Title: Florida Entomologist
Full Citation
Permanent Link: http://ufdc.ufl.edu/UF00098813/00021
 Material Information
Title: Florida Entomologist
Physical Description: Serial
Creator: Florida Entomological Society
Publisher: Florida Entomological Society
Place of Publication: Winter Haven, Fla.
Publication Date: 2004
Copyright Date: 1917
Subject: Florida Entomological Society
Entomology -- Periodicals
Insects -- Florida
Insects -- Florida -- Periodicals
Insects -- Periodicals
General Note: Eigenfactor: Florida Entomologist: http://www.bioone.org/doi/full/10.1653/024.092.0401
 Record Information
Bibliographic ID: UF00098813
Volume ID: VID00021
Source Institution: University of Florida
Holding Location: University of Florida
Rights Management: Open Access

Full Text

Michaud: Cotton as Host Plant for Brown Citrus Aphid


University of Florida, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL 33850

Current Address: Kansas State University, Agricultural Research Center-Hays, 1232 240th Ave., Hays, KS 67601


Seven populations of Toxoptera citricida (Kirkaldy) were sampled in central Florida sweet
orange groves in 2001. All populations contained individuals that accepted cotton seedlings
as a host in a no-choice situation; many of these matured and deposited nymphs that also de-
veloped and became reproductive on the same plant. Significant differences were noted
among populations with respect to the proportion of nymphs accepting, maturing, and ulti-
mately reproducing on cotton. Differences in aphid survival were largely a function of differ-
ences in host plant acceptance, rather than differential mortality on the plant. A significant
proportion of the apterous adults maturing on cotton abandoned the plant without reproduc-
ing. Second and third instars transferred from laboratory colonies maintained on sweet or-
ange were more accepting of cotton than were either first or fourth instars. Apterous adults
accepted cotton at rates similar to second and third instars. Alate adults settled on cotton
seedlings in greenhouse choice experiments and probed the plants, but none deposited
nymphs. Alatae that matured on cotton readily accepted citrus for feeding and reproduction.
It is concluded that cotton may be useful as a factitious host plant for rearing T citricida in
the laboratory, but field planted cotton is unlikely to serve as a reservoir of the aphid in re-
gions where citrus is grown.

Key Words: Gossypium hirsutum, host plants, reproduction, survival, Toxoptera citricida.


Siete poblaciones de Toxoptera citricida (Kirkaldy) fueron muestreadas en huertos de naran-
jas dulces en Florida central en 2001. Todas las poblaciones tenian individuos que aceptaron
plantulas de algod6n como una hospedera en una situaci6n de una sola opci6n; muchas de
estas maduraron y depositaron ninfas que tambien se desarrollaron y se reproducieron en la
misma plant. Diferencias significativas fueron notadas entire las poblaciones con respeto a
la proporci6n de las ninfas que aceptaron, maduraron, y finalmente se reproducieron en el
algod6n. Las diferencias en la sobrevivencia de los afidos fueron mayormente en funci6n de
las diferencias en aceptar la plant como una hospedera, y no debido a la mortalidad dife-
rential en la plant. Una proporci6n significant de los adults apteros maduraron en el al-
god6n y abandonaron la plant sin reproducirse. Las ninfas en el segundo y tercero estadio
transferidos de las colonies de laboratorio mantenidos en naranjas dulces fueron mas recep-
tivas al algod6n que las ninfas en el primero o cuatro estadio. Los adults apteros aceptaron
el algod6n en las proporciones similares de las ninfas en el segundo y tercero estadio. Los
adults alados posaron sobre las plantulas de algod6n en experiments de selecci6n en el in-
vernadero y probaron las plants, pero ninguno deposit ninfas. Los adults alados que ma-
duraron sobre el algod6n aceptaron con rapidez el citrico para alimentarse y reproducirse. Se
concluye que el algod6n puede ser util como una plant hospedera facticiosa para criar T cit-
ricida en el laboratorio, pero es poco possible que el algod6n sembrado en el campo servird
como un refugio del afido en regions donde se siembra los citricos.

The brown citrus aphid, Toxoptera citricida United States including Louisiana, Texas, Arizona
(Kirkaldy) (BCA), is the primary vector of citrus and California. Although it has been present in
tristeza virus (CTV), one of the important diseases Belize, Central America, since 1996 (Halbert 1996),
of citrus world-wide (Meneghini 1946). Its impor- the Yucatan Peninsula was not infested until 1999
tance as a pest of citrus derives from its high effi- (Michaud & Alvarez 2000). Northerly movement
ciency in transmitting this virus, rather than from of the BCA has been also slow along the eastern
any direct damage (Michaud 1998). The BCA has seaboard of Mexico, and the major citrus-growing
been present in Florida since 1995, but remains states of Tabasco and Veracruz remain uninfested
absent from other citrus-growing regions of the to date. If and when further northerly movement

Florida Entomologist 87(2)

occurs, citrus plantings as far north as Texas could
be heavily impacted as most citrus in the region,
both north and south of the border, is planted on
sour orange rootstock. Various strains of CTV
cause "quick decline" of trees on sour orange and
this rootstock must be abandoned wherever the vi-
rus and its vector are present together. An area-
wide effort to identify and eliminate CTV-infected
trees prior to the arrival of BCA is the best strat-
egy for ameliorating the inevitable impact on the
citrus industry.
The performance of BCA has been compared
on various citrus varieties (Komazaki 1989) and
related species of Rutaceae (Tang et al. 1999), but
its ability to utilize non-rutaceous plants has not
previously been explored. Although a substantial
number of plants have been recorded as potential
hosts for BCA (Michaud 1998), the actual role of
these plants in supporting BCA populations in
the field is unknown. It is suspected that many
plants listed as hosts may represent mis-identifi-
cations of the aphid due to its similarity to the
black citrus aphid, Toxoptera aurantii (Boyer de
Fonscolombe), a related species with a very broad
host range (Halbert & Brown 1996). Observations
of BCA behavior suggest that anomalous host
plant associations may arise when high-density
populations 'overflow' from heavily infested citrus
trees. Crowding in BCA colonies stimulates alate
production (Michaud 2001), and large numbers of
reproductive apterae also emigrate from crowded
colonies (Michaud & Belliure 2000). These dis-
persing apterae ascend almost any other green
plant adjacent to the source tree and often settle
to feed. Residual nutrition acquired from the orig-
inal host plant may then permit some limited re-
production to continue on the colonized plant,
creating the semblance of host suitability. Thus,
discrete field observations of host plant associa-
tions can be misleading and careful laboratory
studies are required to determine whether a par-
ticular plant is truly a potential or suitable host.
The cotton plant, Gossypium hirsutum L., was
first reported as an occasional host plant of BCA
in southern Africa (Symes 1924) and later in Aus-
tralia (Carver 1978). The present study was un-
dertaken to evaluate the potential suitability of
cotton as a host plant for BCA for two reasons.
First, laboratory studies of the BCA and its bio-
logical control agents are hampered by the contin-
uous requirement for citrus trees with new
growth suitable for aphid colony growth and de-
velopment. These are expensive to acquire and
maintain, demand warm temperatures and in-
tense supplementary lighting in order to produce
new growth, and are susceptible to many other
pests in a greenhouse environment. If the BCA
could be reared effectively on a herbaceous host
plant that could be planted from seed as required,
laboratory studies of BCA biology and ecology
would be greatly facilitated. Second, the close

proximity of cotton plantings to citrus groves in
many regions of Texas and California raises the
question of whether or not cotton fields could po-
tentially support reservoir BCA populations that
could reinfest citrus, just as they now serve as a
reservoir for Aphis gossypii Glover, another vector
of CTV (Cisneros & Godfrey 2001).
The present study had three objectives: (1) to
assess the general acceptability and suitability of
cotton for various BCA populations in central
Florida, (2) to test whether acceptance of cotton,
and subsequent developmental performance, var-
ies with the growth stage of the aphid colonizing
the novel host, and (3) to determine whether alate
aphids developing on citrus would colonize cotton
and vise versa.


Variation among Populations in Acceptance of Cotton
in No-Choice Experiments

Preliminary work conducted by Dr. A. Chow in
Immokalee, FL indicated that BCA could be in-
duced to feed on cotton provided that very young
plants were provided and that relatively cool tem-
peratures were maintained. Seeds of cotton, Gos-
sypium hirsutum L., var "Suregrow", were
planted individually in plastic cones (20 cm ht x 4
cm diam) filled with Metromix 500 potting soil.
The cones were held at 24 2C in a climate-con-
trolled greenhouse under natural light until ger-
mination. Following germination of the cotton,
and before expansion of the first pair of true
leaves, cones were individually labeled and a
coating of Tanglefoot (The Tanglefoot Company,
Grand Rapids, MI 40504) was placed around the
inner rim of each.
Seven populations of BCA were sampled in
sweet orange groves in seven distinct locations in
Polk County, FL between 25-IX-2001 and 4-XI-
2001 by collecting a single, heavily-infested citrus
terminal from each grove and transporting it to
the laboratory in a 500-ml ventilated plastic con-
tainer. A series of 60 apterous, BCA 4th instars
were selected from each sample under a low
power stereo microscope and transferred with a
sable hair brush in groups of 5 to each of 12 cotton
seedlings. The seedlings were then placed in a
growth chamber set to 16:8, L:D period, 75% RH,
and a constant temperature of 20.0 1C. Each
replicate was examined once every 24 h and the
number of nymphs remaining on the seedling was
recorded, as was the number dying in the Tangle-
foot barrier. In addition, data were recorded on
the number of nymphs maturing to the adult
stage, the number of adults that reproduced, and
the number of second generation nymphs that
matured. The data were analyzed by one-way
ANOVA (SPSS 1998) followed by an LSD test for
separation of means (a = 0.05).

June 2004

Michaud: Cotton as Host Plant for Brown Citrus Aphid

Variation among Instars in Acceptance of Cotton
in No-Choice Experiments

A stock colony of BCA was initiated from mate-
rial field-collected in Polk County, FL, in March,
2002 and maintained on potted sweet orange, Cit-
rus sinensis L., var. "Pineapple" at 24 2C in a cli-
mate-controlled greenhouse under natural light.
Cotton seeds were planted individually in plastic
cones, germinated in the greenhouse, and Tangle-
foot was applied to the rim of the cone as above.
Colonies of BCA were removed from the stock lab-
oratory culture and the aphids separated accord-
ing to stage (nl-n4 and apterous adults) under a
10x stereo microscope. For each growth stage, five
aphids were transferred individually with a sable
hair brush to a single cotton seedling in each of 20
replicates. Any aphid suspected to have sustained
injury in the process of transfer was immediately
replaced. Since adults are not reproductive for
least 24 h following their last molt (Michaud
2001), pre-reproductive apterous adults were ob-
tained by removing all adults from a stock BCA
colony and, the next morning, harvesting all those
that molted to adult overnight. All experimental
replicates were maintained in a climate-controlled
growth chamber under the same conditions as de-
scribed above. Replicates were examined daily
and the following information was recorded: the
number of nymphs that settled and remained
feeding on the plant after 24 h, the number matur-
ing to the adult stage, and the number of adults
that became reproductive. For transferred adults,
only the number reproducing on the cotton was re-
corded. The data were analyzed by one-way
ANOVA (SPSS, 1998) followed by an LSD test for
separation of means (a = 0.05). Survival of first in-
stars was compared between experiments with a
Chi-square, Goodness-of-fit test.

Alate Acceptance of Host Plants in Choice Experiments

Alate aphids were produced in high-density
BCA colonies grown on potted sweet orange trees
in the greenhouse (as above). Sweet orange seed-
lings and cotton seedlings were planted individu-
ally in plastic cones as above. Orange seedlings
ca. 6 mo old with a single growing terminal were
used in experiments; cotton seedlings were 2-4
days old. The experiments were performed in the
greenhouse in wood frame cages (120 cm long by
65 cm wide by 80 cm high). Each cage was
screened with white muslin on the side panels
and had a clear plexiglass roof. In each trial (n =
12) the alate source consisted of a single 15-cm di-
ameter pot containing a sweet orange plant with
a mature BCA colony producing alatae. This alate
source was placed in the center of a cage with 4
trap plants in plastic cones (2 cotton seedlings
and 2 sweet orange seedlings) arranged equidis-
tant (40 cm) in an alternating sequence around

the source plant. After 24 h, the numbers of ala-
tae settling and feeding on each of the trap plants
were counted and the plants were replaced with
cotton and citrus in reversed positions in the cage.
In cases where alatae settled on a seedling, the
seedling was isolated in another cage and exam-
ined on subsequent days to determine whether or
not reproduction occurred.
Alate BCA were produced on cotton by trans-
ferring large numbers of reproductive apterous
adults from the stock colony to the potted cotton
seedlings and then moving them into a climate-
controlled growth chamber under the same condi-
tions as described above. The adult aphids were
left to reproduce for a period of 48 h whereupon
all adults were removed and first instar nymphs
were left in situ to complete development. A total
of 25 alatae produced on potted cotton seedlings
were caged individually on flushed sweet orange
terminals in the greenhouse in a muslin bag fas-
tened with a twist-tie at the base of the twig. Ob-
servations were then made at 24 and 48 h to
determine whether or not alates accepted the ter-
minal and deposited nymphs.


Variation among Populations in Acceptance of Cotton
in No-Choice Experiments

All seven populations of BCA sampled con-
tained some apterous fourth instars that accepted
cotton as a host plant (Fig. 1), but there was sig-
nificant variation among populations in the pro-
portion of aphids that accepted the cotton
seedling within the first 24 h (F = 3.708; 6,76 df;
P < 0.01). There were also significant differences
among populations in the number of individuals
molting to adult (F = 4.868; 6,76 df; P < 0.001) and
the number of adults reproducing (F = 3.706; 6,76
df; P < 0.01). Overall, a mean SEM of 21.4
1.8% of aphids accepted cotton, 16.4 1.6%
molted to adult, and 10.2 1.6% became repro-
ductive. Population 2 had the highest proportion
of individuals accepting, maturing and reproduc-
ing on cotton; only populations 1 and 6 had as
many individuals accepting cotton, but their suc-
cess in maturing and reproducing was signifi-
cantly lower than population 2. A total of 69
aphids matured to the adult stage and 43 of these
(63.2%) deposited at least one nymph before
abandoning the plant. Apterous adults that be-
came reproductive produced a mean (SEM) of
13.9 1.82 progeny (Fig. 2a) with no significant
difference among populations in adult fecundity
(F = 0.904; 6,26 df; NS) or in the number of prog-
eny maturing (F = 1.109; 6,26 df; NS). The mean
reproductive rate ranged from 0.4-2.2 nymphs/
adult/day of reproduction (mean = 0.94 0.23)
and the overall maturation rate of 2nd generation
nymphs was 33.8% (Fig. 2b).

Florida Entomologist 87(2)


40 -

30 -

20 -

10 -

S% accepting
S% maturing
*% reproducing






Fig. 1. Performance data for 4th instar Toxoptera citricida obtained from seven different populations in central
Florida and transferred to cotton seedlings (N = 12), five per plant. "% accepting" = percentage of aphids feeding on
the cotton seedling after 24 h, "% maturing" = percentage molting to adult,"% reproducing" = percentage depositing
at least one nymph following molt to adulthood. Means in columns bearing the same letter are not significantly dif-
ferent among populations in a one-way ANOVA followed by LSD (a = 0.05).

Variation among Instars in Acceptance of Cotton
in No-Choice Experiments
There were significant differences among in-
stars in the proportion maturing to adulthood
when transferred from citrus to cotton seedlings
(F = 8.622; 5,72 df; P < 0.001) and also significant
differences in the proportion becoming reproduc-
tive (F = 5.755; 5,92 df; P < 0.001). Second and
third instars were more likely to remain on a cot-
ton seedling, survive to adulthood, and become re-
productive than were either first or fourth instars
(Fig. 3). Experiment-wide, a total of 266 of the
aphids transferred as immatures molted to adult-
hood, and 190 of these (71.4%) deposited nymphs
before abandoning the plant. Pre-reproductive
adults transferred to cotton seedlings remained
to reproduce with a frequency similar to second
and third instars (Fig. 3).

Alate Acceptance of Host Plants in Choice Experiments
The proportion of nymphs maturing into ala-
tae versus apterae on the source plants varied
considerably under the conditions of these exper-

iments, largely due to variation in both the num-
ber of apterous adults accepting the cotton, their
distribution among the plants, and their repro-
duction during the 48-h period. High aphid den-
sity within colonies is the primary environmental
factor influencing wing development in BCA
(Michaud 2001), but high density colonies were
difficult to achieve on cotton seedlings, leading to
much lower rates of alate production than were
achieved on citrus. A total of 186 alate aphids set-
tled on plants and began feeding in the 12 repli-
cations of this experiment. Of these, 181 settled
and fed on a sweet orange seedling and 5 settled
and fed on a cotton seedling (Chi-square = 83.269,
P < 0.001). Since observations were made only
once every 24 h, it is possible that additional
alates settled on cotton seedlings for shorter peri-
ods without remaining to feed. Whereas 98.3% of
alates remained on orange seedlings long enough
to initiate reproduction, all five that settled on
cotton abandoned the plant within the following
24 h without depositing any nymphs. All 25 ala-
tae that were reared on cotton and then caged in-
dividually on sweet orange terminals accepted
the plant and began reproduction within 48-72 h.


June 2004

Michaud: Cotton as Host Plant for Brown Citrus Aphid


3 4


2 3

4 5

6 7

Fig. 2. Mean fecundities (+SEM) of Toxoptera citricida from seven field population that matured on cotton seed-
lings following transfer from citrus in the 4th instar (a), and proportions of the second generation nymphs that ma-
tured (b). There were no significant differences among populations (ANOVA, P > 0.05).


