Citation
Development and Evaluation of Biorational Dips for Ornamental Cuttings Infested with the Madeira Mealybug, Phenacoccus Madeirensis Green

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Title:
Development and Evaluation of Biorational Dips for Ornamental Cuttings Infested with the Madeira Mealybug, Phenacoccus Madeirensis Green
Creator:
Guerrero, Sarahlynne C
Place of Publication:
[Gainesville, Fla.]
Florida
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University of Florida
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english
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1 online resource (124 p.)

Thesis/Dissertation Information

Degree:
Master's ( M.S.)
Degree Grantor:
University of Florida
Degree Disciplines:
Entomology and Nematology
Committee Chair:
HODGES,AMANDA C
Committee Co-Chair:
OSBORNE,LANCE S
Committee Members:
LEPPLA,NORMAN C
HARDER,AMY MARIE
RODA,AMY LOUISE
Graduation Date:
5/3/2014

Subjects

Subjects / Keywords:
Family names ( jstor )
Host plants ( jstor )
Hypochromic anemia ( jstor )
Insecticides ( jstor )
Leaves ( jstor )
Mortality ( jstor )
Pests ( jstor )
Phytotoxicity ( jstor )
Pyrethrins ( jstor )
Soaps ( jstor )
Entomology and Nematology -- Dissertations, Academic -- UF
biorational -- invasives -- mealybug -- phytotoxicity
City of Davenport ( local )
Genre:
bibliography ( marcgt )
theses ( marcgt )
government publication (state, provincial, terriorial, dependent) ( marcgt )
born-digital ( sobekcm )
Electronic Thesis or Dissertation
Entomology and Nematology thesis, M.S.

Notes

Abstract:
In order to prevent the spread of cryptic invasive pests, a model on-site biorational dip protocol was developed by evaluating the phytotoxicity and efficacy of Naturl Oil (soybean oil), WetcitTM(alcohol ethoxylate), Publix Soap (nonylphenol ethoxylate), and Vapor Gard (di-1-p-menthene) at varying concentrations on coleus cuttings infested with Phenacoccus madeirensis Green. Efficacy was achieved with 1.0% Naturl Oil (76.09%) and 0.1% WetcitTM (74.99%) by Day 3 and reached highest mortality by Day 14 with 1.0% Naturl Oil (90.97%) and 0.1% WetcitTM (90.47%). Six of the key foliage phytotoxicity ratings for 0.1% WetcitTM were also higher than 1.0% Naturl Oil: chlorosis, chlorotic flecking, necrotic flecking, holes, tip chlorosis, and tip necrosis. Subsequently, 1.0% Naturl Oil was selected as the model dip treatment and compared to other Naturl Oil concentrations for determining exposure time and host plant efficacy. Exposure time efficacy for 1.0% was attained by Day 7 with the 30 (71.81%), 60 (72.54%), and 120 (80.24%) second dip and reached highest mortality with the 30 (87.82%), 60 (88.21%), and 120 (92.00%) second dip by Day 14.Verbena efficacy was achieved by Day 7 for 1.0% (80.74%) and 1.5% (85.85%) and attained highest mortality by Day 14 with 1.0% (80.74%) and 1.5% (85.85%). Mint efficacy for 1.0% (82.71%) and 1.5% (96.89%) was also reached by Day 7 but 1.5% (96.89%) had the highest mortality by Day 14. Overall, 1.0% Naturl Oil as a 30 second dip was determined as the model on-site dip protocol for treating infested ornamental cuttings. ( en )
General Note:
In the series University of Florida Digital Collections.
General Note:
Includes vita.
Bibliography:
Includes bibliographical references.
Source of Description:
Description based on online resource; title from PDF title page.
Source of Description:
This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Thesis:
Thesis (M.S.)--University of Florida, 2014.
Local:
Adviser: HODGES,AMANDA C.
Local:
Co-adviser: OSBORNE,LANCE S.
Electronic Access:
RESTRICTED TO UF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE UNTIL 2014-11-30
Statement of Responsibility:
by Sarahlynne C Guerrero.

Record Information

Source Institution:
UFRGP
Rights Management:
Applicable rights reserved.
Embargo Date:
11/30/2014
Classification:
LD1780 2014 ( lcc )

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DEVELOPMENT AND EVALUATION OF BIORATIONAL DIPS FOR ORNAMENTAL CUTTINGS INFESTED WITH THE MADEIRA MEALYBUG, PHENACOCCUS MADEIRENSIS GREEN By SARAHLYNNE GUERRERO 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 2014

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2014 Sarahlynne Guerrero

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To my loving family and friends. Your unconditional love and support will always guide me.

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4 ACKNOWLEDGMENTS I wish to thank my co advisors: Dr. Amanda Hodges, for funding my graduate research assistantship and providing constant guidance and support through my MS program, and Dr. Lance Osborne, for supporting my extensive research and extension internship endeavors at the UF/IFAS Mid Florida Research and Education Center in Apopka, FL. To the Biosecurity Research and Extension Laboratory, thank you for all their overall graduate program advice, guidance, and he lp with running my Gainesville experiments: Stephanie Stocks, Ashley Poplin, Hashmet Eke, Abby Griffin, and most especially Dr. Gupreet Brar and Eric Leveen. I willingne ss to help me conduct my Apopka experiments: Katherine Houben, Irma Herrera, Luis Aristizabal, Younes Belmourd, and most especially Fabieli Irizarry. Dr. Norman Leppla gave me research and life changing advice over coffee breaks. Dr. Amy Harder inspired me to always improve and facilitate change. Dr. Amy Roda provided outstanding feedback and advice from a regulatory perspective and as a friend. Dr. Mihai Girucanu provided exceptional assistance with understanding SAS and my statistical analysis. Jessica Ka lina helped me navigate through Microsoft Word and Excel on several late night occasions. The faculty and staff of the Entomology and Nematology Department helped me throughout my degree program. Additionally, I thank all the County Extension Agents and Master Gardeners who gave me the privilege to work with them during my extension internship.

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5 TABLE OF CONTENTS page ACKNOWLEDGMENTS ................................ ................................ ................................ ........ 4 LIST OF TABLES ................................ ................................ ................................ .................. 7 ABSTRACT ................................ ................................ ................................ ............................ 8 CHAPTER 1 LITERATURE REVIEW OF THE MADEIRA MEALYBUG ( PHENACOCCUS MADEIRENSIS GREEN) ................................ ................................ ................................ 10 Ornamental Industry Overview ................................ ................................ ......................... 10 Impact of Invasive Species on Ornamental Plants ................................ ............................. 12 Model Invasive Pest of Ornamental Plants ................................ ................................ ........ 14 Biology ................................ ................................ ................................ ..................... 15 Taxonomy ................................ ................................ ................................ ................ 17 Distribution ................................ ................................ ................................ .............. 18 Chemical Control Methods ................................ ................................ ............................... 18 Biorational Control Methods ................................ ................................ ............................ 23 Oils ................................ ................................ ................................ .......................... 23 Soaps and Surfactants ................................ ................................ ................................ 25 Antitranspirants ................................ ................................ ................................ ........ 26 Research Objectives ................................ ................................ ................................ ......... 27 2 DEVELOPMENT OF THE MODEL BIORATIONAL DIP ................................ .............. 61 Introduction ................................ ................................ ................................ ..................... 61 Materials and Methods ................................ ................................ ................................ ..... 64 Rearing Colonies of P. madeirensis ................................ ................................ ........... 64 Host Plant Maintenance ................................ ................................ ............................. 64 Cutting Preparation ................................ ................................ ................................ ... 65 Assessment of Biorational Phytot oxicity on Cuttings ................................ .................. 65 Assessment of Biorational Effects on Plants ................................ ............................... 66 Efficacy of Biorationals on P. madeirensis Mortality ................................ .................. 68 Statistics ................................ ................................ ................................ .......................... 70 Results ................................ ................................ ................................ ............................ 71 Efficacy Bioassay ................................ ................................ ................................ ..... 71 Phytotoxicity Assessment ................................ ................................ .......................... 72 Model Dip Treatment Determination ................................ ................................ ......... 75 Discussion ................................ ................................ ................................ ....................... 75 Efficacy Bioassay ................................ ................................ ................................ ..... 75 Phytotoxicity Assessment ................................ ................................ .......................... 78 Model Dip Treatment Determination ................................ ................................ ......... 80 Model Dip Treatment Implications ................................ ................................ ............ 81

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6 3 EVALUATION OF THE MODEL BIORATIONAL DIP ................................ ................. 85 Introduc tion ................................ ................................ ................................ ..................... 85 Materials and Methods ................................ ................................ ................................ ..... 86 Rearing Colonies of P. madeirensis ................................ ................................ ........... 86 Ev P. madeirensis Mortality ........................ 87 P. madeirensis Mortality on Various Host Plants ............ 88 Statistics ................................ ................................ ................................ .......................... 89 Results ................................ ................................ ................................ ............................ 90 Discussion ................................ ................................ ................................ ....................... 91 4 IMPLICATIONS AND FUTURE DIRECTIONS ................................ ........................... 104 LIST OF REFERENCES ................................ ................................ ................................ ...... 109 BIOGRAPHICAL SKETCH ................................ ................................ ................................ 124

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7 LIST OF TABLES Table page 1 1 Invasive ornamental pests encountered in Florida (Frank and Thomas 2004). ................ 29 1 2 Reported host plants of Phenacoccus madeire nsis. ................................ ....................... 30 1 3 Reported distribution of Phenacoccus madeirensis. ................................ ...................... 50 1 4 Natural predators and parasitoids of Phenacoccus gossypii and Ph enacoccus madeirensis (Chong 2005). ................................ ................................ .......................... 53 1 5 Reported insecticides used to control Phenacoccus madeirensis for homeowner and ornamental industry use. ................................ ................................ .............................. 56 2 1 Cumulative average percent of dead P. madeirensis per coleus cutting dipped for one minute in selected biorational treatments at each time period ................................ ........ 82 2 2 Average coleus cuttin g phytotoxicity rating per biorational material concentration for key phytotoxicity characters. ................................ ................................ ....................... 83 2 3 Average coleus cutting root length per biorational material. ................................ .......... 84 3 1 Dip treatment efficacy evaluated on various arthropod ornamental and agricultural pests. ................................ ................................ ................................ .......................... 95 3 2 Dip treatment efficacy evaluated on various ornamental and agr icultural plants. ............ 98 3 3 Cumulative average percent of dead P. madeirensis per coleus cutting dipped in varying dip exposure times for each time period. ................................ ........................ 101 3 4 Cumulative average percent of dead P. madeirensis per verbena cutting dipped for ............................. 102 3 5 Cumulative a verage percent of dead P. madeirensis per mint cutting dipped for one ................................ .... 103

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8 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 DEVELOPMENT AND EVALUATION OF BIORATIONAL DIPS FOR ORNAMENTAL CUTTINGS INFESTED WITH THE MADEIRA MEALYBUG, PHENACOCCUS MADEIRENSIS GREEN By Sarahlynne Guerrero M ay 2014 Chair : Amanda Hodges Co Chair : Lance Osborne Major : Entomology and Nematology In order to prevent the spread of cryptic invasive pests, a model on site biorational dip protocol was developed by evaluating the phytotoxicity and efficacy of l Oil (soybean oil) Wetcit TM (alcohol ethoxylate), Publix Soap (nonylphenol ethoxylate) and Vapor Gard (di 1 p menthene) at varying concentrations on coleus cuttings infested with Phenacoccus madeirensis Green. Efficacy was achieved with (76.09%) and 0.1% Wetcit TM (74.99%) by Day 3 and reached highest mortality by Day 14 with Wetcit TM (90.47%). Six of the key foliage phytotoxicity ratings for 0.1% Wetcit TM were also chlor osis chlorotic fle cking necrotic flec king holes tip chlor osis and tip model dip treatment and efficacy. Exposure time efficacy for 1.0% was attained by Day 7 with the 30 (71.81%), 60 (72.54%), and 120 (80.24%) second dip and reached highest mortality with the 30 (87.82%), 60 (88.21%), and 120 (92.00%) second dip by Day 14.Verbena efficacy was achieved by Day 7 for 1.0% (80.74%) and 1.5% (85.85%) and attained highest mortality by Day 14 with 1.0%

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9 (80.74%) and 1.5% (85.85%). Mint efficacy for 1.0% (82.71%) and 1.5% (96.89%) was also reached by Day 7 but 1.5% (96.89%) had the highest mortality by Day 14. Overall Oil as a 30 second dip was determined as the model on site dip protocol for treating infested ornamental cuttings.

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10 CHAPTER 1 LITERATURE REVIEW OF THE MADEIRA MEALYBUG ( PHENACOCCUS MADEIRENSIS GREEN) Ornamental Industry Overview Nursery a nd greenhouse ornamental crops are integral to United States agriculture. The total value of sales generated from 50,784 farm s devoted to floriculture, propagation, and sod production exceeds $16 billion (United States Department of Agriculture National Ag ricultural Statistics Service 2007). Plant production from propagation operations and floriculture crops are one of the most important components of the ornamental industry. Floriculture crops include bedding plants, garden plants, cut flowers, cut florist greens, foliage plan ts, and potted flowering plants whereas propagation operations refer to the production of cuttings, seedlings, liners, and plugs. T he total value of United States sal es for floriculture crops is approximately $6.5 billion while propaga tion operations generate $ 440 million (United States Department of Agriculture National A gricultural Statistics Service 2007). Among 47 states surveyed in the 2007 Census of Agriculture, the five leading floriculture crop states were California ($1.2 billi on), Florida ($909 million), Michigan ($439 million), Texas ($326 million), and North Carolina ($260 million) while the top five pro pagation operation states were Florida ($91 million), California ($84 million), Washington ($29 million), Michigan ($28 mill ion) and Colorado ($28 million). r and humid climate provides an ideal environme nt for ornamental crop production and sale. Although the ornamental industry is ranked fourth in Florida cash receipts, foliage and floriculture still crea te a sizeable share amounting $695 million in comparison to vegetables and melons ($1.7 billion) an d citrus ($1.5 billion) (Florida Department of Agriculture and Consumer Services 2013). Approximately 1,750 farms; 9,958 open acres of production area; and 264 million square feet of greenhouse or shaded production area are

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11 devoted to Florida propagation operations and floriculture crops (United States Department of Agriculture National Agricultural Statistics Service 2007). For floriculture sales, Florida ge nerates 21% of United States sales estimated at $3.94 billion (Florida Department of Agriculture and Consumer Services 2013). Additionally, Florida also creates 23% of the United States propagation sales valued at $355 million (Florida Department of Agricu lture and Consumer Services 2013). Overall, Florida produces 72% of United States foliage plant production sales worth $433 million and is recognized as the most profitable foliage plant production state in the United States (Florida Department of Agricult ure and Consumer Services 2013). Internal, inter regional, and international trade of all nursery products contribute to the total whole value of ornamental sales in Florida. Resu lts from Hodges (2011) indicate that 67% of customer sales occurred within t he state. Inter regional sales to states such as North Carolina (11.5%), Connecticut (4.8%), Georgia (2.3%), a nd Texas (2.1%) also contribute towards 0.2%) and Europe (0.1%) provide the s mallest contribution. Nursery products commonly produced or traded in Florida include tropical foliage (31%), deciduous and flowering trees (31%), miscellaneous plant types (10%), flowering potted plants (8%), and propagated liners, cuttings, and plugs (7% ) (Hodges 2011). Results from Hodges (2011) which surveyed 556 Florida growers in 2008 showed that the purchasing of propagation materials, such as cuttings, seedlings, whips, grafts, and liners mostly occurred wi thin the state of Florida (75%); however su bstantial purchases were made from California (10%), Oregon (3.5%), Washington (1.2%), Georgia (1.1%), and Costa Rica (2.7%) and brought to Florida. Other international purchases and shipments to Florida nursery growers involved vendors from the Bahamas, D ominican Republic, Brazil, Belize, Mexico, Canada,

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12 Belgium, Germany, Netherlands, China, Australia, and New Zealand. Additionally, the Florida Department of Agriculture and Consumer Services (2013) reported nursery and greenhouse product import and export shipp ing routes to Japan, Netherlands, and Sweden Impact of Invasive Species on Ornamental Plants The National Invasive Species Council (2001) define s an invasive species as a species that is not native to the ecosystem under consideration and whose intr oduction causes or is likely to cause economic or environmental harm or harm to human health However, another definition described by Hodges and Stocks ( 2010) defines an invasive species as any species that competes with humans by consuming or damaging food, fiber, or other materials intended for human consumption or use Nevertheless, t h e introduction of invasive species threatens the United States ornamental industry. Although further research is needed to determine invasive pest damage costs to growe rs within Florida and the United States, Oetting et al. (2006) indicated that Georgia suffered major orn amental pest damage costs from scales and mealybugs ($20 million), mites ($22.3 million), thrips ($7.5 million), whiteflies ($4.2 million), aphids ($2 m illion), and caterpillars ($1.3 million). Hodges et al. (1998) also reported that Florida growers also target mites, aphids, whiteflies, scales and mealybugs, and caterpillars. Overall, the most detrimental Florida ornamental pests include h emi pterans, thr ips, and mites (Table 1 1) (Buss 1993, Frank and Thomas 2004). Source regions for non native species traveling to Florida include the Caribbean, Central America, and South America and as much as 24% of arthropods entered Florida as plant cargo contaminants (Frank and Thomas 2004, Jenkins et al. 2014) In Florida, up to two new non native arthropod s have been reported every month ( Thomas 2004, Dr. Amanda Hodges, personal communication). Increased movement of plant material heightened by the r elaxation of tra de barriers contribute to the increasing immigration rate of non native arthropods from the Old

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13 World to the New World (Pimentel et al. 2000, Dr. Amy Roda, personal communication). Additional contributing factors that may also influence the rate of invasiv e pest immigration include an increasing human population, rapid movement of people and materials, alteration of the environment, and climate change (Pimentel et al. 2000, Frank and Thomas 2004, Devitt et al. 2012). On a national scale, Pimentel et al. (2 005) estimated 4,500 arthropod species and nearly 100 aquatic invertebrate species were introduced to the United States. Approximately 95% of these introductions were accidental and facilitated by contaminated plants, soil, or water ballast from ships and other human mediated transport vectors, such as airplanes, trains, and cars (Pimentel et al. 2000). Exact national or state figures on the number of non native species or invasive pests immigrating as plant cargo contaminants are unknown since the United States Department of Agriculture, Animal and Plant Health Inspection Services inspect only 2% of imported plant stock (Brasier 2008). However, at United States ports of entry and border crossings, t he Port Information Network (PIN) database records inv asiv e pest interceptions on plant stocks during inspections. McCullough et al. (2006) evaluated PIN from 1984 2000 and found 73% to 84% of interceptions were insects from the orders Hemiptera, Lepidoptera, and Diptera. Most invasive pest interceptions were mad e while inspecting baggage (62%), cargo (30%) and plant propagated material (7%) and occurred at airports (73%), U nited States Mexico land border crossings (13%), and marine ports (9%). Similarly, Jenkins et al. (2014) reported 77% of intercepted arthropod s entering Puerto Rico and the United States Virgin Islands by freight or luggage originating within the Caribbean. The following orders were commonly intercepted during inspection: Hemiptera (52%), Diptera (16%), Coleoptera (10%), Lepidoptera (8%), Thysan optera (5%), Acari (4%), and Hymenoptera (2%).

