas was due solely to







47

evaporation, and ranged from 2.59 0.19 mg for dextrose to 4.87 + 0.68 mg for plasma.

In contrast, fluid loss in test arenas ranged from 5.20 0.58 mg for blood to 50.80 3.46

mg for dextrose (Fig. 4-3, Table 4-2). Temperatures in test arenas (27.36 + 0.31C) were

statistically significantly higher (F=4.58; df = 1, 24; P = 0.0428) than in control arenas

(26.38 + 0.340C), probably due to the presence of ants.

Consumption

There were significant differences (F = 29.97; df = 4, 95; P < 0.0001) in

consumption of the fluids by Pharaoh ants. Consumption of 5% dextrose (49.14 + 3.51

mg) was significantly higher than consumption of all other fluids. Consumption of

Ensure (16.05 + 6.23 mg) was significantly greater than consumption of plasma (9.31 +

3.17 mg ), saline (3.09 + 1.14 mg), and blood cells (2.46 + 0.48 mg) (Table 4-2, Fig. 4-3).

Baseline mean consumption of water (0.48 + 0.07 mg) was lower than the consumption

of all test fluids. None of the fluids in any of the microcapillary tubes in either the test or

control arenas were completely depleted.

Discussion

Number of Ants

Numbers of ants remaining in microcapillary tubes at conclusion of test period did

not correlate to heaviest consumption or to observed foraging behavior. Ants remaining

in microcapillary tubes appeared to be feeding on remaining fluid. Foraging activity

observed during the first hour of the test period was of largest recruitment to the

microcapillary tubes containing 5% dextrose.

In contrast, the Ensure had the highest number of ants remaining in the tubes after

24 h. The mean number of ants remaining in microcapillary tubes after 24 h within each

arena comprised 6% of colony workers (n=250) (Fig. 4-1).









Fluid Measurements

The significant differences in mass between different fluids probably reflects the

different densities of the fluids used in this experiment, since all microcapillary tubes

were filled to the same volume of -21.2 tl. Higher initial masses for both Ensure@ and

blood were associated with the higher viscosity observed in these two fluids. Similarly,

saline solution contained only 0.9% electrolytes, making it the least viscous fluid used.

Differing amounts of provided fluid did not affect consumption since no fluid was

depleted. Differing amounts of provided fluid (Table 4-2) might have resulted in

different rates of fluid loss for different fluids. Although Ensure@ had significantly more

fluid mass provided in both control and test arenas than other fluids, evaporation of

Ensure@ in control arenas did not differ from that of other test fluids.

Fluid loss in test arenas (Table 4-2) was comprised of both consumption and

evaporation; therefore, the amount of fluid lost to evaporation had to be subtracted in

order to determine consumption values.

Although temperatures within test arenas were statistically significantly higher

than in control arenas, both means were within 1C of temperatures reported as ideal for

Pharaoh ants (Peacock and Baxter 1949, Peacock et al. 1955c). Hence, relative to

temperature, foraging was probably optimal in this experiment. Humidity did not differ

between control and test arenas; however, the humidity levels inside arenas of -52%

were well below the 80% recommended by (Peacock and Baxter 1949, Peacock et al.

1955c) and also lower than the 65% humidity recommended by (Haack 1987) as optimal

for the Pharaoh ant. Low humidity probably increased the moisture needs of the test

colonies, and probably increased the rate of evaporation of all fluids. If low humidity







49

resulted in an increased moisture requirement, then the consumption of fluids may have

increased.

Consumption

(Mailleux et al. 2000) determined that ant foraging behavior is a response to the

nutritional demands of the ants and to the characteristics of the food resources, such as

the density and spatial distribution, distance to food, and the quality of food.

Consumption of high-carbohydrate and high-protein fluids was expected, since a

nutritional need was assumed to be induced by a 3 d starvation period. Food

characteristics affecting ant foraging behavior varied only in the nutritional composition

and quality of the different fluids offered, since all microcapillary tubes were placed

randomly in replications (Mailleux et al. 2000).

In this experiment, all fluids were consumed in significant proportions, indicating

that the ants consumed a wide variety of foods when provided with a choice. This agrees

with observations that Pharaoh ants change food selections based on satiety and

preference, and that they will even alternate between high-carbohydrate and high-protein

food sources (Granovsky and Howell 1983, Edwards and Abraham 1990).

Consumption also can be influenced by colony needs, and can change depending

on a variety of factors such as colony size, proportion of larvae to workers, and numbers

of egg-laying queens. For example, egg-laying by queens and larger numbers of larvae

both increase the colony's need for proteins. In the red imported fire ant, Solenopsis

invicta (Buren), even worker size or crop contents can have an impact on food flow

within a colony's members (Cassill and Tschinkel 1999).









Dextrose

Pharaoh ants have been reported to foraging most strongly to fluids with highest

sugar concentrations (Sumpter and Beekman 2003), such as sucrose solutions. While the

Pharaoh ants consumed all fluids, 5% dextrose was consumed significantly more than all

other liquids (Fig. 4-3). Mean consumption of 5% dextrose (49.14 + 3.51 mg) was

consumed 3 times more than Ensure@, than the next most consumed fluid (Table 4-3).