The fact that all sampled BCA populations
contained fourth instars able to feed, and ulti-
mately reproduce, on cotton suggests that the
physiological ability to utilize cotton as a host
plant is probably a general characteristic of T cit-
ricida populations. While it is not surprising that
considerable variation exists among populations
with respect to the acceptance of cotton, the po-
tential significance of this variation remains ob-
scure, given that reports of BCA attacking cotton
in the field are evidently rare (Symes 1924;

Carver 1978). However, BCA will also readily col-
onize Murraya paniculata (L.) Jack and Mal-
pighia punicifolia L. under laboratory conditions
(J. P. Michaud, unpublished) but rarely, if ever,
utilizes these plants in nature.
If alatae are more selective of their host than
are apterae, this could provide a partial explana-
tion of why potential host plants such as cotton
are rarely, if ever, utilized in the field. Alatae are
physiologically very different from apterae in
many ways. Their lower reproductive rate and
longer lifespan (Takanashi 1989) may afford
them more opportunity to be selective among host

0.4 I

0.2 -

0 4-



1 t I 1


Florida Entomologist 87(2)


b b E Maturing
80 Reproducing b

b b

c a

0 --



1st 2nd 3rd 4th Adult

Fig. 3. Performance data for Toxoptera citricida transferred from sweet orange to cotton seedlings at various life
stages. "% maturing" = percentage of aphids molting to adults, "% maturing" = percentage molting to adult, "% re-
producing" = percentage depositing at least one nymph as an adult. Means in columns bearing the same letter are
not significantly different among life stages in a one-way ANOVA followed by LSD (a = 0.05).

plants. Apterae may often be constrained to ac-
cepting sub-optimal plants when dislodged from
their primary host. Therefore, when reporting un-
usual host records for aphids it might be useful to
distinguish between alate-founded versus apter-
ous-founded colonies. Alate aphids are known to
settle and probe on many non-host plants. For ex-
ample, BCA alates probing soybean can contrib-
ute to transmission of soybean mosaic virus
without ever colonizing the plant (Halbert et al.
1986). Similarly, many apterous-founded colonies
on anomalous host plants may be chance events
without ecological significance for the aphid pop-
ulation. An alate-founded colony foundresss with
nymphs) is likely the best indicator of recurrent
host plant utilization in nature.
It is important to note that aphid death in the
first two experiments was almost invariably the re-
sult of aphids leaving the cotton seedling and dying
in the Tanglefoot barrier, rather than simply expir-
ing on the plant. Thus the differences observed in
'survival' and'maturation' are largely a function of
differential host plant acceptance, rather than dif-
ferential mortality on the plant. Of all nymphs re-
maining on the cotton seedling for the first 24 h in

the first experiment, more than three quarters ma-
tured and almost half became reproductive.
The variation in acceptance of cotton among
different BCA instars was neither positively nor
negatively correlated with aphid growth stage. If
aphid nymphs increasingly 'acclimated' to cotton
over the course of their development, one might
expect early instars to perform better than later
instars, but this was clearly not the case. Inter-
mediate instars had higher acceptance and better
performance on cotton than did either first or
fourth instars. Better acceptance and survival of
later instars was initially predicted on the as-
sumption that more time spent feeding on the
high quality host would yield better nutritional
status and greater survival when transferred to a
lower quality host. This would seem to adequately
explain the results for early instars, but not for
later instars. It is also possible that migration
tendency is age- or size-dependent to some de-
gree, since size and nutritional status could
strongly influence survival during migration.
Therefore, the pattern of acceptance observed in
Fig. 3 is likely a function of various factors acting
at different stages of development.

June 2004

Michaud: Cotton as Host Plant for Brown Citrus Aphid

The foraging decisions of adult aphids neces-
sarily concern the placement of their offspring,
rather than being exclusively concerned with food
consumption. A large proportion of the apterae
maturing on cotton left the plant immediately
upon molting to the adult stage (37.7% in the first
experiment, and 28.6% in the second). Migration
of reproductive apterae from BCA colonies has
been documented in response to crowding
(Michaud & Belliure 2000, 2001), but pre-repro-
ductive apterae were not observed to emigrate
under these conditions. In the present experi-
ments, the emigration of many apterae immedi-
ately following the adult molt might reflect a
decision to seek a more suitable host plant for
progeny while adequate resources are still avail-
able. Although the majority of maturing apterae
opted to allocate a fraction of their (potential) off-
spring to the cotton seedling before emigration,
virtually all ultimately opted to abandon the
plant. In the first experiment, only three repro-
ductive apterae died on the seedling and re-
mained hanging by their stylets; the remaining
individuals were all recovered from the Tangle-
foot barrier. Thus the estimate of fecundity is
more reflective of the length of time apterous
adults tolerated feeding on the cotton, rather
than their intrinsic reproductive potential on the
plant. Furthermore, the observed reproductive
rate was only a fraction of that typically observed
on citrus at a comparable temperature (Taka-
nashi 1989) and is indicative of the relatively low
suitability of cotton as a host for BCA.
Alate BCA frequently landed on cotton seed-
lings in the greenhouse but never remained on
them long enough to deposit nymphs under the
conditions of these experiments. While alatae
placed directly on cotton seedlings and main-
tained at 20C in a growth chamber will ulti-
mately deposit some nymphs (J. P. Michaud,
unpublished), this is not a meaningful observa-
tion since alatae seldom fly at this temperature, if
they are able to fly at all. Thus colonization of cot-
ton seedlings in the field by BCA alatae seems un-
likely even under cool temperature conditions.
These experiments demonstrate that cotton
seedlings may be colonized by apterous morphs of
BCA, that BCA can develop and reproduce suc-
cessfully on cotton under certain conditions, and
that alatae developing on cotton will readily re-
turn to citrus. Cotton may, therefore, be useful as
a factitious host plant for rearing BCA for pur-
poses of scientific study, although colony growth
rates are slower than on citrus. However, given
that cotton is only acceptable during the seedling
stage, and only to apterae under conditions of rel-
atively low ambient temperature, it seems un-
likely that there is much risk of cotton serving as
a pest reservoir for BCA under field conditions.


The author thanks Drs. C. Childers and R. Stuart for
reviewing the manuscript. Dr. A Chow provided valu-
able initial insights. This research was supported by the
Florida Agricultural Experiment Station and a grant
from USDA, CSREES and approved for publication as
Journal Series No R-09317.


CARVER, M. 1978. The black citrus aphids, Toxoptera cit-
ricidus (Kirkaldy) and T aurantii (Boyer de Fonsco-
lombe) (Homoptera: Aphididae). J. Aust. Entomol.
Soc. 17: 263-270.
CISNEROS, J. J., AND L. D. GODFREY. 2001. Midseason
pest status of the cotton aphid (Homoptera: Aphid-
idae) in California cotton: Is nitrogen a key factor?
Environ. Entomol. 30: 501-510.
HALBERT, S. B. 1996. Entomology Section: Citrus. Tri-
ology 34:8. Fla. Dept. Agric. & Cons. Serv., Div. Plant
Industry, Gainesville, FL.
HALBERT, S. B., G. X. ZHANG, AND Z. Q. PU. 1986. Com-
parisons of sampling methods for alate aphids and
observations on epidemiology of soybean mosaic vi-
rus in Nanjing, China. Ann. Appl. Biol. 109: 473-483.
HALBERT, S. B., AND L. G. BROWN. 1996. Toxoptera citri-
cida (Kirkaldy), brown citrus aphid-identification,
biology and management strategies. Fla. Dept. Agr.
& Cons. Serv., Div., Entomol. Cir. No. 374.
KOMAZAKI, S. 1989. Growth and reproduction in the
first two summer generations of two citrus aphids,
Aphis citricola Van der Groot and Toxoptera citrici-
dus (Kirkaldy) (Homoptera: Aphididae) on Citrus.
Appl. Entomol. Zool. 23: 220-227.
MENEGHINI, M. 1946. Sobre a naturaleza e tranmissibil-
idade de docena "Tristeza" dos citricos. Biologico 12:
MICHAUD, J. P. 1998. A review of the literature on the
brown citrus aphid, Toxoptera citricida, (Kirkaldy).
Florida Entomol. 81: 37-61.
MICHAUD, J. P. 2001. Colony density and wing develop-
ment in Toxoptera citricida (Homoptera: Aphididae).
Environ. Entomol. 30: 1047-1051.
MICHAUD, J. P., AND R. ALVAREZ. 2000. First collection
of brown citrus aphid from Quintana Roo, Mexico.
Florida Entomol. 83: 357-358.
MICHAUD, J. P., AND B. BELLIURE. 2000. Consequences
of foundress aggregation in the brown citrus aphid,
Toxoptera citricida. Ecol. Entomol. 25: 307-314.
MICHAUD, J. P., AND B. BELLIURE. 2001. Impact of syr-
phid predation on production of migrants in colonies
of brown citrus aphid, Toxoptera citricida (Ho-
moptera: Aphididae). Biol. Control 21: 91-95.
SPSS. 1998. SPSS 8.0 for Windows. SPSS Inc., Chicago, IL.
SYMES, C. B. 1924. Notes on the black citrus aphis. Rho-
desia Agric. J. 11: 612-626.
TAKANASHI, K. 1989. The reproductive ability of apter-
ous and alate viviparous morphs of the citrus brown
aphid, Toxoptera citricidus (Kirkaldy). Jap. J. Appl.
Entomol. Zool. 33: 266-269.
HUNTER. 1999. Effects of host plant and temperature
on the biology of Toxoptera citricida (Homoptera:
Aphididae). Environ. Entomol. 28: 895-900.

Florida Entomologist 87(2)

June 2004


Department of Crop Protection, Ghent University, Coupure Links 653, B-9000 Ghent, Belgium


Insecticidal effects of an encapsulated formulation of lambda-cyhalothrin on the southern
green stinkbug Nezara viridula (L.) and one of its predators, Podisus maculiventris (Say),
were investigated in the laboratory. Both pentatomids were exposed to the insecticide via
contaminated drinking water and by residual contact. Nymphs and adults of N. viridula
were more susceptible to the insecticide than nymphs of P. maculiventris, both by ingestion
and contact exposure. For the respective ways of exposure, LC5s values calculated for P. ma-
culiventris fourth instars were 30-190 times and 3-13 times higher than those ofN. viridula
fourth instars. Insecticidal activity of the pyrethroid by ingestion was 6-10 times greater
against nymphs of N. viridula than against adults of the pest. In both the ingestion and re-
sidual contact experiments, nymphs of P. maculiventris recovered from initial knockdown.
LC5s values for predator nymphs increased 1.7- to 2.7-fold between 24 and 48 h after the
start of the experiment. Recovery from knockdown was not observed in N. viridula. The data
from the current laboratory study suggest that encapsulated lambda-cyhalothrin may be ef-
fective for controlling the southern green stinkbug with little adverse effects on the predator
P. maculiventris, but field experiments are needed to confirm this. Possible reasons for the
differential toxicity of the insecticide to both pentatomids are discussed.

Key Words: lambda-cyhalothrin, pyrethroid, Nezara viridula, Podisus maculiventris, Pen-
tatomidae, non-target effects


Los efectos insecticides de una mezcla encapsulada de lambda-cyhalothrin en la chinche he-
dionda verde de sur (southern green stink bug), Nezara viridula (L.) y uno de sus depreda-
dores, Podisus maculiventris (Say), fueron investigados en el laboratorio. Ambos
pentat6midos fueron expuestos al insecticide por medio de agua para beber contaminada y
por el contact del residue del insecticide. Las ninfas y los adults de N. viridula fueron mas
susceptibles al insecticide que las ninfas de P. maculiventris, por la ingesti6n y por la expo-
sici6n por contact. Para las respectivas formas de exposici6n, los valores LC5, calculados por
las ninfas de P. maculiventris en el cuatro estadio fueron 30-190 veces y 3-13 veces mas altos
que los valores para las ninfas de N. viridula en el cuatro estadio. La actividad de la insec-
ticida piretroide por ingesti6n fu6 6-10 veces mayor contra las ninfas de N. viridula que con-
tra los adults de esta plaga. En ambos experiments de ingesti6n y por contact del residuo,
las ninfas de P. maculiventris se recuperaron del derribo inicial. Los valores LC5, para las
ninfas del depredador aumentaron 1.7 al 2.7 veces entire las 24 y 48 horas despu6s de empe-
zar el experiment. La recuperaci6n del derribo inicial no fue observada en N. viridula. Los
datos del studio de laboratorio actual sugerieron que el lambda-cyhalothrin encapsulada
puede ser efectivo para controlar la chinche hedionda verde de sur con pocos efectos adversos
sobre el depredador P. maculiventris, pero se necesita llevar a cabo experiments en el campo
para confirmarlo. Se discuten las razones posibles para la toxicidad differential del insecti-
cida para ambos pentat6midos.

The southern green stinkbug, Nezara viridula
(L.), is a highly polyphagous pest that is widely dis-
tributed in the tropical and subtropical regions of
the world (Todd 1989; Panizzi et al. 2000). This pen-
tatomid causes important economic damage to var-
ious field crops, including soybean, beans, rice, corn,
cotton and tobacco. In Europe, it has been found in-
creasingly in greenhouses, where it attacks vegeta-
ble crops like tomato, sweet pepper, and eggplant.
Control of this pest is based largely on the intensive
use of chemical pesticides, including carbamates,

organophosphates and some pyrethroids (Jackai
et al. 1990; Ballanger & Jouffret 1997; Panizzi et al.
2000). For instance, in Brazil it was estimated that
in the mid 1990s over 4 million liters of insecticides
were used annually to control stinkbugs in soybean
(Correa-Ferreira & Moscardi 1996). Such massive
use of insecticides not only increases production
cost, it may also affect populations of beneficial in-
sects and trigger pest resurgence problems.
In several regions, efforts have been made to
develop integrated pest management (IPM) pro-

Vandekerkhove & De Clercq: Effects of Lambda-cyhalothrin on Two Pentatomids 113

grams against N. viridula (Panizzi et al. 2000).
Biological control of the pest has focused mainly
on the potential of parasitoids (Jones 1988). Re-
leases of the scelionid egg parasitoid Trissolcus
basalis (Wollaston) have successfully suppressed
outbreaks of the southern green stinkbug in soy-
bean (Corr6a-Ferreira & Moscardi 1996). Several
arthropod predators also have an important im-
pact on N. viridula populations. De Clercq et al.
(2002) reported high predation rates by nymphs
and adults of the predatory pentatomid Podisus
maculiventris (Say) on the different life stages of
the southern green stinkbug. This generalist
predator is native to North America where it is
commonly found in a variety of natural and agri-
cultural ecosystems (De Clercq 2000). Although it
appears to have a preference for larvae of lepi-
dopterous and coleopterous insects, the predator
frequently has been found in association with N.
viridula in the southern United States (Drake
1920; Ragsdale et al. 1981; Stam et al. 1987). Po-
disus maculiventris has been used in European
greenhouses since 1997 for augmentative biologi-
cal control of caterpillar outbreaks, and predation
on N. iridula may be an additional asset here for
this beneficial insect (De Clercq et al. 2002).
In France, Ballanger & Jouffret (1997) re-
ported effective control ofN. viridula in soybean
with foliar sprays of lambda-cyhalothrin. This
contact and stomach insecticide belonging to the
pyrethroid group has been used against a broad
spectrum of pests in a variety of crops (Anony-
mous 2000). In 1999, a micro-encapsulated for-
mulation (Zeon TechnologyTM) of this pyrethroid
was commercialised, offering reduced health haz-
ards and an improved environmental profile
(Ham 1999; Anonymous 2000). In the current
study, insecticidal activity of an encapsulated for-
mulation of lambda-cyhalothrin to N. viridula
and its predator P. maculiventris was assessed in
the laboratory. Both pentatomids were exposed to
the insecticide via ingestion and residual contact.
The implications of our findings for the control of
the southern green stinkbug and the use of the
predator P maculiventris in IPM programs tar-
geted against this and other agricultural pests
are discussed.


Insect Cultures

A laboratory colony of N. iridula was estab-
lished in 1999 with insects originating from field
collections in France, Spain and Italy. Stinkbugs
were fed on green bean pods (Phaseolus vulgaris
L.) and sunflower seeds (Helianthus annuus L.).
A culture of P maculiventris was started in 1999
with specimens originating from a field collection
in 1996 near Beltsville, Maryland, USA. The
predators were fed mainly larvae of the greater

wax moth, Galleria mellonella L. Colonies of all
insects were maintained in growth chambers at
23 1C, 75 + 5% RH and a 16:8 h light:dark pho-


A commercial formulation of micro-encapsu-
lated lambda-cyhalothrin (Karate Zeon, 100 g/l)
was obtained from Syngenta, Ruisbroek, Belgium.