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14 Inspections for invasive species are long and tedious as invasive species are difficult to detect due to their appearance, biology, and behavior (Jenkins et al. 2014, McCullough et al. 2006). Adult and immatu re specimen screenings take additio nal time since primary or secondary screening characteristic s may be missing or difficult to interpret G enetalia dissection under a dissecting microscope may be necessary for making a final identification on adult or imm ature specimens. Extensive dissecting microscope use may also be required for egg or small immature final identification. Model Invasive Pest of Ornamental Plants Phenacoccus madeirensis is an invasive cosmopolitan pest that is cryptic, polyphagous in nat ure, and one of the most difficult to manage with synthetic insecticides (Chong 2005, Ludwig 2009) Phenacoccus madeirensis is considered a major plant pest particularly throughout the southeastern United States (Miller et al. 2002 Chong 2005 ). More than 60 plant families include known hosts for P. madeirensis, including several important agricultural and ornamental commodities like tomatoes ( Solanum lycopersicum ), soybeans ( Glycine max ), citrus ( Citrus sp.), hibiscus ( Hibiscus sp.), chrysanthemums ( Chrysa nthemum sp.) and coleus ( Solenostemon sp.) (Table 1 2). Adult female mealybugs are traditionally used to identify mealybugs ( Williams and Granara de Willink 1992 ). The adult P. madeirensis female is wingless, oval, elongate, flattened dorsoventrally, and 2 3.5mm (0.08 0.14in) in length (Green 1923). When squashed, the body exudes green fluid. Thick white mealy wax covers the gray body and red legs. Females have bare intersegmental areas on the thorax that form a pair of dark longitudinal lines on the dorsu m. Additionally, females have eighteen pairs of lateral wax filaments that encompass the body with the posterior pair of wax filaments projecting furthest from the body but less than the entire length of its body. Eleven days after mating, gravid females c an oviposit up to 530 eggs (Chong

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15 et al. 2003). Female longevity typically decreases as temperature increases which accounts for ovipositing females living up to 19 days and virgin females surviving for 38 days in controlled, moderate temperatures (Longo e t al. 1995, Chong et al. 2003). Plant damage caused by P. madeirensis is similar to other mealybug species and can be severe during high infestation. Crawlers appear to collectively feed on the host plant with piercing sucking mouthparts and cause dimpling and yellowing. F eeding leads to serious direc t damage on ornamental crops such as premature leaf drop, curling, deformation, and stem and growth shoot infestation resulting in retarded plant growth. However, honeydew excretion inflicts the most aesthetic damage on host plants as b lack sooty mold develops from honeydew droplets left on the host plant. H oneydew production also attracts nearby ants which provide predator protection in exchange for honeydew as a food source (Bethke 2009). Biology Sexual dimorp hism and behavioral differences are apparent during the life cycle of P. madeirensis Newly emerged first instars or crawlers appear yellow, lack a white mealy wax on the body, and are 0.5 to 0.75mm (0.02in 0.03in) in length (Green 1923, Townsend et al. 20 00). Crawlers undergo a wandering stage where they disperse from the ovisac either by crawling or catching the wind. Crawlers roam around the host plant trunk, twigs, leaves, and fruit in search for an ideal feeding site. Crawlers feed on the host plant us ing thei r piercing sucking mouthparts and are commonly found feeding on the undersides of leaves or tender shoots. Honeydew production begins shortly after feeding. Through each successive instar, honeydew excretion increases as nymph mobility decreases (S inacori 1995). Second instar body color and behavior are used to determine gender (Townsend 2000). Both male and female nymphs continue to feed and exude honeydew on the host plant while developing a thin white mealy wax on the body which varies in length from 1.0 1.5mm (0.04

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16 0.06in). Males turn pale to bright pink and construct a white narrow filamentous cocoon on the undersides of leaves in which they encase themselves and molt three times before becoming an adult male (Sinacori 1995, Williams 1985, Town send et al. 2000). Females remain yellow, feed collectively, and become sessile on the host plant. In addition to crawlers, male and female second instars also overwinter (Sinacori 1995). Gender is also distinguished by third and fourth instar appearance a nd behavior. Third instar m ales remain pale to bright pink in body color and are fully enclosed within their cocoon as they enter the pre pupal stage. In contrast, the third instar female body color turns gray and develops a thicker layer of white mealy wa x. Female body length also expands and ranges between 1.75 2mm (0 .07 0.08in). Additionally females become increasingly sessile and st art feeding alone along leaf midribs. In the fourth instar females reach reproductive maturity as wingless adults while m ales enter the pupal stage within their cocoons (Townsend et al. 2000) Adult f emales release the sex pheromone trans 1R, 3R chrysanthemyl R 2 methylbutanoate to eventually mate with newly emerged adult males (Ho et al. 2011). Males in the fifth instar e merge from their cocoons as reproductive, winged adults and search to mate with an adult female. Males appear gnat like with a single pair of wings and have vestigial mouthparts; stout, truncate genital capsules; a thin white mealy wax covering their red b rown body ; and a pair of white p osterior waxy filaments protruding from the body but less than its entire length. Adult males immediately search and mate with adult females as they only live for 3 6 days due to their vestigial mouthparts (Sinacori 1995, Ho et al. 2011, Chong et al. 2003). G ravid females oviposit up to 530 eggs within a white filamentous ovisac and die immediately afterwards (Chong et al. 2003). Ovisacs contain a cluster of yellow eggs that turn darker over time (Longo et al. 1995). All egg s are oval in shape and less than 0.5mm (0.02in) in

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17 length. Fecundity generally decreases as temperature increases which accounts for females ovipositing as few as 67 eggs and up to 530 eggs depending on the temperature (Yeh et al. 2006, Chong et al. 2003) Taxonomy Most scientific literature references P. madeirensis as indigenous to the neotropics (Williams and Granara de Willink 1992). However, Williams (1987) suggests that P. madeirensis is native to the Nearctic area of northern Mexico due to his work on the taxonomic revision of Phenacoccus Until the late 1980s, P. madeirensis was frequently mistaken for P. gossypii since taxonomic descriptions of Phenacoccus species previously relied on Myers (1928), Ferris (1950) and McKenzie (1967) (Chong 2005) The revised taxonomic description of P. madeirensis and P. gossypii by Williams (1987) ultimately clarified the distribution of both species within the United States. As a result, P. madeirensis became a commonly recognized and encountered pest in the sou thern United States, whereas P. gossypii became a rare and localized pest within Mexico, California, Texas and Florida. At the field identification level, P. madeirensis cannot be differentiated from the native Mexican mealybug, Phenacoccus gossypii Towns end and Cockerell, and the franseria mealybug, Phenacoccus franseriae Ferris. Both mealybugs are commonly intercepted at U.S. ports of entry from Mexico (Williams 1987, Williams and Watson 1988). Several identification characteristics viewed by slide mount ing are used for final identification On the thorax, P. madeirensis lacks multilocular disk pores from the mediolateral areas while P. gossypii has dorsal mediolateral multilocular disk pores. Phenacoccus franseriae only has dorsomedial cerarrii on abdomi nal segments VI and VII. Phenacoccus madeirensis also can be distinguished by its lanceolate, tiny cerarian setae and dorsal setae (Williams 1987, Williams & Watson 1988).

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18 Distribution Although native to the New World, the first record of P. madeirensis wa s from the coast of Africa in Madeira Island (Green 1923). Williams and Granara de Willink (1992) inferred that P. madeirensis was introduced to Africa through the sale and distribution of infested host plants on Atlantic triangular slave trade. Crawler in festation on host plants has been difficult to detect since crawlers are small, pale, and evade detection by hiding on the undersides of leaves or burrowing within plant crevices or nodes. Additionally, crawlers do not readily attract ants or induce the gr owth of black sooty mold since they exude minimal honeydew. During the late 1980s to early 1990s, the distribution of P. madeirensis included temperate to tropical regions within North America, South America, the Caribbean and Africa. However, P. madeiren sis spread to the Mediterranean, Southeast Asia, Oceania, and Pacific Islands as the international trade and transportation of infested host plants expanded from more tropical to temperate regions around the world (Table 1 3). Phenacoccus madeirensis event ually became recognized as a cosmopolitan pest. The most current publications cite the spread of P. madeirensis in Europe. Muniappan (2011) and Papadopoulou and Chryssohoides (2012) reported n ew P. madeirensis populations from Thailand and Greece Chemic al Control Methods Florida greenhouse and nursery growers undergo immense pressure to produce high quality nursery products in a warm, humid climate under high invasive pest pressure due to international trade and transportation. As a result, the most prev alent control method is the use of synthetic pesticides (Hodges 2011). Nearly 98% of Florida growers use at least one pesticide (Hodges et al. 1998) (Table 1 5). The Florida statewide average number of pesticides used per nursery was 5.8 with no indication of using different modes of action. South Florida scored higher than the statewide average with regard to treating bedding plants, floriculture crops, and

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19 all crops for larger firms. The high pesticide use may be influenced by the zero tolerance of any me alybug species on shipped nursery stock to California and other southern states (Dr. Amy Roda, personal communication). Mealybug biology and behavior impact the efficacy type, and application of current synthetic insecticides. Crawlers generally lack a wa xy or filamentous covering which makes them the most susceptible life stage to contact based synthetic insecticides (Buss and Turner 1993). However, crawlers can hide in crevices which may be an issue with low residual pesticides and foliage spray applica tions (Dr. Amy Roda, personal communication). Nymphs gradually increase their protective covering through successive instars and become subsequently harder to control with contact based synthetic insecticides as their protective covering grows thicker. Adu lts are naturally protected since they are well covered with smooth or filamentous wax. Ovipositing mealybug species such as P. madeirensis protect their eggs in a waxy, filamentous ovisac which prevents direct contact with synthetic insecticides. Overall, each life stage moves to, feeds on, or hides in parts of the plant that may be difficult for contact based synthetic pesticides to reach such as the undersides of leaves, notches created by plant nodes or buds on the stems, or crevices on the leaves, frui t, or stems (Hollingsworth and Hamnett 2009). Insecticidal dip treatments are considered a viable option for treating infested foliage cuttings. A foliage cutti ng is an excised stem and leaf that can be transplanted into a suitable growing medium for prod ucing new roots, stems, and leaves independent from the parent plant. Conover and Poole (1970) and Osborne (1986) described why foliage cuttings cannot be reliably treated with current synthetic insecticide application practices: (1) r oot application of sy stemic insecticides is relatively ineffect ive since cuttings are rootless; (2) w hen cuttings are placed under mist for root stimulation and moisture low volume spray application does not ensure

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20 adequate coverage ; and (3) high volume spray application can uproot the cuttings from the pot (Osborne 1986). A l imited number of studies ev a luated dip ping as an effective control for pests on ornamental cuttings Only three studies directly assess ed synthetic dip treatment efficacy on mealybugs and scales infesting propagation commodities (Osborne 1986, Hata et al. 1992 and Hansen et al. 1992). Synthetic dip treatments include the use of insecticides in the following classes: organophosphates, carbamates, and pyrethroids. The most commonly tested synthetic insectici de dip treatment for ornamental cuttings or flowers infested with mealybugs or scales is a type of pyrethroid called fluvalinate. Osborne (1986) was the first to use fluvalinate as a dip treatment for ornamental pests. The solanum mealybug Phenacoccus sol ani was one of several pest species tested on Hoya carnosa (L. f.) and Gynura procumbens (Lour.) cuttings. The first dip treatment test for P. solani had two replicates. Approximately 22 cuttings of Hoya carnosa infested with a density of 19 P. solani per cutting were assigned at random to either a control (water dip for 1 minute) or one minute dip in the recommended rate of fluvalinate (1.68g per 100 liters of water). Mortality was assessed 3, 7, and 14 days after treatment. Results from the first dip tre atment test showed at least 70% P. solani mortality 14 days after treatment. The second dip treatment test for P. solani also had two replicates. Ten cuttings of Gynura procumbens infested with a density of 30 P. solani per cutting were randomly assigned to four different treatments: 1) control (twelve minute dip in water and placed in water filled vials to root), 2) one minute dip in the recommended rate of fluvalinate and placed in water filled vials, 3) two minute dip in the recommended rate of fluvalin ate and placed in water filled vials, and 4) one minute dip in the recommended rate of fluvalinate, potted in soil, and placed under

PAGE 21

21 mist that actuated every 30 minutes for 15 seconds from 0800 to 2000 hours. Mortality was assessed 3, 7, and 14 days after treatment. Fluvalinate treated cuttings had significantly fewer mealybugs, averaging 8.3 P. solani in comparison to 58.8 on the control cuttings. N o significant difference was found between the different dipping times fo r fluvalinate treated cuttings and n o phytotoxicity was reported with fluvalinate. However, fluvalinate treated cuttings showed longer root development than the water control Hata et al. (1986) went a step further and investigated the potential synergistic effect of fluvalinate with insecti cidal soap (0.1g per liter of water + 9.6mLper liter of water by volume) as a postharvest dip treatment for harvest sprayed and non harvest sprayed red ginger flowers. Harvest sprayed red ginger flowers were treated with the recommended rate for chlorpyrif os and a spreader thicker, Triton B 1956, mixed together and applied until runoff. At foliage harvest, the plants were sprayed at 347 liters per hectare at two week intervals totaling six applications. All red ginger flowers in the Hata et al. (1986) stu dy were infested with several different ornamental mealybug pests, including the citrus mealybug, longtailed mealybug, and the obscure mealybug, Pseudococcus affinis (Maskell). All sprayed and non sprayed flowers harvested from the field were divided into two groups. Half were treated with water (control) and half were dipped in the postharvest dip treatment. Mortality was assessed 24 and 48 hours post treatment for flowers that were harvest sprayed and dipped and flowers that were only dipped. Results show ed 0% mean percentage of flowers infested with mealybugs for sprayed and dipped flowers while only dipped flowers showed 3% to 17% mealybugs present. Hansen et al. (1992) tested and compared the synergistic effect of fluvalinate and cyfluthrin mixed with i nsecticidal soap. The maximum label concentrations for fluvalinate (0.5mL per 100 liters of water), cyfluthrin (0.3mL per 100 liters of water), and insecticidal soap

PAGE 22

22 (20mL solution) were used both separately and in combination with soap as postharvest dip treatments for controlling the coconut mealybug, Nipaecoccus nipae (Maskell). Two cuttings of red ginger flowers were infested with either nymphs or adults and randomly assigned to a dip treatment. After counting the number of mealybugs present on each cu tting, the infested two flowers were submerged in each dip treatment for 0, 1, 5, and 15 minutes. Adult and nymph mortality were assessed a day after treatment and varied greatly. Flowers dipped for 0 minutes in all treatments showed less than 14% adult m ortality and 20% nymph mortality. Flowers treated with fluvalinate for 1 minute showed the highest nymph mortality (63.4%) compared to all other test times for fluvalinate. For the other treatments, highest nymph mortality was more than 97% as a 15 minute dip. Highest adult mortality for fluvalinate (94.2%) and fluvalinate with insecticidal soap (94.5%) occurred during the 1 minute dip. Cyfluthrin with insecticidal soap achieved the overall highest adult mortality as a 15 minute dip (98.4%) compared to all other treatments at the same dip time. Overall, Osborne (1986), Hata et al. (1992) and Hansen et al. (1992) indicated fluvalinate as an effective mealybug dip treatment. However, the current use of fluvalinate is limited due to environmental and worker he alth issues. The United States Environmental Protection Agency classifies fluvalinate as a moderately toxic compound in EPA toxicity class II (Extension Toxicology Network 1996). Fluvalinate is a broad spectrum insecticide known to be highly toxic to fish, aquatic invertebrates, and non target insects, such as beneficial arthropods used in IPM programs (United States Environmental Protection Agency 1986). Additionally, workers have reported coughing, sneezing, throat irritation, itching, or burning sensatio ns on the arms or face with or without a rash, headache or nausea (United States Environmental Protection Agency 1986). As a result, fluvalinate has been and continues to be a Restricted Use Pesticide that only

PAGE 23

23 certified applicators are legally allowed to purchase and apply. The alternative synthetic insecticide approved for use as an ornamental dip for cuttings is tau fluvalinate, also known as (2R) fluvalinate. Tau fluvalinate is also a broad spectrum insecticide with r estricted use (United States Environ mental Protection Agency 2005). Biorational Control Methods Biorational insecticides, such as oils, surfactants, and anti transpirants, may be viable dip treatment alternatives for controlling mealybugs and scales on ornamental cuttings. Stansly et al. (19 96) defined biorational insecticides as any type of insecticide active against target pest populations, relatively innocuous to non target organisms, and therefore non disruptive to biological control methods A wide variety of insecticides with differen t modes of action can fit this definition and therefore are considered biorational insecticides. Schuster and Stansly (2003) listed the following as biorational insecticides: chemical controls, such as oils, surfactants, neem, Bacillus thuringiensis produc ts, and new chemical classes, such as insect growth regulators (pyriproxyfen, bupro fezin, tebufenozide, novaluron) and miscellaneous insecticides (pymetrozine, spinosad, indoxacarb, emamectin, benzoate, rynaxypyr, metaflumizone, spinetoram, flubendiamide, pyridalyl). In particular, oils and surfactants are known to be efficacious against ornamental and agricultural pests. Anti transpirants currently are being explored for potential insecticidal activity. Oils Oils were one of the first types of chemicals us ed to control agricultural pests (Liu and Stansly 2000). For over 100 years, resistance to oils has never been recorded and this may be due in part to several different modes of action. Capinera (2008) lists the following modes of action for oils: mortalit y by starvation when oils prevent plant pests from using their piercing sucking mouthparts, death by suffocation or desiccation when oils halt respiration via spiracle blockage,

PAGE 24

24 or toxicity from muscle or nerve damage caused by oils penetrating and degradi ng the tracheae. Overall, oils have very short residual activity which is ideal when used in conjunction with biological controls. The most important types of oil used for insecticides today include narrow range horticultural oil, essential oil, and vegeta ble oil. Both narrow range horticultural oil and essential oils are efficacious, cost effective controls for a variety of plant pests but easily can cause phytotoxicity on agricultural and ornamental products, thus reducing their marketability. Narrow rang e horticultural oils are made of highly refined petroleum oil, while essential oils are composed of hydrophobic liquids containing volatile aromatic compounds from plants that are typically extracted by distillation (Capinera 2008, Hollingsworth and Hamnet 2009). Use of narrow range horticultural oil requires vigilant care and a delicate balance between application rate and plant management, since phytotoxicity has been reported on plants that were weakened or under moisture stress (Liu and Stansly 2000, Mi ller 1989, Capinera 2008). Phytotoxicity damage also has been documented on herbaceous and foliage plant material sprayed with limonene or other essential oils known to be effective against mealybugs and scales (Isman 1999, Isman 2000, Ibrahim et al 2001, Hollingsworth 2005, Cloyd et al. 2009). In comparison to narrow range horticultural oils and essential oils, vegetable oils consist of extracted triglycerides from plants (Sharma and Mudhoo 2011). Vegetable oils, such as soybean oil and cotton seed oil, di ffer from narrow range horticultural oil and essential oils since they tend to show only slight phytotoxicity. Soybean oil and cotton seed oil inflict minimal phytotoxicity on collards and tomatoes infested with whiteflies, and are known to perform as well if not better than horticultural oils (Liu and Stansly 2000). Several studies reported only slight phytotoxicity caused by cotton seed oil or soybean oil on agricultural or ornamental plants

PAGE 25

25 (Rock and Crabtree 1987, Lancaster et al. 1999). Additionally, P less et al. (1995) recommended soybean oil as an effective dip and spray treatment for controlling plant feeding pests like San Jose scale, terrapin scale, and European red mite. Overall, vegetable oils tend to provide good control on agricultural pests wi th minimal phytotoxicity, thus showing further promise as dip treatments for ornamental cuttings infested with plant feeding pests like P. madeirensis Soaps and Surfactants Surfactants and soaps are a type of chemical adjuvant meant to reduce the surface tension of water, thereby enhancing the biological activity of pesticides by modifying pesticide spray droplet size, retention, and spreading on leaf surfaces (Katagi 2008). Various types of surfactants are derived from plants and petroleum oils which can be further modified into different molecular weights or ionic character. The chemical structures of typical surfactants used during a pesticide application include anionic, cationic, and non ionic surfactants. Non ionic surfactants with a polyethoxy chain as a hydrophilic part are the most popular for pesticide use to date. Examples of non ionic surfactants also commonly found in household cleaning products include: alcohol ethoxylate, octylphenoxy ethoxylate, alkylphenoxy ethoxylate, sorbitan alkylate, an d novel silicone derivatives (Katagi 2008). Historically, surfactants have been used as wetting, spreading, emulsifying, or sticking agents to improve the effectiveness and coverage of many pesticides (Liu and Stansly 2000). As such, further research is n eeded to determine the surfactant mode of action on insects. Only Coret and Chamel (1995) reported the non ionic surfactant mode of action on plant cuticles. Nevertheless, non ionic surfactant insecticidal activity has been observed. Several decades of re search report insecticidal effects of surfactants on ornamental and agricultural pests (Liu and Stansly 2000, Imail et al. 1994, Davidson et al. 1991, Hesler and Plapp 1986,

PAGE 26

26 Tattersfield and Gimingham 1927, Wolfenbarger et al. 1967, Imail and Tsuchiya 1995 Cory and Langford 1935). Select growers near Apopka, FL have recently used dish detergents as cheap cost effective insecticides for controlling ornamental pests (Dr. Lance Osborne, personal communication). Antitranspirants Antitranspirants are chemicals which create a physical or physiological barrier capable of reducing transpiration and therefore water loss when applied to plant foliage (Davenport et al. 1969). Chemicals used to create a physical barrier include spray emulsions of latex, wax, or acrylic which form a film over the leaf surface and reduce water loss (Davenport et al. 1969). Physiological barriers involve chemicals that act as plant growth regulators by closing the stomata and inhibiting plant growth (Davenport et al. 1969). Plant growth in hibition has been extensively reported after antitranspirant application for preventing water loss (Gale and Hagan 1966, Rowan 1988, Davenport et al. 1969, McConnel 1985, Mmbaga and Sheng 2002). Insecticidal effects of antitranspirants also have been obser ved, although the mode of action for insecticidal antitranspirants has not been described in the scientific literature (French 1988). Additional efficacy studies on ornamental pests are needed to evaluate antitranspirants as viable biorational insecticides From a nursery manager standpoint, there are several advantages to using biorational insecticides, such as soaps, surfactants, oils, and anti transpi rants : (1) safety to human and environmental health, (2) apparent lack of resistance mechanisms among in sects and mites, (3) reliable efficacy on other important plant feeding ornamental pests such as the silverleaf whitefly, Bemisia argentifolii Bellows and Perring (4) relatively low cost with no applicator certification required (5) compatibility with usin g biological controls within an integrated pest management program, and (6) inexpensive to purchase (Butler et al. 1993, Stansly et al. 2006,

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27 Chapman 1967, Davidson et al. 1991, Liu and Stansly 1995, Liu and Stansly 2000, Agnello et al. 1994, Pless et al. 1995). Research Objectives Propagative horticultural products such as cuttings are highly profitable for the Florida ornamental industry. Ornamental cuttings are constantly being imported and exported among the southeastern states and throughout the world The likelihood of invasive pests evading detection on plant cargo or being resistant to postharvest pesticides is high but oftentimes mitigated since many countries and states have a zero tolerance for mealybugs and will refuse entry of shipments and com modities if a single pest is found (Dr. Amy Roda, personal communication). As a result, nursery growers need a safe, effective, easy to implement, and low cost application method and treatment to prevent invasive species from entering their greenhouses and nurseries. Using insecticides as a dip treatment is an effective application method in comparison to spraying and systemic insecticide use for cuttings. However, further research is needed on utilizing dips as a treatment method for infested ornamental c uttings. From the late 1980s to mid 1990s, fluvalinate was the most commonly tested dip treatment until the Environmental Protection Agency labeled fluvalinate as a restricted use pesticide. Subsequently biorational d ip treatments such as oils, surfactants and anti transpirants, show promising potential as a cost effective, safe, and efficacious treatment for ornamental pests infesting imported cuttings. Overall, infested host plant sale and distribution between states or across continents on airplanes, s hips, and trucks successfully contribute to the disp ersal and establishment of invasive species outside of greenhouse conditions. Further research on this invasive pathway requires the use of a model ornamental invasive pest to assess overall efficacy of b iorationals as viable dip treatment for controlling infested cuttings. In this study, the Madeira mealybug, Phenacoccus madeirensis Gre en (Hemiptera: Pseudococcidae), was used as the model ornamental plant pest as

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28 it is already established in Florida but h as traveled worldwide as an invasive cryptic species Additionally, P. madeirensis is an extensively studied invasive ornamental pest that is difficult to treat with synthetic insecticides and can easily evade detection during plant inspection. For the pur poses of this study, P. madeirensis was used to accomplish the following research objectives: 1. Develop a model biorational dip treatment by assessing phytotoxicity and subsequent biorational dip treatment efficacy on coleus cuttings infested with P. madeire nsis 2. Evaluate the model biorational dip treatment at diff erent exposure times, rates, and on three ornamental cuttings infested with P. madeirensis.