These results were similar to those obtained in the no-choice test (Chapter 3).

Haack et al. (1995) reported a similarly high Pharaoh ant recruitment to simple

carbohydrates: he observed that -100% of workers foraged at carbohydrate source within

-3 h of introduction of food into arena, while only 2% foraged to proteins within 24 h,

and -80% foraged to lipids after 24 h. In hospitals, 5% dextrose is often squirted onto

floors when nurses or doctors clear intravenous tubing prior to inserting tubing into

patients. The amount of time that might elapse between spillage and cleanup is

unknown; if dextrose is present in the rooms, this experiment clearly indicates that

Pharaoh ants will recruit to this fluid.

Ensure@

When given a choice of fluids after 3 d starvation, Pharaoh ants preferred high-

sugar fluids most; the high concentration of sugars in Ensure@ probably helped make it

the second most-preferred fluid in this experiment, despite its relatively high evaporation

rate (Fig. 4-4) and tendency to dry out and partially solidify, thus making it more difficult

to extract than the less viscous dextrose.

The relatively high consumption of Ensure@ in this choice test contrasted with the

lesser consumption of Ensure@ in the no-choice test (Chapter 3). Pharaoh ants preferred

fluids with high concentrations of sugar. Although Pharaoh ant consumption of Ensure@







51

was second only to consumption of dextrose, it was significantly less than that of

dextrose. Ensure@ has fewer simple sugars, and more complex sugars than dextrose,

making it a more complex fluid. Insects rapidly convert ingested glucose in trehalose for

storage in hemolymph, and must take an additional, high-energy step of converting

complex sugars into glucose first (Nation 2003). Ensure@ dietary supplement is further

complicated since it contains high concentrations of proteins and lipids in addition to

sugars.

Finally, Ensure@ had more ants stuck in it than all other fluids (Table 4-1, Fig. 4-

2). Some ants remaining in fluid were dead during final weighing; Sudd (1957a)

observed Pharaoh ants respond in a negative manner to dead Pharaoh ants. He reported

that when a Pharaoh ant encountered dead of its own species, it became alarmed and ran

away when within 1 cm of dead and; however, those ants were killed by crushing. In this

experiment, most ants remaining in the fluid were still alive; ants which were dead had

not been crushed. Any chemicals emitted via crushing that may have served as an alarm

pheromone were probably not present in this experiment. Therefore, the presence of

dead ants in the tubes was not expected to influence feeding behavior. Visual

observation of ants in microcapillary tubes did not note any forage reluctance to enter

tubes containing ants. Ultimately, the nutritional complexity and dead ant accumulation

in Ensure@ did not prevent foragers from removing significant quantities of fluid.

Plasma

Plasma evaporated at a significantly higher rate than all other fluids, and was the

only fluid with a higher evaporation rate than Ensure@. While plasma evaporated at the

highest rate, consumption of plasma (9.312 + 3.17) was not significantly different than

that of saline (3.093 + 1.14). The high evaporation rate of plasma probably increased the







52

concentration of ions within the solution more rapidly than occurred in saline. The

presence of heparin anticoagulant may have influenced ant consumption of plasma, and

may have influenced the rate of evaporation as well.

Both human plasma and saline solution are very aqueous fluids which are high in

sodium and chlorine electrolytes. In fact, the electrolyte content of 0.9% NaCl solution

and human plasma are identical; saline is often used as a fluid replacement to increase

blood volume (AABB 2002). So it was not surprising that Pharaoh ant preference for

these two fluids was not statistically different.

NaCI

Saline solution is a very aqueous solution, composed mostly of water, with 0.9%

sodium chloride ions. Evaporation of saline was the second lowest of tested fluids;

evaporation of saline was higher than that of dextrose. Both fluids were highly aqueous

solutions with low-density so that ant ingestion was easy. Consumption of saline was

significant, but was less than that of high-sugar fluids.

Many electrolyte solutions are repellent to red imported fire ants (Vinson 1970).

However, Vinson determined that the addition of salt increased the acceptance of sugars

by red imported fire ants. Thus, the available choice of NaCl solution in test arenas may

have resulted in increased consumption of high-sugar fluids, dextrose and Ensure. In

this experiment, Pharaoh ants consumed 0.9% NaCl solution (3.093 + 1.14) despite the

ad libitum presence of water. This clearly indicated that the Pharaoh ants were interested

in saline solution for nutritional reasons, especially when baseline water consumption test

results (0.48 + 0.07 mg) were taken into account.









Blood

Blood cells were the least consumed of all tested fluids. Blood was the only fluid

which had a higher rate of evaporation (3.32 + 0.25) than of consumption (2.046 + 0.48),

although blood loss in the test arena (5.20 + 0.58) was greater than evaporation (Table 4-

2). Very little foraging activity was observed on microcapillary tubes containing blood

cells. The ants did not recruit to it very strongly as a food source, although they did

consume it in significant amounts. Solidification of the blood cells was observed. The

small inner diameter of the microcapillary tube and a hardened, solidified surface

probably made extraction very difficult for foragers.