Toxicity Bioassays

Exposure via ingestion. In these experiments,
nymphs and adults ofN. viridula and nymphs of P
maculiventris were exposed to insecticide through
treated drinking water. Both pentatomids take up
moisture in the absence of food and are regularly
seen drinking even when food is available. Mois-
ture can be supplied via plant materials, like green
beans, but can also be provided as free water. Us-
ing gravimetric methods, we estimated that unfed
newly molted fourth instars of P maculiventris
and N. viridula take up about 2 and 4.5 pl of free
water, respectively, during a 24-h period.
Newly molted fourth instars and reproduc-
tively active adult female N. viridula were ran-
domly collected from stock cultures and
transferred to plastic petri dishes (9 cm diam)
lined with absorbent paper. A replicate consisted
of four insects in a dish for N. viridula nymphs,
whereas adults were placed singly in dishes.
Newly molted fourth instars of P maculiventris
were placed in petri dishes (9 cm diam) in groups
of three. Each dish was supplied with a moisture
source, consisting of a paper plug fitted into a
plastic dish (2.5 cm diam). The paper plug was
saturated with 2 ml of the insecticide in tap wa-
ter. Control groups were supplied with tap water
alone. At least 20 nymphs or 10 adults were
tested with each of at least 10 concentrations.
Choice of concentrations was based on prelimi-
nary range-finding tests. Test concentrations
ranged from 0 to 200 mg a.i./l (11 concentrations)
and from 0 to 100 mg a.i./l (10 concentrations) for
N. uiridula nymphs and adults, respectively, and
from 0 to 800 mg a.i./l (13 concentrations) for P.
maculiventris nymphs. Both pentatomids were
exposed to the contaminated moisture source dur-
ing a 1-wk period. To stimulate drinking behavior,
the insects were not provided with food during
the first 24 h. From the second day on, nymphs
and adults ofN. viridula were supplied with sun-
flower seeds as needed; predator nymphs were fed
greater wax moth larvae ad libitum.
In range-finding tests, the ability of P. macu-
liventris in particular to recover from initial poi-
soning became apparent. Therefore, mortality
counts were performed 1, 2, and 7 d after the ini-
tial treatment. Mortality percentages included
dead and affected individuals. Insects were scored

Florida Entomologist 87(2)

as affected when they were incapable of coordi-
nated movement upon prodding with a fine brush.
Residual exposure. To evaluate residual contact
activity of lambda-cyhalothrin, fourth instars of
both pentatomids were exposed by tarsal contact to
dry residues on filter paper. Whatman No. 41 filter
papers were fitted into petri dishes (9 cm diam) and
the dishes were sprayed in a Cornelis spray cham-
ber (Van Laecke & Degheele 1993). Each dish was
sprayed with 2 ml of insecticide suspension in wa-
ter, yielding a homogeneous spray deposit on the
filter paper of approximately 5 mg/cm2. For the con-
trols, dishes were sprayed with 2 ml of water. The
plates were left to dry for about 1 h before introduc-
ing the insects. A replicate consisted of a petri dish
containing three P. maculiventris or four N. virid-
ula nymphs. Twenty to 40 insects were tested per
concentration, with a minimum of 10 concentra-
tions. Concentrations were chosen on the basis of
range-finding tests and ranged from 0 to 200 mg
a.i./l (12 concentrations) for N. viridula nymphs
and from 0 to 400 mg a.i./l (10 concentrations) forP.
maculiventris nymphs. To stimulate food searching
by the nymphs and maximize contact with the
treated surface, no food or moisture were provided
during the first 24 h. From the second day on, the
insects were supplied with water and food. Water
was supplied via a soaked paper plug in a 2-cm-
diam cup. Nymphs of P maculiventris were given
freshly killed wax moth larvae and those ofN. viri-
dula were offered sunflower seeds. Mortality
counts were made after 1, 2, and 7 d, allowing for
the assessment of recovery after initial knockdown.

Data Analysis

Mortality of the test insects after 1, 2, and 7 d
was corrected for control mortality by Abbott's for-
mula (Abbott 1925). Lethal concentration values
and their 95% confidence limits were calculated
from probit-regressions with POLO PC (LeOra
Software 1987). All concentrations tested were used
for LC-calculations; numbers (n) given in the foot-
notes of Tables 1 and 2 thus reflect actual numbers
of insects tested and used in probit-regressions.


In our laboratory setup, there was no indica-
tion of a repellent or antifeedant effect for encap-
sulated lambda-cyhalothrin. In ingestion assays,
both P. maculiventris and N. viridula were regu-
larly observed to suck on moisture sources con-
taminated with varying concentrations of the
compound. In the residual contact experiment,
nymphs of both pentatomids were usually on the
treated surfaces (filter paper, petri dish walls)
and only occasionally on the untreated lid.
Nymphs and adults of N. viridula were more
susceptible to the insecticide than nymphs of
their predator P. maculiventris, both by ingestion

(Table 1) and contact exposure (Table 2). For the
respective routes of exposure, LC5s values calcu-
lated for P. maculiventris fourth instars were 30-
190 times and 3-13 times higher than those ofN.
viridula fourth instars. Further, insecticidal ac-
tivity of the pyrethroid by ingestion was 6-10
times greater against nymphs ofN. viridula than
against adults of the pest. Lethal concentration
values calculated for N. viridula nymphs were
lower for ingestion exposure than for contact ex-
posure, suggesting that lambda-cyhalothrin was
more active by ingestion than by tarsal contact
with dry residues. However, some of the nymphs
in the ingestion bioassays were partially or fully
in tarsal contact with the moist plug when drink-
ing, so these insects may have been exposed to the
insecticide both via ingestion and via contact with
wet residues. Differences in biological activity
due to exposure routes were less apparent in the
predatory pentatomid P. maculiventris.
In both the ingestion and residual contact ex-
periments, nymphs of P maculiventris recovered
from initial knockdown. LC5s values for predator
nymphs increased 1.7- to 2.7-fold between 24 and
48 h after the start of the experiment. Recovered
predators were able to attack prey and feed nor-
mally. Comparisons of the LC5, values for P mac-
uliventris fourth instars after 2 and 7 d, however,
show that some of the individuals that appeared
to have recovered after 2 d died before reaching
the fifth stadium when they were continuously ex-
posed to contaminated drinking water or filter pa-
per during a 7 d period. At 23C, optimally fed
fourth instars of both pentatomids usually reach
the next stadium in 4-5 d (De Clercq & Degheele
1992). There was little or no recovery from knock-
down in N. viridula. Here, LC5, values after 1 and
2 d were generally similar and further decreased
with exposure time. Likewise, slopes were equal
after 1 and 2 d (x2, P > 0.05), but were significantly
increased after 7 d of exposure. Only in the inges-
tion study with adults, LCg, values suggest there
may have been some recovery at the upper end.


In our experiment, both nymphs and adults of
N. viridula were susceptible to encapsulated
lambda-cyhalothrin. However, nymphs suffered
about 5 times greater mortality when exposed by
ingestion than by residual contact. For the control
of N. viridula, the practical significance of inges-
tion exposure may be limited given that pyre-
throids have no systemic properties. It is unlikely
that the insect would ingest the compound in the
field, except when it would drink from fresh spray
deposits. Although lambda-cyhalothrin is widely
recommended for the control of a broad range of
insect pests, including stinkbugs, there are few
studies reporting on the insecticidal effects of this
pyrethroid on the southern green stinkbug. Ac-

June 2004




LC value' D

Insect LC10 LC50 LCo0 X2 (df) Slope SE

1 day
Nezara fourth instar 0.32 (0.12-0.61) 4.61 (3.12-6.50) 66.98 (39.35-147.46) 8.46 (8) 1.10 + 0.11
Nezara female adult 2.79 (0.42-6.06) 28.70 (15.29-82.31) 295.72 (96.63-646.70) 8.03 (7) 1.26 0.25
Podisus fourth instar 20.85 (12.67-29.68) 144.53 (116.50-181.80) 1001.83 (671.00-1776.99) 7.56 (10) 1.52 + 0.15 o

2 days
Nezara fourth instar 0.30 (0.10-0.60) 3.83 (2.47-5.7) 49.43 (28.48-116.13) 10.72 (8) 1.15 + 0.11
Nezara female adult 2.41 (0.13-6.19)* 40.04 (18.14-253.32)* 663.79 (141.99-1,798,890)* 13.84 (7) 1.05 0.23
Podisus fourth instar 28.00 (16.29-40.8) 248.97 (193.15-340.43) 2214.20 (1279.44-5112.59) 7.77 (10) 1.35 + 0.15

7 days
Nezara fourth instar 0.12 (0.05-0.22) 0.97 (0.64-1.32) 7.64 (5.57-11.63) 2.14 (8) 1.41+ 0.16
Nezara female adult 1.36 (0.05-3.66)* 10.26 (3.92-26.16)* 77.23 (29.10-2164.97)* 24.40 (7) 1.46 0.25
Podisus fourth instar 33.25 (20.83-46.08) 183.28 (146.88-234.81) 1010.36 (673.73-1838.99) 12.09 (10) 1.73 + 0.17.

LC values and slopes in mg a.i./l; LC-values are followed by 95% fiducial limits except when marked with an asterisk (*) where 90% fiducial limits are given.
n = 456, 137, and 411 for N. viridula fourth instars, N. viridula females and P. maculiventris fourth instars, respectively. i

Florida Entomologist 87(2)

Cl I- Chl I CO L-
C11 X o&- coct-
Cl '

0. 0


0 1

t- 0

i 0
1C '-U


r 1
CO ^
^- (


^, '-

o1 C1
10] CO]
In CO]


00 0


^ ci

'-' t^-


0 00
CO cc



^ C
g a

cording to Ballanger & Jouffret (1997) and Gouge
et al. (1999), the pest can be adequately controlled
in soybean with non-encapsulated lambda-cyhal-
othrin at 20-30 g a.i./ha. In topical application ex-
periments on third instars of N. viridula,
Baptista et al. (1995) reported LD5, values of 0.82-
0.25 pg/g for a technical grade formulation of
lambda-cyhalothrin 3-24 h after treatment; the
pyrethroid was 20-40 times more active against
the pest than monocrotophos.
Susceptibility of the spined soldier bug, P ma-
culiuentris, to classical and novel insecticides has
been studied to some extent (see De Clercq 2000
for a review). Besides direct and residual contact,
predatory pentatomids can be poisoned by drink-
ing contaminated free water or plant sap (in case
of systemic compounds) or by feeding on contam-
inated prey (De Clercq et al. 1995). In the current
study, P. maculiuentris was less vulnerable to
lambda-cyhalothrin than N. viridula, both in the
ingestion and contact exposure bioassays. Based
on lethal concentration values, nymphs of the
predator were somewhat more susceptible to the
compound by residual contact than by ingestion.
The LC5s value for fourth instars of P maculiven-
tris exposed to lambda-cyhalothrin via ingestion
was similar to that found in an earlier study for
nymphs treated similarly with deltamethrin (158
mg a.i./l, Mohaghegh et al. 2000). In a number of
studies, P. maculiventris and related asopines
have demonstrated a better tolerance to pyre-
throids than their lepidopterous prey (Yu 1988;
Zanuncio et al. 1993; Picanco et al. 1996). Yu
(1988) hypothesized that thickness and lipid con-
tent of the cuticle may affect the penetration rate
of the lipophilic pyrethroids and may thus be re-
sponsible for differences in toxicity to the heavily
sclerotized pentatomid predators and their soft-
bodied caterpillar prey. The finding of Baptista et
al. (1995) that lambda-cyhalothrin was 5-9 times
more toxic to the velvetbean caterpillar, Anticar-
sia gemmatalis (Hibner) than to N. viridula sup-
ports this hypothesis. It may, however, not
explain the differences found in our study be-
cause P. maculiventris and N. viridula are both
pentatomids with a similarly sclerotized cuticle.
Different drinking rates explain only in part the
observed differences in toxicity of lambda-cyhalo-
thrin to the studied pentatomids by ingestion.
Preliminary gravimetric tests indicated that un-
fed fourth instars ofN. viridula ingested twice as
much free water during a 24-h period than did
those of P. maculiventris, despite similar body
weights (approximately 12.5 mg). Alternatively,
higher detoxification or excretion rates may be re-
sponsible for the lower susceptibility of P macu-
liventris to lambda-cyhalothrin as compared toN.
viridula. Pyrethroids are known to be metabo-
lised in insects mainly by esterases and microso-
mal oxidases (Shono et al. 1979). Yu (1987, 1988)
found, however, that these enzyme activities were

June 2004

,-^ S
C.] '-I
0 0
C; 01

O' 00C
oo' s
c-1 CC

t 01

t^ W

~co o
C(] CO~

C-l CC


o co

(0 CM
C, C.
a In

Q .2


Vandekerkhove & De Clercq: Effects of Lambda-cyhalothrin on Two Pentatomids 117

generally lower in the spined soldier bug than in
its caterpillar prey. Likewise, first tests have
shown about 8-fold higher esterase activities in
the phytophagous pentatomid N. viridula than in
the predatory pentatomid P. maculiventris (un-
published data). Recently, it has been shown that
glutathione S-tranferases also may play a signifi-
cant role in detoxifying pyrethroids (e.g., Ko-
staropoulos et al. 2001). In this context, it is
worth noting that Yu (1987) reported about 2-fold
higher glutathione transferase activity toward
CDNB in the spined soldier bug than in its lepi-
dopterous prey. Further studies on the pharmaco-
kinetics of lambda-cyhalothrin are warranted to
explain the difference in toxicity of the compound
to the studied pentatomids.
A number of field studies have demonstrated
adverse effects of lambda-cyhalothrin on various
beneficial arthropods, including predatory het-
eropterans, although in some cases negative ef-
fects on field populations of natural enemies were
transient (Pilling & Kedwards 1996; Cole et al.
1997; van den Berg et al. 1998; Al-Deeb et al.
2001; Stewart et al. 2001). Encapsulation of
broad-spectrum insecticides, including pyre-
throids, is aimed at improving their selectivity
and suitability for IPM programs (Scher et al.
1998; Ham 1999). However, several studies com-
paring encapsulated and non-encapsulated for-
mulations of various insecticides have shown
highly variable results with a range of beneficial
arthropods (see Pogoda et al. 2001 for references).
Pogoda et al. (2001) reported that a micro-encap-
sulated formulation of lambda-cyhalothrin was as
toxic as an emulsifiable-concentrate formulation
of the compound to the oriental fruit moth, Gra-
pholita molesta (Busck). These workers also
found that the encapsulated formulation of
lambda-cyhalothrin was less toxic than the emul-
sifiable concentrate to a pyrethroid-resistant pop-
ulation of the phytoseiid mite Typhlodromus pyri
Scheuten but more toxic to a pyrethroid-suscepti-
ble population of the predator.
The current laboratory trials indicate that en-
capsulated lambda-cyhalothrin has a good insec-
ticidal activity against the southern green
stinkbug and is relatively safe to the predatory
stinkbug P maculiventris. However, field studies
are in place to test further the selectivity of this
insecticide toward P maculiventris. Further re-
search also is needed to determine if the studied
encapsulated formulation of lambda-cyhalothrin
is more selective toward P. maculiventris and
other natural enemies compared with other for-
mulations of the compound.


The authors thank Thomas Van Leeuwen for his
helpful suggestions and Bea Roos for excellent technical

ABBOTT, W. S. 1925. A method of computing the effective-
ness of an insecticide. J. Econ. Entomol. 18: 265-267.
AL-DEEB, M. A., G. E. WILDE, AND K. Y. ZHU. 2001. Ef-
fects of insecticides used in corn, sorghum, and al-
falfa on the predator Orius insidiosus (Hemiptera:
Anthocoridae). J. Econ. Entomol. 94: 1353-1360.
ANONYMOUS. 2000. Karate with Zeon technology. Tech-
nical bulletin. Syngenta, Basel, Switzerland. 25 pp.
BALLANGER, Y., AND P. JOUFFRET. 1997. La punaise
verte et le soja. Phytoma 50: 32-34.
DAD. 1995. Toxicidade comparative de lambda-cyha-
lothrin a lagarta-da-soja, Anticarsia gemmatalis
Hueb., 1818 (Lepidoptera, Noctuidae) e ao percevejo
verde, Nezara viridula (L., 1758) (Hemiptera, Penta-
tomidae). Sci. Agric. Piracicaba 52: 183-188.
BERSON. 1997. Effects of Karate insecticide on bene-
ficial arthropods in Bollgard cotton, pp. 1118-1120. In
Proceedings 1997 Beltwide Cotton Conferences, New
Orleans. Nat. Cotton Counc. Amer., Memphis, TN.
logical control of soybean stink bugs by inoculative
releases of Trissolcus basalis. Entomol. Exp. Appl.
79: 1-7.
DE CLERCQ, P. 2000. Chapter 32: Predaceous Stinkbugs
(Pentatomidae: Asopinae), pp. 737-789. In C. W.
Schaefer, and A. R. Panizzi [eds.], Heteroptera of
Economic Importance. CRC Press, Boca Raton, FL.
828 pp.
D. DEGHEELE. 1995. Toxicity of diflubenzuron and
pyriproxyfen to the predatory bug Podisus macu-
liventris. Entomol. Exp. Appl. 74: 17-22.
DE CLERCQ, P., AND D. DEGHEELE. 1992. Development
and survival of Podisus maculiventris (Say) and Po-
disus sagitta (Fab.) (Heteroptera: Pentatomidae) at
various constant temperatures. Canadian Entomol.
124: 125-133.
J. KLAPWIJK. 2002. Predation by Podisus maculiven-
tris on different life stages ofNezara uiridula. Flor-
ida Entomol. 85: 197-202.
DRAKE, C. J. 1920. The southern green stink-bug in
Florida. Florida State Plant Board Q. Bull. 4: 41-94.
AND C. PATRICK. 1999. Managing soybean insects.
Texas Agricultural Extension Service, B-1501, 36 pp.
HAM, D. 1999. New technology brings improved insect
control. Australian Cottongrower 20: 102-103.
SRIVASTAVA. 1990. Insect pests of soybean in the
tropics, pp. 91-156 In S. R. Singh [ed.], Insect Pests
of Tropical Food Legumes. John Wiley & Sons,
Chichester, U.K. 451 pp.
JONES, W. A. 1988. World review of the parasitoids of
the southern green stink bug, Nezara viridula (L.)
(Heteroptera: Pentatomidae). Ann. Entomol. Soc.
Amer. 81: 262-273.
MOURKIDOU. 2001. Gluthatione S-transferase in the
defence against pyrethroids in insects. Insect Bio-
chem. Molec. Biol. 31: 313-319.
LEORA SOFTWARE. 1987. Polo-PC: a user's guide to pro-
bit or logit analysis. LeOra Software, Berkeley, CA.