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29 Table 1 1. Invasive ornamental pests encountered in Florida (Frank and Thomas 2004). Pest Group Scien tific Name Common Name Origin Aphids Aphis gossypii Glover Melon aphid Eurasia Myzus persicae (Sulzer) Green peach aphid Asia Hard and Soft Scales Aulacaspis yasumatsui Takagi Cycad aulacaspis scale Asia Bemisia tabaci Gennadius Sweetpotato whitefly Asia Coccus hesperidum L. Brown soft scale Asia Pseudaulacaspis cockerelli (Cooley) False oleander scale Asia Mealybugs Maconellicoccus hirsutus (Green) Pink hibiscus mealybug Asia Planococcus citri (Risso) Citrus mealybug Asia Mites Tetranychus u rticae Koch Two spotted spider mite Europe Thrips Frankliniella occidentalis (Pergande) Western flower thrips Western United States Scirtothrips dorsalis (Hood) Chilli thrips Asia Thrips palmi Karny Melon thrips Asia Whiteflies Aleurodicus dugesii Co ckerell Giant whitefly Mexico Bemisia argentifolii Bellow & Perring Silverleaf whitefly Middle East Dialeurodes citri (Ashmead) Citrus whitefly Asia Singhiella simplex (Singh) Fig or ficus whitefly Asia

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30 Table 1 2. Reported h ost plants of Phenacocc us madeirensi s Family Scientific Name Common Name Reference Acanthaceae Acanthus mollis (Oyster Plant) Ben Dov 2004; Sinacori 1995 Aphelandra Aphelandra Stocks 2012 Crossandra infundibuliformis Crossandra (Firecraker Flower, Kanakambara ) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Dicliptera sexangularis Sixangle Foldwing Stocks 2012 Eranthemum pulchellum Blue Sage Stocks 2012 Hemigraphis alternata Redivy (Red Ivy, Red Flame Ivy) Stocks 2012 Hemigraphis repanda Dragon flame Ben Dov 1994; Ben Dov 2004; Williams 1987 Hypoestes phyllostachya Polka Dot Plant Stocks 2012 Justicia pectoralis Freshcut (Chapantye, Zeb Chapantye, Carpintero, Te Criollo, Curia, Death Angel, Masha Hari) Stocks 2012 Justicia spicigera Mexican Honeysuckle Stocks 2012 Odontonema strictum Firespike (Cardinal Guard, Scarlet Flame) Stocks 2012 Pachystachys coccinea (=Jacobinia coccinea) Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Pachystach ys lutea Lollipop Plant (Golden Shrimp Plant) Stocks 2012 Peristrophe hyssopifolia Marble Leaf Peristrophe Stocks 2012 Pseuderanthemum fasciculatum Falseface Stocks 2012

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31 Table 1 2. Continued Family Scientific Name Common Name Reference Acanthaceae Pseuderanthemum laxiflorum Shooting Star (Star Flower, Purple Flase Erantheum, Dazzler) Stocks 2012 Ruellia brittoniana (Mexican Petunia, Common Ruellia, Wild Petunia) Stocks 2012 Ruellia elegans St ocks 2012 Thunbergia battiscombei Scrambling Sky Flower Stocks 2012 Thunbergia erecta Bush Clockvine Stocks 2012 Thunbergia grandiflora Bengal Trumpet (Bengal Clock Vine, Clock Vine, Sky Flower) Stocks 2012 Verbesina virginica F rostweed (White crownbeard, Iceplant, Iceweed, Virginia Crownbeard, Indian Tobacco, Richweed, Squawweed) Ben Dov 1994; Ben Dov 2004; Williams 1987 Amaranthaceae Amaranthus Amaranth Ben Dov 1994 ; Ben Dov 2004; Williams 1987 Iresine Bloodleaf Ben Dov 1 994; Ben Dov 2004; Williams 1987 Amaryliidoideae Narcissus Daffodil Ben Dov 1994; Ben Dov 2004; Williams 1987 Anacardiaceae Mangifera indica Mango Ben Dov 1994; Ben Dov 2004; Kondo et al. 2001; Williams 1987 Annonaceae Annona muricata Soursop Ben Dov 2004; Kondo et al. 2001 Annona Montana Kondo et al. 2001 Apiaceae Eryngium foetidum Spiritweed Stocks 2012 Petroselinum hortense Parsley Ben Dov 2004; Mazzeo et al. 1994 Apocynaceae Allamanda cathartica Golden Trumpet Stocks 2012 Mandevilla amabilis Thai Rose Stocks 2012

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32 Table 1 2. Continued Family Scientific Name Common Name Reference Apocynaceae Mandevilla laxa Chilean Jasmine Ben Dov 1994; Ben Dov 2004; Williams 1987 Mandevilla splendens (=Dipladenia splendens) lamanda Ben Dov 1994 ; Ben Dov 2004; Stocks 2012; Williams 1987 Plumeria Plumeria (Frangipani) Stocks 2012 Trachelospermum difforme Climbing Dogbane Stocks 2012 Trachelospermum jasminoides Confederate Jasmine Stoc ks 2012 Aquifoliaceae Ilex vomitoria Yaupon (Yaupon Holly, Cassina) Stocks 2012 Araceae Dieffenbachia maculata Dumbcane Ben Dov 1994; Ben Dov 2004; Williams 1987 Schismatoglottis calyptrata (=Schismatoglottis neoguineensis) Stocks 2012 Araliaceae Aralia Spikenard Ben Dov 1994; Ben Dov 2004; Fatsia japonica Paperplant (Fatsi, Japanese Aralia) Stocks 2012 Hedera helix English Ivy (Common Ivy) Stocks 2012 Polyscias Ben Dov 1994; Ben Dov 2004; Williams 1987 Schefflera actinophylla Octop us Tree (Umbrella Tree, Amate) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987

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33 Table 1 2. Continued Family Scientific Name Common Name Reference Araliaceae Schefflera arboricola Dwarf Umbrella Tree Ben Dov 1994; Ben Dov 2004; Williams 1987 A recaceae Dypsis lutescens Yellow Butterfly Palm (Golden Cane Palm, Areca Palm) Stocks 2012 Wodyetia Wodyetia Stocks 2012 Asclepiadaceae Hoya carnosa Porcelain Flower (Wax Plant) Ben Dov 2004; Mazzeo et al. 1994; Mazzeo et al. 2008; Stocks 2012 Hoy a purpurea fusca Silver Pink Wax Plant Stocks 2012 Asparagaceae Agave Agave (Century plant) Ben Dov 2004; Mazzeo et al. 1994; Mazzeo et al. 2008 Liriope muscari Big Blue Lilyturf (Lilyturf, Border Grass, Monkey Grass) Stocks 2012 Asteraceae Ageratin a adenophora (=Eupatorium adenophorum) Eupatory (Sticky snakeroot, Crofton Weed, Mexican Devil) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams and Granara de Willink 1992 Ageratum houstonianum Bluemink (Flossflower, Garden Ageratum, Blueweed, Pussy F oot) Ben Dov 2004; Granara de Willink 2003 Ambrosia Ragweeds (Bitterweeds, Bloodweeds) Ben Dov 1994; Ben Dov 2004; Stocks 2012 Artemisia californica California sagebush Ben Dov 1994; Ben Dov 2004; Williams 1987 Aster Aster Ben Dov 1994; Ben Dov 20 04 Bidens alba Romerillo Stocks 2012 Bidens pilosa Spanish Needles Kondo et al. 2001; Stocks 2012 Borrichia Seaside Tansies Stocks 2012 Calendula Pot Marigold Ben Dov 1994; Ben Dov 2004; Williams 1987

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34 Table 1 2. Continued Family Scientific Name Common Name Reference Asteraceae Chrysanthemum frutescens Marguerite Stocks 2012 Chrysanthemum morifolium Stocks 2012 Chrysopsis Golden Aster Ben Dov 1994; Ben Dov 2004; Williams 1987 Chromolaena odorata Christmas Bush (Com mon Floss Flower, Siam Weed) Stocks 2012 Cineraria Cineraria Ben Dov 1994; Ben Dov 2004; Williams 1987 Cynara scolymus Globe Artichoke (Alcachofra, Alcachofera, Artichaut, Tyosen Azami) Stocks 2012 Dahlia Dahlia Stocks 2012 Eclipta prostrata Fa lse Daisy Stocks 2012 Emilia fosbergii Florida Tasselflower Stocks 2012 Erigeron philadelphicus Philadelphia Daisy Plantain) Ben Dov 1994; Ben Dov 2004; Williams 1987 Euryops chrysanthemoides (=Gamolepis chrysanthemoides) (African Bush Daisy) Stocks 2012 Eupatorium capillifolium Dogfennel Ben Dov 1994; Ben Dov 2004; Williams 1987 Eupatorium odoratum Jack in the Bush Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granar a de Willink 1992 Eupatorium serotinum Lateflowering Thoroughwort (Late Boneset) Ben Dov 1994; Ben Dov 2004; Williams 1987 Gaillaria pulchella Firewheel (Indian Blanket, Indian Blanket Flower, Sundance) Stocks 2012

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35 Table 1 2. Continued Family Sc ientific Name Common Name Reference Asteraceae Gazania African daisy Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Gerbera jamesonii Barberton Daisy (African Daisy) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Gynura aurantiaca V elvetplant (Purple Passion Plant, Purple Passion Vine) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Haplopappus Haplopappus Ben Dov 1994; Ben Dov 2004; Williams 1987 Helianthus annuus Sunflower Ben Dov 1994; Ben Dov 2004; Williams 1987 H elianthus tephrodes Algodones Sunflower Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Helichrysum Helichrysum Ben Dov 2004; Stocks 2012; Williams 2004 Leucanthemum vulgare (= Chrysanthemum leucanthemum) Oxeye Daisy (Common Daisy, Common Dais y, Dog Daisy, Margarite, Moon Daisy, and Ox eye Daisy) Ben Dov 2004; Granara de Willink 2003 Mikania micrantha Climbing Hempweed (Mile a minute, Chinese creeper, Bittervine) Ben Dov 1994; Ben Dov 2004; Williams 1987 Mikania scandens Climbing Hempvine (Climbing Hempweed, Climbing Boneset, Guaco) Stocks 2012 Osteospermum South African Daisy (African Daisy, Cape Daisy, Blue Eyed Daisy) Stocks 2012 Parthenium hysterophorus Santa Maria Feverfew (Whitetop Weed) Ben Dov 1994; Ben Dov 2004 Pluchea ca mphorata Camphor Pluchea Stocks 2012 Pluchea odorata Sweetscent (Salt Marsh Fleabane, Shrubby Camphorweed) Ben Dov 1994; Ben Dov 2004

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36 Table 1 2. Continued Family Scientific Name Common Name Reference Asteraceae Rudbeckia fulgida Orange Coneflower Stocks 2012 Rudbeckia hirta Black Eyed Susan (Brown Eyed Susan, Brown Betty, Brown Daisy, Gloriosa Daisy, Golden Jerusalem, Poorland Daisy, Yellow Daisy, Yellow Ox Eye Daisy) Stocks 2012 Senecio hybridus Cineraria Stocks 2012 Solidago Goldenrod s Ben Dov 1994; Ben Dov 2004; Williams 1987 Sphagneticola trilobata Bay Biscayne Creeping Oxeye Stocks 2012 Stevia rebaudiana Candyleaf (Sweetleaf, Sugarleaf) Stocks 2012 Stokesia Cornflower Aster Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Tagetes Marigold Stocks 2012 Tanacetum parthenium Feverfew Stocks 2012 Taraxacum officinale Dandelion (Common Dandelion) Ben Dov 2004; Mazzeo et al. 1994 Tithonia diversifolia Tree Marigold (Mexican Tournesol, Mexican Sunfl ower, Nitobe Chrysanthemum) Stocks 2012 Vernonia Vernonia Stocks 2012 Wedelia trilobata Bay Biscane Creeping oxeye Ben Dov 1994; Ben Dov 2004; Williams 1987 Blechnaceae Blechnum Blechnum (Hard fern) Stocks 2012 Begoniaceae Begonia Begonia Ben Dov 1994; Ben Dov 2004; Mazzeo et al. 1994; Williams 1987

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37 Table 1 2. Continued Family Scientific Name Common Name Reference Bignoniaceae Jacaranda Jacaranda Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Borginaceae Cordia curassavi ca Black Sage (Wild Sage) Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Symphytum officinale Boneset (Common Comfrey, Quaker Comfrey, Cultivated Comfrey, Knitbone, Consound, Slippery root) Ben Dov 1994; Ben Dov 2004; Wi lliams 1987 Brassicaeae Brassica oleraceae Kondo et al. 2001 Brassica campestris Kondo et al. 2001 Bromeliaceae Ananas comosus Pineapple Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Tillandsia Tillandsia Ben Dov 2004 Burs eraceae Bursera simaruba Gumbo Limbo Stocks 2012 Cactaceae Hatiora salicornioides Dancing Bones Stocks 2012 Opuntia Nopales (Paddle Cactus) Stocks 2012 Cactaceae Hylocereus undatus Dragonfruit (Pitaya, Red P itaya) Ben Dov 2004; Mazzeo et al. 1994; Mazzeo et al. 2008 Campanulaceae Lobelia cardinalis Cardinalflower Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Caprifoliaceae Viburnum odoratissimum Sweet Viburnum Stocks 2012 Viburnum suspensum Vi burnum (Snadanqua Viburnum, Sandandwa Viburnum) Stocks 2012 Celtidaeae Trema micrantha Jamaican Nettletree (Florida Trema, Guacimilla) Stocks 2012

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38 Table 1 2. Continued Family Scientific Name Common Name Reference Commenlinaceae Commelina diffusa C limbing Dayflower (Spreading Dayflower) Stocks 2012 Compositae Ligularia tussilaginea Leopard Plant (Aureo Maculata) Ben Dov 1994; Ben Dov 2004; Williams 1987 Convovulaceae Ipomea setifera Morning Glory Ben Dov 2004; Matile Ferrero and Germain 2004 Jacquemontia blanchetti Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Crassulaceae Adromischus cristatus Crinkle leaf plant Ben Dov 1994; Ben Dov 2004; Williams 1987 Kalanchoe beharensis (Kalanchoe) Stocks 2012 Cucu rbitaceae Cucurbita peop Pumpkin (Field Pumpkin) Stocks 2012 Cupressaceae Juniperus chinensis Chinese juniper Ben Dov 1994; Ben Dov 2004; Williams 1987 Dioscoreaceae Tacca Bat Flower (Arrowroot) Ben Dov 1994; Ben Dov 2004; Williams 1987 Ebanaceae Di ospyros duclouxii Beltra and Soto 2011; Ben Dov 2004 Ericaceae Arbutus unedo Cane Apple (Strawberry Tree, Apple of Cain) Ben Dov 2004; Mazzeo et al. 1994 Euphorbiaceae Acalypha godseffiana Beefsteak Plant (Copperleaf, Fire Dragon, Jacobs Coat, Match Me If You Can, Three Seeded Mercury) Stocks 2012 Acalypha hispida Fox Tail Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Acalypha wilkesiana Fire Dragon) Ben Dov 1994; Ben Dov 2004; Kondo et al. 2001; Stocks 2012;

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39 Table 1 2. Continued Family Scientific Name Common Name Reference Euphorbiaceae Chamaesyce Sandmat Stocks 2012 Cnidoscolus Ben Dov 1994; Ben Dov 2004; Williams 1987 C odiaeum variegatum Croton (Garden Croton, Varigated Croton) Stocks 2012 Croton glandulosus Vente Conmigo Ben Dov 1994; Ben Dov 2004; Williams 1987 Croton punctatus Gulf Croton Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Euphor bia pulcherrima Poinsettia (Noche Buena) Ben Dov 1994; Ben Dov 2004; De Lotto 1977 Jatropha curcas Barbados Nut (Purging Nut, Physic Nut, JCL) Stocks 2012 Jatropha diversifolia (=Manihot diversifolia) Ben Dov 1994; Ben Dov 2004; Williams 1987 Ja tropha integerrima Peregrina (Spicy Jatropha) Stocks 2012 Manihot aesculifolia Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Manihot esculenta Cassava (Yuca, mogo, manioc, mandioca, kamoting kahoy) Ben Dov 1994; Ben Dov 2004; Ko ndo et al. 2001; Stocks 2012; Williams 1987; Williams and Granara de Willink 1992 Manihot glaziovii Ceara Rubbertree Ben Dov 1994; Ben Dov 2004; Couturier et al. 1985 Manihot michaelis Ben Dov 1994; Ben Dov 2004

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40 Table 1 2. Continued Family Sci entific Name Common Name Reference Euphorbiaceae Manihot rhomboidea Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Phyllanthus amarus Carry Me Seed (Sanskrit, Bahupatra) Stocks 2012 Phyllanthus debilis Kondo et al. 2001 Ricinus communis Castor oil plant Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Fabaceae Acacia flexuosa Acacia (Thorntrees, Whistling Thorns, Wattles, Yellow Fever Acacia, Umbrella Acacia) Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Arachis hypogaea Peanut (Groundnut) Stocks 2012 Cajanus cajan Pigeonpea (Tropical Garden Pea, Kadios, Congo Pea, Gungo Pea, Gunga Pea, No Eye Pea) Ben Dov 1994; Ben Dov 2004; Williams 198 7; Williams and Granara de Willink 1992 Calliandra haematocephala Calliandra Stocks 2012 Cassia imperialis Cassias (Cassia) Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Coronilla Coronilla Beltra and Soto 2011; Ben Dov 2004 Desmodium tortuosum Dixie Ticktrefoil (Florida Beggarweed) Ben Dov 1994; Ben Dov 2004; Williams 1987 Erythrina bogotensis Beltra and Soto 2011; Ben Dov 2004

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41 Table 1 2. Continued Family Scientific Name Common Name Reference Fabaceae Erythrina ca ffra Coast Coral Tree Ben Dov 1994; Ben Dov 2004; Williams 1987 Erythrina viarum Bucayo Ben Dov 2004; Sinacori 1995 Glycine max Soybean (Soya Bean) Ben Dov 2004; Kondo et al. 2001 Mimosa pudica Shameplant (Sensitive Plant, Touch me not) Ben Dov 19 94; Ben Dov 2004; Williams and Granara de Willlink 1992 Phaseolus aureus Kondo et al. 2001 Sophora secundiflora Mescal Bean (Texas Mountain Laurel) Ben Dov 1994; Ben Dov 2004; Williams 1987 Sophora tomentosa Yellow Necklacepod (Silver Bush, Neck lace Pod) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams and Granara de Willink 1992 Trifolium Clover (Trefoil) Stocks 2012 Vigna radiata Mung Bean (Mungbean, Mung, Mungo, Green Gram, Golden Gram) Ben Dov 2004; Kondo et al. 2001 Garryaceae Aucub a Aucuba Stocks 2012 Geraniaceae Geranium Cranesbills Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Pelargonium hortorum Zonal Geranium (Garden Geranium, Malva, Malvon) Ben Dov 2004; Granara de Willink 2003; Stocks 20 12 Pelargonium pelatum Ivyleaf Geranium (Ivyleaf Pelargonium, Cascading Gernamium, Kolsuring) Stocks 2012 Pelagaonium zonale Kondo et a. 2001

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42 Table 1 2. Continued Family Scientific Name Common Name Reference Gesneriaceae Chrysothemis pulchel la (=Tussacia pulchella) Squarestem (Sunset bells, Copper Leaf, Black Flamingo, Chrysothemis) Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Episcia decurrens Ben Dov 1994; Ben Dov 2004; Williams 1987 Nematanthus wettsteinii Goldf ish Plant (Candy Corn Plant) Ben Dov 1994; Ben Dov 2004; Williams 1987 Saintpaulia African Violet Stocks 2012 Hamamelidaceae Loropetalum chinense Chinese Fringe Flower (Chinese Witchhazel, Loropetalum) Stocks 2012 Lamiaceae Anisomeles indica (=Epime redi indicus) Indian catmint Ben Dov 1994; Ben Dov 2004; Kondo et al. 2001;Williams 1987; Williams and Granara de Willink 1992 Coleus Coleus Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987; Williams and Granara de Willink 1992 Melissa officin alis Common Balm (Lemon Balm) Stocks 2012 Mentha Mint Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Ocimum basilicum Basil (Sweet basil) Ben Dov 2004; Kondo et al. 2001; Mazzeo et al. 1994; Stocks 2012 Orthosiphon aristatus s Whiskers (Java Tea, Kumis Kucing, Misai Kucing) Stocks 2012 Plectranthus australis Little Spurflower (Swedish Ivy, Swedish Begonia, Creeping Charlie) Stocks 2012