Blood was one of the two most viscous fluids in this experiment, and

consumption of very dense and highly-nutritious blood cells may have resulted in

satiation at lower levels of consumption. Alternatively, blood cells are high in protein,

but are not high in sugars, so the lower consumption of blood cells, as compared to that

of high-sugar fluids dextrose and Ensure, might have indicated that the Pharaoh ants'

nutritional need for carbohydrates was higher than for protein during the test period.

It is clear that in this study, dextrose was preferred over all other fluids. It is

possible that high sugar (sole nutrient) concentration is favorable for Pharaoh ant fluid

consumption. Sumpter and Beekman (2003) reported Pharaoh ants preferentially recruit

to foods with highest sucrose concentration. When ants are placed in test arena, they

consumed all patient-care fluids. Knowing that Pharaoh ants are attracted to and will

consume fluids used in patient-care, especially dextrose, hospital staff should not spread

these fluids in hospital; if these fluids are present (especially near patient), they should be

cleaned up quickly to decrease potential for Pharaoh ant attraction to patients within

hospital and other health-care environments.







54

Table 4-1. Mean number of ants remaining (either feeding, stuck or dead) in fluids at end
of 24 h choice test feeding period.


Liquid Food


Blood

5% Dextrose

Ensure@

0.9% NaCl

Plasma


Water


Test arenas
Mean number ants SE


0.71 + 0.49b

1.06 + 0.34b

5.77 + 0.96a

1.94 + 0.56b

1.12 + 0.42b


2.77 + 1.10


Means followed by same letter are not significantly different by Tukey's HSD-test (P =
0.05; SAS Institute 2000).
a Water analysis done separately as baseline study; water data not part of test fluid
analysis.












Table 4-2. Mean (choice test) initial weights of fluid quantity provided (mg) and mean fluid loss (mg) for each of five different
patient-care fluids used in hospital environments (24 h) for starved (3 d) pharaoh ant colonies.

Mean (mg) SEM

Control arenas Test arenas
Patient-care
Fluid Initial wt Fluid Lossa Initial wta Fluid Lossb


Blood 75.81 + 0.28b 3.32 + 0.25ab 76.03 + 0.42b 5.20 + 0.58c

5% Dextrose 70.47 + 0.17cd 2.59 + 0.19b 70.51 + 0.25cd 50.80 + 3.46a

Ensure@ 76.92 + 0.18a 4.00 + 0.24ab 77.25 + 0.35a 19.62 + 6.08b

0.9% NaCl 69.76 + 0.13d 2.98 + 0.48b 69.99 + 0.18d 5.82 + 1.16c

Plasma 71.15 + 0.24c 4.87 + 0.68a 71.48 + 0.17c 13.07 + 3.19bc

Water 69.48 + 0.12 3.21 + 0.19 69.54 + 0.16 5.64 + 0.59


Means within a column followed by the same letter are not significantly different by Tukey's HSD-test (P = 0.05).
a Fluid Loss in control arenas is to evaporation only.
bFluid loss in test arenas due to evaporation and consumption combined.
c Water analysis done separately as baseline study; water data not part of test fluid analysis.












Table 4-3. Mean (choice test) consumption (mg) for each of five different patient-care fluids used in hospital environments (24 h) for
starved (3 d) pharaoh ant colonies.


Liquid Food Test arenas df t P
Mean consumption SEM (mg)


Blood 2.046 + 0.48c 38 4.23 0.0001

5% Dextrose 49.140 + 3.51a 38 14.02 <0.0001

Ensure@ 16.054 + 6.23b 38 2.58 0.0056

0.9% NaCl 3.093 + 1.14bc 38 2.71 0.0101

Plasma 9.312 + 3.17bc 38 2.94 0.0056


Water 0.48 + 0.07 16 7.25 <0.0001


Mean consumption within a row (Ho: t = 0) analyzed via Student's t-test (P = 0.05; SAS Institute 2000).
Mean consumption of fluids (mg) listed in columns analyzed via Tukey's means separation test (P = 0.05).
Means within a column followed by same letters are not significantly different by Tukey's HSD-test (P = 0.05).
a Water analysis done separately as baseline study; water data not part of test fluid analysis.


















6 5.77a


5 -


u. 4


"3 3 2.77
z
1.94b
2

1.06b 1.12b
1 0.71b


0 -I I.I...
Water Blood 5% Dextrose Ensure 0.9% NaCI Plasma


Figure 4-2. Mean number of ants (either dead or feeding) remaining in fluid at end of 24 h choice test feeding period. Means
followed by same letter are not significantly different by Tukey's HSD-test (P = 0.05; SAS Institute 2000). Water analysis
done separately as baseline study; water data not part of test fluid analysis.
