Florida Entomologist 87(2)

Toxicity of selected insecticides to the spined soldier
bug, Podisus maculiventris (Heteroptera: Pentato-
midae). Biocontr. Sci. Technol. 10: 33-40.
HERY, AND R. M. MCPHERSON. 2000. Chapter 13:
Stink bugs (Pentatomidae), pp. 421-474 In C. W.
Schaefer, and A. R. Panizzi [eds.], Heteroptera of Eco-
nomic Importance. CRC Press, BocaRaton, FL. 828 pp.
P. CAMPOS. 1996. Toxicity of insecticides to Dione
juno juno (Lepidoptera: Heliconidae) and selectivity
to two of its predaceous bugs. Trop. Sci. 36: 51-53.
PILLING, E. D., AND T. J. KEDWARDS. 1996. Effects of
lambda-cyhalothrin on natural enemies of rice insect
pests, pp. 361-366. In Proceedings 1996 Brighton
Crop Protection Conference: Pests & Diseases, Brigh-
ton, UK. British Crop Prot. Counc., Farnham, UK.
Effects of encapsulation on the toxicity of insecti-
cides to the Oriental fruit moth (Lepidoptera: Tortri-
cidae) and the predator Typhlodromus pyri (Acari:
Phytoseiidae). Canadian Entomol. 133: 819-826.
1981. Quantitative assessment of the predators of
Nezara viridula eggs and nymphs within a soybean
agroecosystem using an ELISA. Environ. Entomol.
10: 402-405.
SCHER, H. B., M. RODSON, AND K-S. LEE. 1998. Microen-
capsulation of pesticides by interfacial polymeriza-
tion utilizing isocyanate or aminoplast chemistry.
Pestic. Sci. 54: 394-400.
SHONO, T., K. OHASAWA, AND J. E. CASIDA. 1979. Metab-
olism of trans- and cis-permethrin, trans- and cis-

cypermethrin, and decamethrin by microsomal en-
zymes. J. Agric. Food Chem. 27: 316-325.
1987. Predation and food as factors affecting sur-
vival ofNezara viridula (L.) (Hemiptera: Pentatomi-
dae) in a soybean ecosystem. Environ. Entomol. 16:
VANDERBERG. 2001. Combining exclusion tech-
niques and larval death-rate analyses to evaluate
mortality factors of Spodoptera exigua (Lepidoptera:
Noctuidae) in cotton. Florida Entomol. 84: 7-22.
TODD, J. W. 1989. Ecology and behavior ofNezara uiri-
dula. Annu. Rev. Entomol. 34: 273-292.
Evaluation of pesticide effects on arthropod predator
populations in soya bean in farmers' fields. Biocontr.
Sci. Technol. 8: 125-137.
VAN LAECKE, K., AND D. DEGHEELE. 1993. Effect of in-
secticide synergist combinations on the survival of
Spodoptera exigua. Pestic. Sci. 37: 283-288.
YU, S. J. 1987. Biochemical defense capacity in the
spined soldier bug (Podisus maculiventris) and its
lepidopterous prey. Pestic. Biochem. Physiol. 28:
Yu, S. J. 1988. Selectivity of insecticides to the spined
soldier bug (Heteroptera: Pentatomidae) and its lep-
idopterous prey. J. Econ. Entomol. 81: 119-122.
A. RODRIGUES. 1993. Impact of two formulations of
deltamethrin in aerial application against Eucalyp-
tus caterpillar pests and their predaceous bugs.
Meded. Fac. Landbouww. Univ. Gent 58: 477-481.

June 2004

Gunawardene et al.: Lubber Grasshopper Reproductive Tactics


'Department of Biological Sciences, Behavior, Ecology, Evolution, & Systematics Section
Illinois State University, Normal, IL, USA, 61790-4120

2Present address: Department of Biological Sciences, University of North Florida
4567 St. Johns Bluff Rd., S. Jacksonville, FL 32224

We tested whether reproductive tactics of a univoltine insect can be predicted by local ecol-
ogy, specifically mean length of the frost free period (FFP) as a measure of the potential ac-
tive season. We measured reproductive tactics and longevity for populations of the lubber
grasshopper Romalea microptera (Beauvois) from Miami, Florida (FL; 365 days FFP), Lydia,
Louisiana (LA; 280 days FFP), and Athens, Georgia (GA; 224 days FFP). Differences in local
climate led us to predict that GA grasshoppers will have shorter interclutch intervals, fewer
clutches, and shorter lifespan than FL grasshoppers, with LA grasshoppers intermediate in
these traits. When reared in a common laboratory environment, longevity, total reproductive
period, and number of clutches produced were not clearly related to FFP. Longevity and re-
productive period of LA grasshoppers were significantly less than those of FL grasshoppers,
and number of clutches produced by LA grasshoppers was less than that for the FL or GA
grasshoppers. First interclutch interval was significantly greater for LA than for GA grass-
hoppers. Our data suggest that phylogenetic relationships among populations may be a bet-
ter predictor of reproductive tactics in this species.
Key Words: age at reproduction; climate; clutch size; grasshopper; life history; longevity;

Probamos si las tactica reproductivas de un insecto del univoltine se pueden predecir por
ecologia local, specificamente longitud del period libremente mala de la helada (FFP) como
media de la estaci6n active potential. Medimos tactica y la longevidad reproductivas para
tres poblaciones del saltamontes, Romalea microptera (Beauvois), Miami, Florida (FL; 365
dias FFP), Lydia, Louisiana (LA; 280 dias FFP), y Athens, Georgia (GA; 224 dias FFP). Estas
diferencias en clima local conducen a la predicci6n que los saltamontes de GA tendran
period entire las hornadas del huevos mas cortos, pocos horadas del huevos, y esperanza de
vida mas corta que saltamontes del FL, con los saltamontes del LA intermedios en estos ras-
gos. Cuando estaba alzada en un ambiente comun del laboratorio, la longevidad, el period
reproductive del total, y el numero de los horadas del huevos producidos no fueron relacio-
nados claramente con FFP. La longevidad y el period reproductive de los saltamontes del LA
eran perceptiblemente menos que los de los saltamontes del FL, y el numero de los horadas
producidos por los saltamontes de LA era menos que eso para los saltamontes del FL o de GA.
El primer intervalo del hornadas era perceptiblemente mayor para el LA que para los salta-
montes de GA. Nuestros datos sugieren que las relaciones phylogenetic entire poblaciones
puedan ser un predictor mejor de estos aspects de tactica reproductivas en esta especie.
Translation provided by the author.

Latitudinal variation in life histories can be re-
lated to adaptation to local climate (Rowe & Lud-
wig 1991; Temte 1993; Hemborg et al. 1998;
Johansson & Rowe 1999; Berkenbusch & Rowden
2000; Hatle et al. 2002). For a univoltine organ-
ism, age at first reproduction and duration of in-
terclutch intervals are likely to be positively
related to the duration of the active season (Roff
1992; Forsman 2001), because of time-constraints
in areas with shorter active seasons. There is of-
ten a tradeoff between early reproduction and
longevity (e.g., De Souza Santos & Begon 1987;

Rowe & Scudder 1990; Kaitala 1991; Stearns
1992; Leroi et al. 1994; Rowe et al. 1994;
Miyatake 1997; Frankino & Juliano 1999). This
trade-off yields a prediction of reduced longevity
and late-life reproduction in populations from ar-
eas with short active seasons, where early repro-
duction is advantageous.
Hatle et al. (2002) examined latitudinal varia-
tion and trade-offs in reproductive tactics during
the first oviposition cycle for three populations of
the univoltine Eastern lubber grasshopper, Ro-
malea microptera (Beauvois), testing for the joint

Florida Entomologist 87(2)

relationships of latitude to age at first reproduc-
tion, somatic storage (body mass immediately af-
ter oviposition relative to initial mass), and clutch
mass. All three populations differed in their mul-
tivariate responses for the three reproductive tac-
tics we studied. This difference across
populations was due primarily to age at first re-
production, secondarily to somatic storage, and
less so to clutch mass. Age at first reproduction
was least in Georgia (GA) (34.5 1.2 days; mean
+ SE), and significantly greater for Louisiana
(LA) (38.5 1.4 days) and Florida (FL) (41.5 1.4
days) grasshoppers. Estimated somatic storage
was greatest in FL and LA, and least in GA grass-
hoppers. Clutch mass was greatest in LA and GA,
and least in FL grasshoppers. Thus, allocation of
resources among these reproductive tactics is dif-
ferent across populations, in ways that could be
adaptive for each local climate.
In the present study, we investigate interpopu-
lation differences in number of egg clutches, in-
terclutch intervals, period of reproduction, and
longevity using the same lubber grasshopper pop-
ulations used by Hatle et al. (2002). Differences in
climate and potential active season duration for
these populations are indicated by the differences
in mean duration of the frost free period (FFP) for
these locations: Miami, Florida (FL, 365 days
FFP); Lydia, Louisiana (LA, 280 days FFP); and
Athens, Georgia (GA, 224 days FFP) (Koss et al.
1988). Because of the shorter period potentially
suitable for reproduction, we predict GA grass-
hoppers will produce clutches faster, with shorter
interclutch intervals than FL grasshoppers. Be-
cause of the putative tradeoff of longevity and
early reproduction, we also predict that GA grass-
hoppers should have a shorter lifespan. Based on
climate, the number of clutches and longevity for
LA grasshoppers should be intermediate between
those for GA and FL grasshoppers.


Grasshoppers were shipped as young nymphs
from our three source populations to our labora-
tory at Illinois State University (Normal, IL,
USA). Each population was reared on a 14L:10D
photoperiod and a corresponding 32:24C ther-
mocycle. This photoperiod was chosen to approxi-
mate those observed at each of the sites in mid-
active season for the adult grasshoppers. Photo-
phases of 14 h occur at Athens at approximately
25 July, at Lydia at approximately 7 July, and at
Miami at approximately 26 June (the longest pho-
tophase observed at Miami is 13.75 h) (US Naval
Observatory 2003). A 14-h photophase was used
by Hatle et al. (2002) in a previous comparison of
reproductive tactics of these same populations.
Photoperiod affects reproduction in R. microp-
tera, with females from south Florida (Luker
et al. 2002) and north Georgia (R. Homeny &

S. Juliano, unpubl.) altering reproductive tactics
in response to short photoperiods (11.5 and 12.0
h, respectively) associated with autumn. Thus, a
14 h photophase, typical of mid-summer at all
sites, provides a reasonable point of comparison.
All grasshoppers were offered Romaine lettuce
and oatmeal ad libitum throughout the experi-
ment. For a laboratory colony of lubbers from
south Florida, the first oviposition cycle (~35 d)
involves first somatic growth and then reproduc-
tive growth. During the first ~10 d the primary
oocytes are not vitellogenic, despite a ~50% in-
crease in somatic mass (Sundberg et al. 2001).
Hence, the nymphal stages appear to be relatively
unimportant for acquiring nutritional resources
for egg production, and we are justified in con-
ducting a common garden experiment beginning
with newly molted adult females.
After adult eclosion, males and females were
reared separately. Every other day, males and fe-
males were randomly paired for mating. Mated fe-
males were placed on 1.0 kg of sand with ~7%
water (by mass) for oviposition. The calendar date
of first oviposition, and all subsequent ovipositions,
was recorded for each female. We maintained
mated females until death or until 25 September
2002, when we terminated the experiment.
Data on interclutch interval, number of
clutches, and period of reproduction were ana-
lyzed by one-way ANOVA with multiple compari-
sons (REGWQ method PROC GLM, SAS Inst.,
Inc. 1990a) among population means when the
overall ANOVA test was significant. Assumptions
of normality and homogeneity of variances were
met. Proportions of experimental females in the
three populations remaining alive at the end of
the experiment were compared by Fisher's exact
test (PROC FREQ, SAS Inst., Inc. 1990b). Lon-
gevity for the three populations was analyzed by
nonparametric survival analysis (PROC
LIFETEST, SAS Inst. Inc. 1990b, Allison 1995).
Pairs of populations were compared for propor-
tions alive and for survival time distributions
with two-group Fisher's exact tests and two-
group nonparametric survival analyses, respec-
tively, with a sequential Bonferroni correction at
experimentwise a = 0.05 (Rice 1989).


Clutch Production

Mean number of clutches produced differed
significantly among populations (F2,54 = 13.50, P =
0.0001). LA grasshoppers produced significantly
fewer clutches than did GA or FL grasshoppers,
whereas GA and FL grasshoppers produced simi-
lar numbers of clutches (Fig. 1A).
The interclutch intervals between first and
second, second and third, and third and fourth
ovipositions were determined for each population

June 2004

Gunawardene et al.: Lubber Grasshopper Reproductive Tactics





3 B

2 A


3 10 3 13 10 6 16 13 7
SIstovp 2ndovp
30 A a 02nd op- 3r o-p
25 3rd o-p 4thO p AB
20 B a
0 -


Fig. 1. Reproductive tactics of three populations of
lubber grasshoppers, reared in a common environment.
A. Number of clutches produced by each population.
Means ( SE) for LA (N = 24), GA (N = 15), and FL (N =
18) grasshoppers associated with the same letters are
not significantly different at a = 0.05. B. Clutch inter-
vals for three populations. Sample sizes are given at the
top of the graph, above the corresponding mean. Within
each interval, means ( SE) associated with the same
letters are not significantly different at a = 0.05. C. Time
from each grasshopper's 1st clutch until its last clutch.
Means ( SE) for LA (N = 10), GA (N = 13), and FL (N =
16) grasshoppers associated with different letters are
significantly different at a = 0.05.

(Fig. 1B). None of the interclutch intervals was
significantly different at P = 0.05, but the interval
from first to second oviposition for the LA vs. GA
grasshoppers came close (F2,36 = 3.12, P = 0.0564).

Reproductive Period and Longevity

Reproductive period was quantified as the
time from the first clutch until the last clutch
(Fig. 1C). This period differed significantly among

populations (F2,36 = 4.86, P = 0.0136) and was sig-
nificantly less for LA grasshoppers than for FL
grasshoppers. GA grasshoppers were intermedi-
ate and statistically indistinguishable from the
other two populations.
The proportion alive at the end of the experi-
ment differed significantly among the three popu-
lations (P < 0.0001). Pairwise tests indicated that
the proportion alive for FL (0.63, N = 19) was sig-
nificantly greater than that for LA (0.12, N = 25)
and for GA (0.24, N = 17). Proportions alive for
GA and LA did not differ significantly.
Survival distributions were quantified as the
time from first clutch until death or the end of the
experiment on 25 September. Individuals alive at
the end of the experiment yielded censored observa-
tions, which are accounted for by PROC LIFETEST
(see Allison 1995 for details). Survival analysis in-
dicated significant differences in longevity among
populations (Fig. 2). Pairwise tests indicated that
longevity for FL was significantly greater than that
for LA, and that GA was intermediate, and statisti-
cally indistinguishable from both FL and LA (Fig.
2). The majority of the FL individuals were still
alive at the end of the experiment (Fig. 2). The early
survivorship curves for FL and GA were very simi-
lar, indicating lower mortality than that for LA (Fig.
2), but later, mortality for GA accelerated and was
more similar to that for LA (Fig. 2).


Our prediction, based on local climate, that GA
grasshoppers should have quicker clutch produc-
tion and shorter lifespan than FL grasshoppers,
and that LA grasshoppers would be intermediate,
was not supported. Most reproductive tactics
were roughly equal for FL and GA populations
and longevity did not differ significantly. The
most striking result in the data is the difference
in reproductive tactics (number of clutches, repro-
ductive period) and longevity between FL and LA
We used the period from first clutch until
death to estimate longevity. Hatle et al. (2002)
found that these populations varied in the period
from adult molt to first oviposition in a pattern
partially consistent with variation of the frost-
free interval (i.e., GA < LA = FL; see Introduction
for means). In the present experiment, our mea-
sure of longevity did not include the period from
adult molt to first clutch. Because of this, we have
underestimated the difference in longevity be-
tween GA grasshoppers and LA and especially FL
grasshoppers, and we have also underestimated
the difference in longevity between LA grasshop-
pers and FL grasshoppers. Although the estimate
of longevity we used is incomplete, the fact that it
underestimates the difference between FL and
LA grasshoppers strengthens our inference that
these two populations differ in longevity.