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43 Table 1 2. Continued Family Scientific Name Common Name Reference Lamiaceae Plectr anthus scutellarioides Coleus Stocks 2012 Plectranthus nummularius Whorled Plectranthus Ben Dov 1994; Ben Dov 2004; Williams 1987 Salvia coccinea Blood Sage (Texas Sage, Scarlet Sage, Tropical Sage) Ben Dov 1994; Ben Dov 2004; Williams 1987 Salvi a guaranitica Anise Scented Sage (Hummingbird Sage) Stocks 2012 Salvia officinalis Kitchen Sage (Garden Sage, Common Sage) Stocks 2012 Salvia splendens Scarlet Sage (Tropical Sage) Kondo et al. 2001; Stocks 2012 Loasaceae Petalonyx thurberi Thurber Ben Dov 1994; Ben Dov 2004; Peterson 1965, Williams 1987 Lythraceae Cuphea llavea (=Cuphea speciosa) Cuphea Stocks 2012 Cuphea ignea Cigar Flower (Cigar Plant, Firecracker Plant, Mexican Cigar) Stocks 2012 Malvaceae Abutilon Abuti lon (Chinese Bell Flower, Chinese Lantern, Mallow, Indian Mallow, and Flowering Maple) Stocks 2012 Althaea Althaea (Mashmallow plant) Ben Dov 1994; Ben Dov 2004; Williams 1987 Gossypium Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987; Willia ms and Granara de Willink 1992 Hibiscus acetosella False Roselle Stocks 2012

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44 Table 1 2. Continued Family Scientific Name Common Name Reference Malvaceae Hibiscus cannabinus Kenaf Ben Dov 1994; Ben Dov 2004; Williams 1987 Hibiscus esculentus Ok ra Ben Dov 1994; Ben Dov 2004; Williams 1987 Hibiscus mutabilis Confederate Rose (Cotton Rosemallow) Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Hibiscus rosa sinensis Chinese Hibiscus (China Rose, Shoe Flower) Ben Dov 1994; Ben Dov 2004; Matile Ferrero and Germain 2004; Jansen et al. 2010; Stocks 2012; Hibiscus tiliaceus Coast Hibiscus (Sea Hibiscus, Beach Hibiscus, Coast Cottonwood, Green Cottonwood, Native Hibiscus, Native Rosella, Cottonwood Hibisicus, Kurraj ong, Sea Rosemallow, Norfolk Hiiscus, Hau, Purau) Stocks 2012 Jute Jute Ben Dov 2004; Kondo et al. 2001 Malva Malva Ben Dov 2004; Mazzeo et al. 1994; Stocks 2012 Malvaviscus arboreus Wax Mallow Purse) Ben Dov 1994; Ben Dov 2004; Kondo et al. 2001; Williams 1987 Sida Sida Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987; Williams and Granara de Willink 1992 Talipariti tiliaceum Sea Hibsicus (Beach Hibiscus, Cottontree, Mahoe) Stocks 2 012 Theobroma cacao Cacao Tree (Cocoa Tree) Ben Dov 2004; Donald 1956

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45 Table 1 2. Continued Family Scientific Name Common Name Reference Malvaceae Urena lobata Ceasarweed Ben Dov 1994; Ben Dov 2004; Williams 1987 Menispermaceae Clyclea insularis Kondo et al. 2001 Moraceae Artocarpus communis Breadfruit Ben Dov 1994; Ben Dov 2004; Williams 1987 Ficus benghalensis Banyan (Banian) Ben Dov 2004; Williams 2004 Ficus pumila Climbing fig (Creeping Fig) Stocks 2012 Myrtaceae Eugenia uniflora Brazilian Cherry (Surinam Cherry, Cayenne Cherry) Stocks 2012 Oleaceae Ligustrum japonicum Japanese Privet (Wax Leaf Privet) Ben Dov 1994; Ben Dov 2004; Williams 1987 Onagraceae Ludwigia octovalvis Mexican Primrose Willow (Narrow Leaf Water Primrose, S eedbox) Stocks 2012 Passifloraceae Passiflora edulis Passion Fruit (Purple Grandilla, Passionfruit, Maracuja) Ben Dov 2004; Kondo et al. 2001 Passiflora incarnata Purple Passionflower (Maypop, True Passionflower, Wild Apricot, Wild Passion Vine) Stock s 2012 Primulaceae Primula Primula Ben Dov 1994; Ben Dov 2004; Marotta and Tranfaglia 1990 Poaceae Aventa sativa Oats Ben Dov 2004 Polygonaceae Rumex Sorrel (Dock) Ben Dov 2004; Williams 2004 Portulacaceae Portulaca oleracea Little Hogweed (Verdola ga, Pigweed, Common Purslane, Pusley) Stocks 2012 Ranunculaceae Clematis tashiroi Kondo et al. 2001 Rubiaceae Gardenia jasminoides Cape Jasmine (Common Gardenia, Cape Jessamine) Ben Dov 2004; Granara de Willink 2003; Stocks 2012

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46 Table 1 2. Conti nued Family Scientific Name Common Name Reference Rubiaceae Ixora West Indian Jasmine (Rangan, Kheme, Ponna, Chann Tanea, Techi, pan, Santan, Jarum Jarum, Jungle Flame, Jungle Geranium) Stocks 2012 Pentas lanceolata Egyptian Starcluster Stocks 2012 Rutaceae Citrus Citrus Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams and Granara de Willink 1992 Citrus limonium Lemon Ben Dov 2004; Mazzeo et al. 1994; Stocks 2012 Ruta graveolens Common Rue (Herb of Grace) Stocks 2012 Zanthoxylum fagara Lim e Pricklyash (Wild Lime, Colima, Una de Gato, Corriosa) Stocks 2012 Rosaceae Malus domestica Apple Ben Dov 2004; Mazzeo et al. 1994 Malus sylvestris European Wild Apple Ben Dov 1994; Ben Dov 2004; Williams 1987 Rosa Rose Ben Dov 1994; Ben Dov 2004 ; Williams 1987 Rubus Rubus Stocks 2012 Sapindaceae Cupaniopsis anacardioides Carrotwood (Tuckeroo, Beach Tamarind, Green Leaved Tamarind) Ben Dov 2004; Stocks 2012 Dodonaea viscosa Florida hopbush Stocks 2012 Nephelium lappaceum Rambutan Ben Do v 2004; Williams 2004 Scrophulariaceae Capraria biflora Goatweed Stocks 2012 Leucophyllum frutescens Texas Barometer Bush (Texas Sage, Texas Ranger, Silverleaf, Cenizo) Stocks 2012 Smilacaeae Smilax Catbriers (Greenbriers, Prickly Ivys, Smilaxes) Be n Dov 1994; Ben Dov 2004; Williams 1987 Solanaceae Brunfelsia Brunfelsia Stocks 2012

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47 Table 1 2. Continued Family Scientific Name Common Name Reference Solanaceae Capsicum annuum Cayenne Pepper (Chili pepper) Ben Dov 1994; Ben Dov 2004; Kondo et al 2001; Mazzeo et al. 1994; Stocks 2012; Williams 1987; Williams and Granra de Willink 1992 Cestrum diurnum Day Blooming Cestrum (Day Blooming Jessamine) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Cestrum nocturnum Night Blooming Cestrum (Night Blooming Jessamine, Lady of the Night, Raat ki Rani) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987, Williams and Granara de Willink 1992 Datura metel Ben Dov 1994; Ben Dov 2004; Stocks 2012; Willi ams 1987 Solanaceae Physalis viscosa Starhair Groundcherry (Grape Groundcherry, Arrebenta Cavalo, Balaozinho, Camambu) Stocks 2012 Solanum diphyllum Twoleaf Nightshade Stocks 2012 Solanum integrifolium Kondo et al. 2001 Solanum lycopersicum (= Lycopersicon esculentum) Tomato (Garden Tomato) Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Solanum melongena Eggplant (Aubergine, Melongene, Brinjal, Guinea Squash) Ben Dov 1994; Ben Dov 2004; Mazzeo et al. 1994; Wil liams 1987, Williams 2004; Williams and Granara de Willink 1992 Solanum nigrum Kondo et al. 2001 Solanum pseudocapsicum Jerusalem Cherry (Madeira Winter Berry, Winter Cherry) Ben Dov 1994; Ben Dov 2004; Williams 1987

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48 Table 1 2. Continued Family Scientific Name Common Name Reference Solanaceae Solanum torvum Turkey Berry shoo Bush, Wild Eggplant, Pea Eggplant, Pea Aubergine) Stocks 2012 Solanum tuberosum Potato Ben Dov 1994; Ben Dov 2004; Williams 1987 ; Williams and Granara de Willink 1992 Solanum wendlandii Giant Potato Creeper (Costa Rica Nightshade) Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Solanum wrightii Giant Potato Tree (Brazilian Potato Tree) Stocks 2012 Tillaceae Corchorus olitorius Kondo et al. 2001 Triumfetta semitriloba Sacramento burbark Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams and Granara de Willink 1992 Urticaceae Boehmeria Smallspike False Nettle Stocks 2012 Parietaria floridana Florida p ellitory Ben Dov 1994; Ben Dov 2004; Williams 1987 Pilea Pilea Ben Dov 1994; Ben Dov 2004; Williams 1987 Pouzolzia zeylanica Graceful Pouzolzsbush Stocks 2012 Urera Urera Ben Dov 1994; Ben Dov 2004; Williams and Granara de Willink 1992 Urtica N ettle Ben Dov 1994; Ben Dov 2004; Williams 1987 Verbenaceae Callicarpa americana American Beautyberry Stocks 2012 Citharexylum spinosum Florida Fiddlewood (Spiny Fiddlewood) Ben Dov 1994; Ben Dov 2004; Stocks 2012

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49 Table 1 2. Continued Family Scie ntific Name Common Name Reference Verbenaceae Clerodendrum bungei Rose Glorybower (Bagflower, Bleeding Heart) Stocks 2012 Clerodendrum paniculatum Pagoda Flower (Bagflower, Bleeding Heart) Stocks 2012 Duranta erecta Golden Dewdrop (Pigeon Berry, Sk yflower, Xcambocoche) Stocks 2012 Lantana camara Lantana (Spanish Flag, West Indian Lantana, Red Wild Sage, Red Yellow Sage) Ben Dov 1994; Ben Dov 2004; Kondo et al. 2001; Stocks 2012; Williams 1987; Williams and Granara de Willink 1992 Lantana monte vidensis Trailing Shrubverbena (Weeping Lantana, Creeping Lantana, Small Lantana, Purple Lantana, Trailing Verbena) Ben Dov 1994; Ben Dov 2004; Williams 1987; Williams and Granara de Willink 1992 Stachytarpheta jamaicensis Light Blue Snakeweed (Blue Por terweed, Jamaica Vervain, Indian Snakeweed, Nettle Leaved Vervain) Stocks 2012 Verbenaceae Verbena canadensis Rose Mock Vervain (Purple Verbena) Stocks 2012 Verbena hybrida Verbenas (Vervains) Ben Dov 1994; Ben Dov 2004; Stocks 2012; Williams 1987 Vi taceae Vitis vinifera Common Grapevine (Wine Grape) Ben Dov 2004; Mazzeo et al. 1994 Zingiberaceae Curcuma longa Kondo et al. 2001 Zingiber mioga Japanese Ginger (Myoga Ginger) Ben Dov 2004; Kondo et al. 2001

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50 Table 1 3. Reported distribution of Phenacoccus madeirensis Region Country State Reference North America Mexico CABI 2000; Williams and Granara de Willink 1992 United States Alabama Ben Dov 1994; CABI 2000 California Ben Dov 1994; CABI 2000; Williams 1987 Florida Ben Dov 1994; CA BI 2000; Williams 1987 Georgia Chong 2005; Townsend et al. 2000 Illinois Ben Dov 1994; CABI 2000 Louisiana Ben Dov 1994; CABI 2000 Maryland Ben Dov 1994; CABI 2000 Minnesota Ben Dov 1994; CABI 2000 Mississippi Ben Dov 1994; CABI 2000 N ew York Ben Dov 1994; CABI 2000 North Carolina Ben Dov 1994; CABI 2000 Texas Ben Dov 1994; CABI 2000; Williams 1987 Virginia Ben Dov 1994; CABI 2000 Wisconsin Ben Dov 1994; CABI 2000 Caribbean Antigua and Barbuda Ben Dov 1994; CABI 2000 Ba hamas CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Barbados CABI 2000; Williams 1992 Bermuda CABI 2000; Williams 1987 British Virgin Islands CABI 2000; Williams and Granara de Willink 1992 Cayman Islands CABI 2000; Willi ams 1987; Williams and Granara de WIllink 1992 Costa Rica CABI 2000; Williams and Granara de Willink 1992 Cuba CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Dominican Republic CABI 2000; Williams and Granara de Willink 1992 G renada CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Guadeloupe CABI 2000; Williams and Granara de Willink 1992 Guatemala CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Haiti CABI 2000; Williams and Granara de Willink 1992 Jamaica CABI 2000; Williams and Granara de Willink 1992 Montserrat CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Panama CABI 2000; Williams and Granara de Willink 1992 Puerto Rico CABI 2000; Williams and Grana ra de Willink 1992 St. Kitts Nevis Ben Dov 1994; CABI 2000; Williams and Granara de Willink 1992 St. Lucia CABI 2000; Williams and Granara de Willink 1992

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51 Table 1 3. Continued Region Country State Reference Caribbean Trinidad and Tobago Ben Dov 1994; CABI 2000; Williams 1987; Williams and Granara de Willink 1992 South America Bolivia CABI 2000; Williams and Granara de Willink 1992 Brazil CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Colombia CABI 2000; William s and Granara de Willink 1992 Ecuador CABI 2000; Williams and Granara de Willink 1992 Guyana CABI 2000; Williams and Granara de Willink 1992 Paraguay CABI 2000; Williams and Granara de Willink 1992 Peru CABI 2000; Williams and Granara de Wi llink 1992 Venezuela CABI 2000; Williams and Granara de Willink 1992 Africa Angola Ben Dov 1994; CABI 2000 Benin CABI 2000 Cameroon Ben Dov 1994; CABI 2000; Williams 1987 Cape Verde Ben Dov 1994; Williams 1987; Williams 1987 Congo Domi nican Republic CABI 2000 Ivory Coast Ben Dov 1994; CABI 2000 Gabon CABI 2000 Gambia Ben Dov 1994; CAB 2000; Williams 1987 Ghana CABI 2000 Liberia Ben Dov 1994; CABI 2000 Mozambique Ben Dov 1994; CABI 2000; Williams 1987 Nigeria Ben Dov 1994; CABI 2000; Williams 1987 Sao Tome and Principe CABI 2000 Senegal Ben Dov 1994; CABI 2000 Sierra Leone Ben Dov 1994; CABI 2000; Williams 1987 Togo CABI 2000; Williams 1987 Zaire Williams 1987 Zimbabwe Ben Dov 1994; CA BI 2000; Williams 1987 Mediterranean Crete Ben Dov 2004; Jansen 2010 France Ben Dov 2004; Matile Ferrero and Germain 2004 Greece Papadopoulou and Chryssohoides 2012 Italy Ben Dov 1994; CABI 2000; Longo et al. 1995; Mazzeo et al. 1994; Sina cori et al. 1995; Sinacori and Tsolakis 1994 Pakistan Ben Dov 2004; Williams 2004 Portugal Ben Dov 1994; CABI 2000; Williams 1987; Williams and Granara de Willink 1992 Spain Ben Dov 2004; Beltra 2011 Southeast Asia, Oceania, and Pacific Island s Guam CABI 2000 Japan Kondo 2001 Miconesia CABI 2000

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52 Table 1 3. Continued Region Country State Reference Southeast Asia, Oceania, and Pacific Islands Taiwan Yeh et al. 2006 Thailand Muniappan et al. 2011 Philippines Williams 2004 Vietnam Williams 2004

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53 Table 1 4 N atural predators and parasitoids of Phenacoccus gossypii and Phenacoccus madeirensis (Chong 2005) Order Family Species Prey/Host Species References Diptera Syrphidae Toxomerus marginata Macquart P. gossypii Heming 1936 Coleoptera Coccinellidae Cryptolaemus montrouzieri Mulsant Many Chong 2005 Coccinellidae Diomus austrinus Gordon P. madeirensis Chong 2005 Neuroptera Chrysopidae Chrysopa oculata Say P. gossypii Heming 1936 Chrysopidae sp. P. gossypi i Aguilar and Lamas 1980 Chrysopidae Dichochrysa sp. P. madeirensis Sinacori and Tsolakis 1994; Miller et al. 2004 Hemerobiidae Sympherobius californicus Banks P. gossypii Aguilar and Lamas 1980 Hemerobiidae Sympherobius fallax Navs P. madeirensis S inacori and Tsolakis 1994; Miller et al. 2004 Hemerobiidae Sympherobius pygmaeus (Rambur) P. madeirensis Sinacori and Tsolakis 1994; Miller et al. 2004 Hymenoptera Aphelinidae Coccophagus gurneyi Compere P. gossypii Gordh 1979; Peck 1963; Thompson 1953 Encrytidae Acerophagus coccois Smith P. gossypii Ashmead 1900; Van Driesche et al. 1986, 1987; Noyes and Hayat 1994; Noyes 2003 Acerophagus coccois Smith P. madeirensis Castillo and Bellotti 1990; Noyes 2003; Lhr; Rosen 1969; Beardsley 1976; Van Dri esche et al. 1987 Acerophagus pallidus Timberlake P. gossypii Flanders 1935; Thompson 1953; Simmonds 1957; Peck 1963; Herting 1972; De Santis 1989; Noyes and Hayat 1994 Acerophagus pallidus Timberlake P. madeirensis Herting 1972; Noyes and Hayat 1994 ; Aenasius flandersi Kerrich (= Aenasius phenacocci Bennet) P. gossypii Herting 1972; De Santis 1979; Noyes and Hayat 1994; Noyes 2003 A. flandersi P. gossypii Bennett 1957 A. flandersi P. madeirensis Noyes 2000 Aenasius masii Domenichini P. g ossypii De Santis 1979; Noyes and Hayat 1994; Coquis and Salazar 1976 Anagyrus sp. P. gossypii Herting 1972; Noyes and Hayat 1994; Noyes 2003; Salazar 1972

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54 Table 1 4 Continued Order Family Species Prey/Host Species References Hymenoptera Encrytida e Anagyrus sp. P. madeirensis Lhr et al. 1990; Boussienguet and Neuenschwander 1989; Neuenschwander et al. 1987; Noyes and Hayat 1994 Anagyrus diversicornis (Howard) P. gossypii Kerrich 1982; Van Driesche et al. 1987; De Santis 1989 Anagyrus diversi cornis (Howard) P. madeirensis Noyes 2000 Anagyrus elgeri (Kerrich) P. madeirensis De Santis 1989; Kerrich 1982; Noyes and Hayat 1994 Anagyrus fusviventris Girault P. gossypii Viggiani and Battaglia 1983; Noyes and Hayat 1994; Noyes 2000 Anagyrus loecki Noyes & Menezes P. madeirensis Noyes 2000 Anagyrus pseudococci (Girault) P. gossypii De Santis 1979; Noyes and Hayat 1994 Anagyrus sinope Noyes & Monezes P. gossypii Noyes 2000 Anagyrus sinope Noyes & Monezes P. madeirensis Noyes 2000 B lepyrus insularis (Cameron) P. madeirensis Boussienguet and Neuenschwander 1989; Noyes and Hayat 1994; Noyes 2000 Cheiloneurus carinatus Compere P. madeirensis Herting 1972 Chrysoplatycerus ferrisi Timberlake P. gossypii Kerrich 1978; Noyes and Hayat 1994 Coccidoxenoides perminutus Girault P. madeirensis Herting 1972 Dicarnosis ripariensis Kerrich P. gossypii Kerrich 1978; Noyes and Hayat 1994; Noyes 2003 Ericydnus lamasi (Domenichini) P. gossypii Salazar 1972; De Santis 1979; De Santis 1983; Noyes and Hayat 1994; Noyes 2000 Gryranusoidea sp. P. madeirensis Boussienguet and Neuenschwander 1989; Noyes and Hayat 1994 Gryranusoidea phenacocci (Beardsley) P. gossypii Beardsley 1969; Noyes and Hayat 1994

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55 Table 1 4 Continued Order Family Sp ecies Prey/Host Species References Hymenoptera Encrytidae Holcencyrtus sp. P. gossypii Salazar 1972; Noyes and Haya 1994 Holcencyrtus myrmicoides (Compere & Zinna) P. madeirensis Herting 1972 Leptomastidea sp. P. gossypii Coquis and Salazar 1976; No yes and Hayat 1994 Leptomastidea abnormis (Girault) P. gossypii Dozier 1932; Heming 1936; Thompson 1954; Peck 1963; Gordh 1979; Trjapitzin 1989; Noyes and Hayat 1994; Noyes 2000 Leptomastix sp. P. madeirensis Boussienguet and Neuenschwander 1989; Noy es and Hayat 1994 Leptomastix dactylopii Howard P. gossypii Bess 1939; Fullaway 1946 Leptomastix dactylopii Howard P. madeirensis Peck 1963; Gordh 1979; Noyes and Hayat 1994; Noyes 2000 Metanotalia madeirensis (Walker) P. madeirensis Zuparko 1995 Prochiloneurus sp. P. gossypii Coquis and Salazar 1976; Noyes and Hayat 1994 Prochiloneurus bolivari Mercet P. madeirensis Boussienguet and Neuenschwander 1989; Noyes and Hayat 1994 Prochiloneurus insolitus (Alam) P. madeirensis Neuenschwander et al. 1987 Prochiloneurus seini (Dozier) P. gossypii Salazar 1972; Noyes and Hayat 1994 Pseuaphycus angelicus (Howard) P. gossypii Flanders 1935; Thompson 1954; Peck 1963; Herting 1972; Gordh 1979 Pseuaphycus angelicus (Howard) P. madeirensis Hertin g 1972 Pseudaphycus mundus Gahan P. gossypii Gahan 1946; Peck 1963; Herting 1972; Gordh 1979; Noyes and Hayat 1994; Noyes 2003 Zarhopalus zancles Noyes P. madeirensis Noyes 2000 Pteromalidae Pachyneuron eros Girault P. gossypii De Santis 1979 Sig niphoridae Chartocerus sp. P. madeirensis Noyes 2000 Signiphoridae Chartocerus dactylopii (Ashmead) P. gossypii Gordh 1979; Noyes 2003 Signiphoridae Chartocerus niger (Ashmead) P. gossypii Herting 1972