6.00


5.00 4.87a


4.00ab
4.00 -
E= 3.32ab
0 2.98b
3.00
1 2.59b
0

LL
2.00 -



1.00



0.00 ....
Blood 5% Dextrose Ensure@ 0.9% NaCI Plasma




Figure 4-3. Mean evaporation (choice test) of fluids used in hospital environments for patient-care ([tg) (24 hours). Data was
collected from the control arenas: evaporation was calculated by subtracting final mass of fluid remaining at the end of the
24 h test period from the amount of fluid initially provided. Means followed by same letter are not significantly different
by Tukey's HSD-test (P = 0.05; SAS Institute 2000).



















49.14a
/I


5% Dextrose


16.05ab


Ensure@


50



40
E
c-
0
a. 30
E
c-
0
20



10


3.09bc


0.9% NaCI


Plasma


Figure 4-4. Mean (choice test) consumption (mg) for tested fluids used during test period (24 h) for starved (3 d) pharaoh ant
colonies. Means followed by same letter are not significantly different by Tukey's HSD-test (P = 0.05; SAS Institute
2000).


9.31 bc
M


2.05c
| -'1_ I


Blood














CHAPTER 5
NO-CHOICE DEHYDRATED FLUID TEST

Introduction

Hospitals, nursing homes and other medical treatment facilities are often infested

with Pharaoh ants (Green et al. 1954, Edwards and Baker 1981a, Eichler 1990), a tropical

pest species that has worldwide distribution (Hblldobler and Wilson 1990, Fowler et al.

1994). Pharaoh ants have been discovered in sensitive patient-care areas, such as bum

units where they were observed feeding on open wounds (Weidner 1982), in neonatal

units where they were observed feeding upon the intact skin of newborns (Steinbrink

1978) as cited by Eichler (1990). Burn victims and newborns typically lie on beds

containing sheets, and are provided with various fluids to meet hydration and nutritional

needs. Fluids can spill and desiccate on sheets and might attract Pharaoh ants to the

patient.

Consumption and preference tests were performed for 5 patient-care fluids used in

hospitals to determine if Pharaoh ants actually consumed each fluid (Chapter 3) and to

determine which fluids they preferred (Chapter 4) in their liquid state when given a

choice. While they consumed all fluids, consumption of liquid dextrose was significantly

higher than all other fluids and was the preferred fluid in both the no-choice and the

choice tests.

These five fluids are typically administered directly to the patient via inserted

tubes. Saline, 5% dextrose, red blood cells and serum are used to replace lost fluids

(AABB 2002); saline and dextrose can also be injected peritoneally (into the







61

abdomen/stomach) (Wenzel et al. 2002). EnsureTM dietary supplement is unique among

these five fluids since it is administered solely through injection into the patient's

stomach (Wenzel et al. 2002). Regardless of the method used to provide these fluids to

patients, any of these fluids might easily spill onto the patient bed and solidify upon the

bed-sheets.

It was determined that these five fluids needed to be provided to the Pharaoh ants

in their desiccated state (i.e., as solids) to determine if the ants demonstrated interest in

spilled patient-care fluids in hospitals. The potential availability of these substances

might be substantially greater in their dried form than in their liquid form, and could be a

factor in attracting the ants to patient beds and to the patient. The objective of this

experiment was to determine if Pharaoh ants would forage to desiccated patient-care

fluids used in hospital using a no-choice test.

Materials and Methods

Ants

Pharaoh ant test colonies were extracted from large maintenance/parent colonies

housed and reared at the Urban Entomology Laboratory, University of Florida,

Gainesville, FL. See Appendix A for full details on maintenance of ants. Colonies

consisted of 250 workers, 3 dealate queens and approximately 50 mg brood.

One hundred twenty-five workers were selected from cells to ensure presence of

nurses and 125 workers were selected from outside cells to ensure presence of foragers

(Haack 1987, Haack and Vinson 1990, Haack et al. 1995). Brood included eggs, all

larval stages and pupae. See Appendix B for full details on selection of ants and

preparation of experimental test colonies.









Foraging Arena and Nest

Foraging arenas (Fig. 5-1) consisted of clear plastic boxes (27 cm by 19 cm by 10

cm deep; Catalogue No. 195C, Pioneer Plastics, Dixon KY) which were coated with

FluonTM (Catalogue No. ADI, Asahi Glass Fluoropolymers, Downington, PA) on interior

sides to prevent ant escape. One nest cell was placed within one comer of each arena.

Cells consisted of sterile, covered plastic Petri dishes (50 mm diameter by 15 mm depth;

Falcons Model No. 351007, Becton Dickinson and Co., Franklin Lakes NJ) filled

halfway with Castone dental plaster (Model No. 99045, Dentsply International, York

PA). One entrance hole (-5 mm diameter) was melted into the upper side wall of the

Petri dish base via insertion of the metal tip of a hot glue gun (sans glue) just above the

level of the hardened plaster. The Petri dish lid was placed on top of the base and the

entire nesting cell was placed into the foraging arena with the entrance hole facing the

opposite diagonal comer of the arena.