Florida Entomologist 87(2)

1 00~ -south LA. In north GA, lubbers may be present
1 into September (D. W. Whitman, pers. comm.), but
07rida () are clearly declining in abundance during August
075' Forida (a)
07 (M. Brown, pers. comm.). It is unclear why this LA
population of lubbers senesces in September, but
So 050 based on our laboratory data, we propose that de-
0 creased survivorship of LA grasshoppers in Sep-
2 G or- i () tember is a result of intrinsic factors that bring on
f5 2 1a ab senescence, rather than a result of increased dis-
P= 0018 Louisiana (b) ease or predation in this natural environment.
o We find no evidence that interpopulation dif-
S 10 20 30 40 50 60 70 80 ferences in interclutch intervals correlate with
the duration of the FFP at these sites. The time
Time since 1 opposition (d) required to produce the first clutch seems likely to
Fig. 2. Survivorship curves for three populations, be- be the most critical period with respect to laying
ginning at first oviposition. Open points on the curves multiple clutches before the end of the favorable
represent one or more censored observations. Overall X2 season. The calendar dates of laying the second
is for a nonparametric log-rank test of the null hypoth- and third clutches are likely to be earlier if the
esis of equivalent survivorship curves for FL (N = 18), first clutch is shorter. This may explain why we
GA (N = 15), and LA (N = 25). Curves associated with failed to find interpopulation differences inter-
the same letters are not significantly different by pair- clutch intervals that correlate with the FFP,
wise log-rank tests (a = 0.05). correlate with 1 FFP
whereas Hatle et al. (2002) found interpopulation
differences in the time required to produce the
first clutch that did correlate with the FFP.
An estimate of total longevity for these three If local climate is not strongly related to these
populations can be obtained by adding mean reproductive tactics, what does determine these
times to produce the first clutch reported by Hatle fitness-related traits? Sequence analysis of mito-
et al. (2002, see Introduction for values) and mean chondrial DNA yielded a 69% probability that GA
longevities from first oviposition recorded in the and FL populations are more closely related to
present study. Because of censoring, these mean each other than either is to the LA population
longevities underestimate actual longevity, par- (Mutun and Borst 2004). Thus, if reproductive
ticularly for FL. Mean longevities from first ovipo- tactics in this grasshopper are primarily associ-
sition for FL, GA, and LA are 51.5, 43.2, and 35.2 ated with phylogenetic lineage, and not readily
days, respectively, which yield estimates of lon- modified by local climate-driven natural selection
gevities from eclosion of 93.0, 77.7, and 73.7 days, (e.g., because genetic variation for these reproduc-
respectively. Thus, these estimates suggest it is tive tactics is limited) we obtain alternative pre-
FL that is unusual in its longevity and that de- dictions: GA and FL grasshoppers will have
spite a considerably greater apparent active sea- similar reproductive tactics and longevity, and LA
son, LA grasshoppers have a lifespan as short as grasshoppers will differ from GA and FL grass-
that for GA. These differences in longevity appear hoppers. One of our results is consistent with this
to be correlated with differences in first clutch hypothesis. Grasshoppers from GA and FL pro-
mass (FL < GA = LA, Hatle et al. 2002), suggest- duced a similar number of clutches, whereas LA
ing that longevity is indeed negatively related to grasshoppers produced a smaller number of
early reproductive effort across populations. clutches. Differences in interclutch intervals, time
Five years of field observations suggest that from first to last clutch, and longevity are not ob-
this population of LA grasshoppers senesces dur- viously consistent with either of these hypotheses.
ing the first week of September (J.D. Hatle, pers. Our results suggest that if these life history
obs.). Senescence occurs despite the fact that the tactics are related to ecological conditions at each
mean temperature in Lydia, LA for 07 September site, those conditions must involve more than ac-
is 27C and mean rainfall for September is 144 tive season duration, at least as it can be quanti-
mm (Weather.com 2003). Indeed, conditions seem fied by a crude measure like FFP. Alternatively,
to be ideal for lubbers during September in LA, some differences in these life history tactics may
and vegetation is still lush at this season. The in fact not reflect current adaptation, but rather,
mean temperature in Miami, FL for 07 September the phylogenetic constraints derived from the his-
is 28C and the mean rainfall is 160 mm. In Ath- stories of different lineages. Because we have only
ens, GA the mean temperature on 07 September is examined three populations, our ability to corre-
24C and the mean rainfall is 98 mm. In contrast late reproductive tactics with phylogeny is quite
to their September senescence in LA, lubbers are limited. Thus, at present we cannot distinguish
present nearly the entire year in south FL, and all between the hypotheses of more complex ecologi-
but the coldest months in north FL (Capinera cal determinants of reproductive tactics or phylo-
et al. 2001), which has a climate very similar to genetic constraints on reproductive tactics.

June 2004

Gunawardene et al.: Lubber Grasshopper Reproductive Tactics


We thank J. Spring, R. Robichaux, M. R. Brown, and
P. Phares for collecting and shipping grasshoppers, C.
Wagner, M. B. Thake, T. Barry, J. Smith, R. Kesinger, S.
Rowe, S. Karle, and S. Yi for feeding grasshoppers, and
D. W. Borst, D. W. Whitman, and two anonymous refer-
ees for suggestions on the manuscript. This research
was supported by DBI-9978810 to S. A. Juliano, D. W.
Borst and D. W. Whitman.


ALLISON, P. D. 1995. Survival analysis using the SAS
System: A practical guide. SAS Institute, Inc., Cary,
BERKENBUSCH, K., AND A. A. ROWDEN. 2000. Latitudi-
nal variation in the reproductive biology of the bur-
rowing ghost shrimp Callianassa filholi (Decapoda:
Thalassinidea). Marine Biology 136:497-504.
2001. Grasshoppers of Florida. University Press of
Florida. 143 pp.
vival costs of reproduction in grasshoppers. Func-
tional Ecology 1:215-221.
FORSMAN, A. 2001. Clutch size versus clutch interval:
life history strategies in the colour-polymorphic
pygmy grasshopper Tetrix subulata. Oecologia
FRANKINO, W. A., AND S. A. JULIANO. 1999. Costs of re-
production and geographic variation in the repro-
ductive tactics of the mosquito Aedes triseriatus.
Oecologia 120:59-68.
JULIANO. 2002. Geographic variation of reproductive
tactics in lubber grasshoppers. Oecologia 132:517-523.
Trade-off between reproduction and moult-a com-
parison of three Fennoscandian pied flycatcher pop-
ulations. Oecologia 117:374-380.
JOHANSSON, F., AND L. ROWE. 1999. Life history and be-
havioral responses to time constraints in a dam-
selfly. Ecology 80:1242-1252.
KAITALA, A. 1991. Phenotypic plasticity in reproductive
behaviour of waterstriders: trade-offs between re-
production and longevity during food stress. Func-
tional Ecology 5:12-18.
EZELL. 1988. Freeze/Frost Data. Climatology of the

US, No. 20, Suppl. No. 1. National Climate Data
Center, NOAA, Asheville, NC.
1994. Long-term laboratory evolution of a genetic
life-history trade-off in Drosophila melanogaster. 1.
the role of genotype-by-environment interaction.
Evolution 48:1244-1257.
LUKER, L. A., J. D. HATLE, AND S. A. JULIANO. 2002. Re-
productive responses to Photoperiod by a South Flor-
ida Population of the Grasshopper Romalea
microptera (Orthoptera: Romaleidae). Environmen-
tal Entomology 31:702-707.
MIYATAKE, T. 1997. Genetic trade-off between early fe-
cundity and longevity in Bactrocera cucurbitae
(Diptera: Tephritidae). Heredity 78:93-100.
MUTUN, S., AND D. W. BORST. 2004. Intraspecific mito-
chondrial DNA variation and historical biogeography
of the lubber grasshopper, Romalea microptera. An-
nals of the Entomological Society of America. In press.
RICE, W. R. 1989. Analyzing tables of statistical tests.
Evolution 43:223-225.
ROFF, D. A. 1992. The Evolution of Life Histories: The-
ory and Analysis. Chapman & Hall, New York.
ROWE, L., AND D. LUDWIG. 1991. Size and time of meta-
morphosis in complex life cycles: time constraints
and variation. Ecology 72:413-427.
ROWE, L., AND G. G. E. SCUDDER 1990. Reproductive
rate and longevity in the waterstrider, Gerris buenoi.
Canadian Journal of Zoology 68:399-402
condition, and the seasonal decline of avian clutch
size. American Naturalist 143:698-722.
SAS INSTITUTE, INC. 1990a. SAS/STAT@ User's Guide,
vol. 2, Version 8 Edition. SAS Institute Inc., Cary NC.
SAS INSTITUTE, INC. 1990b. SAS/STAT@ User's Guide,
vol. 1, Version 8 Edition. SAS Institute Inc., Cary NC.
STEARNS, S. C. 1992. The Evolution of Life Histories.
Oxford University Press, Oxford.
WHITMAN. 2001. Morphology and development of oo-
cyte and follicle resorption bodies in the Lubber
grasshopper, Romalea microptera (Beauvois). J. Or-
thoptera Res. 10:39-51.
TEMTE, J. L. 1993. Latitudinal variation in the birth
timing of captive California sea lions and other cap-
tive North Pacific pinnipeds. Fisheries Bulletin
US NAVAL OBSERVATORY. 2003. http://aa.usno.navy.mil/
Weather.com. 2003. http://www.weather.com/weather/

Florida Entomologist 87(2)

June 2004


1Department of Entomology, Michigan State University, East Lansing, MI 48824

2Entomology & Nematology Department, University of Florida, Gainesville, FL 32611


The host foraging behavior of the larval parasitoid Diachasma alloeum (Muesebeck) (Hy-
menoptera: Braconidae) from natural populations was directly observed in a highbush blue-
berry, Vaccinium corymbosum L., plantation. More D. alloeum were observed alighting on
blueberry fruit clusters infested with Rhagoletis mendax Curran larvae than were observed
alighting on uninfested blueberry fruit clusters 80 cm away. Approximately equal numbers
of D. alloeum alighted on uninfested blueberries that were mechanically damaged versus
undamaged. The majority of D. alloeum females were attracted to host-infested blueberries
15 to 21 days after R. mendax females had oviposited into fruit. Female D. alloeum spent
more time alighting on R. mendax-infested blueberry fruit clusters than on uninfested blue-
berry clusters 80 cm away. There was no difference in the duration of time spent by D. al-
loeum on mechanically damaged versus undamaged uninfested blueberries. The data herein
are an initial step toward elucidating the cues mediating microhabitat selection by D. al-
loeum in blueberries.

Key Words: Conservation biological control, alighting behavior, Diachasma alloeum, Rhago-
letis mendax


El comportamiento del parasitoide larval Diachasma alloeum (Muesebeck) (Hymenoptera:
Braconidae) por la busqueda del hospedero para alimentarse en poblaciones naturales fue
observado directamente en plantaciones de mora azul, Vaccinium corymbosum L. Se obser-
varon un mayor numero de D. alloeum posando sobre los racimos de la fruta de la mora azul
infestados con larvas de Rhagoletis mendax Curran de los que fueron observados posando so-
bre los racimos de fruta de mora azul no infestados separados por 80 cm de distancia. Aproxi-
madamente numeros iguales de D. alloeum posaron sobre las moras azules no infestadas
que fueron daiadas por la maquinaria agricola versus las no daiadas. La mayoria de las
hembras de D. alloeum fueron atraidas a las moras azules infestadas con el hospedero 15 a
21 dias despu6s que las hembras de R. mendax ovipositaron en la fruta. Las hembras de D.
alloeum pasaron mas tiempo posando sobre racimos de fruta de la mora azul infestados con
R. mendax que en los racimos de la mora azul no infestadas separados por 80 cm de distan-
cia. No habia una diferencia en la duraci6n del tiempo que paso el D. alloeum sobre las moras
azules daiadas por la maquinaria agricola versus las moras azules no daiadas y no infesta-
das. Los datos presentados aqui son un paso inicial hacia el aclaramento de las seiales me-
diadoras para la selecci6n del microhabitat hecho por el D. alloeum en la mora azul.

Hosts of insect parasitoids are often character-
ized by complex and patchy distributions making
successful host location a major challenge for in-
sect natural enemies (Hoffmeister & Gienapp
1999). Exploitation of chemical or visual cues as-
sociated with plants utilized by herbivorous hosts
is known to increase host-searching efficiency of
insect parasitoids (Vet & Dicke 1992; Godfray
1994; Vet et al. 1995). In addition, parasitoids are
often attracted to damaged plants with cases of
heightened attraction to plant damage created
specifically by the herbivore host (Turlings et al.
1991; McAuslane et al. 1991, Henneman et al.

2002). The chemical cues released by herbivore-
damaged plants and exploited by parasitoids in-
clude systemically released plant-volatile com-
pounds (Dicke et al. 1993; Rose et al. 1996).
Several braconid species from the subfamily
Opiinae are known to parasitize larval stages of
Tephritidae (Wharton & March 1978).Diachasma
alloeum (Muesebeck) occurs on hawthorn, Cratae-
gus mollis, and apple, Malus domestic
Borkhausen, in the northeastern U.S.A. and bor-
dering regions of Canada and was thought to spe-
cifically attack the apple maggot fly, Rhagoletis
pomonella (Walsh) (Glas & Vet 1983). Recently, D.

Stelinski et al: D. alloeum Attracted to Host Infested Blueberries

alloeum has also been reported attacking another
member of the Rhagoletis sibling species complex,
the blueberry maggot fly, Rhagoletis mendax Cur-
ran (Liburd & Finn 2003). Parasitization percent-
ages of R. mendax larvae by D. alloeum collected
from abandoned blueberry plantings in Michigan
were extremely high, ranging from 30-50%. These
rates of parasitization are higher than those
known for R. pomonella, which range from 0.1 to
20.1% (Rivard 1967; Cameron & Morrison 1977;
Maier 1982).
Only three detailed studies have been pub-
lished on the behavior of D. alloeum attacking R.
pomonella in hawthorns or apples. Boush & Baer-
wald (1967) reported on the courtship behavior
and suggested the presence of a female-produced
sex pheromone. Prokopy & Webster (1978) and
later Glas & Vet (1983) analyzed the oviposition
behavior of D. alloeum with specific interest in
elucidating the stimuli involved in host-searching
behavior. Visual orientation was found to play an
important role for location of picked hawthorn
fruit in laboratory assays and no difference in at-
tractiveness was found between uninfested and
R. pomonella-infested hawthorn fruit (Glas & Vet
1983). However, ovipositor probing activity and
duration of stay were strongly influenced by the
presence and movement of R. pomonella larvae
feeding inside hawthorn fruit. The authors con-
cluded that host movement within hawthorns
was the prime stimulus for the location of host-in-
fested fruit by D. alloeum (Glas & Vet 1983).
Recent studies with Diachasmimorpha juglan-
dis (Muesebeck) have shown that females can dis-
tinguish between host-infested and uninfested
walnut fruits before alighting (Henneman 1996,
1998). As they approach fruit, females hover close
to the fruit surface for up to 1 sec before they
alight or fly away, possibly assessing volatiles in
order to decide whether to land (Henneman et al.
2002). Presence of fruit damage, however, rather
than presence of larval infestation by R. juglan-
dis larvae appears to produce the necessary cues
for fruit choice by D. juglandis females (Henne-
man et al. 2002). Furthermore, both olfactory and
visual cues are used by D. juglandis females to
distinguish between infested and uninfested wal-
nuts (Henneman et al. 2002).
To our knowledge, nothing has been published
about the biology of D. alloeum in blueberry
plantings. The current communication describes
observations of the behavioral interactions of
D. alloeum females with uninfested, mechani-
cally damaged, and R. mendax-infested blueber-
ries in an abandoned blueberry plantation. The
specific objectives were to 1) determine whether
blueberries infested with R. mendax larvae are
more attractive to D. alloeum females than unin-
fested fruit, 2) determine whether mechanically
damaged and uninfested blueberries are more at-
tractive to D. alloeum females than undamaged

and uninfested fruit, 3) document duration of vis-
its and associated behaviors of D. alloeum on R.
mendax-infested and uninfested blueberries in
the field.


Research Site

Observational studies were conducted in the
summer of 2001 in an abandoned plantation of
highbush blueberry, Vaccinium corymbosum L. in
Fennville, MI. The abandoned plantation was
highly infested by R. mendax with approximately
45% of picked berries containing developing lar-
vae in 1999 and 2000. In addition, this plantation
was known to harbor a substantial population of
D. alloeum. Parasitization rates ofR. mendax col-
lected from this plantation were above 50% in
1999 and 2000.

Insect Source

Rhagoletis mendax were reared from larvae
collected from fruit of unsprayed blueberries (var.
Jersey) from the plantation described above and
from an organically managed plantation 3.2 km
away. Flies were reared according to the protocol
outlined in Liburd et al. (2003). Prior to testing,
flies were maintained in aluminum screen-Plexi-
glas cages (30 x 30 x 30 cm) (BioQuip, Gardenia,
CA) and supplied with water and food (enzymatic
yeast hydrolysate and sucrose) (ICN Biomedicals,
Inc., Costa Mesa, CA). Adults were kept at 24C,
55-60% RH, under a 16:8 (L:D) photocycle.
Three weeks after removal of R. mendax pu-
paria from 4C (diapause), D. alloeum began
emerging from more than 50 and 2% of puparia
collected from the abandoned and organically
managed sites, respectively. The parasitoids were
identified by R. A. Wharton (Texas A&M Univer-
sity) and voucher specimens were deposited at
Michigan State University (A. J. Cook Arthropod
Research Collection).