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56 Table 1 5. Reported insecticides used to contro l Phenacoccus madeirensis for homeowner and ornamental industry use. Chemical Name Or Common Name Registered Product Or Trade Name Chemical Class References Acephate (C) 1300 Orthene TR (F,G) Organophosphate Osborne et al. 2006 Acephate Pro 75 (WSP) Or ganophosphate Buss and Turner 2006; Osborne et al. 2006 Acephate 97UP Organophosphate Bethke 2010 Orthene 75 S Organophosphate Townsend et al. 2000 Orthene TT&O Spray 97 (WSP) Organophosphate Buss and Turner 2006; Bethke 2010 Orthene Turf (F,G) Org anophosphate Osborne et al. 2006 PT 1300 Orthene TR Organophosphate Bethke 2010 Tree and Ornamental Spray 97 (F,G) Organophosphate Osborne et al. 2006 Acetamiprid (SY,C) Tristar 30 SG (F,G) Neonicotinoid Price et al. 2001; Osborne et al. 2006 TriSta r 70 (F,G,WSP) Neonicotinoid Price et al. 2001; Buss and Turner 2006; Osborne et al. 2006; Bethke 2010; Ludwig 2009 Azadirachtin (SY) Azatin XL Biological Insecticide (F,G) Insect Growth Regulator Price et al. 2001; Osborne et al. 2006; Bethke 2010 Azat in XL Plus (EC) Insect Growth Regulator Buss and Turner 2006; Bethke 2010 Azatin XL 0.26 (EC) Insect Growth Regulator Townsend et al. 2000; Bethke 2010 Azatrol (EC) Insect Growth Regulator Price et al. 2008 Ornazin 3% EC Insect Growth Regulator Price et al. 2001; Osborne et al. 2006; Bethke 2010 Beauveria bassiana ATCC 74040 (C) Naturalis L Biological Price et al. 2008 Beauveria bassiana Strain GHA (C) Botanigard 22 WP Biological Price et al. 2001; Bethke 2010 Botanigard ES Mycoinsecticide Biologi cal Price et al. 2001; Bethke 2010 Mycotrol O Biological Price et al. 2001 Bendiocarb (SY,C) Turcam 76 (WP) Pyrethroid Townsend et al. 2000 Bifenthrin (C) Attain TR Pyrethroid Price et al. 2001; Bethke 2010 Attain TR Micro (F,G) Pyrethroid Price et a l. 2001; Osborne et al. 2006

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57 Table 1 5. Continued Chemical Name Or Common Name Registered Product Or Trade Name Chemical Class References Bifenthrin Bifenthrin PRO Multi Insecticide (EPA Reg. No. 51036 391)* Pyrethroid Price et al. 2001; Bethke 2010 Talstar Flowable (F, G) Pyrethroid Price et al. 2001; Buss and Turner 2006; Osborne et al. 2006 Talstar GC* (F,G) Pyrethroid Buss and Turner 2006 Talstar Nursery Flowable* (F) Pyrethroid Price et al. 2001; Buss and Turner 2006 TalstarOne Multi Inse cticide Pyrethroid Price et al. 2001 Talstar T&O 10 (WP) Pyrethroid Townsend et al. 2000 Buprofezin (C) Talus 40 (SC) Insect Growth Regulator Price et al. 2001 Talus Insect Growth Regulator (F,G) Insect Growth Regulator Price et al. 2001; Osborne et a l. 2006 Carbaryl (C,SY) Sevin (SL) Carbamate Buss and Turner 2006 Sevin 50W Carbamate Townsend et al. 2000 Sevin 80 (WSP) Carbamate Buss and Turner 2006 Chlorpyrifos (C) Chlorpyrifos Pro 2 (F) Organophosphate Osborne et al. 2006 Chlorpyrifos Pro 4 (F) Organophosphate Osborne et al. 2006 Duraguard ME* (F) Organophosphate Price et al. 2001; Osborne et al. 2006; Bethke 2010 Dursban 50 (WP) Organophosphate Townsend et al. 2000; Osborne et al. 2006 Chlorpyrifos (C) & Cyfluthrin (C) Duraplex TR* (G) Organophosphate & Pyrethroid Price et al. 2001; Osborne et al. 2006; Bethke 2010 Clothianidin (SY) Celero 16 WSG (F) Neonicotinoid Price et al. 2001; Buss and Turner 2006; Osborne et al. 2006 Cyfluthrin (C) Bayer Advanced Garden Power Force Multi Insect Killer** (SL) Pyrethroid Buss and Turner 2006 Decathlon 20 WP (F,G) Pyrethroid Price et al. 2001; Townsend et al. 2000; Osborne et al. 2006; Bethke 2010 Tempo 20 WP GC* (WSP) Pyrethroid Buss et al. 2006 Cyfluthrin Temp 20 Power Pak (WSP) Pyrethroid Bu ss et al. 2006

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58 Table 1 5 Continued Chemical Name Or Common Name Registered Product Or Trade Name Chemical Class References Cyfluthrin Tempo Ultra (SC, WP, WSP) Pyrethroid Buss et al. 2006 Tempo 2 (EC) Pyrethroid Buss et al. 2006 Cyfluthrin (C) & Imidacloprid (SY,C) Discus (F) Pyrethroid & Neonicotinoid Osborne et al. 2006 Deltamethrin (C) DeltaGard GC 5SC* (SC) Pyrethroid Buss and Turner 2006; Bethke 2010 DeltaGard T&O 5SC (SC) Pyrethroid Buss and Turner 2006; Bethke 2010 Diazinon (C) Diazinon 25 (EC) Organophosphate Townsend et al. 2000 Dimethoate (SY,C) Dimethoate 400 (F) Organophosphate Osborne et al. 2006 Dinotefruan (SY, C) Safari 20 SG Neonicotinoid Price et al. 2001; Bethke 2010; Ludwig 2009 Safari 2G Neonicotinoid Price et al. 2001 ; Bethke 2010 Disulfoton (SY) Bayer Advanced Garden 2 in 1 Systemic Azalea, Camellia & Rhododendon Care** (G) Organophosphate Buss and Turner 2006 Fenoxycarb (C) Preclude TR Insect Growth Regulator Price et al. 2001 Precision 25 (WP) Insect Growth Reg ulator Townsend et al. 2000 Fenpropathrin (C) Tame 2.4 EC Spray* (EC) Fenopropathrin Price et al. 2001; Townsend et al. 2000 Fenpyroximate (C) Akari 5 SC Pyrazole Price et al. 2001 Fish Oil (C) Organocide** (EC,F,G) Biorational Buss and Turner 2006; Osb orne et al. 2006 Flonicamid (SY) Aria (G) Pyridincarboxamids Price et al. 2001; Osborne et al. 2006 Imidacloprid (SY) Bayer Advanced Garden Tree & Shrub Insect Control** (F) Neonicotinoid Buss and Turner 2006 Marathon 1% (F,G) Neonicotinoid Price et al 2001; Buss and Turner 2006; Osborne et al. 2006; Bethke 2010 Marathon 60 G & N in WSP (F,G,WP) Neonicotinoid Price et al. 2001; Osborne et al. 2006; Bethke 2010 Marathon II (F, G) Neonicotinoid Price et al. 2001; Osborne et al. 2006; Bethke 2010; Lud wig 2009 Merit 2 (F) Neonicotinoid Buss and Turner 2006 Merit 75 (WP, WSP) Neonicotinoid Buss and Turner 2006

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59 Table 1 5 Continued Chemical Name Or Common Name Registered Product Or Trade Name Chemical Class References Isaria fumosorosea (C) PFR 97 Biological Dr. L. S. Osborne, personal communication Lambda cyhalothrin (C) Lambda Cy (EC) Pyrethroid Price et al. 2001 Scimitar GC* Pyrethroid Price et al. 2001 Scimitar (CS) Pyrethroid Buss and Turner 2006 Scimitar (WP, WSP) Pyrethroid Buss and Turner 2006 Malathion (C) Malathion 5 (SL) Organophosphate Buss and Turner 2006 Malathion 5EC (F,G) Organophosphate Osborne et al. 2006 Malathion 57 (EC) Organophosphate Buss and Turner 2006 Malathion 8 (SL) Organophosphate Buss and Turner 2006 Ma lathion 8 E (EC) Organophosphate Buss and Turner 2006 Malathion 8F (EC) Organophosphate Buss and Turner 2006 Malathion 8 Spray (SL) Organophosphate Buss and Turner 2006 Methiocarb (C) Mesurol 75WP Carbamate Townsend et al. 2000 Naled (C) Dibrom 8 Em ulsive Carbamate Price et al. 2001 Mineral Oil (C) JMS Stylet Oil Biorational Bethke 2010 SafTSide Biorational Bethke 2010 Sunspray Ultra Fine (F,G) Biorational Townsend et al. 2000; Osborne et al. 2006 Sunspray 6E (EC) Biorational Buss and Turner 2 006 Sunspray 11E (EC) Biorational Buss and Turner 2006 Ultra Fine Oil (F,G) Biorational Osborne et al. 2006; Bethke 2010 Volck** (EC) Biorational Buss and Turner 2006 Neem Oil (C) Triact 70 (EC,F,G) Biorational Price et al. 2001; Buss and Turner 200 6; Osborne et al. 2006; Bethke 2010 Triact 80 Biorational Townsend et al. 2000 Permethrin (C) Astro Pyrethroid Price et al. 2008; Bethke 2010 Perm UP 3.2 (EC) Pyrethroid Price et al. 2008 Permethrin E Pro Pyrethroid Price et al. 2008 Permethrin Pr o Termite Turf Ornamental (EC) Pyrethroid Buss and Turner 2006 Potassium Salts of Fatty Acids AllPro Insecticidal Soap 40% Insecticidal Soap Price et al. 2008

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60 Table 1 5 Continued Chemical Name Or Common Name Registered Product Or Trade Name Chemical Class References Potassium Salts of Fatty Acids Insecticidal Soap 49.52 CF (F, G) Insecticidal Soap Buss and Turner 2006; Osborne et al. 2006 M Pede (F,G) Insecticidal Soap Price et al. 2001; Townsend et al. 2000 Insecticidal Soap Bu ss and Turner 2006 Pyrethrin (C) PyGanic Crop Protection EC 1.4 (EC) Pyrethroid Price et al. 2001 PyGanic Crop Protection EC 5.0 (EC) Pyrethroid Price et al. 2001 Spectracide Rose & Flower Insect Spray** (F) Pyrethroid Price et al. 2001 Pyrethrin (C) & Piperonyl Butoxide (SYN) 1100 Pyrethrum TR Pyrethroid Price et al. 2001 EverGreen EC 60 6 (EC) Pyrethroid Price et al. 2001 PT Pyrethrum TR Pyrethroid Bethke 2010 Pyrethrum TR Micro Pyrethroid Price et al. 2001 Pyrenone Crop Spray Pyrethroid Pri ce et al. 2001 Pyrethrin (C) & Rotenone (C) and other associated resins Pyrellin EC Pyrethroid Price et al. 2001; Bethke 2010 Pyriproxyfen (C) Distance Insect Growth Regulator (EC,F,G) Insect Growth Regulator Price et al. 2001; Buss and Turner 2006; Osbo rne et al. 2006 Distance 0.86 (EC) Insect Growth Regulator Townsend et al. 2000 S Kinoprene (C) Enstar II (G) Insect Growth Regulator Price et al. 2001; Osborne et al. 2006 Enstar II 5 (EC) Insect Growth Regulator Townsend et al. 2000; Bethke 2010 Sp irotetramat (C) Kontos Ketoenole Ludwig 2009 Tau fluvalinate (C) Mavrik Aquaflow Pyrethroid Price et al. 2001; Bethke 2010 Mavrik 2EC Pyrethroid Townsend et al. 2000 Thiamethoxam (SY,C) Flagship 0.22 G Neonicotinoid Price et al. 2001 Flagship 25 W G (F,G) Neonicotinoid Price et al. 2001; Buss and Turner 2006 C = Contact; EC = Emulsifiable concentrate; F = Water dispersible liquid; G = Granule; SC = Water soluble liquid; SL = Water soluble liquid; SY = Systemic; SYN= Synergist; WG = Wettable granule s; WP = Wettable powder; WSP = Water solute powder, *Restricted use product; **Homeowner product

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61 CHAPTER 2 DEVELOPMENT OF THE MODEL BIORATIONAL DIP Introduction Importation of cryptic, invasive pests as plant contaminants on ornamental cuttings has posed a serious threat to Florida. The United States Department of Agriculture (2007) cited Florida as the leading ornamental propagation state with an estimated annual revenue of $91 million. Approximately 89.4% of United States flower imports and 82% of all ai r imports in the United States and Latin American/Caribbean Region pass through the Miami Plant Protection Quarantine Inspection Station at the Miami International Airport (Miami Dade Aviation Department Marketing Division 2013). However, limited resources and trade demands result in the inspection of only 2% of imported plant stock by the United States Department of Agriculture, Animal and Plant Health Inspection Services (Brasier 2008). Additionally, the humid, warm climate of Florida is suitable for the e stablishment of invasive pests such as aphids, scales, mealybugs, mites, thrips, and whiteflies listed in Table 1 1 (Frank and Thomas 2004). Effective control measures for limiting the spread of cryptic, invasive species as plant contaminants on ornamenta l cuttings were made apparent after the invasion of the B biotype and pesticide resistant Q biotype of the sweetpotato whitefly, Bemisia tabaci (Gennadius) on poinsettia cuttings. In the 1990s, the B biotype whitefly was introduced into the United States a s a plant contaminant on poinsettia cuttings moving from Israel to Florida to California, and farmers (Dalton 2006, De Barro et al. 2008, McKenzie et al. 2009). Similarly, in the mid 2000s, the Q biotype was imported into the United States as a plant contaminant on poinsettia cuttings from Guatemala, Honduras, and southern Mexico that w ere shipped and grown into infes ted poinsettia plants found in Encinitas, Calif ornia (Dalton 2006, Bethe et al. 2009).

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62 Treatment methods for controlling ornamental cutting pests have been limited to on site synthetic insecticide dips as cuttings lack roots, thus rendering high volume sprays, soil based systemic insecticides, and dre nches ineffective (Osborne 1986, Conover and Poole 1970, Hansen et al. 1992). In contrast to synthetic insecticide dips (e.g. organophosphates, carbamates, and pyrethroids), biorational dips (e.g. surfactants, anti transpirants) have multiple modes of acti on and insecticide resistance has not been reported (Capinera 2008, Gunning et al. 1984, Cahill et al. 1995, Kranthi et al. 2001). Organic oils and surfactants such as detergents have been known to induce mortality in several ways: (1) starvation by preven ting plant pests from using their piercing sucking mouthparts, or (2) death by suffocation or desiccation by halting respiration via spiracle blockage, or (3) toxicity from muscle or nerve damage caused by penetrating and damaging the tracheae (Capinera 20 08). Observations have shown that biorational treatments tended to induce less severe phytotoxicity than synthetic insecticides when applied at the recommended label rate. Popular biorational treatments such as soybean oil and cotton seed oil have been ob served to cause slight chlorotic spotting and marginal chlorosis on collard and tomato leaves (Liu and Stansly 2000). Anti transpirants have only been known to inhibit plant growth on soybean, pine, and corn seedlings (Gale and Hagan 1966, Rowan 1988, Dave nport et al. 1969, McConnel 1985, Mmbaga and Sheng 2002). Additionally, surfactants such as Silwet L 77 and Wetcit TM have shown variation in phytotoxicity among different plant types, including stunted plant growth, leaf drop, chlorosis, or necrosis (Beyer et al. 1988, Sun 1996). In comparison to biorational treatments, Hata and Hara (1988) described severe phytotoxicity on the flower anthurium, Anthurium andraeanum Andre following synthetic insecticide application. Organophosphates such as chloropyrifos (Dursban) and acephate

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63 (Orthene 75 and Isotox Insect Killer) induced chlorotic and necrotic spotting on mature leaves, bronzed abaxial surfaces, leaf curling, stem deformation, and spathe discoloration. Oxamyl, a carbamate active ingredient, caused chlorot ic spotting and marginal chlorosis on new leaf growth and chlorotic spotting and leaf curling on mature leaves. Pyrethroids such as bifenthrin (Talstar 10) and cyfluthrin (Tempo 2) varied in causing phytotoxicity. Implementation of a biorational on site d ip may fit well within a Florida nursery and greenhouse operation. In addition to effectively controlling ornamental pests while inducing minimal plant phytotoxicity, biorational treatments have been known to be less expensive than synthetic dips or sprays (Liu and Stansley 2000, Dr. Lance Osborne, personal communication). Additionally, biorational treatments have limited worker, community, and environmental concern as biorationals may either leave no residues or quickly decompose after use (Weinzierl and H enn 1991, United States Environmental Protection Agency 2013, Dr. Eileen Buss, personal communication). The purpose of this study was to develop an on site biorational model dip treatment in order to prevent the dissemination of invasive species as plant contaminants on ornamental cuttings being imported and exported from the United States. Such knowledge is important because invasive species threaten the United States agricultural and ornamental industry as contaminants on imported and exported plant carg o. In this study, the Madeira mealybug, Phenacoccus madeirensis Green was selected as the model invasive ornamental pest while coleus, Solenostemon sp., was designated as the model ornamental cutting. A foliage phytotoxicity rating system redesigned after Hansen et al. (1992) and ornamental cutting health analysis from Osborne et al. (1986) was used to evaluate phytotoxicity for several concentrations for each type of biorational dip. Based on the phytotoxicity results, the highest concentration

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64 from each type of biorational dip with acceptable phytotoxicity was used to determine efficacy against P. madeirensis Afterwards, t he treatment with the highest P. madeirensis mortality was determined as the model biorational dip treatment. Materials and Methods R earing Colonies of P. madeirensis Separate rearing rooms used for two P. madeirensis colonies reared on Solenostemon Acalypha sp p (copperleaf) were maintained at a 14L:10D photoperiod and standard room temperature (27 0 C) (Chong 2003). In order to prevent a complete loss of colony due to the development of entomopathogenic fungal spores, separate P. madeirensis rearing rooms were maintained at 20% and 60% relative humidity. Manual watering of host plants for P. madeirensis colonies occurred three times per week. Specimens from each colony at UF MREC were sent to the Florida Department Agriculture and Consumer Services Division of Plant Industry for mealybug identification and stored afterwards at the Florida State Collec tion of Arthropods as voucher specimen. Host Plant Maintenance Solenostemon enclosed greenhouse from January to July 2013 with a 22.8 0 C average daily temperature and 68% mean relative humid ity at the University of Florida, Mid Florida Research and Education Center (UF MREC) in Apopka, FL. Plants were rooted in Faford Growing Mix 2/C 2 soil composed of Canadian sphagnum peat moss, perlite, vermiculite, dolomitic limestone, and wetting agent (2:1:1:1:1, vol:vol:vol:vol:vol) and wer e hand watered daily until soil was evenly moistened Plants over 30 inches (72.6cm) in height were spaced at least one foot apart and flowering parts were pruned to encourage further branching.