Water was provided ad libitum in a 15 ml vial (Model No. 55-9, Thornton

Plastics, Salt Lake City UT) with three large cotton balls inserted into the top to prevent

leakage and to provide easy access for the ants. The water vial was placed in the opposite

diagonal comer of the test unit arena across from the cell.

Hospital Sheets

Hospital sheets (Forest General Hospital, Newberry Road, Gainesville, FL) were

washed in extremely hot temperatures and underwent a rigorous sterilization process

(John Bennet, personal communication). Freshly laundered hospital sheets were obtained




























hospital sheet square (25 by 25
cm) on top of aluminum foil
square (also 25 by 25 cm)


dehydrated fluid;

inside penciled
circle (20 cm
diam.)


27 cm


m- M


Figure 5-1. Arenas for dehydrated liquid food no-choice experiment. Arenas contained
125 inside workers, 125 outside workers, 3 dealate queens, and -50 mg brood
(all stages). Fluids drops placed on hospital sheets desiccated for 4 d and
placed in center of arena, resting on aluminum foil. Fluids placed within
penciled circle (20 cm diameter).


p







64

from a local hospital and were cut into small square (25 by 25 cm). Circles (20 by 20

cm) were penciled onto hospital sheets and were centered so that a 2.50 cm border

existed between the penciled circle and the exterior border of the sheet. Pencil notations

were made along the edge of the hospital sheet, outside of the circled border to identify

the fluid applied to a sheet.

Heavy-duty aluminum foil was cut into small squares (25 by 25 cm) and placed

under the hospital sheets. Each square of aluminum foil with its respective hospital sheet

square was placed into a large (40 by 52 by 12 cm deep) plastic tray (Model No. 400-5N,

Panel Controls, Greenville, SC) and suspended -0.3 m above a portable room heater in

preparation for fluid application and dehydration.

Dehydration of Fluids

Fluids used in this experiment consisted of tap water and 5 liquid patient-care

fluids: human blood cells, human plasma, 5% dextrose and 0.9% NaCl. Each fluid was

placed in a sterile test tube. Clean, dry plastic 15 ml disposable pipettes were used to

withdraw fluid and place a drop of -1 ml volume in the center of the 25 by 25 cm

hospital sheet, within the penciled circle's 20 cm boundaries. A separate pipette was

used for each fluid, and fresh pipettes were used each time. No mixing of pipettes

occurred. Drops were left to air-dry for 1-2 h until visual observation indicated thorough

desiccation. Drops were added in this manner until a total of 5 drops were placed on each

hospital sheet. After the final drop dried, the large tray was placed on a lower wall-rack

in the experimental laboratory in away from the light, and remained there for 4 d until

used in the experimental test. The tray was not covered.

Room temperature was maintained at 26.70 C 28.90 C and relative humidity

fluctuated between 40-56%. Fluids were permitted to dehydrate during the same period







65

that ants were starved. Standardized Pharaoh ant colonies were starved (only water

provided) for 72 h after being placed in the arenas. The hospital sheet square containing

the dried substance was placed in the center of the test unit, halfway between the nest cell

and the water. Each arena was covered with a clear plastic lid to maintain humidity and

temperature within arena.

Experimental Design

The experiment was designed as a randomized complete block, blocking on

colony. Each block consisted of Pharaoh ant colonies (125 inside workers, 125 outside

workers, 3 dealate queens and -50 mg brood) obtained from the same parent colony and

randomly assigned to each treatment (dehydrated fluid type).

Ants were selected from 6 different parent colonies. A single fluid was offered to

each test colony. Control arenas located adjacent to test arenas were identical except that

they lacked ants.

Timed Observations

Observations of foraging behavior were noted upon immediate placement of

fluids into arena, and the number of ants that appeared to be feeding on the dried fluids

began 15 min after initial exposure and continued at 15 min intervals for 2 h. Ants were

considered to be feeding if they were standing stationary and assumed a feeding posture

with antennae and/or palps touching dried fluids that were contained within the circled

portion of the hospital sheet, regardless of how much of their body was within the circle.

Ants feeding on fluid that spilled beyond the penciled circle were not counted.

Similarly, ants engaging in trophallaxis were not counted, even if this activity occurred in

the center of the circle. Ants that were in a resting posture within the circle were also not

counted. Ants that were walking or otherwise actively moving during the timed







66

observation intervals were not counted, even if antennae were in contact with dehydrated

fluids.

Statistical Analysis

Four analyses were done using analysis of variance (ANOVA) with means

separated using Tukey's HSD (SAS Institute 2000). First, counts of feeding ants over all

dried fluids among the 15 min intervals were compared. Second, the total number of ants

feeding on each dried fluid for the entire 2 h test period was compared.

Third, for each time interval, the mean numbers of feeding ants were compared

among fluid types. Finally, for each fluid, the mean numbers of ants feeding were

compared among each of the 15 min intervals.