Observational Study

Forty pairs of blueberry fruit clusters were se-
lected for observation on 12 June before R.
mendax emergence. Each pair of clusters was ap-
proximately 80 cm apart and each individual
cluster contained 20-35 blueberries. All clusters
were approximately 15-cm from the uppermost
bush; this location within the blueberry bush can-
opy has been found to be the most effective posi-
tion for trapping blueberry maggot (Liburd et al.
2000). At this stage of the season, blueberry fruit
was still green and unripe. Experimental bushes
were flagged and selected clusters were individu-
ally enveloped with 1 L translucent plastic bags
that had been punctured with a pin multiple

Florida Entomologist 87(2)

times. Bags were positioned around blueberry
fruit clusters such that berries did not directly
contact the bag surface. The purpose of this bag-
ging was to prevent native R. mendax from ovi-
positing into the selected berry clusters. On 19
June, we captured the first R. mendax on moni-
toring traps in the abandoned plantation. Twenty
of the 40 bagged berry clusters were monitored
from 25 June until 15 July to determine whether
this bagging method interfered with normal berry
development and to determine whether this tech-
nique successfully prevented R. mendax from ovi-
positing into berries. On each day, a single bagged
cluster was randomly chosen for inspection. All
fruit within that cluster were dissected and in-
spected for R. mendax larvae. In addition, on each
day, two randomly selected clusters (15-20 ber-
ries) that had not been previously enveloped with
a plastic bag were dissected for R. mendax larvae.
No R. mendax larvae were found in berries that
were enveloped by our plastic bags. In addition,
berry size and color did not differ between bagged
and unbagged berries. Among the unbagged clus-
ters that were dissected, R. mendax infestation
was first detected on 14 July.
The remaining 20 pairs of bagged clusters
were used for the observational study. On 16 July,
10 of the 20 pairs of bagged blueberry clusters
were randomly chosen for R. mendax infestation.
Ten laboratory-reared and mated R. mendax fe-
males (10-15 days old) were introduced into one of
the bagged blueberry clusters from each pair at
1200 hours. Introduced R. mendax were left in the
bags for 24 h and then removed. Four of the 10
bags containing introduced R. mendax were ob-
served for 1 h to confirm that flies were oviposit-
ing into berries.
The other 10 pairs of bagged blueberry clusters
were chosen for mechanical damage. The blueber-
ries on one cluster of each bagged pair were me-
chanically damaged by making three equally
spaced punctures in the skin of the berries with a
0-size insect pin. These manipulations resulted in
10 replicates of two paired treatments: 1) unin-
fested and undamaged berries versus R. mendax-
infested berries, and 2) uninfested and undam-
aged berries versus uninfested and mechanically
damaged berries. All other blueberries within a
1.5 m radius of each experimental pair were re-
moved from bushes. The paired treatment clus-
ters were 80 cm apart in a two-by-two design with
the two treatments placed in alternate positions;
each pair of treatment clusters was separated by
at least 4 m.
Direct visual observations began 24 h after ini-
tial treatment manipulations were made and con-
tinued thereafter on every second day.
Observations were conducted between 1230 and
1530 h. Two or more observers rotated among the
ten replicates of each treatment pair conducting
approximately 20-min observational bouts per lo-

cation. Observations were terminated 31 days af-
ter the treatment manipulations were conducted.
During each period of observation, the plastic
bags enveloping blueberry fruit clusters were re-
moved and replaced immediately after observa-
tions were terminated. Also, native R. mendax
were prevented from alighting on experimental
clusters during observations. Observed events
were spoken into a hand-held microcassette audio
recorder by an investigator sitting or standing
0.75 m from the paired treatment clusters. Data
recorded were: 1) landing by D. alloeum on berry
clusters, 2) duration of visits on berry clusters, 3)
oviposition into berries by D. alloeum. We at-
tempted to collect observed D. alloeum with an as-
pirator after they oviposited and before they left
experimental berry clusters. We estimate to have
captured 70% of all visitors. The captured D. al-
loeum were taken to the laboratory and their
identity was confirmed.

Statistical Analysis

Results of all dual-choice tests were analyzed
by paired t tests (SAS Institute 2000). In all cases,
significance level was P < 0.05. All values are


Significantly more D. alloeum alighted per day
on blueberry clusters that were infested with R.
mendax larvae than on blueberry clusters that
were uninfested (2.6 0.5 and 0.3 + 0.09, respec-
tively). There was no significant difference be-
tween the number ofD. alloeum alighting per day
on blueberry clusters containing mechanically
damaged and uninfested fruit compared with the
number alighting on clusters containing undam-
aged and uninfested fruit (0.5 0.8 and 0.5 + 0.9,
respectively). Blueberries that were infested with
R. mendax larvae attracted the majority (64%) of
D. alloeum females between 15 and 21 days after
R. mendax females had oviposited into fruit (Fig.
1). There was no noticeable difference in attrac-
tiveness of mechanically damaged and undam-
aged fruit over time.
Female D. alloeum spent significantly more
time alighting on R. mendax-infested blueberry
clusters than on uninfested blueberry clusters
(10.0 1.1 min and 2.3 1.2 min, respectively).
There was no significant difference in the dura-
tion of time spent by D. alloeum on mechanically
damaged and uninfested blueberries compared
with undamaged and uninfested blueberries (1.1
0.1 min and 0.9 + 0.2 min, respectively). Of the
41 D. alloeum observed alighting on R. mendax-
infested blueberry clusters, 34 were observed
making a single ovipositional probe into blueber-
ries. All of the D. alloeum that were observed ovi-
positing into berries performed "excreting"

June 2004

Stelinski et al: D. alloeum Attracted to Host Infested Blueberries

R. mendax-infested versus uninfested berries

S n it II_.

1 3 5 7 9 11 13 15 17

E Infested

N Uninfested

Ir H,

19 21 23 25 27 29 31

Day after blueberry manipulation
Fig. 1. Numbers of D. alloeum observed alighting on R. mendax-infested and uninfested blueberries spaced 80
cm apart every other day after R. mendax oviposition.

behavior directly thereafter as previously de-
scribed by Glas & Vet (1983). Specifically, after
ovipositing, these females walked on the blue-
berry dragging and dabbing their ovipositors on
the fruit surface and excreting a clear fluid. None
of the D. alloeum observed alighting on unin-
fested fruit attempted to oviposit.

More host-infested blueberry fruit were visited
by female D. alloeum than uninfested fruit sug-
gesting that females have the capability of distin-
guishing R. mendax-infested berries prior to
alighting. D. juglandis have also been shown to
distinguish host-infested from uninfested fruits
prior to alighting (Henneman 1996, 1998), relying
on both visual and olfactory cues to make their
decision (Henneman et al. 2002). D. juglandis dis-
tinguish host-infested fruit in the early stages of
infestation (3-4 d after fly oviposition) as eggs are
beginning to hatch (Henneman et al. 2002). In
contrast to our results, previous laboratory stud-
ies comparing the attractiveness ofR. pomonella-

infested hawthorn fruit with uninfested fruit,
showed that D. alloeum did not exhibit a prefer-
ence and landed equally on both types of fruit
(Glas & Vet 1983). However, the hawthorn fruit
used in that study were field collected and in-
fested by R. pomonella under laboratory condi-
tions following a period of cold storage (Glas & Vet
1983). Thus, it is possible that the volatile profiles
released by such picked and stored fruits may
have differed from those of unpicked and R.
pomonella-infested hawthorn fruit. It will be in-
formative to determine whether D. alloeum dis-
tinguishes between R. pomonella-infested and
uninfested hawthorn fruit under field conditions
using unpicked fruit as was done in this study.
The behavior ofD. alloeum documented in the
current study varied in some respects from that
previously reported in hawthorns. The majority of
D. alloeum visits and ovipositions into R. mendax-
infested blueberries occurred 15-21 days after fe-
male R. mendax had oviposited into the fruit. At
this stage, the majority ofR. mendax were likely in
the second instar (Lathrop & Nickels 1932; Neun-
zig & Sorensen 1976). After the twenty-first day,



Florida Entomologist 87(2)

there was a dramatic reduction in the number ofD.
alloeum approaching and alighting on R. mendax-
infested blueberries (Fig. 1). At this point, the ma-
jority ofR. mendax larvae should have reached the
third instar and were likely beginning to exit dry-
ing fruit to pupariate in the soil (Lathrop & Nick-
els 1932). In hawthorns, D. alloeum is known to
attack the third (final) instar ofR. pomonella (Glas
& Vet 1983). In addition, D. alloeum spent less to-
tal time on blueberries during oviposition com-
pared with hawthorns. On average, D. alloeum
spent approximately 10 min on blueberries after
alighting, while they spent anywhere from 18 to
140 min on hawthorn fruit during probing and ovi-
position bouts (Glas & Vet 1983). Furthermore, D.
alloeum were observed making only one oviposi-
tional probe per blueberry, while 1 to 5 oviposi-
tional probes have been observed per individual R.
pomonella-infested hawthorn fruit (Glas & Vet
1983). These differences in behavior ofD. alloeum
in blueberries versus hawthorns are possibly due
to the differences in size and skin rigidity between
blueberries and hawthorns. Given the smaller size
and comparatively less rigid fruit skin of blueber-
ries, it may be easier forD. alloeum to find and ovi-
posit into a younger R. mendax larva in less time
in blueberries than a comparably sized R.
pomonella larva in hawthorns.
In the current study, more D. alloeum females
landed on R. mendax-infested blueberries com-
pared with uninfested berries, but not on me-
chanically damaged blueberries compared with
the undamaged ones. This activity peaked 15-21 d
after R. mendax had oviposited into blueberries.
In contrast, D. juglandis females chose walnuts
based on the presence of fruit damage rather than
the presence of R. juglandis larvae inside the
fruit (Henneman et al. 2002). However, in that
study, mechanically damaged walnuts took on a
distinctly different appearance (darkened) com-
pared with undamaged walnuts, which was
shown to influence fruit selection by D. juglandis.
Color of host-infested walnuts is known to be an
important visual cue mediating searching behav-
ior of D. juglandis (Henneman 1998). In the cur-
rent study, mechanically damaged blueberries did
not appear different from undamaged berries.
Furthermore, R. mendax-infested blueberries re-
mained morphologically indistinguishable from
uninfested berries for more than 20 days after R.
mendax oviposition. It has been documented that
certain female parasitic wasps exhibit an innate
attraction to plant-released volatiles (Geervliet et
al. 1996). Also, parasitic wasps are known to ex-
hibit attraction to the host marking pheromone of
tephritid fruit flies (Hoffmeister & Gienapp
1999). Based on the current results, we postulate
that plant volatile compounds released by R.
mendax-infested blueberries, but not mechani-
cally-damaged fruit, provide an olfactory cue that
attracts female D. alloeum. However, it is also

possible that acoustic signals given off by chewing
and tunneling R. mendax larvae within infested
blueberries provide D. alloeum with an oviposi-
tional stimulus.
Although labor intensive, our approach of con-
ducting direct visual observations of D. alloeum
responding to R. mendax-infested blueberries un-
der authentic field conditions was indeed possi-
ble. Moreover, the data produced are an initial
step toward elucidating the cues mediating mi-
crohabitat selection by D. alloeum in blueberries.
The next step will be to determine whether R.
mendax-infested blueberries release volatile pro-
files that differ quantitatively or qualitatively
from those released by uninfested fruit. Finally,
we hope to identify the relevant volatiles that
may be involved in mediating attraction ofD. al-
loeum to R. mendax-infested blueberries as has
been done for other parasitoids (Turlings et al.
1991). Identification of plant volatiles attractive
to D. alloeum may allow for recruitment of these
beneficial insects in blueberry plantations,
thereby improving biologically based manage-
ment tactics for R. mendax.


We thank the owners of the blueberry plantings used
in this study, who wished to remain anonymous. We
thank Dan Young and Jessie Davis for assistance in col-
lecting R. mendax-infested blueberries and with con-
ducting field observations. We also thank Gary Parsons
(Assistant curator, A.J. Cook Arthropod Research Col-
lection, MSU) for his assistance in preparation of D. al-
loeum specimens before species identification and
deposition of vouchers.


BOUSH, G. M., AND R. J. BAERWALD. 1967. Courtship be-
havior and evidence of a sex pheromone in the apple
maggot parasite Opius alloeus (Hymenoptera; Bra-
conidae). Ann. Entomol. Soc. Am. 60: 865-866.
CAMERON, P. J., AND F. O. MORRISON. 1977. Analysis of
mortality in the apple maggot, Rhagoletis
pomonella, in Quebeck. Can. Entomol. 109: 769-787.
MAN. 1993. Herbivory induces systemic production of
plant volatiles that attract predators of the herbi-
vore: extraction of endogenous elicitor. J. Chem.
Ecol. 19: 581-599.
GEERVLIET, J. B. F., L. E. M. VET, AND M. DICKE. 1996.
Innate responses of the parasitoids Cotesia glomer-
ata and C. rubecula (Hymenoptera: Braconidae) to
volatiles from different plant-herbivore complexes.
J. Insect. Behav. 9: 525-538.
GLAS, P. C. G., AND L. E. M. VET. 1983. Host-habitat lo-
cation and host location by Diachasma alloeum
Muesebeck (Hym.; Broconidae), a parasitoid of
Rhagoletis pomonella Walsh (Dipt.; Tephritidae).
Neth. J. Zool. 33: 41-54.
GODFRAY, H. C. J. 1994. Parasitoids. Behavioral and
Evolutionary Ecology. Princeton University Press,

June 2004

Stelinski et al: D. alloeum Attracted to Host Infested Blueberries

HENNEMAN, M. L. 1996. Host location by the parasitic
wasp Biosteres juglandis (Hymenoptera: Braconi-
dae) under field and greenhouse conditions. J. Kans.
Entomol. Soc. 69: 76-84.
HENNEMAN, M. L. 1998. Maximization of host encoun-
ters by parasitoids foraging in the field: females can
use a simple rule. Oecologia 116: 467-474.
AND R. A. RAGUSO. 2002. Response to walnut olfac-
tory and visual cues by the parasitic wasp Diachas-
mimorpha juglandis. J. Chem. Ecol. 28: 2221-2244.
HOFFMEISTER, T. S., AND P. GIENAPP. 1999. Exploitation
of the host's chemical communication in a parasitoid
searching for concealed host larvae. Ethology 105:
LATHROP, F. H., AND C. B. NICKELS. 1932. The biology
and control of the blueberry maggot in Washington
County Maine. U.S. Dept. Agric. Tech. Bull. 275: 77 p.
CASAGRANDE. 2000. Effects of trap size, placement and
age on captures of blueberry maggot flies (Diptera: Te-
phritidae). J. Econ. Entomol. 93: 1452-1458.
2003. Response of blueberry maggot fly (Diptera: Te-
phritidae) to imidacloprid-treated spheres and se-
lected insecticides. Can. Entomol. 135: 427-438.
LIBURD, O. E., AND E. M. FINN. 2003. Effect of overwin-
tering conditions on the emergence ofDiachasma al-
loeum reared from the puparia of blueberry maggot.
In VanDriesche, R. G. (ed.) Proceedings of the Inter-
national Symposium on Biological Control of Arthro-
pods. Honolulu, Hawaii, 14-18 January 2002, USDA,
Forest Servive, Morgantown, WV.
MAIER, C. T. 1982. Parasitoids emerging from puparia
of Rhagoletis pomonella (Diptera: tephritidae) in-
festing hawthorn and apple in Connecticut. Can. En-
tomol. 113: 867-870.

1991. Stimuli influencing host microhabitat location
in the parasitoid Campoletis sonorensis. Entomol.
Exp. et Appl. 58: 267-277.
NUENZIG, H. H., AND K. A. SORENSEN. 1976. Insect and
mite pests of blueberries in North Carolina. N.C. Ag-
ric. Exp. Stn. Bull. 427: 39 p.
PROKOPY, R. J., AND R. P. WEBSTER 1978. Oviposition-
deterring pheromone of Rhagoletis pomonella a
kairomone for its parasitoid Opius lectus. J. Chem.
Ecol. 4: 481-494.
RIVARD, I. 1967. Opis lectus and 0. alloeus (Hymenop-
tera: Braconidae), larval parasites of the apple mag-
got, Rhagoletis pomonella (Diptera: Tiphritidae), in
Quebec. Can. Entomol. 99: 896-897.
TUMLINSON. 1996. Volatile semiochemicals released
from undamaged cotton leaves. Plant Physiol. 111:
SAS INSTITUTE. 2000. SAS/STAT User's Guide, version
6, 4th ed., vol. 1. SAS Institute, Cary, NC.
PROVEAUX, AND R. E. DOOLITTLE. 1991. Isolation
and identification of allelochemicals that attract the
larval parasitoid, Cotesia marginiventris (Cresson),
to the microhabitat of one of its hosts. J. Chem. Ecol.
17: 2235-2251.
VET, L. E. M., AND M. DICKE. 1992. Ecology and info-
chemical use by natural enemies in a tritrophic con-
text. Annu. Rev. Entomol. 37: 141-172.
VET, L. E. M., W. J. LEWIS, AND R. T. CARD. 1995. Par-
asitoid foraging and learning, pp. 65-101 In R. T
Card6 and W. J. Bell (eds.) Chemical Ecology of In-
sects 2. Chapman and Hall, New York.
WHARTON, R. A., AND P. M. MARSH. 1978. New world
Opiinae (Hymenoptera: Braconidae) parasitic on Te-
phritidae (Diptera). J. Wash. Acad. Sci. 68: 147-167.

Florida Entomologist 87(2)


'Department of Zoology and Entomology, University of Pretoria, Pretoria 0002, Republic of South Africa

2North Carolina State University, Department of Entomology, Box 7613, Raleigh, NC 27695-7613

The population dynamics (in terms of seasonal development) of Gryllotalpa africana Palisot
de Beauvois was documented for the first time in South Africa. An irritant drench (soapy wa-
ter solution) was used to quantify life stage occurrence on turfgrass over a one-year period.
Oviposition took place from early October (spring), with eggs incubating for approximately
three weeks. Nymphs reached the adult stage from March (late summer) and most individ-
uals overwintered in this stage. Adult numbers peaked in early September (early spring),
declining through spring. G. africana was therefore univoltine in the study area. The adult
population was female biased in spring. The smallest nymphs and adults (in relation to
mean length) were collected in December (early summer), while the smallest nymphs (in re-
lation to mean length) occurred in November (late spring).