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65 Cutting Preparation Coleus tip cuttings were prepared by randomly selecting and excising the stem of a stock plant at a slight angle in order to prevent stem damage. All tip cuttings were at least 3 inches (7.62cm) in height and consisted of four mature leaves on the top port ion of the plant. Excess basal leaves on the stem and flowering parts on the tip were removed. Tip cuttings with blemishes that resembled phytotoxicity characteristics were discarded. A total of thirty blemish free tip cuttings were made for each experimen t. Cuttings were made once the stock plant was over 30 inches (72.6cm) in height. New stock plants were grown using the same methods described above. Assessment of Biorational Phytotoxicity on Cuttings Dip t reatment p rotocol Four biorational materials we vegetable oil, Stoller Enterprises, Inc, Houston, Texas); Wetcit TM (8.92% alcohol ethoxylate with selected adjuvants, AGRI Inc, Trophy Club, Texas); Publix Mild & Gentle Ultra Dish Detergent (select anionic and nonionic surfa ctants, stabilizers, quality control agents, perfume and colorant, Publix Super Markets, Inc, Lakeland, Florida); and Vapor Gard (96% di 1 p Methene with 4% select inert ingredients, Miller Chemical & Fertilizer Corporation, Hanover, Pennsylvania). Four c oncentrations for each biorational material were prepared and thoroughly mixed with a magnetic stirrer for five minutes: 0.10% (1mL of treatment per 999mL of de ionized water), 0.50% (5mL of treatment per 995mL of de ionized water), 1.00% (10mL of treatmen t per 990mL of de ionized water), and 1.50% (15mLof treatment per 985mL of de ionized water). The treatment control was 1000mL of de ionized water. Another 1000mL of de ionized water was used to treat a second set of control plants to evaluate the effect o f the mist irrigation watering system. These control plants served as an early detection measure for fungal or bacterial growth on cuttings that may be mistaken for phytotoxicity character expression. Each dip treatment was

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66 randomly assigned five coleus ti p cuttings and evenly re mixed with a magnetic stirrer for two minutes prior to usage. All five cuttings for each dip treatment were dipped simultaneously for one minute in a 6.5 quart rectangular plastic bin. During each dip, all tip cuttings were fully submerged, lightly agitated, and rotated vertically after 30 seconds to maximize treatment surface area coverage on the tip cuttings. Dipped cuttings were lightly shaken to remove excess runoff and air dried until runoff ceased. Each cutting was transplan ted into a labeled ounce plastic pot filled with moistened Faford Growing Mix 2/C 2 soil. Two replicates of five cuttings for each biorational material and concentration were conducted on the same day with one replicate completed in the morning and the other in the afternoon. For the phytotoxicity assessment, data collection methodology for foliage phytotoxicity was redesi gned after Hansen et al. (1992). T he methodology for obtaining data on cutting height, root volume, and root length was modele d after Osborne et al. (1986). Cutting o bservation and w atering p ractices For two weeks, all cuttings were kept under a misting system which actuated every 10 minutes for 30 seconds from 0800 to 2000 hours. Mist control cuttings were placed on a separate bench and hand watered twice every day until soil was moist. Afterwards, all cuttings were removed from the mist and manually watered in the morning three times per week until the end of the fourth week. Observations made for each cutting included the foll owing information: presence of any pests, including the identity, possible life stage, and sex, and any non pest or non treatment suspected cutting damage. Assessment of Biorational Effects on Plants Plant h eight Prior to transplanting treated cuttings, cutting length was recorded. Initial length for cuttings treated with Wetcit TM Detergent, Vapor Gard, and the control was taken on the same date cuttings were treated and planted: 2 April 2013, 3 April 2013, 4 April 2013, and 8 April 2013, respectively. Cuttings were

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67 measured from the base to the tip for initial and final cutting height. Final plant height for cuttings treated with Wetcit TM Gard, and the control occurred on 23 April 2013, 24 April 2013, 25 April 2013, and 29 April 2013, respectively. Foliage d amage. Each coleus tip cutting was divided into five sections for phytotoxicity foliage analysis. Each of the four mature leaves on the tip c utting constituted a leaf section. The fifth leaf section was attributed to new flush growth. The initial foliage phytotoxicity ratings for cuttings treated with Wetcit TM Vapor Gard and the control were taken on the same date cuttings were treated and planted: 2 April 2013, 3 April 2013, 4 April 2013, and 8 April 2013, respectively. Similar to the rating system described by Hansen et al. (1992), two raters simultaneously issued independent ratings at the same time dipping occurred but the ratings were not summed. The rating system was modified further in order to characterize the symptoms of phytotoxicity. The symptoms were assigned numerical values ranging from 0 5, with 0 as no damage to stem and leaves to 5 as foliage death. The following modifications were made to the Hansen et al. (1992) rating system to score phytotoxicity of the treatments weekly for four weeks: (1) no injury, (2) slight injury, (3) moderate injury, (4) severe injury, and ( 5) dead; and eleven distinct phytotoxicity characters were rated for each leaf section: chlorosis (C), leaf curling (Y), chlorotic flecking (CF), necrotic flecking (NF), marginal necrosis (MN), marginal chlorosis (MC), chlorotic streaking (CS), necrotic st reaking (NS), tip chlorosis (TC), tip necrosis (TN), and holes (H). Final ratings for cuttings treated with Wetcit TM Gard, and the control occurred on 23 April 2013, 24 April 2013, 25 April 20 13, and 29 April 2013, respectively.

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68 Root v olume and l ength. Soil was gently rinsed and dislodged from the root system of each cutting by using a hose at low pressure and dipping roots in water. Root volume for cuttings treated with Wetcit TM Publix Mild & Gentle Ultra Dish Detergent, Vapor Gard, and the control was recorded on 23 April 2013, 24 April 2013, 25 April 2013, and 29 April 2013, respectively. Root volume was evaluated by excising the root system of the cutting and placing the roots into a known volume of water in a 100mL graduated cylinder (Osborne 1986). The total volume of roots with water was recorded. From that value, the volume of water displaced by the roots was determined and evaluated as the root volume of the cutting. Avera ge root length was assessed in the same manner as Osborne (1986) where three of the longest roots for each cutting were measured from the base to root tip and averaged. Data for each treatment was recorded on 23 April 2013, 24 April 2013, 25 April 2013, an d 29 April 2013, respectively. Efficacy of Biorationals on P. madeirensis Mortality Based on the results of the phytotoxicity assessment, the highest concentration from each type of biorational dip with acceptable damage was selected for the efficacy bioa ssay. A total of TM 1.0% Publix Mild & Gentle Ultra Dish Detergent, and 0.1% Vapor Gard. Five replicates for each biorational material were conducted in a 5 X 5 randomiz ed complete block design. For each replicate, a total of 25 coleus cuttings, each infested with 15 P. madeirensis mealybugs ranging from crawlers to virgin female adults, were evaluated for mortality. Data collection for each replicate began on a separate date and time: 14 March 2013 in the morning, 14 March 2013 in the afternoon, 28 March 2013 in the morning, 1 April 2013 in the morning, and 1 April 2013 in the afternoon. Dip treatment preparation E ach selected bi orational material was prepared and thor

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69 Oil per 990mL of de ionized water), 0.1% Wetcit TM (1mL of Wetcit TM per 999mL of de ionized water), 1.0% Publix Mild & Gentle Ultra Dish Detergent (10mL of Publix Soap per 990mL of de ionized water), and 0.1% Vapor Gard (1mL of Vapor Gard per 999mL of deionized water). One 1000mL of de ionized water was used as a control. Coleus c utting p reparation. To i nfest cuttings with more than 15 mealybugs, coleus plants were re infested by exposure to P. madeirensis from the colony maintained at UF MREC one week before the trial. Infested cuttings were made by excising the stems at a 45 o angle to avoid stem damage. Cuttings were at least 3 inches (7.62cm) in height with four mat ure leaves on top. Excess basal leaves on the stem and flowering parts on the tip were removed. Exactly 15 mealybugs were randomly selected to remain on the cutting. The mealybugs left on cuttings ranged from crawlers to the adult females. Excess mealybugs oviscas, male adults, or male cocoons were removed using a moistened camel hair paintbrush. If there were less than 15 mealybugs present on a cutting, mobile mealybugs from different colony leaves that varied in size and ranged from crawlers to adult fem ales were randomly selected and transferred onto the cutting using a moistened camel hair paintbrush All transferred mealybugs were left to settle on the cutting for at least fifteen minutes. All cuttings had a random number of active mealybug life stages present. A total of 15 mealybugs were left on each cutting. Each cutting was carefully sealed in a clear Ziploc bag to maintain cutting moisture and reduce the risk of losing mealybugs when transporting the cutting to the greenhouse for treatment. Dippin g m ethod and p lanting p rocedure. The dipping method and planting procedure described in the phytotoxicity assessment section was used during each efficacy bioassay. Cutting o bservation and w atering p ractices For two weeks, all cuttings were kept under a m isting system that activated every 10 minutes for 30 seconds from 0800 to 2000 hours.

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70 Afterwards, all cuttings were removed from mist and manually watered in the morning three times per week. Observations made at each mortality time period for each cutting in cluded : mealybug feeding and movement post trea tment; presence of other arthropods, ovisacs, male cocoons or male adults; phytotoxicity symptoms and severity; and handling damage Mealybug m ortality e valuation. Two raters simultaneously recorded the nu mber of live and dead mealybugs immediately after dipping and on the third, seventh, and fourteenth day. If the mealybug cadaver was present, the life stage of the dead mealybug was recorded. For the analysis, the following time periods were used to determ ine mealybug mortality: immediately after dipping (Day 1), between Day 1 and the third day (Day 3), between the third day and seventh day (Day 7), and between the seventh and fourteenth day (Day 14). The baseline for an effective biorational treatment was determined at 70% mortality (Dr. Lance Osborne, personal communication). Mealybug mor t ality was modified from Hata et al. (1992): (1) no movement after gently touching with a moistened camel hair paint brush at least three times and (2) a shriveled, hollow, or blackened appearance. Living mealybugs were distinguished by the following: (1) leg movement, (2) mouthpart movement, (3) visible mealybug resistance or displacement when brushed gently with a moistened camel hair paint brush at least three times and ( 4) detachment from its original position after or prior to being touched Statistics Efficacy bioassay and phytotoxicity assessment summary statistics were prepared using a fit linear model in the GLIMMIX Procedure (General Linear Model for Mixture Distrib utions) to fit binary outcomes and account for non normality and non homogenous variances in SAS 9.3 (SAS Institute Inc., 2002 2010, Cary, NC, USA). For the efficacy bioassay, five replicates were conducted in a 5 X 5 random complete block design. For each replicate, a total of 25 cuttings,

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71 each infested with 15 P. madeirensis mealybugs, were evaluated for mortality. D ifferences between cumulative percentages of dead P. madeirensis per selected treatment at each time period were analyzed by using Tukey Kram er Least Squares Means for multiple comparisons The p hytotoxicity assessment analyzed cutting height, root volume, average root length, and each of the eleven phytotoxicity characteristics for foliage phytotoxic ity. For each biorational material, two repl icates were conducted on the same day in a random complete block design with one replicate completed in the morning and the other in the afternoon. A total of 30 cuttings were used for each replicate and evaluated for phytotoxicity. Tukey Kramer Least Squa res Means for multiple comparisons was used to determine significant differences between the following: average ratings per rater, average ratings for each week per foliage phytotoxicity characteristic, average ratings for each biorational material for eac h foliage phytotoxicity character, average ratings for each concentration of Wetcit TM Vapor Gard, Publix Soap, and respectively, and average ratings across all biorational material concentrations (SAS Institute Inc., 2002 2010, Cary, NC, USA ). Additionally, Tukey Kramer Least Squares Means for multiple comparisons was also utilized to verify differences between the following: average measurements for each biorational material average measurements for each concentration of Wetcit TM Vapor Gar d, Publix Soap, and respectively, average measurements across all concentrations, and average measurements in initial and final cutting height, respectively. Results Efficacy Bioassay The c umulative average percent of dead P. madeirensis was significantly different between the following selected treatments for each time period : 0.1% Wetcit TM 0.1% Vapor Table

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72 2 1). During each day, the highest percentage mor 0.1% Wetcit TM concentration treatments followed by 0.1% Vapor Gard and 1.0% Publix Soap. Vapor Gard at 0.1% and 1.0% Publix Soap showed no significant difference in mortality In comparison to all other treat ments, the control had the lowest mortality for each day. None of the selected treatments achieved 100% mortality by Day 14. Phytotoxicity Assessment Foliage ratings significantly differed within concentrations for each biorational material over a period o f three weeks. Comparisons between the different Wetcit TM concentration treatments showed that 0.1% had the lowest ratings for all characters while 1.0% and 1.5% scored the highest for all of the following characters: chlorosis (F=83.94; df= 4; p<0.0001); chlorotic flecking (F=50.09; df= 4; p<0.0001), necrotic flecking (F=90.03; df= 4; p<0.0001), holes (F=28.52; df= 4; p<0.0001), chlorotic streaking (F=78.31; df= 4; p<0.0001), necrotic streaking (F=11.88; df= 4; p=1.000), leaf curling (F=1.64; df= 4; p=0.16 10), marginal chlorosis (F=7.11; df= 4; p<0.0001), marginal necrosis (F=79.99; df= 4; p=0.0534), tip chlorosis (F=3.18; df= 4; p=0.0127), and tip necrosis (F=2.75; df= 4; p=0.0266). Comparisons within Vapor Gard concentration treatments also showed that 0.1% had the lowest ratings for 10 characters while 1.0% and 1.5% had the highest ratings for all of the following characters: chlorosis (F=283.50; df= 4; p<0.0001); chlorotic flecking (F=177.11; df= 4; p<0.0001), necrotic flecking (F=144.50; df= 4; p<0.00 01), holes (F=25.26; df= 4; p<0.0001), chlorotic streaking (F=64.45; df= 4; p<0.0001), necrotic streaking (F=0.00; df= 4; p=1.000), leaf curling (F=6.62; df= 4; p<0.0001), marginal chlorosis (F=4.93; df= 4; p=0.0006), marginal necrosis (F=69.55; df= 4; p<0 .0001), tip chlorosis (F=9.46; df= 4; p<0.0001), and tip necrosis (F=52.41; df= 4; p<0.0001).

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73 Differences within Publix Soap concentration treatments included 0.1% with the lowest ratings for the following 6 characters: chlorosis chlorotic flecking, necro tic flecking, holes, chlorotic streaking, and tip chlorosis. Publix Soap at1.5% had the highest ratings for all of the following characters: chlorosis (F=56.71; df= 4; p<0.0001); chlorotic flecking (F=51.72; df= 4; p<0.0001), necrotic flecking (F=12.81; df = 4; p<0.0001), holes (F=12.14; df= 4; p<0.0001), chlorotic streaking (F=111.14; df= 4; p<0.0001), necrotic streaking (F=0.00; df= 4; p=1.000), leaf curling (F=12.55; df= 4; p<0.0001), marginal chlorosis (F=2.76; df= 4; p=0.0263), marginal necrosis (F=3.96 ; df= 4; p=0.0033), tip chlorosis (F=27.85; df= 4; p<0.0001), and tip necrosis (F=19.93; df= 4; p<0.0001). had the lowest ratings for 4 characters: chlorosis chlorotic flecking, necrotic flecking, and holes. 1.5% had the highest ratings for all of the following characters: chlorosis (F=49.55; df= 4; p<0.0001); chlorotic flecking (F=47.41; df= 4; p<0.0001), necrotic flecking (F=17.02; df= 4; p<0.0001), hol es (F=4.68; df= 4; p=0.0009), chlorotic streaking (F=40.40; df= 4; p<0.0001), necrotic streaking (F=0.00; df= 4; p=1.000), leaf curling (F=4.91; df= 4; p=0.0006), marginal chlorosis (F=5.17; df= 4; p=0.0004), marginal necrosis (F=3.50; df= 4; p=0.0074), ti p chlorosis (F=7.05; df= 4; p<0.0001), and tip necrosis (F=2.65; df= 4; p=0.0314). When all biorational material concentrations were compared together, Wetcit TM at 1.0% and 1.5% had the highest ratings for 6 characters followed by Vapor Gard at 1.5% (4), Publix Table 2 showed no significant difference but had the lowest r atings for 9 characters : chlorosis, chlorotic flecking, necrotic flecking, holes, tip chlorosis, chlo rotic streaking, necrotic streaking, marginal chlorosis, and marginal necrosis Overall, foliage ratings for all biorational material

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74 concentrations compared together for each phytotoxicity character were significantly different: chlorosis (F=33.00; df= 1 0; p<0.0001), chlorotic flecking (F=20.08; df= 10; p<0.0001), necrotic flecking (F=25.72; df= 10; p<0.0001), holes (F=8.92; df= 10; p<0.0001), tip chlorosis (F=11.58; df= 10; p<0.0001), tip necrosis (F=12.25; df= 10; p<0.0001), chlorotic streaking (F=12.00 ; df= 10; p<0.0001), necrotic streaking (F=2.97; df= 10; p=0.0004), marginal chlorosis (F=2.01; df= 10; p=0.0196), marginal necrosis (F=21.33; df= 10; p<0.0001), and leaf curling (F=6.50; df= 10; p<0.0001). For each phytotoxicity character, Rater 1 issued higher ratings than Rater 2 for 9 out of 11 characters but both raters showed no significant difference between necrotic streaking and marginal necrosis ratings The following characters showed significant difference between ratings: chlorosis (F=365.12; df= 1; p<0.0001), chlorotic flecking (F=280.42; df= 1; p<0.0001), necrotic flecking (F=91.38; df= 1; p<0.0001), holes (F=578.15; df= 1; p<0.0001), chlorotic streaking (F=152.46; df= 1; p<0.0001), necrotic streaking (F=0.34; df= 1; p=0.5577), leaf curling ( F=3.81; df= 1; p=0.0509), marginal chlorosis (F=346.39; df= 1; p<0.0001), marginal necrosis (F=3.73; df= 1; p=0.0534), tip chlorosis (F=328.83; df= 1; p<0.0001), and tip necrosis (F=21.71; df= 1; p<0.0001). p (F=1.88; df= 4; p=0.1133), Wetcit TM (F=2.71; df= 4; p=0.0296), and Vapor Gard (F=1.17; df= 4; p=0.03212) demonstrated no significant difference in cutting height withi n concentrations for each biorational material Similarly, root volume within concentr ations for each biorational material exhibited no significant difference p=0.6226), Wetcit TM (F=1.24; df= 4; p=0.2935), Vapor Gard (F=1 .39; df= 4; p=0.2380). However, root length was sign ificantly different within concentrations for each biorational

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75 material ( Table 2 1.0%, and 1.5% had the longest root length (F=3.79; df= 4; p=0.0026). Publix Soap concentrations sho wed no significant difference (F=1.95 df= 4; p=0.0879). Differences within Wetcit TM concentrations included 1.0% with the shortest root length in comparison to all other Wetcit TM concentrations (F=3.55; df= 4; p=0.0041). In contrast to all other Vapor Gard concentrations, Vapor Gard at 1.5% also had the shortest root length (F=7.88; df= 4; p<0.0001). Model Dip Treatment Determination At each time period, TM concentration treatments had the highest mortality (F=48.51; df= 4; p<0.0001) ( Table 2 1). However, in this study, 0.1% Wetcit TM the following key phytotoxicity characters: chlorosis (F=33.00, df= 12, p<0.0001), chlorotic flecking (F= 20.08, df= 12, p<0.0001), necrotic flecking (F=25.72, df= 12, p<0.0001), holes (F=8.92, df= 12, p<0.0001), tip chlorosis (F=11.58, df= 12, p<0.0001), and tip necrosis (F=12.25, df= 12, p<0.0001) ( Table 2 2). Discussion Efficacy B ioassay The most effectiv e treatments over time for controlling P. madeirensis Oil and 0.1% Wetcit TM respectively. Efficacy for both treatments surpassed the baseline, 70% mortality, by Day 3. For both treatments, mortality already exceeded 50% by Day 1 in compa rison to 0.1% Vapor Gard and 1.0% Publix Soap which had less than 50% mortality. By Day 7, 0.1% Vapor Gard and 1.0% Publix Soap finally reached 70% mortality The highest Wetcit TM showed 90% mortality in comparison to 1.0% Publix Soap and 0.1% Vapor Gard with 85%

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76 mortality. Differences in P. madeirensis mortality over time between treatments may be attributed to the mechanical effects of dipping, insecticidal properties o f each treatment, or treatment susceptibility of P. madeirensis at various life stages. The mechanical effects of dipping may have influenced the number of P. madeirensis remaining on each treated cutting The dipping technique was conducted by one desig nated assistant through the experiment which reduced variation in agitation. However, gentle cutting agitation during each dip may have dislodged P. madeirensis During each dip, cuttings were agitated against the sides and bottom of the dipping container. As a result, mobile or feeding P. madeirensis may have been displaced from the cutting or squished altogether by the agitation process. Additionally, the proboscis or legs of P. madeirensis may be broken or harmed by the agitation process thus inducing ev entual death by starvation or secondary infection. The number of P. madeirensis remaining on each treated cutting over time may have varied due to the active ingredient of each treatment. Although different concentrations were l and 0.1% Wetcit TM were the most effective treatments over each time period Active ingredients differed Soap (nonylphenol ethoxylate), Vapor Gard (di 1 p menthene), and Wetcit TM (alcohol ethoxylate). Several studies cited exceptional insecticidal control with soybean oil applied at the recommended dosage rate (Liu and Stansly 2000, Pless et al. 1995, Butler et al. 1993, Amer et al. 2001). Additionally, several scientific studies cited alcohol ethoxyla te insecticidal activity (Liu and Stansly 2000, Imail et al. 1994, Davidson et al. 1991, Hesler and Plapp 1986, Tattersfield and Gimingham 1927, Wolfenbarger et al. 1967, Imail and Tsuchiya 1995, Cory and Langford 1935). However, few studies cited insectic idal activity with di 1 p menthene and nonylphenol ethoxylate. Only Allen et al. (1993) reported that di 1 p menthene reduced Western flower

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77 thrips, Frankliniella occidentalis (Pergande) feeding activity by 40%. Nonylphenol ethoxylate has only been reporte d as an inert ingredient within pesticide products (U.S. Environmental Protection Agency 2007). The treatment susceptibility of P. madeirensis at various instars may have also directly impacted mortality rates for treated cuttings In this study, varying l ife stages of P. madeirensis ranging from crawlers to females adults were used for the efficacy bioassay. Each instar of P. madeirensis has been known to vary in white mealy wax cover production. Townsend (2000) observed that first instar P madeirensis l ack a waxy coating on the body and only develop a mealy wax cover after the second instar. Additionally, Buss and Turner (2003) confirmed that crawlers were more susceptible to synthetic insecticides. As a result, cuttings which had more early instars of P madeirensis may have had a higher mortality over time in comparison to cuttings that had more female adults which produce a thick mealy wax cover for protection (Buss and Turner 2003, Townsend 2000, Green 1923). Overall, the efficacy bioassay may be im proved in several ways High water control mortality was observed throughout this experiment and may be prevented by reducing the impact of mechanical dipping and restricting the number of times mealybugs may be touched. In future studies, mealybug mortali ty could be evaluated by gently touching the mealybug with a moistened camel hair paint brush three times. Mealybugs on water control cuttings were consistently touched more than three times by both raters during the mortality evaluation and continuous tou ching may have broken or pierced the proboscis, legs, or thorax thus inducing eventual death by starvation or secondary infection. To reduce the impact of mechanical dipping, mortality by insecticidal activity may be better analyzed by dipping cuttings in a large basin where cuttings do not touch the sides or base when fully submerged and agitated. Additionally,