Results

Usually 1 or 2 ants were foraging in the arenas during placement of dehydrated

fluids into arenas. Foragers in arenas containing Ensure and dextrose exhibited an

immediate change in behavior they rapidly increased their walking speed and began

waving antennae in front and above their heads in an excited manner. They took fast

running steps in short bursts towards the dehydrated fluid, and located the fluid within a

few seconds; recruitment of additional ants in arenas containing Ensure and dextrose

frequently occurred in less than 5 min. In arenas with dehydrated dextrose, foragers

exhibited a tendency to feed for several minutes before recruiting other ants. In arenas

with dehydrated Ensure, initial foragers tended to engage in extensive antennal tapping of

dehydrated fluid while rapidly encircling the entire hospital sheet; after completely

circling the fluid they rapidly ran to the nesting cell. Shortly afterwards, several ants

were observed proceeding in a hurried manner towards the Ensure, where most engaged

in feeding behavior for several minutes. A few returned to the nest, again resulting







67

immediately in an increased number of foragers emerging from the nest. Ant behavior in

arenas in which hospital sheets soaked with blood cells, plasma, saline or water did not

elicit any increased foraging activity or immediate recruitment behavior. Foragers in

these arenas continued to walk around the arena in a slow, unhurried manner and did not

seem to respond to dehydrated blood cells, plasma, saline or water as a food source.

Occasionally, a forager was observed feeding on blood cells, plasma, or saline, but no

foragers were observed on sheets that had been soaked with water. Foragers did not

exhibit the same type of excited behavior in these arenas as was observed in arenas with

high-sugar dehydrated fluids. Even after feeding for several minutes, ants were not

observed running back to the nest, and no mass recruitment was observed in arenas

lacking sugar.

Peak feeding of all fluids simultaneously occurred at 45 min and steadily decreased

afterwards. However, ant counts were not significantly different amongst time intervals

(F = 0.97, df =7, 1144; P = 0.4499) (Fig. 5-2). The number of Pharaoh ants feeding

upon Ensure (13.89 + 0.53) was significantly higher than on all other dehydrated fluids.

Feeding upon 5% dextrose was significantly higher than on plasma, blood cells and saline

(F = 617.75, df = 5, 1146; P < 0.0001) (Fig. 5-3). Numbers of ants feeding upon

Ensure was significantly higher at each time interval than on all other fluids (Fig. 5-4

through 5-11). There was no significant difference in numbers of ants feeding on all

other fluids per time interval. Only Ensure and dextrose had significant feeding among

time intervals during the 2 h experiment. Feeding of Ensure peaked at 45 min (18.0 +

2.0 ants) (Fig. 5-12) and feeding of dextrose also peaked at 45 min (2.4 + 0.4 ants) (Fig







68

5-13). Feeding on the saline, blood cells, and plasma were virtually non-existent as mean

counts at any time interval was never higher than 0.2 ants (Fig. 5-14, 5-15, 5-16).

Discussion

Pharaoh ants were starved for 72 h in this no-choice solid food test this was 24 h

longer than starvation period of 48 h used in my liquid consumption tests (Chapters 3, 4).

In contrast, a 72 h starvation period resulted in observable foraging behavior for nearly

all test arenas. Starvation periods >72 h were not attempted because Pharaoh ant

recruitment to carbohydrates can be very high after extensive starvation, due to a more

rapid depletion of carbohydrate stores (Nation 2003). Haack (1987) reported that 7 d

starvation made it difficult to count numbers of foraging Pharaoh ants because they

overwhelmed high-sugar foods upon initial exposure. Haack et al. (1995) reported that 2

to 3 d starvation was sufficient to deplete protein stores enough to elicit strong foraging

behavior for proteins. Ensure@ was the only fluid with significant consumption

throughout the experiment, and it was notable that it was the only complex fluid

provided in this experiment. It has high concentrations of all three major nutrients, with

17% protein, 17% lipid, and 26% carbohydrate.

The number of ants recorded feeding at both Ensure@ and dextrose peaked at the

45 min time interval, although is was not significantly different from ant counts of most

other time intervals. These data differ slightly from that collected by Haack (1987) who

reported that 93% of workers fed within the first 30 min of food placement in the arena.

The Ensure@ and dextrose were the only fluids to have significant numbers of ants

feeding upon them, and they shared a similar trait: high sugar content. Although the

types of sugars presented differed and were present in their solid form, they elicited

foraging and feeding behavior on a significant number of foraging ants. Foraging to







69

solid, dehydrated sugar here differs from the results of Haack et al. (1995) who observed

that Pharaoh ants did not respond to solid sugar crystals as a food source. In fact, they

observed the ants filling tiny cracks in the nesting cell walls with the sugar crystals.

Since the solid sugars used by Haack et al. (1995) were manufactured and sold as table

sugar, both enantiomers of glucose were probably present in those crystals. In contrast,

the solid sugar present in my experiment was crystalline structures of D-glucose, the

biologically active enantiomer which does not require energy for digestion, since it is

passively absorbed through the midgut wall and stored primarily as glycogen (Nation

2003).