Key Words: Univoltine, spring oviposition, life stage, turfgrass, mole cricket

La dinamica de la poblaci6n (en terminos del desarrollo estacional) de Gryllotalpa africana
Palisot de Beauvois fu6 documentada por la primera vez en Africa del Sur. Una mojada irri-
tante (una soluci6n de aqua con jab6n) fu6 utilizada para cuantificar la ocurrencia de los es-
tadios de vida en el c6sped durante un period de un aio. La oviposici6n occuri6 desde el
principio de octubre (la primavera), incubando los huevos por aproximadamente tres sema-
nas. Las ninfas llegaron la 6tapa adulta desde marzo (al final del verano) y la mayoria de los
individuos pasaron el invierno en este estadio. El numero mas alto de adults se obtuvo en
el principio de septembre (al principio de la primavera), y decline atrav6z de la primavera.
Desde entonces, el G. africana fue univoltino en la area del studio. Habian una inclinaci6n
viciada hacia las hembras en la primavera. Las ninfas y los adults mas pequeios (en rela-
ci6n al promedio de la longitud) fueron recolectados en diciembre (al principio de verano),
mientras que las ninfas mas pequeias (en relaci6n del promedio de la longitud) fueron reco-
lectados en noviembre (al final de la primavera).

Gryllotalpa africana Palisot de Beauvois (Afri-
can mole cricket) occurs only in Africa (Townsend
1983). The only account concerning the life cycle
of this species is from Zimbabwe (Sithole 1986).
Some notes on the species in South Africa were
provided by Schoeman (1996) and Brandenburg
et al. (2002).
Females lay 30-50 oval, white eggs in hardened
chambers in the soil (Sithole 1986). Incubation pe-
riod is temperature dependent, varying from 15-
40 days. Nymphs feed on earthworms and roots of
plants and under favorable conditions, develop
through six instars with wing bud development
visible in later instars (Sithole 1986). The
nymphal period lasts three to four months. One
generation per year is known (Sithole 1986). Ac-
cording to Schoeman (1996), there are approxi-
mately 10 instars of G. africana in South Africa
and research by Brandenburg et al. (2002) showed
that burrows of the African mole cricket are typi-
cally Y-shaped and range from 100 mm to 230 mm
in length. The life and seasonal cycle of G. afri-
cana has not been investigated in South Africa

and no reports on the seasonal development of
G. africana on African turfgrass are available.
Life cycle, seasonal development and behavior
documentation under the name G. africana in-
clude reports by the United States Department of
Agriculture (1974) (U.S.A. potential introduction
from Asia), Kim (1993, 1995) (Asia), Muraliran-
gan (1979) (Asia), Tindale (1928) (Australia) and
Goodyer (1985) (Australia). It is unknown if these
studies refer to "true" G. africana from Africa.
Life cycle and seasonal development reports
(including voltinism) for similar mole cricket spe-
cies may vary significantly between geographical
areas (Hudson 1987). In a specific area, different
species and even different genera may show gen-
eral life cycle similarity (including voltinism)
(Frank et al. 1998). Therefore analysis of varia-
tions in life cycle, seasonality, and other factors
for species occurring in climates similar to an
area of interest may be more useful in estimating
these parameters for a population than multiple
studies of that species under a range of environ-
mental conditions.

June 2004

de Graaf et al.: Seasonal Development of G. africana


Infested kikuyu grass (Penisetum clandestinum
Hochst ex Chiou) areas at Pretoria Country Club
(2547'16"S; 2815'28"E) were flushed with soapy
water (50 ml Sunlight (Lever Ponds Pty Ltd.,
Durban) dishwashing soap/5 liters H O n this is
a simple, inexpensive but effective surveillance
technique (Short & Koehler 1979). Flushes started
at noon with varying numbers of samples per sam-
pling date (number necessary to collect ca. 100
crickets) with equal sampling intensity (10 liters
soapy water) at each site over an annual period
(Oct 2001-Nov 2002). Sampling was conducted ev-
ery two weeks. Flushed areas were chosen at ran-
dom within each site with the exception that no
area was sampled twice over the duration of the
experiment. Emerging crickets were captured,
counted and total length measured from the poste-
rior of the abdomen (excluding cerci) to the distal
end of the labrum. Pronotal and abdominal
lengths were also measured and recorded. Adults
were sexed and females dissected to determine egg
and oocyte presence for each sampling date. Oo-
cytes were deemed mature (eggs) when covered by
an egg shell (vitelline membrane and chorion). The
long axis of mature eggs was generally longer than
2.5 mm. All sampled areas were under similar ir-
rigation programs and soap flushing efficiency was
assumed to be homogenous for adults and nymphs
between and within sites throughout the study pe-
riod. Immigration and emigration (especially
through flight) were also assumed to be at equilib-
rium and not to affect absolute cricket sizes and
life stage percentages during the study.
Deviation from an equal sex ratio was investi-
gated by the two-tailed binomial distribution
[Sokal & Rohlf 1997; "Statistica" Version: 5 (Stat-
soft, Inc. 1995)]. The Bonferroni method was used
to lower the type one error probability for each
comparison, resulting in an overall significance
level not exceeding 0.05 in the entire series of
tests (Sokal & Rohlf 1997).


The life cycle of G. africana for each ontogenic
stage as a percentage of the total population over
an annual period is graphically presented in Fig.
1. Percentages were calculated by using adult and
nymphal counts for a specific sampling date. Eggs
were not sampled in the field therefore an 'esti-
mate' of egg percentage on that date was calcu-
lated as equal to the mean first instar population
percentage three weeks (mean egg hatch time) af-
ter that date. The egg percentage over time there-
fore only refers to fertilized eggs and may be
subject to considerable variation, as incubation
period is temperature dependant (Frank et al.
1998; Potter 1998). Life stage percentages were
subsequently determined from the ontogenic ratio

obtained. To obtain an annual presentation (from
Nov 2001 to Oct 2002), data were therefore needed
from Oct 2001 to Nov 2002. Fig. 1 shows 61%
adults and 39% nymphs comprised the overwin-
tering population (Jun-Aug). Patchy, relatively
small samples (<40 individuals) were obtained
during winter, which may contribute to the incon-
sistent results obtained during that period (Fig.
1). After overwintering, adult numbers (as a popu-
lation percentage) peaked at 64% and diminished
to 1% during Sep/Oct (spring) and Nov/Dec
(spring/summer), respectively (Fig. 1). The egg
population peaked at the end of Oct (spring) at
41.52%, further following the adult percentage in-
clination, but with some eggs laid until late Feb
(Fig. 1). Oviposited eggs ranged from 2.5-3.5 mm
in length. The graph of nymphal percentages
showed an approximate direct inverse relation-
ship with the adult-percentage-graph when no
eggs were present (Fig. 1). High egg percentages
were associated with the lowest nymphal percent-
ages (Fig. 1). G. africana had a univoltine life cycle
in the study area (Fig. 1). There is a lack of com-
plete percentage overlap for each ontogenic stage
at the beginning and end of the period (Fig. 1).
Mean monthly nymph and overall (adult and
nymph) total length of G. africana for 12 months
are shown in Fig. 2. First instars were 5.95 0.218
mm (mean SD) long, with a midline pronotal
length of 1.52 0.054 mm (data not shown). The
mean monthly nymphal length varied from 6.6
2.56 mm to 25.8 3.70 mm from Nov 2001 (first
and second instars present) to Oct 2002 (late in-
stars present), respectively (Fig. 2). Nymphs over-
wintered from early Jun 2002 when they were 23.0
+ 4.16 mm in length (data not shown), averaging
22.1 3.9 mm over the month (Fig. 2). The mean
monthly overall (adult and nymph) length was at a
minimum (10.3 6.51 mm) and maximum (31.1
5.53 mm) in Dec 2001 and Oct 2002, respectively
(Fig. 2). The mean monthly length of sampled
nymphs and the total (nymphs and adults) popula-
tion showed a relative decline during the winter
(Fig. 2)). No females were sampled in Jan and Feb
2002, when one male in each month was flushed
(resulting in no standard deviation values). Adult
males and females were not distinctly segregated
by mean length over monthly intervals, except for
spring and early summer months, when females
tended to be longer (data not shown). Males and fe-
males measured (mean SD) 35.91 2.16 mm and
36.11 2.40 mm, respectively, in Sep 2002, 31.75
2.38 mm and 34.52 + 3.94 mm, respectively, in Oct
2002, and in Dec 2001 30.83 2.11 mm and 32.33
1.84 mm, respectively. Males and females were at
a maximum length of 36.7 2.33 mm and 37.2
1.85 mm, respectively in Nov 2001 and at a mini-
mum of 30.8 1.61 mm and 30.2 1.27 mm, re-
spectively in July 2002. The mean adult length
over one year was 34.1 3.87 mm, with a midline
pronotal length of 7.8 0.31 mm (data not shown).

Florida Entomologist 87(2)










0 -

- 80



- 20

Fig. 1 The ontogenic stage population percentage of Gryllotalpa africana at Pretoria Country Club, Pretoria,
South Africa from November 2001 to October 2002. Winter period.

Pronotal length was within the ranges reported by
Townsend (1983). Development may be measured
by other parameters than total length, but this
study is also concerned with management, where a
total length measurement is more practical and
easily interpreted by turf managers. Management
related sizes for other mole cricket species have
also been reported in total length (Potter 1998;
Brandenburg & Williams 1993).
Table 1 summarizes female reproductive activ-
ity and the sex ratio ofG. africana per month over
an annual period. Female oocytes started to de-
velop in Apr and the percentage females with oo-
cytes peaked in the winter months (Table 1).
During Jul 2002, 92.3 + 10.13% of females con-
tained oocytes, a figure which was 20.0 42.16%
in Dec 2001. The mean percentage oocytes per fe-
male was highly variable in Dec 2001, but ap-
peared to fit a declining pattern. Oocytes smaller

than one mm in length were found in females
from Apr 2002 to Aug 2002 they increased to 1.5-
2.0 mm in Sept 2002 and 2.0-2.5 mm during Oct
2002, Nov 2001 and Dec 2001 (data not shown).
Females containing eggs (2.5 mm to 3.5 mm long)
were sampled regularly in Sep 2002, Oct 2002,
Nov 2001 and Dec 2001, but peaked in Oct 2002 at
43.0 0.00% of the female population. The highest
number of internal eggs per female was found in
Sep 2002 (38.4 8.55), progressively declining to
Dec 2001 (12.3 9.78). The significance level for
each sample was calculated as P > 0.00217 (P >
0.05/23 comparisons). Table 1 summarizes the
mean ( SD) monthly percentage males of the
adult population over 12 months. The adult field
sex ratio was male biased one sampling date in
May 2002 (date 1: 82.22% males,P > 0.00002, N =
45, date 2: 51.61% males, P > 0.89908, N = 62). Fe-
male bias (in the adult population) was found in

June 2004

de Graaf et al.: Seasonal Development of G. africana


E 30.00

.c 20.00


C 10.00

C- r C0 N C1 C4 C4 CN 4C
p p o p p o p p o o o
0 M C a 5 O
Z>2 < 5 a < 0

Fig. 2 The monthly mean total length ( SD) (from the posterior of the abdomen (excluding cerci) to the distal
end of the labrum) of the nymph and total (adult + immature) population of Gryllotalpa africana at Pretoria Coun-
try Club, Pretoria, South Africa from November 2001 to October 2002. Total = black, nymphs = white.

both Aug 2002 samples (date 1: 12.12% males, P >
0.00001, N = 33, date 2: 24.53% males, P >
0.00027, N = 53). The first Sep 2002 adult sample
was also female biased (date 1: 25% males, P >
0.00023, N = 56, date 2: 30.65% males, P >
0.00316, N = 62). The statistical results also indi-
cated a female bias for the first Oct 2002 adult
sample (date 1: 18.87% males, P > 0.00001, N =
53, date 2: 27.5% males, P > 0.00643, N = 40).
Field (Table 1) sex ratio data (as a male per-
centage, respectively) were normally distributed
in the linear scale (Sokal & Rohlf 1997) for com-
parable months (Kolmogorov-Smirnov test, P >
0.05) ("Statistica" Version: 5 (Statsoft, Inc. 1995)).


During the study period, vitellogenesis was ob-
served from Sep and G. africana females laid fer-
tilized eggs from Oct (mid spring). The highest
number of fertilized eggs in the field was calcu-
lated as occurring during the end of Oct with some
fertilized eggs laid until early Mar. Oviposition
was in clutches (pers. obs.) but the subterranean
nature of egg laying and clutches per female are
unknown. The number of eggs per female and the
adult population started declining from late Sep
and reached a minimum in mid Dec (early sum-
mer). The monthly spring oviposition period was
characterized by the longest females over an an-
nual period that also comprised a significant pro-
portion of the adult population. Female abdomen

length appeared to increase with egg contain-
ment, as females were on average longer than
males only at this time. However, female abdomen
length did not appear to be linearly related to egg
numbers. Absolute length may therefore not be
the best measure to quantify adult size. Gender
behavioral changes may also have influenced
sampling results (lengths) over this period, but
were assumed not to cause significant prejudice.
The data suggest mortality among males was
high during late winter/early spring (causing a fe-
male bias). Migration through flight was not re-
sponsible for temporal gender bias in the field, as
the monthly flight sex ratio was not significantly
different to the monthly field sex ratio and also
showed similar patterns. High male mortality af-
ter mating has been reported for other mole crick-
ets (Scapteriscus spp.) with a univoltine life cycle
(Brandenburg & Williams 1993; Buss et al. 2002),
which suggests, if G. africana males show a simi-
lar tendency, that mating of G. africana occurred
before spring in the present study. Mating may
have occurred in autumn, which has been reported
for univoltine S. borellii (Walker & Nation 1982),
which also oviposit during spring (Frank et al.
1998). Further research (examining female sper-
mathecae for sperm) will confirm mating periodss.
The majority of adults were presumed dead (none
recovered by soap flushing from the soil) by Dec
(early summer), when the sex ratio approached
60:40. This suggests that high female mortality oc-
curred after the oviposition period as reported for

Florida Entomologist 87(2)


Percentage females Percentage females containing
containing oocytes eggs (mean SD) (number of Percentage males in
Date (mean SD) eggs per female) (mean SD) population (mean SD)
November 2001 51.9 16.94 35.71 12.83 (23.4 8.20) 36.1 1.78
December 2001 20.0 42.16 40.0 21.09 (12.3 9.78) 40.0 16.24
January 2002 No females No females 100 a
February 2002 No females No females 100 a
March 2002 0.0 0.0 (0.0) 65.5 11.92
April 2002 22.1 14.67 0.0 (0.0) 53.0 5.08
May 2002 36.6 7.11 0.0 (0.0) 64.5 15.18*
June 2002 91.8 7.06 0.0 (0.0) 50.5 0.63
July 2002 92.3 10.13 0.0 (0.0) 36.6 9.32
August 2002 80.0 17.58 0.0 (0.0) 19.8 6.07*
September 2002 62.7 3.87 32.7 8.60 (38.4 8.55) 28.0 2.83*
October 2002 45.6 7.95 43.0 0.00 (31.3 9.15) 22.58 0.04*
"Only one male and no females sampled (insufficient number for an inference).
*P < 0.001 in at least one sample (see results) (Two tailed binomial distribution, Bonferroni correction (P = 0.05/23 = 0.002)).

other mole crickets with a univoltine life cycle
(Brandenburg & Williams 1993; Buss et al. 2002).
Eclosion (egg hatch) began in November, when
distinctive first and second instars were abundant,
and continued up to mid March. First instars were
dorsally black with thin, white, horizontal, abdom-
inal stripes, apterous and from personal observa-
tions, were the only active jumpers. Their total
length was approximately 7 mm. Second instars
were dorsally brown, apterous and up to 9 mm to-
tal length. All following instars were dorsally grey-
ish-brown (adults and nymphs are light yellow on
the ventral side) and resembled adults in appear-
ance but were smaller and only developed wing
buds in later instars. The relatively long oviposi-
tion period caused some instar overlap, as evident
from standard deviation values for mean nymph
absolute length. The overall (adult and nymph)
mean absolute length was highly variable in No-
vember, but length was shorter with less variabil-
ity in December, as a result of adult mortality over
the two months. Nymphal development rate in-
creased with relative warmer temperatures and
the new generation adults appeared from late
summer/early autumn. Adults have fully devel-
oped tegmina and hind wings and are capable of
flight. The new generation adults consisted of more
males during autumn, with a significant male in-
clination in May (although May samples were sub-
ject to relatively high variance). This suggests that
males may eclose before females and then subse-
quently die earlier. The data indicate a minimum
period of five months from oviposition to adult. The
life cycle may, however, have been as long as eight
or nine months if oviposition took place in late
summer. The seasonal ontogenic stage occurrence
was relatively similar in flush samples from across
the Pretoria region (unpublished data).