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78 the impact of insecticidal activity may also be evaluated better by testing each life stage independent from one another. Additional experiments ma y be conducted for each life stage important for nursery and greenhouse owners to control: ovisacs, crawlers, and gravid females. Phytotoxicity Assessment Root volume, average root length, cutting height, and foliage phytotoxicity were factors used to det ermine the least phytotoxic concentration for each biorational material However, the root volume and cutting height for each biorational material showed no significant difference so root length and foliage phytotoxic ity were analyzed In this study, co leus phytotoxicity characters and patterns were similar to those described in Hansen et al. (1992) on the uluhe fern and pothos treated with insecticidal soap and synthetic dips. For example, leaf curling and marginal chlorosis indicated initial cutting fr agility during the first rating week. Other characters, such as tip chlorosis, would climax in intensity within the first two weeks and then diminish by the fourth week, thus indicating positive plant health and development. Most foliage characters such as chlorosis, chlorotic flecking, necrotic flecking, holes, marginal necrosis, and tip necrosis were fully expressed by the second week when cuttings were rooted. Wetcit TM concentrations showed significant differences in ratings where higher ratings were iss ued to higher concentrations. O bservations from Pypekamp (2008) confirmed that Wetcit TM sprayed at 0.5% induced necrotic lesions compared to 0.25% concentration which caused necrotic spotting. Wetcit TM at 0.1% and 0.5% showed no significant difference in r oot volume, average root length, and root height. However, w ithin Wetcit TM concentrations, 0.1% had the lowest ratings for each phytotoxicity character. Due to these results, 0.1% was determined as the highest Wetcit TM concentration with acceptable damage

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79 Significant differences in Vapor Gard concentrations also indicated that higher concentrations resulted in higher phytotoxicity In general, 0.1% was determined as the highest Vapor Gard concentration with acceptable damage While 0.1% and 0.5% showed no significant difference in average root volume, root length, and cutting height, 0.1% had the lowest ratings for 10 characters compared to 0.5%. Publix Soap concentrations also showed significant differences in ratings where higher concentrations were ma rked with higher ratings. However, 1.0% was designated as the highest Publix Soap concentration with acceptable damage Upon final examination on the fourth week, cuttings treated with 1.0% Publix Soap and 0.1% Publix Soap showed similar character expressi on. Differences in rater perception over time may have attributed to significant differences in ratings where Rater 1 was stricter than Rater 2. concentration increased, higher ratings were issued. However, observations from Butler et al. Overall, the highest concentration with acceptable damage was 1.0%. When all bio rational material concentrations were compared, ratings between 0.1%, 0.5%, and 1.0% showed no significant difference and had the lowest ratings for 8 out of 11 icant difference in average root volume, root length, and cutting height. Future improvements for rating foliage may include more rating training for each rater, averaging ratings between the raters, and issuing ratings underneath a well lit lamp inside th e greenhouse. Other improvements may also be implemented for measuring average root volume,

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80 root length, and cutting height. Roots may be cleaned by dunking and lightly agitating when suspended in water as opposed to being pressure rinsed under a hose. Mod el Dip Treatment Determination Final determination of the model biorational dip was a two step process. First, the highest concentration with acceptable phytotoxicity from each biorational material was selected and designated as a treatment in the efficacy bioassay. Second, the most effective treatment for controlling P. madeirensis over time was determined as the model dip treatment. Although 1.0% TM showed no significant difference in mortality and had the highest mortality comp treatment because of its lower phytotoxicity. Several scientific studies cited Wetcit TM Oil as an effective treatment for seve ral pests. In this study, 0.1% Wetcit TM had higher foliage a s pray at the recommended rate was an effective treatment with over 86% nymph mortality of sweetpotato whitefly, Bemisia tabaci Gennadius. In this study, insecticidal soaps, petroleum based oil, and 15 other surfactants were evaluated simultaneously under gr eenhouse conditions. spray treatment for egg and adult stages of the two spotted spider mite, Tetranychus urticae Koch. Several studies have cited Wetcit TM inse cticidal activity (Liu and Stansly 2000, Imail et al. 1994, Davidson et al. 1991, Hesler and Plapp 1986, Tattersfield and Gimingham 1927, Wolfenbarger et al. 1967, Imail and Tsuchiya 1995, Cory and Langford 1935).

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81 Model Dip Treatment Implications As a prev entative measure for importing invasive species through cuttings, the one time site one minute dip provides Florida growers an efficacious treatment option for cuttings at an acceptable phytotoxicity level. Use of the dip a lso provides growers an environmentally safe treatment while preventing synthetic insecticide development within a Florida greenhouse operation. Within a systems approach, integration of the dip treatment ultimately may help prevent the establishment of im ported mealy bugs and other invasive species. R esults from this study confirmed that the dip effectively controlled P. madeirensis within three days. In future studies, s everal application techniques c ould be modified to develop a dip protocol within a syst ems approach. Factors which impact efficacy c ould be evaluated and include various host plants and invasive species, dip exposure time, dipping location, and dipping application within a synthetic insecticide treatment program.

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82 Table 2 1. Cumulative ave rage percent of dead P. madeirensis per coleus cutting dipped for one minute in selected biorational treatments at each time period Biorational Material Time Period (Days) 1 (Mean + SE) 3 (Mean + SE) 7 (Mean + SE) 14 (Mean + SE) Control 20.26% + 1.629 %c 37.17% + 2.246%c 49.25% + 2.371%c 65.19% + 2.206%c 0.1% Wetcit 56.29% + 2.491%a 74.99% + 1.967%a 83.10% + 1.525%a 90.47% + 0.9958%a 0.1% Vapor Gard 44.90% + 2.503%b 65.48% + 2.322%b 75.68% + 1.932%b 85.72% + 1.357%b 1.0% Publix Soap 45.82% + 2.311%b 66.32% + 2.099%b 76.36% + 1.759%b 86.17% + 1.253%b 57.75% + 2.422%a 76.09% + 1.870%a 83.92% + 1.441%a 90.97% + 0.9376%a Within a column, means followed by the same letter are not significantly different (P> 0.05; Tukey Kramer Grouping fo r Least Squares Means; F=48.51; df=4, 1867; p<0.0001)

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83 Table 2 2 Average coleus cutting phytotoxicity rating per biorational material concentration for key phytotoxicity characters Biorational Material Treatment Chlorosis Chlorotic Flecking Necrotic F lecking Holes Tip Chlorosis Tip Necrosis Control 1.4937i 1.4887j 1.1650hi 1.1500ghi 1.0775cde 1.0263g 0.1% 1.4325i 1.4325j 1.1600hi 1.1075ghi 1.0225f 1.0400fg 0.5% 1.5200ghi 1.5100ij 1.1600hi 1.0900hi 1.0275f 1.0200g 1.0% 1.5425ghi 1.535 0jhi 1.2125fghi 1.0750i 1.0200f 1.2100bcd 1.5% 1.9211d 1.8806de 1.4982bc 1.1828fgh 1.0597cdef 1.0884efg Publix Soap Control 1.5000hi 1.4987j 1.1675hi 1.1938fgh 1.0313ef 1.0513fg 0.1% 1.5350ghi 1.5300jhi 1.2025fghi 1.2125efg 1.0575cdef 1.1600cde 0.5% 1.6525fg 1.6475gh 1.4075cde 1.2450def 1.0725cdef 1.2025bcd 1.0% 1.8925de 1.8500de 1.6125ab 1.3050cde 1.1350ab 1.0025g 1.5% 1.9025d 1.8600de 1.6500a 1.3575bc 1.1600a 1.2300bcd Wetcit Control 1.8425de 1.8187ef 1.2475fgh 1.4300b 1.0275f 1.2313bc 0.1% 1.7600ef 1.7075fg 1.2900efg 1.4100bc 1.0350def 1.2900ab 0.5% 2.0700c 1.9550cd 1.4250cd 1.3975bc 1.0675cdef 1.2500bc 1.0% 2.2950b 2.1275b 1.6075ab 1.6525a 1.0625cdef 1.2575bc 1.5% 2.2600b 2.0925b 1.6725a 1.5575a 1.0300ef 1.3025ab Vapor Gard Control 1.5300ghi 1.5262ij 1.2013ghi 1.1163ghi 1.0238f 1.0538fg 0.1% 1.6200fgh 1.6150ghi 1.1300i 1.1575fghi 1.0650cdef 1.0250g 0.5% 1.8125de 1.7850ef 1.3075def 1.1950fgh 1.0975bc 1.0325fg 1.0% 2.1875bc 2.0225bc 1.5200bc 1.3325cd 1.0900bcd 1.1325def 1.5% 2 .5775a 2.3100a 1.5200bc 1.3150cde 1.0675cdef 1.3650a Statistics F 33.00 20.08 25.72 8.92 11.58 12.25 df 12, 9408 12, 9408 12, 9408 12, 9408 12, 9408 12, 9408 P <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 <0.0001 Within a row, means followed by the same l etter are not significantly different (P> 0.05; Tukey Kramer Grouping for Least Squares Means).

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84 Table 2 3. Average coleus cutting root length pe r biorational material Treatment (Mean + SE) Publix Soap (Mean + SE) Wetcit TM (Mean + SE) Vapor Gard (Mean + SE) Control 17.7900 + 0.5303a 17.2900 + 0.5303a 15.4300 + 0.5303ab 18.1200 + 0.5303a 0.1% 15.4000 + 0.5303b 16.3300 + 0.5303a 16.6100 + 0.5303a 18.7900 + 0.5303a 0.5% 15.8900 + 0.5303b 17.0900 + 0.5303a 14.7000 + 0.5303ab 18.6400 + 0.5303a 1.0% 17.3400 + 0.5303ab 16.9900 + 0.5303a 14.1400 + 0.5303b 17.2100 + 0.5303ab 1.5% 16.3000 + 0.5303ab 15.8900 + 0.5303a 14.5600 + 0.5303ab 15.2300 + 0.5303b F 3.79 1.95 3.55 7.88 df 5, 216 5, 216 5, 216 5, 216 P 0.0026 0.0879 0.0041 <0.0001 Within a row, means followed by the same letter are not significantly different (P> 0.05; Tukey Kramer Grouping for Least Squares Means).

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85 CHAPTER 3 EVALUATION OF THE MODEL BIORATIONAL DIP Introduction From the late 1980s to early 1990s, several studies explored the efficacy of fluvalinate and other synthetic insecticides as an adoptable post harvest treatment for mealybug infested ornamental products in Florida and Hawaii, respectively (Osborne 1986, Hata et al. 1986, Hansen et al. 1992). Several additional stud ies evaluated factors that impacted the ease and efficacy of synthetic dip treatments for ornamental and agricultural products infested with other cryptic, invasive pests. Aside from pesticide type and concentration, Mann et al. (1995) and Hara et al. (199 6) evaluated dipping temperature and Hansen et al. (1992), Hata et al. (1993), Tenbrink et al. (1992), Osborne (1986) studied dipping technique, which encompassed the length of dipping time, submersion depth, or agitation intensity. Dip efficacy was also evaluated across numerous ornamental and agricultural pests, various types of plants, and dip treatment characteristics, such as the type of treatment used and treatment concentration (Table 3 1, Table 3 2). The timing of when a dip treatment occurred wit hin a season or treatment schedule was also analyzed by Hansen et al. (1992), Hata and Hara (1993) and Hata et al. (1993). Overall, fluvalinate has been the most extensively reviewed treatment for use on various pests, plants, and concentrations. However, further research is needed to determine how a model biorational dip treatment may serve as an effective and reliable treatment for infested cuttings. As determined from the was determined as a model dip treatment due to low phytotoxicity and high P. madeirensis mortality. examination include dipping time and impact of various ho st plants. Determining how these

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86 factors impact dipping efficacy elucidates a more effective method for greenhouse and nursery growers to treat cuttings infested with cryptic, invasive pests such as P. madeirensis Materials and Methods Rearing Colonies of P. madeirensis The procedure for rearing colonies of P. madeirensis was described in the materials and methods section from the previous chapter and was used for the exposure time and host plant efficacy bioassays conducted at the University of Florida Mi d Research and Education Center (UF MREC) in Apopka, FL. Host p lant m aintenance The host plant maintenance protocol for Solenostemon spp. exposure time bioassa y. For the host plant efficacy bioassay, Mentha spp. (spearmint) and Verbena spp. (verbena) stock plants were grown at UF MREC. Mint and verbena stock plants were grown from cuttings in an enclosed greenhouse from January to May 2013 with a 22.8 0 C average daily temperature and 68% mean relative humidity at UF MREC. Cuttings were rooted in 64 count cutting trays filled with Faford Growing Mix 2/C 2 soil composed of Canadian sphagnum peat moss, perlite, vermiculite, dolomitic limestone, and wetting agent (2: 1:1:1:1, vol:vol:vol:vol:vol). Mint and verbena stock plants were hand watered daily until soil was evenly moistened and flowering parts were pruned to encourage further branching. Cutting p reparation Coleus cuttings were prepared using the same procedur e described in the materials and methods section from the previous chapter. However, mint and verbena cuttings were made by randomly selecting and excising the stem of a stock plant at a slight angle in order to prevent stem damage. All mint and verbena cu ttings were at least 3 inches (7.62cm) in height. Mint or verbena cuttings with blemishes that resembled phytotoxicity characteristics

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87 were discarded. Additionally, mint and verbena cuttings were made once the stock plant was over 6 inches (15.24cm) in hei ght from the 64 count cutting tray. P. madeirensis Mortality 1 second, 15 seconds, 30 seconds, 60 seconds, and 120 seconds Three replicates were conducted control, respectively. For each replicate, a total of 30 cuttings, each infested with 15 P. madeirensis mealybugs ranging from crawlers to virgin female adults, were evaluated for mortality. Data collection for each replicate began on a separate date and time: 11 April 2013, 19 April 2013, and 3 May 2013. Dip t reatment p reparation. 0mL of ionized water) were prepared and thoroughly mixed with a magnetic stirrer for five minutes. Five 1000mL of de ionized water was used as a control. Coleus c utting p reparation. The protocol for preparing coleus cuttings was described in the evaluation of biorationals on P. madeirensis mortality section from the previous chapter. Dipping m ethod and p lanting p rocedure Five infested coleus cuttings were randomly ol, respectively. Cuttings were dipped simultaneously in a 6.5 quart rectangular plastic bin. For the duration of each dip treatment, cuttings were fully submerged, lightly agitated, and rotated vertically at the middle of the designated exposure time to m aximize surface area coverage. Dipped cuttings were lightly shaken to remove excess runoff and air dried until runoff ceased. Each cutting was transplanted into a labeled twelve inch plastic pot filled with moistened Faford Growing Mix 2/C 2 soil.

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88 Cuttin g o bservation and w atering p ractices. The procedure for observing and watering cuttings was described in the evaluation of biorationals on P. madeirensis mortality section from the previous chapter. Mealybug m ortality e valuation. The method for evaluating P. madeirensis mortality was also explained in the evaluation of biorationals on P. madeirensis mortality section from the previous chapter. P. madeirensis Mortality on Various Host Plants Two types of host plants were used for the host plant efficacy bioassays: mint and 1.0%, and 1.5%. Three replicates for each host plant was conducted in a 5 X 5 randomized complete block design. F or each replicate, a total of 25 cuttings, each infested with 15 P. madeirensis mealybugs ranging from crawlers to virgin female adults, were evaluated for mortality. Data collection for each verbena replicate began on 8 May 2013 in the morning, 8 May 2013 in the afternoon, and 9 May 2013 in the morning. Data collection for each mint replicate began on 9 May 2013 in the afternoon, 10 May 2013 in the morning, and 10 May 2013 in the afternoon. Dip t reatment p reparation For each host plant replicate, four c Oil were prepared and thoroughly mixed with a magnetic stirrer for five minutes: 0.10% (1mL of ionized L of de Oil per 985mL of de ionized water). One 1000mL of de ionized water was used as a control. Mint and v erbena c utting p reparation. To insure that cuttings had more than 15 mealybugs, mint and verbena plants w ere re infested by exposure to P. madeirensis from the colony maintained at UF MREC one week before the trial. Infested cuttings were made by

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89 excising the stems at a 45 o angle to avoid stem damage. Cuttings were at least 3 inches (7.62cm) in height. Exactl y 15 mealybugs were randomly selected to remain on the cutting. Mealybugs left on cuttings ranged from crawlers to adult females. Excess mealybugs, oviscas, male adults, or male cocoons were removed using a moistened camel hair paintbrush. If there were le ss than 15 mealybugs present on a cutting, mobile mealybugs from different colony leaves that varied in size and ranged from crawlers to adult females were randomly selected and transferred onto the cutting using a moistened camel hair paintbrush. All tran sferred mealybugs were left to settle on the cutting for at least fifteen minutes. All cuttings had a random number of active mealybug life stages present. A total of 15 mealybugs were left on each cutting. Each cutting was carefully sealed in a clear Zipl oc bag to maintain cutting moisture and reduce the risk of losing mealybugs when transporting the cutting to the greenhouse for treatment. Dipping m ethod and p lanting p rocedure. The method for evaluating P. madeirensis mortality was explained in the phyto toxicity assessment section from the previous chapter. Cutting o bservation and w atering p ractices. The procedure for observing and watering cuttings was described in the evaluation of biorationals on P. madeirensis mortality section from the previous chapt er. Mealybug m ortality e valuation. The protocol for determining mealybug mortality was explained in the evaluation of biorationals on P. madeirensis mortality section from the previous chapter. Statistics Summary statistics for the exposure time and host p lant efficacy bioassay were prepared using a fit linear model in the GLIMMIX Procedure (General Linear Model for Mixture Distributions) to account for non normality and non homogenous variances in SAS 9.3 (SAS Institute Inc., 2002 2010, Cary, NC, USA). For the exposure time bioassay, three replicates were

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90 Oil and the control, respectively. For each replicate, a total of 30 cuttings, each infested with 15 P. madeiren sis mealybugs, were evaluated for mortality. For the host plant efficacy bioassay, three replicates for each host plant was performed in a 5 x 5 randomized complete block design. Each replicate had a total of 25 cuttings where each cutting was infested wit h 15 P. madeirensis mealybugs and evaluated for mortality. Significant difference between each time period the average number of dead P. madeirensis per treatment over each time period and the percentage of dead P. madeirensis per treatment over each tim e period were analyzed using Tukey Kramer Least Squares Means for multiple comparisons (SAS Institute Inc., 2002 2010, Cary, NC, USA). Results Exposure t ime b ioassay. The cumulative average percent of dead P. madeirensis was significantly different between each exposure time ( F=2.02; df=4; p=0.0887) ( Table 3 3 ) .Across each time period for 1.0% exposure time had the highest percentage of dead P. madeirensis All control exposure times had a lower percentage of dead P. madeirensis Efficacy bioassay using v erbena. showed a significant difference in the cumulative average percent of dead P. madeirensis between each post treatment time period after bein g used as a dip for one minute ( F=53.22; df=4; p<0.0001 ) ( Table 3 4 ). At each time period percentage of dead P. madeirensis Efficacy bioassay us ing m int. The cumulative average percent of dead P. madeirensis was significantly different post treatment time period after being used as a one minute dip ( F=78.50; df=4; p<0.0001 ) ( Table 3 5). at 1.5 %

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91 had the highest percentage of dead P. madeirensis concentrations at each time period P. madeirensis than 0.1% Na The contro l consistently had the lowest percentage of dead P. madeirensis at each time period Discussion Exposure t ime b ioassay exposure times for controlling P. madeirensis on coleus was from 30 to 120 seconds since efficacy for each exposure time surpassed 70% mortal ity by Day 7. Initially on Day 1 mortality exceeded 35% mortality for each aforementioned exposure time whereas dips achieved significantly less than 30% mortality. The 15 and 1 second dip only exceeded 70% mortality by Day 14. The highest mortality for all exposure times occurred by Day 14 where all 80% mortality Differences in P. madeirensis mortality over time between ex posure times may be attributed to the mechanical effects of dipping, susceptibility of P. madeirensis at various life stages, and most importantly, In the previous chapter, the mechanical act of dipping and tre atment susceptibility differences at various instars of P. madeirensis were described in depth as potential factors that may have influenced differences in P. madeirensis mortality and caused high mortality in the water controls. These factors have also re mained relevant to the exposure time bioassay. Phenacoccus madeirensis mortality most likely differed between treatments as a result of amplified the likelihood wax removal, suffocation, or cell membrane disruption faster than shorter dip exposure times (Butler et al. 1993, Hodgson and Kuhr 1990, Larew and Locke 1990).