Although Pharaoh ants appeared to feed upon dextrose, they aggregated around and

appeared to feed upon Ensure at much higher levels. Ensure was themost viscous

fluid used in this experiment and was the only food that contained multiple nutrients. It

took longer to desiccate than other fluids, and when finally dry, appeared as a brown-

colored textured glob that had a three-dimensional texture. In contrast, all clear fluids

were difficult to see. Repeated applications of dextrose were visible because they

resulted in a noticeable stiffening of the hospital sheet, and often adhered to the sheet to

the underlying aluminum foil. Plasma was observable because it left a straw-colored

stain on the hospital sheet. Blood was easy to see, but it was difficult to apply to the

sheet in controlled amounts since the blood cells were very sticky. Blood cells dried in

globs that were relatively large in comparison to clear fluids, although blood cells and

Ensure had almost the same consistency. Saline and water were nearly impossible to see

visually although the application left a wet spot, so that application could be confirmed







70

immediately after application. Desiccation of saline resulted in a crispy hospital sheet

while application of water did not appear to have any effect.

Significant feeding upon high-protein substances or water was not expected.

Higher feeding of dextrose solids and lower feeding upon Ensure@ were anticipated. In

previous consumption studies, Pharaoh ants preferentially consumed 5% dextrose above

all other fluids, so it was interesting that when dried, Pharaoh ants feeding was less than

the Ensure dietary supplement.

Pharaoh ant recruitment to Ensure@ occurred after 4 d desiccation, so this fluid

still elicited foraging after several days. Thus if spilled, Ensure@ may attract Pharaoh

ants at least 4 d after drying out. The complete nutritional status of Ensure, available to

them after 3 d of starvation eliminated the need for resource switching, since it contained

all three major nutrients. Additionally, the workers may have used their 4-toothed (Ogata

et al. 1998) mandibles to scrape and extract solid particles of the lumpy-textured Ensure

residue. Adult workers do not eat solid food, although they can digest solids as larvae

(Le Masne 1953) as cited in (Petti 1998). Instead, when consuming solutions or semi-

solids containing liquids, adults filter the liquid portion through their buccal tube (Petti

1998) and store filtered solids in the infrabuccal pouch. The resulting compacted solids

are later ejected as a pellet, by version of the infrabuccal pocket, after the ant's return to

the nest containing the developing larvae (Petti 1998). Also, worker ants can pick up

larger particles in their mandibles to transport those back to the larvae. Solenopsis invicta

(Buren), another myrmicine species, has even demonstrated the use of tools to assist in

food transportation back to the nest (Barber et al. 1989). High foraging numbers of ants

at the desiccated Ensure fluid probably indicated that this desiccated fluid contained







71

nutrients needed by the larvae, or was suitable for larval consumption and subsequent

larvae to worker trophallaxis. Due to its high sugar and lipid content, Ensure fluid

contained large energy reserves, despite its dehydrated state. Thus, it may have attracted

the foragers with its high nutritional value, even though the food still had to be

transported back to the nest and biologically modified by the larval trophallaxis.

Nutritional requirements of ant colonies may vary with colony size; natural

Pharaoh ant colonies tend towards 1:1:1 ratios of workers:eggs:larvae (Peacock et al.

1955b). Although the experimental colonies used in this experiment were smaller than

natural colonies discovered in human structures, such as that reported anecdotally by

Bellevoye (1889), they were representative of the ratios, and were similar to experimental

colony sizes used by Haack and Vinson (1990).

The attractiveness of dried Ensure@ after at least 4 d portends possible problems

by attracting ants to patients due to spillage on hospital linens. However, extended

attractiveness potentially indicates that this fluid might be useful as part of bait matrix for

Pharaoh ant control. Of course, further testing would be required to assess this potential

relative to other bait attractants.


















3.5 3.42


3 2.90 2.97

2.45 2.51
u) 2.5 2.33 2.45 2.29


S2 1.85
C

1.5 -4


1


0.5


0 -- --- ----- --- F---
15 30 45 60 75 90 105 120
Minutes after Fluid Introduction


Figure 5-2. Mean number of ants feeding on all dehydrated fluids among 15 min intervals during 2 h test period. No significant
differences among time intervals.

































1.49b
7-i


5% Dextrose


0.03c


0.9% NaCI


13.89a
-I-


Ensure@


0.OOc


Water


0.07c


Plasma


0.06c


Blood


Figure 5-3. Mean number of foraging ants exhibiting feeding behavior at dehydrated fluids over entire 2 h test period. Means
followed by same letter are not significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.
















14.0


12.2a
12.0 -



10.0-



E 8.0 -

z

S6.0 -



4.0 -



2.0 -


0.Ob
0.0 -
Water Ensure


Figure 5-4. Mean number of ants feeding on
significantly different (P = 0.05; S


1 .7b


OI I .Ob O.Ob O.Ob

@ 5% Dextrose Plasma Blood 0.9% NaCI


at the 6 dried fluids at 15 min. time interval. Means followed by same letter are not
AS Institute 2000) and Tukey's HSD test.