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92 Overall, the 30 second dip exposure time was determined as the ideal dipping time due to differences in phytotoxicity and P. madeirensis mortality. Only slight chlorosis, chlorotic flecking, and tip curling were readily observed for the 1, 15, and 30 second exposure times Howev er, the 60 and 120 second dip induced slight chlorosis, chlorotic flecking, necrotic flecking, tip curling, and holes. Additionally, P. madeirensis mortality between the 30, 60, and 120 second dip exhibited no significant difference The exposure time bio assay may be improved in several ways As described in the previous chapter, adjustments regarding the impact of mechanical dipping and evaluating each P. madeirensis life stage independent from one another remain applicable for future exposure time studie s. Additionally, each dip exposure time may be evaluated on separate days to reduce the frequency of transferring mobile mealybugs from various mealybug col ony leaves High water control mortality may also be prevented by having both raters only touch the mealybugs three times with a damp camel hair brush during the mealybug mortality evaluation Efficacy b ioassays u sing m int and v erbena concentrations for controlling P. madeirensis on verbena cuttin gs were 1.5% and 1.0%. On Day 1 slightly surpassed 70% mortality by Day 7. The highest mort ality for all treatments occurred by treatments that surpassed 90% mortality by Day 14. controlling P. madeirensis on mint cuttings. On Day 1 1.5% reached over 60% mortality while all other

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93 followed by 1.0% with 70% mortality. The highest mortality for all treatments occurred by Day exceeded 95% mortality by Day 14. P henacoccus madeirensis mortality between treatments for the verbena and mint efficacy bioassay may mostly be influenced by differences in concentration as opposed to the mortality evaluation and the act of agitating cuttings via the dipping or transplanting technique described in the efficacy bioassay discussion from the previous chapter. Increase in mo rtality with higher concentrations mirror results from Liu and Stansly (2000) where the 1.0% and 0.5% vegetable oil dip induced the silverleaf whitefly, Bemisia argentifolii Bellow & Perring, 53% and 60% mortality, respectively, on collard and tomato dippe d leaves. The mint and verbena host plant surface in addition to the repellant activity of mint may have also impacted differences in P. madeirensis mortality between host plants. Mint and verbena surface area varied in texture and plant hair presence thu different between host plants and may be attributed towards differences in efficacy (Odenwald and Turner 2006, Gilman 1999). Additionally, species in the genus Mentha have been known to exhibit repellant activity to several in sect species and therefore may have impacted the number of P. madeirensis found on each cutting over each time period (Ansari et al. 2000, Mekuaninte et al. 2011, Kumar et al. 2011). T he model dip concentration for various host plants was due to differences in phytotoxicity. Several phytotoxicity characters were present on mint and verbena flecking, holes, tip chlorosis, and eventually tip necr osis. Few phytotoxicity characters were

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94 As a result, the most practical application fro acceptable phytotoxicity expression. Thus greenhouse productivity may be higher due to t he Several improvements may be implemented towards prospective host plant studies. As described in the previous chapter, modifications regarding the impact of mechanical dipping and e valuating each P. madeirensis life stage independent from one another also remain applicable to future host plant studies. The addition of coleus as a host plant control in the mint and verbena host plant efficacy bioassay may also be implemented Model d ip p rotocol i mplications Preventing the importation of invasive species on cuttings has driven the need for using of an effective dip protocol with acceptable phytotoxicity. Results from this study showed that the dip protocol c s a 30 second dip. Reduction in exposure time from one minute to 30 seconds showed no significant difference in mortality and lower phytotoxicity. Additionally, more than 70% P. madeirensis mortality was achieved within seven days and expressed marginal ph ytoxicity. In future studies, the dip protocol c ould be evaluated on additional host plants, invasive species, and various dipping locations.

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95 Table 3 1. Dip treatment efficacy evaluated on various arthropod ornamental and agricultural pests. Pest Groups Scientific Name Common Name Dip Treatments Reference Ants Technomyrmex difficilis Forel White footed ant Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Aphids Aphi s gossypii Glover Cotton aphid Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Myzus persicae (Sulzer) Green peach aphid Mavrik Aquaflow Osborne 1986 Pentalonia n igronervosa Coquerel Banana aphid Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Fluvalinate, insecticidal soap, hot water treatment Hara et al. 1996 Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Mavrik Aquaflow, Tempo 2, Safer insecticidal Soap Hansen et al. 1992 Beetles Diaprepes abbreviatus L. Diaprepes root weevil Mavrik Aquaflow, Tempo 2, Safer insecticidal Soap Simanton and Bullock 1973 Phyllotocus ustulatus Blanch Agral, deltamethrin, fluvalina te, petroleum oil, bifenthrin Seaton et al. 1993 Earwigs Chelisoches morio (Fabricius) Black earwig Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Hard and Soft Scales Coccus viridis (Green) Green scale Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Mesolecanium nigrofasciatum (Pergande) Terrapin scale Soybean oil, petroleum oil Pless et al. 1995

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96 Table 3 1. Continued Pest Groups Scientifi c Name Common Name Dip Treatments Reference Hard and Soft Scales Pseudaulacaspis cockerelli (Cooley) False oleander scale Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Quadraspidiotus perniciosus Comstock San Jose scale Soybean oi l, petroleum oil Pless et al. 1995 Mealybugs Nipaecoccus nipae (Maskell) Coconut mealybug Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Phenacoccus solani Ferris Solanum mealybug Mavrik Aquaflow Osborne 1986 Planococcus citri (R isso) Citrus mealybug Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Pseudococcus affinis (Maskell) Obscure mealybug Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Pseudococcus lon gispinus (Targioni Tozzetti) Longtailed mealybug Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Mavrik Aquaflow, chlorpyrifos Hata et al. 1992 Mites Panonych us ulmi (Koch) European red mite Soybean oil, petroleum oil Pless et al. 1995 Sunspray Ultra Fine Agnello et al. 1994

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97 Table 3 1. Continued Pest Groups Scientific Name Common Name Dip Treatments Reference Mites Tetranychus urticae Koch Two spotted spider mite Mavrik Aquaflow Osborne 1986 Moths Epiphyas postvittana Walker Lightbrown apple moth C15 Ampol CPD, C23 Ampol DC Tron NR Taverner et al. 1999 Thrips Frankliniella occidentalis (Pergande) Western flower thrips Insecticidal soap, isopropyl alco hol, insecticidal fog, hot water immersion Mann et al. 1995 Abamectin, chlorpyrifos, oil soap Hata et al. 1993 Scirtothrips cardamoni (Ramakr) Cardamom thrips Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Thrips palmi Karny Melo n thrips Insecticidal soap, isopropyl alcohol, insecticidal fog, hot water immersion Mann et al. 1995 Thrips Thrips palmi Karny Melon thrips Abamectin, chlorpyrifos, oil soap Hata et al. 1993 Whiteflies Bemisia argentifolii Bellow and Perring Silverleaf whitefly Sunspray oil, M Pede, extract of Nicotiana, bifenthrin Liu and Stansly 1995 M Pede, Sunspray oil, Margosan O, bifenthrin Liu and Stansly 1995 Tialeurodes vaporariorum (Westwood) Greenhouse whitefly Mavrik Aquaflow Osborne 1986

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98 Table 3 2. Dip treatment efficacy evaluated on various ornamental and agricultural plants. Scientific Name Dip Treatment Reference Alinia purpurata (Vieill.) K. Schum Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Orthene T, T&0, Talstar F, Tempo 2EC, Tame 2.4EC Hata and Hara 1993 Fluvalinate, insecticidal soap, hot water treatment Hara et al. 1996 Mavrik Aquaflow, chlorpyrifos, insecticidal soap Hata et al. 1992 Anthurium sp. L. Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992 A nthurium andraeanum Linden Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Arundina graminifolia (D. Don) Hochr Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Calathea insignis Petersen Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Calathea lancifolia Boom Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992 Chamelaucium uncinatum Schuaer Agral, deltamethrin, Mavrik Aquaflow, pe troleum oil, bifenthrin Seaton et al. 1993 Cissus rhombifolia Vahl Mavrik Aquaflow Osborne 1986. Citrus sp. L. C15 Ampol CPD, C23 Ampol DC Tron NR Taverner et al. 1999 Mavrik Aquaflow, Tempo 2, Safer insecticidal Soap Simanton and Bullock 1973 Codiaeu m variegatum (L.) Blurne Mavrik Aquaflow Osborne 1986. Cordyline terminalis (L.) Kunth Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992 Mavrik Aquaflow Osborne 1986. Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Curculig o capitulate (Lour.) Kuntze Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Cycas circinalis L. Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992

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99 Table 3 2. Continued Scientific Name Dip Treatment Reference Cyc as revolute Thunb. Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Dendrobium phalaenopsis Fitzg. Insecticidal soap, isopropyl alcohol, insecticidal fog, hot water immersion Mann et al. 1995 Abamectin, chlorpyrifos, oil soap Hata et al. 1993 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Dicranopteris linearis (Burm.f.) Underw. Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992 Dieffenbachia maculate (Lodd.) G. Don. Mavrik Aquaflow Osborne 1986. Dracae na fragans (L.) Ker Gawl Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Dracaena marginata Lam. Mavrik Aquaflow Osborne 1986. Epipremnum aureum (Linden & Andre) Bunt. Mavrik Aquaflow Osborne 1986. Mavrik Aquaflow, Tempo 2, Safer I nsecticidal Soap Hansen et al. 1992 Ficus pumila L. Mavrik Aquaflow Osborne 1986. Gleichenia linearis (Brum. f.) Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Glycine max L. Soybean oil, petroleum oil Pless et al. 1995 Hedera hel ix L. Mavrik Aquaflow Osborne 1986. Hoya carnosa (L.f.) R. Br. Mavrik Aquaflow Osborne 1986. Lycopodium cernuum L. Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992 Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Maranta leu coneura E. Morr. Kerchoviana Mavrik Aquaflow Osborne 1986. Monstera deliciosa Liebm. Mavrik Aquaflow, Tempo 2, Safer Insecticidal Soap Hansen et al. 1992 Perperomia obtusifolia (L.) A. Dietr. Mavrik Aquaflow Osborne 1986. Philodendron scandens oxycardiu m (Schott) Bunt. Mavrik Aquaflow Osborne 1986.

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100 Table 3 2. Continued Scientific Name Dip Treatment Reference Solanum lycopersicum L. Sunspray oil, M Pede, extract of Nicotiana, bifenthrin Liu and Stansly 1995 M Pede, Sunspray oil, Margosan O, bifenth rin Liu and Stansly 1995 Strelitzia reginae Aiton Insecticidal soap, Mavrik Aquaflow Tenbrink et al. 1992

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101 Table 3 3 Cumulative average percent of dead P. madeirensis per coleus cutting dipped in varying dip exposure times for each time period. Treat ment Pesticide Time Period (Days) 1 (Mean + SE) 3 (Mean + SE) 7 (Mean + SE) 14 (Mean + SE) 1 second 24.78% + 2.225%c 41.62% + 2.822%c 60.53% + 2.781%c 81.28% + 1.849%c Control 6.296% + 0.8548%e 12.69% + 1.560%e 23.82% + 2.492%e 46.96% + 3.333%e 15 seconds 32.96% + 2.621%bc 51.54% + 2.921%bc 69.59% + 2.512%bc 86.63% + 1.444%bc Control 8.798% + 1.099%d 17.27% + 1.897%d 30.99% + 2.769%d 55.97% + 3.146%d 30 seconds 35.37% + 2.725%abc 54.21% + 2.920%abc 71.81% + 2.420%abc 87.82% + 1.346%abc Control 11.34 % + 1.336%d 21.68% + 2.185%d 37.32% + 2.935%d 62.78% + 2.917%d 60 seconds 36.21% + 2.751%ab 55.12% + 2.918%ab 72.54% + 2.395%ab 88.21% + 1.315%ab Control 6.562% + 0.8432%e 13.19% + 1.524%e 24.63% + 2.398%e 48.07% + 3.143%e 120 seconds 46.59% + 2.937%a 65.37% + 2.677%a 80.24% + 1.928%a 92.00% + 0.9454%a Control 12.87% + 1.514%d 24.21% + 2.410%d 40.73% + 3.112%d 66.06% + 2.892%d Within a column, means followed by the same letter are not significantly diff erent (P> 0.05; Tukey Kramer Grouping for Least Squares Means; F=2.02; df=4, 2239; p=0.0887).

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102 Table 3 4 Cumulative average percent of dead P. madeirensis concentration for each time period Time Period (Days) 1 (Mean + SE) 3 (Mean + SE) 7 (Mean + SE) 14 (Mean + SE) Control 5.123% + 0.7634%d 10.82% + 1.456%d 31.47% + 2.976%d 51.05% + 3.271%d 0.1% 21.82% + 2.349%c 38.54% + 3.187%c 70.35% + 2.847%c 84.35% + 1.920%c 0.5% 28.26% + 2.556%bc 46.95% + 3.036%bc 77.01% + 2.274%bc 88.38% + 1.460%bc 1.0% 32.60% + 2.625%ab 52.08% + 2.881%ab 80.44% + 1.977%ab 90.33% + 1.230%ab 1.5% 41.63% + 2.880%a 61.58% + 2.777%a 85.85% + 1.585%a 93.23% + 0.9172%a Within a column, means follow ed by the same letter are not significantly different (P> 0.05; Tukey Kramer Grouping for Least Squares Means; F=53.22; df=4, 1115; p <0.0001).

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103 Table 3 5 Cumulative average percent of dead P. madeirensis per mint cutting dipped for one minute in each Na concentration for each time period Time Period (Days) 1 (Mean + SE) 3 (Mean + SE) 7 (Mean + SE) 14 (Mean + SE) Control 7.337% + 1.1017%d 17.66% + 2.039%d 33.65% + 2.919%d 58.29% + 3.096%d 0.1% 19.88% + 2.212%c 40.20% + 3.2 11%c 61.39%% + 3.158%c 81.41% + 2.167%c 0.5% 43.64% + 3.135%b 67.71% + 2.822%b 83.22% + 1.922%b 93.18% + 0.9848%b 1.0% 49.07% + 3.011%b 72.30% + 2.477%b 86.06% + 1.621%b 94.45% + 0.8057%b 1.5% 63.85% + 2.994%a 82.71% + 1.964%a 91.88% + 1.113%a 96.89% + 0.5003%a Within a column, means followed by the same letter are not significantly different (P> 0.05; Tukey Kramer Grouping for Least Squares Means; F=78.50; df=4, 1116; p <0.0001)

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104 CHAPTER 4 IMPLICATIONS AND FUTURE DIRECTIONS Florida greenhouse operatio ns are susceptible to invasive pests due to the warm, humid climate and the high movement of people, tourism and product shipments along 30 ports of entry (U.S. Department of Homeland Security 2013). Pest management and control for invasive and native orna mental pests costs Florida growers 16 20% of their production expenses (Hodges 1998). As a result, further research to control invasive species is needed to meet the n eed for clean ornamental commodities. In this study, P. madeirensis w as used for modeling the control of other invasive cryptic species not known to occur in the United States for several reasons. Hemiptera is the most commonly intercepted order at United States ports of entry (McCullough et al. 2006, Jenkins et al. 2014). As an invasive, cosmopolitan mealybug P. madeirensis is one of several successfully established invasive species in Florida (Frank and Thomas 2004). Scales and mealybugs continue to be a top concern in relation to the movement of cryptic species. Crawlers and nymphs are small and easily hidden on the underside of leaves, notches created by plant nodes or buds on the stems, or crevices on the leaves and stems (Buss et al. 1992, Hollingsworth and Hamnet 2009). The use of synthetic insecticide dips as a con trol for scales and mealybugs on ornamental commodities has been extensively analyzed with fluvalinate (Osborne 1986, Hata et al. 1992, Hansen et al. 1992) (Table 3 1). However, mealybugs such as Phenacoccus madeirensis have been one of the most difficult mealybugs to manage with synthetic insecticides (Chong 2005, Ludwig 2009). Resistance and overuse of synthetic insecticides coupled with the demand for using a cost effective, environmentally safe, non phytotoxic and efficacious treatment for

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105 controlling i nvasive species on ornamental stock has led to evaluating biorational insecticides as a viable treatment. To determine a model biorational dip, four insecticides were used as a one minute dip and evaluated for efficacy and phytotoxicity at varying concentr ations on coleus cuttings infested with P. madeirensis Gard (di 1 p menthene), and Wetcit TM (alcohol ethoxylate). Six foliage phytotoxicity characters were key indicators: chlorosis, chlorotic flecking, necrotic flecking, tip chlorosis, tip necrosis, and holes. The highest concentration for each biorational insecticide with acceptable Vapor Gard. F inal determination for the model biorational dip was based on using each selected biorational insecticide for controlling coleus cuttings infested with P. madeirensis Modifications to the mealybug mortality protocol from Hata et al. (1992) were used to de termine that the (90.47%), followed by 1.0% Publix Soap (86.17%), 0.1% Vapor Gard (85.72%), and the control el dip treatment over 0.1% phytotoxicity character and has been known to effectively control various ornamental pests. Liu and Stansly ( 2000 ) Pless et al. ( 1995 ) Butler et al. ( 1993 ) and Amer et al. ( 2001 ) reported soybean oil at the recommended dosage rate as an efficacious insecticidal control for the silverleaf whitefly, Bemisia argentifolii Bellow & Perring, sweetpotato whitefly, Bemisia tabaci (Gennadius), San Jose scale, Quadraspidiotus perniciosus (Comstock), terrapin scale, Mesolecanium nigrofasciatum (Pergande), European red mite, Panonychus ulmi (Koch), and two spotted spider mite, Tetranychus urticae Koch. In comparison several studies only observed

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106 alcohol e thoxylate insecticidal activity (Liu and Stansly 2000, Imail et al. 1994, Davidson et al. 1991, Hesler and Plapp 1986, Tattersfield and Gimingham 1927, Wolfenbarger et al. 1967, Imail and Tsuchiya 1995, Cory and Langford 1935) Dipping time and host plant efficacy were also tested with the model biorational dip evaluated for efficacy at different exposure dip times and on various types of host plant s. Results for the the 30 (87.82%), 60 (88.21%), and 120 (92.00%) second dip. Host plant efficacy results differed between mint and verbena. Results for mint cuttings showed the highest mortality for 1.5% determined as the model dip protocol for treating orname ntal cuttings due to phytotoxicity. At the shortest exposure time, the 30 second dip showed the most acceptable phytotoxicity of the three highest mortality dipping times. Additionally, mint and verbena cuttings treated with 1.5% e phytotoxicity: moderate chlorosis, chlorotic flecking, necrotic flecking, hole, tip chlorosis, and tip necrosis. However, only marginal phytotoxicity was a nd tip chlorosis. Developing a model dip protocol provides Florida growers a viable, on site treatment option for importing clean ornamental commodities. Results from this study showed 1.0% P. madeirensis mortality by Day 7 for c oleus and verbena cuttings and Day 3 for mint cuttings. As a single on phytotoxicity and greatly reduced the number of P. madeirensis within a nursery operation after

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107 fourteen days. As a result, surveys similar t o Hodges et al. (1998) and Hodges (2011) could evaluate Florida growers on the demand and application of using the model dip a s part of a systems approach Future on site dip studies could with other synthe tic insecticide applications in ter ms of percent mortality since 98% of all Florida nurseries use at least one synthetic insecticide (Hodges et al. 1998, Hodges 2011). Modifications to the model dip protocol may be warranted for developing a postharvest di p protocol as described in Hata et al. (1986 ) Several invasive, cryptic ornamental pests should also be evaluated in future efficacy studies with the model dip protocol. Hemiptera is the most commonly intercepted order at United States ports of entry and includes several of the most damaging ornamental pest groups: scales, mealybugs, whiteflies, and aphids (McCullough et al. 2006, Jenkins et al. 2014, Oetting et al. 2006). As a result, model pests for future efficacy studies could include the following: th e pink hibiscus mealybug, Maconellicoccus hisutus (Green), passionvine mealybug, Planococcus minor (Maskell), lobate lac scale, Paratachardina pseudolobata (Kondo and Gullan), melon aphid, Aphis gossypii Glover, green peach aphid, Myzus persicae (Sulzer), sweetpotato whitefly, Bemisia tabaci Gennadius, and citrus mealybug, Planococcus citri (Risso) (Table 1 1) (Frank and Thomas 2004, McCullough et al. 2006, Oetting et al. 2006, Jenkins et al. 2014 ). More work is also needed to assess phytoxicity on other host plant cuttings. Additional surveys similar to Hodges et al. (1998) and Hodges (2011) could also evaluate Florida growers on the type, number, and buyers of host plant cuttings. For future phytotoxicity assessments with P. madeirensis cuttings made from the most commonly infested P. madeirensis host plants in Florida could be evaluated: Hibiscus spp., Acalypha spp., Mandevilla spp., Jatropha spp., Salvia spp., C estrum spp., Plectranthus spp., Ruellia spp., Leucophyllum spp., Bidens spp., Lantana

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108 spp. Schefflera spp., Orthosiphon spp., Crossandra spp., Capsicum spp., Chrysanthemum spp., Sida spp., Pelargonium spp., Thunbergia spp., Pentas spp., and Solanum spp. (Stocks 2012). I n conclusion preventing the spread of invasive species on ornamental cuttings m ay be achieved by c arefully modifying and integrating the d ip protocol w ithin a systems approach However, m ore phytotoxicity and efficacy information is needed to refine the dip protocol for field application. P hytotoxicity assessments on a variety of ornamental cuttings and efficacy a nalysis on several o rnamenta l invasive pests should be conducted in the future Results from bot h studies may ultimate ly be used to develop a comprehensive dosage rate list for acceptable phytotoxicity damage and effective invasive pest control for imported cuttings

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124 BIOGRAPHICAL SKETCH Sarahlynne Guerrero grew up in Palm Bay, FL and graduated from the Sebastian River High School International Baccalaureate Program in May of 2007. After graduation, she enrolled at the University of Florida and majored in Entomology and Nematology under the encouragement of Drs. Carl Barfield and Rebecca Baldwin. During this time, her interest in biosecurity and invasive species blossomed through her United Sta tes Dep artment of Agriculture, Animal and Pla nt Health Inspection Services, Plant Protection and Quarantine student internship, where she proudly served as assistant to the Regional Identifier, Julieta Brambila, and member of the Florida Cooperative Agricultural Pest Survey Program under Dr. Leroy Whilby. While at UF, Sarahlynne also joined the laboratory of Dr. Robert Meagher and worked on evaluating the efficacy of invasive moth traps for her senior project. Sarahlynne was offered a graduate research assistantsh ip at UF beginning January 2012 under the direction of Dr. Amanda Hodges. While at UF, her research project was co advised by Drs. Amanda Hodges and Lance Osborne. Her project mirrored her strong interest in preventing the dissemination of invasive speci spread of invasive pests through her work with Dow AgroSciences. She began working in the structural fumigation market for Dow AgroSciences in California on September 2013.