16.0 15.a
15.0a

14.0 t


12.0


) 10.0 -


Z 8.0


26.0


4.0-
2.3b
2.0 I

O.Ob O.Ob O.Ob 0.2b
0.0 .. '
Water Ensure 5% Dextrose Plasma Blood 0.9% NaCI


Figure 5-5. Mean number of ants feeding on at the 6 dried fluids at 30 min. time interval. Means followed by same letter are not
significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.

















18.0a


2.3b


0.Ob 0.1 b 0.Ob


Ensure 5% Dextrose


Plasma


Blood


0.9% NaCI


Figure 5-6. Mean number of ants feeding on at the 6 dried fluids at 45 min. time interval. Means followed by same letter are not
significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.


20.0

18.0

16.0

14.0

12.0

10.0

8.0


O.Ob

Water















18.0


16.0 15.6a


14.0


12.0
c0
< 10.0
6
z
c 8.0


6.0


4.0

1.9b
2.0 -

O.Ob O.Ob 0.2b O.Ob
0.0 .*.. -_ '-
Water Ensure 5% Dextrose Plasma Blood 0.9% NaCI



Figure 5-7. Mean number of ants feeding on at the 6 dried fluids at 60 min. time interval. Means followed by same letter are not
significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.
















16.0


13.9a
14.0 -


12.0


a 10.0


Z 8.0
c
(0
6.0 00


4.0


2.0
0.6b
0.Ob 0.2b O.Ob O.Ob
0.0 0 --
Water Ensure 5% Dextrose Plasma Blood 0.9% NaCI



Figure 5-8. Mean number of ants feeding on at the 6 dried fluids at 75 min. time interval..Means followed by same letter are not
significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.














16.0


14.0a
14.0 -


12.0 -


U 10.0


z 8.0


6.0


4.0


2.0 1.0b

O.Ob 0.1 b O.Ob O.Ob
0.0 ...
Water Ensure 5% Dextrose Plasma Blood 0.9% NaCI


Figure 5-9. Mean number of ants feeding on at the 6 dried fluids at 90 min. time interval. Means followed by same letter are not
significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.
















12.5a


O.Ob


1.2b


0.Ob 0.1 b 0.Ob


Water Ensure 5% Dextrose Plasma Blood 0.9% NaCI


Figure 5-10. Mean number of ants feeding on at the 6 dried fluids at 105 min. time interval. Means followed by same letter are not
significantly different (P <0.05) by ANOVA and Tukey's HSD test.



















10.Oa
10 -T



8




c
(0 00

4 -



2
1.0b

O.Ob O.Ob O.Ob O.Ob

Water Ensure 5% Dextrose Plasma Blood 0.9% NaCI



Figure 5-11. Mean number of ants feeding on at the 6 dried fluids at 120 min. time interval. Means followed by same letter are not
significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.

















20.0 -
18.0a
18.0 -

15.6ab
16.0 15.0ab 1 5.
14.0 13.9ab 14.0ab
14.0
12.2ab 1 2.5ab
12.0 -
S 1 0.Ob
S10.0 -

00
I 8.0 -

6.0 -

4.0 -

2.0 -

0.0
15 30 45 60 75 90 105 120



Figure 5-12. Mean number of ants feeding on Ensure@ among 15 min intervals over the 2 h test period. Means followed by same
letter are not significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.




















2.4a


2.3ab


0. 1.7abc

< --

01-R
C
Z 1.5 -
C
(0 -L


1.9abc


1.2abc


1.0bc


0.6c


1.Oabc


Figure 5-13. Mean number of ants feeding on 5% dextrose among 15 min intervals over the 2 h test period. Means followed by same
letter are not significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.






























0.0Oc


0.1 7a
M-


0.18

0.16

0.14

0.12

0.10

0.08

0.06

0.04


0.04b


0.0Oc


0.OOc


0.OOc


0.OOc


0.04b


F']


0.02


u.uu


Figure 5-14 Mean number of ants feeding on saline among 15 min intervals over the 2 h test period. Means followed by same letter
are not significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.















0.25


0.20a
0.20 -




0.15

6
z
c
0.10- 00
0.08a 0.08a



0.05 0.04a 0.04a



0.OOa 0.OOa 0.OOa
0.00 ----
15 30 45 60 75 90 105 120



Figure 5-15. Mean number of ants feeding on blood cells among 15 min intervals over the 2 h test period. Means followed by same
letter are not significantly different (P <0.05) by ANOVA and Tukey's HSD test.
















0.18 0.17a


0.16 -


0.14


0.12
C
0.10 -
z 0.08a 0.08a 0.08a
c 0.08 ---


0.06

0.04a 0.04a 0.04a
0.04 -- --


0.02 -


0.00
15 30 45 60 75 90 105 120



Figure 5-16. Mean number of ants feeding on human plasma among 15 min intervals over the 2 h test period. Means followed by
same letter are not significantly different (P = 0.05; SAS Institute 2000) and Tukey's HSD test.