Most nymphs completed their development by
early June, when an over wintering phase was en-
tered to the end of August, during which time indi-
viduals may have moved deeper down in the soil
profile. During this period, small, patchy infesta-
tions (lowest density sampled during late July)
were found in moist turf areas with relatively high
soil temperatures (usually northern exposures).
Sampling bias may have caused relatively high
variability in life stage constitution during over
wintering. Factors including behavioral changes,
relative smaller samples prone to higher variability
and/or destructive sampling may have contributed
to the bias. Total length during winter samples
showed a relative decline and also may have been
due to sampling bias. Smaller (in relation to length)
individuals sampled may have reflected a tendency
of younger (and shorter) adults and nymphs to stay
active as long as possible to attain a larger size
(longer length) to increase their fitness during the
following spring reproductive period. Larger males
of Scapteriscus spp. produce louder calls and attract
more females (Forrest 1980, 1983, 1991), while
larger Scapteriscus spp. females produce three
times more offspring and 1.5 times as many eggs
per clutch than smaller females (Forrest 1986). The
G. africana population became more adult biased
during spring, when the development was com-
pleted. Adult length during spring was variable by
month, but may support a contention by Forrest
(1987), that as the spring reproductive period sea-
son progresses, a greater proportion of smaller indi-
viduals (of both genders) should mature because
costs due to delaying reproduction increase.
There was annual variation (on a constant spa-
tial scale) in the development of G. africana.
Mean egg hatch in 2002 was 2 weeks later than in
2001. Soap flushes should, therefore, be con-

June 2004

de Graaf et al.: Seasonal Development of G. africana

ducted weekly to quantify spatial and temporal
variance (this is especially important to guide
management practices).
The seasonal development of G. africana re-
ported in this study is very similar to that reported
for univoltine Scapteriscus spp. in the southeast-
ern U.S.A. (Brandenburg & Williams 1993).
Preliminary studies indicate peak oviposition
occurred a few weeks later on golf courses in the
cooler, southern regions (Johannesburg), a pattern
followed by some New World species (Branden-
burg 1997; Potter 1998). Temperature therefore
appeared to be an important factor influencing egg
laying period in G. africana. Brandenburg (1997),
however, found that timing and intensity of egg-
laying and egg hatch do not seem to be closely re-
lated to soil temperature or the number ofS. vici-
nus and S. borellii females captured in acoustic
traps. Hertl et al. (2001) found a significant posi-
tive linear relationship between ovipositing fe-
males (number of eggs laid per female were
constant) and soil moisture in S. borellii. Soil mois-
ture may also influence oviposition in G. africana.
Preliminary studies also show that the propor-
tion of adults in the population prior to overwin-
tering might be smaller in the southern areas
(Johannesburg). Adult overwintering proportions
are variable (on a constant spatial scale) for S.
vicinus (Brandenburg 1997), suggesting that val-
ues reported in this study may also be variable
between years.
Some specific behaviors ofG. africana were ob-
served during the course of this study. Adults
were found to be cannibalistic, especially at high
densities. G. africana adults usually expelled a
characteristic non-sticky, pungent smelling, dark
brown fluid when handled, possibly as a deter-
rence or defense mechanism (pers. obs.). Other
genera (Neocurtilla and Scapteriscus) and Gryllo-
talpa species (G. oya) also are known for secreting
fluids that may be smelly and vary from a low to
high viscosity (Baumgartner 1910; Tindale 1928;
Walker & Masaki 1989).

BAUMGARTNER, W. J. 1910. Observations on the Gryl-
lidae: III Notes on the classification and on some
habits of certain crickets. Kansas Univ. Scientific
Bull. 5: 309-319.
BRANDENBURG, R. L. 1997. Managing mole crickets: De-
veloping a strategy for success.Turfgrass Trends 6: 1-8.
plete Guide to Mole Cricket Management in North
Carolina.. N.C. State Univ. Raleigh.
2002. Tunnel architectures of three species of mole
crickets (Orthoptera: Gryllotalpidae). Florida Ento-
mol. 85: 383-385.
Buss, E. A., J. L. CAPINERA, AND N. C. LEPPLA. 2002.
Pest mole cricket management. Univ. of Fla., Gaines-
ville. World Wide Web: http://edis. ifas.ufl.edu/

FORREST, T. G. 1980. Phonotaxis in mole crickets: Its re-
productive significance. Florida Entomol. 63: 45-53.
FORREST, T. G. 1983. Calling songs and mate choice in
mole crickets, pp. 185-204. In D. T Gwynne and G. K.
Morris [eds.], Orthopteran Mating Systems: Sexual
Selection in a Diverse Group of Insects. Westview
Press, Boulder, CO.
FORREST, T. G. 1986. Oviposition and maternal invest-
ment in mole crickets (Orthoptera: Gryllotalpidae):
effects of season, size, and senescence. Ann. Ento-
mol. Soc. Amer. 79: 918-924.
FORREST, T. G. 1987. Insect size tactics and develop-
mental strategies. Oecologia 73: 178-184.
FORREST, T. G. 1991. Power output and efficiency of
sound production by crickets. Behav. Ecol. 2: 327-338.
FRANK, J. H., T. R. FASULO, AND D. E. SHORT. 1998.
Mcricket Knowledgebase. CD-ROM. Institute of Food
and Agricultural Sciences. Univ. Fla., Gainesville.
GOODYER, G. J. 1985. Mole crickets. Agfacts 37: 1-4.
CHECK. 2001. Effect of soil moisture on ovipositional
behavior in the southern mole cricket (Orthoptera:
Gryllotalpidae). Environ. Entomol. 30: 466-473.
HUDSON, W. G. 1987. Variability in development of
Scapteriscus acletus (Orthoptera: Gryllotalpidae).
Florida Entomol. 70: 403-404.
KIM, K. W. 1993. Phonotaxis of the African mole cricket,
Gryllotalpa africana Palisot de Beauvois. Korean J.
ofAppl. Entomol. 32: 76-82 (Abstr.).
KIM, K. W. 1995. Seasonal changes in age structure and
fecundity of the African mole cricket (Gryllotalpa af-
ricana) population in Suwon, Korea. Korean J. Appl.
Entomol. 34: 70-74 (Abstract cited).
MURALIRANGAN, M. C. 1979. On the food preference and
the morphological adaptations of the gut of some
species of Orthoptera. Current Science 49: 240-241
POTTER, D. A. 1998. Destructive Turfgrass Pests. Ann
Arbor Press, MI.
SCHOEMAN, A. S. 1996. Turfgrass insect pests in South
Africa. Turf and Landscape Maintenance 7: 15.
SHORT, D. E., AND P. G. KOEHLER 1979. A sampling
technique for mole crickets and other pests in turf
grass and pasture. Fla. Entomol. 62: 282-283.
SITHOLE, S. Z. 1986. Mole cricket (Gryllotalpa africana).
Zimbabwe Agric. J. 83: 21-22.
SOKAL, R. R., AND F. J. ROHLF. 1997. Biometry. pp. 57,
61-123, 135, 179-260, 392-440, 451-678. W.H. Free-
man and Company, New York.
STATSOFT INCORPORATED. 1995. Statistica. Version 5.0.
TINDALE, N. B. 1928. Australasian mole-crickets of the
family Gryllotalpidae (Orthoptera). Records of the
South Australian Museum 4: 1-42.
TOWNSEND, B. C. 1983. A revision of the Afrotropical
mole-crickets (Orthoptera: Gryllotalpidae). Bull.
Brit. Mus. Nat. His. (Entomology) 46: 175-203.
Insects not known to occur in the continental United
States. African mole cricket (Gryllotalpa africana
Beauvois). Coop. Econ. Insect Report 24: 41-43.
WALKER, T. J., AND S. MASAKI. 1989. Natural history,
pp. 1-42. In F. Huber, T E. Moore and W. Loher [eds.],
Cricket Behavior and Neurobiology. Cornell Univer-
sity Press, Ithaca.
WALKER, T. J., AND J. L. NATION. 1982. Sperm storage
in mole crickets: Fall matings fertilize spring eggs in
Scapteriscus acletus. Florida Entomol. 65: 283-285.

Florida Entomologist 87(2)


'Archbold Biological Station, P.O. Box 2057, Lake Placid, FL 33862

2Department of Entomology, Museum of Comparative Zoology, Harvard University, Cambridge, MA 02138


The ponerine ant Odontomachus relictus n. sp. is described from specimens collected in
scrub and sandhill habitats on several ancient sand ridges in Florida. It appears to be a
relict species from dry periods in the Pleistocene. Workers are similar to the western species
0. clarus Roger, but males of the two species are strongly divergent. Keys and natural his-
tory notes are provided for workers and males of the four Odontomachus species known from
the U.S. Examination of males might help clarify the taxonomic status of Odontomachus of
Central and South America.

Key Words: Florida endemics, arenicolous arthropods, Odontomachus, ants, Formicidae,


La hormiga ponerine Odontomachus relictus n. sp., es descrita de especimenes recolectados
en los habitat de matorrales y de los bancos de arena en varias lomas de arena antiguas en
la Florida. Parece ser una especie reliquia de los periods secos del Pleistoceno. Los obreras
son similares a la especie occidental 0. clarus Roger, pero los machos de los dos species son
fuertemente divergentes. Se provee las claves y notas de la historic natural sobre las obreras
y los machos de las cuatro species de Odontomachus species conocidas en los Estados Uni-
dos. La examinaci6n de los machos puede ayudar a aclarificar el status taxon6mico de los
Odontomachus de centro y de suramerica.

Members of the genus Odontomachus are of-
ten common and conspicuous insects. They are
relatively large for ants (length often around 8
mm), with elongate mandibles whose powerful
snapping action is produced by massive muscles
accommodated in bulging lobes on the head cap-
sule (Fig. 1A, B). Odontomachus has achieved
some fame for the speed of its mandibular snap,
which occurs in 0.33-1.00 millisecond, the fastest
animal movement known (Gronenberg et al.
1993). In spite of these formidable jaws, backed
up by a sting strong enough to elicit a definitive
reaction in humans, these ants are not particu-
larly fierce, and are usually seen stalking slowly
about singly on the surface of leaf litter. As might
be expected, examples of such large and obviously
interesting ants began to accumulate in collec-
tions at an early date, resulting in the naming of
numerous species and forms. This proliferation of
names was based to a large extent on what
seemed to be a good array of useful characters, in-
cluding pilosity, color, surface sculpture, and the
shape of the mandibles and petiole.
Unfortunately, it appears that many of the char-
acters used in diagnoses of Odontomachus species
show intraspecific variation, resulting in large
numbers of synonyms. Bolton's 1995 catalog of For-

micidae includes 161 specific and subspecific
names for extant Odontomachus species, 60 of
which Bolton lists as valid names. Most of the
credit for the simplification of nomenclature should
go to Brown's 1976 review of the genus. Lineages
that include variable species also may include
cryptic species, and this seems to be true of Odon-
tomachus. In the U.S., Odontomachus nomencla-
ture was at its most austere following Brown's
1976 salutary pruning of the genus, resulting in
one recognized species in the Southeast, 0. brun-
neus (Patton), and one recognized species in the
Southwest, 0. clarus Roger. Deyrup et al. (1985)
showed that there were three species in the South-
east: 0. brunneus, 0. ruginodis M. R. Smith (prob-
ably a relatively recent introduction to the area),
and what appeared to be an isolated population of
the southwestern 0. clarus, restricted to arid
dunes in central peninsular Florida. New evidence
reveals that the southeastern species thought to be
0. clarus is a different, undescribed species.
The purpose of this paper is to name this new
species, to present an identification guide to the
four U.S. species, to summarize the known natu-
ral history of all four species, to briefly cover the
nomenclature of the species, and to indicate a few
residual problems.

June 2004

Deyrup & Cover: Odontomachus Ants of United States


Fig. 1. Odontomachus relictus, new species, worker. A. Lateral view. B. Frontal view of head. C: Posterior view
of petiole.

OF Odontomachus

Family Formicidae, subfamily Ponerinae, tribe
Odontomachini (Hdlldobler & Wilson 1990). Od-
ontomachus is a senior synonym of Pedetes Dalla
Torre, Ci..'....... .... Emery, and Myrtoteras
Matsumura (Bolton 1995); these junior synonyms
have not been used for more than 25 years, and
there is no current confusion about these names.

Diagnosis (modified from Holldobler and Wilson 1990)

Mandibles slender, elongate, attached near
middle of anterior margin of head, abruptly bent
inward at apex, widened apices expanded with
three teeth arranged in a vertical series; third ab-
dominal segment not differentiated by a constric-
tion from the rest of the abdomen. Nuchal carina
(the ridge delimiting the occiput) V-shaped, nar-
rowed toward its median into a mid-dorsal groove;
apophyseal lines present as convergent lines from
the vertex of the head up to the nuchal carina.
The shape of the nuchal carina and the presence
of apophyseal lines distinguish Odontomachus
from the somewhat similar genus Anochetus. A
simple character for separating Odontomachus
from Anochetus in the U.S. is the shape of the peti-
ole, ending in a dorsal spine or simple cone in Od-
ontomachus, and in a pair of spines or a pair of
angles in Anochetus mayri Emery, the only repre-
sentative of its genus known from the U.S. (Deyrup
2002).Anochetus mayri, which is native to the West
Indies and exotic in Florida, is smaller (4 to 5 mm
in length) than the Odontomachus species consid-

ered here.Anochetus kempfi Brown (West Indies) is
within the size range of local Odontomachus, but
very slender, and with a two-spined petiole.

Odontomachus relictus Deyrup and Cover,
new species

Diagnosis of worker (Fig. 1).

Distinguished from other U.S. Odontomachus
by the following combination of character states:
conspicuous striae present on basalar lobe (oval
sclerite at dorsal posterior corner of mesopleu-
ron); posterior side of petiole without transverse
striae; appressed hair on first gastral tergite
sparse, short, spaces between hairs often as wide
as the length of hairs.

Description of holotype worker

Features visible in lateral view described from
left side. Measurements in mm. Total length
(length of head excluding mandibles + length of
mesosoma + length of petiole + length of gaster):
7.48; head length: 2.12; width of head at rear mar-
gins of eyes: 1.80; width of head at widest part of
occipital lobes: 1.62; length of left mandible: 1.20;
maximum width of eye: 0.30; maximum width of
clypeal area: 0.30; length of mesosoma: 2.67; length
of petiole: 0.52; length of gaster: 2.17. Head: fine
striae diverging from frontal lobes, covering frontal
aspect of head, disappearing before occipital area,
covering only the upper quarter of the extraocular
furrow; posterior lateral corners, occipital area, un-
derside of head smooth and shining. Mesosoma:

Florida Entomologist 87(2)

pronotum with roughly circular concentric striae,
without longitudinal striae reaching the hind mar-
gin; mesonotum and propodeum with transverse
striae; striae present on basalar lobe; mesopleuron
smooth, shining, with longitudinal striae along
dorsal and ventral margins. Petiole: apically spi-
nose; in profile anterior face convex, posterior face
bisinuate; posterior face in posterior view smooth
and shining, without hairs. Gaster: shining, no sur-
face sculpture except for minute punctures from
which hairs emerge; first tergite sparsely covered
with short, appressed, pale hairs, spaces between
hairs usually as wide as length of hairs; first terg-
ite with sparse, scattered, suberect long hairs, in-
cluding an uneven subapical row. Color: head,
antennae, mesosoma, petiole dark reddish brown,
contrasting with blackish-brown gaster; legs dark
yellow, contrasting with mesosoma.

Diagnosis of queen

Queens of the four U.S. species were examined.
Diagnosis as in worker.

Description of a paratype queen

Measurements in mm. Total length (length of
head excluding mandibles + length of mesosoma
+ length of petiole + length of gaster): 8.37; head
length: 2.15; width of head at rear margins of
eyes: 1.77; width of head at widest part of occipi-
tal lobes: 1.70; length of left mandible: 1.20; max-
imum width of eye: 0.27; maximum width of
clypeal area 0.25; length of mesosoma: 2.75;
length of petiole: 0.52; length of gaster: 2.95.
Structural character states and color as in
worker, except for occurrence of ocelli and expan-
sion of the mesosomal dorsum (pronotum, me-
sonotum, scutellum) associated with flight;
pronotum transversely striate, mesonotum longi-
tudinally striate.

Diagnosis of male

Distinguished from other U.S. Odontomachus
by the following combination of character states:
ocelli very large, wider than the distance between
the lateral ocelli and the eyes (Fig. 2A); body color
medium brown, antennae yellowish.

Description of a paratype male

Measurements in mm. Total length (length of
head excluding mandibles + length of mesosoma +
length of petiole + length of gaster): 6.66; head
length: 1.07; width of head at widest part, includ-
ing eyes:1.45; length of mesosoma: 2.25; length of
forewing: 4.95; length of petiole: 0.47; length of
gaster: 2.57. Head: in frontal view, eyes longer
than the distance between them dorsally; median
ocellus wider than the distance between a lateral

ocellus and the margin of the eye; clypeus in pro-
file not strongly protuberent. Mesosoma: prono-
tum, mesopleural area above and below episternal
suture feebly shining, not striate; mesonotum
finely striate, transversely on anterior quarter, re-
mainder longitudinally; scutellum convex, shin-
ing, lacking a median carina; propodeum without
a raised carina, feebly shining, with weak, fine
striae in the following patterns: a median series of
concentric ovals, posterior portion with transverse
bisinuate lines, obliquely longitudinal lines later-
ally ventral to spiracle; propodeum in profile long
and low, without a declivitous posterior portion;
gaster shining, tergites without surface sculpture
except for fine, hair-bearing punctures, evenly cov-
ered with long, fine, sub-appressed hairs.
Type localities and associated information, as
appear on specimen labels.-Holotype Worker.
FL: Highlands Co., Archbold Biological Station,
15-IV-1996, M. Deyrup, recently burned mature
scrub, Red Hill, nest in sand. Paratype dealate
queen used for description of queen (designated
on label). Same locality, collector as holotype, 18-
1-1983, Florida scrub habitat with Ceratiola eri-
coides, E. side of Tract 7. Paratype male used for
description of male (designated on label). Same lo-
cality, collector as holotype, 8-XI-1990, at window,
main building. Paratype workers and queens (all
from Florida). The following specimens all have
the same locality and collector as the holotype: 18
workers: same data as holotype (nest series with
holotype); 2 workers: 30-IX-1982, sandhill area
with Quercus laevis, SE Tract, at bait; 4 workers:
3-X-1982, in patch of Hypericum, road 19E, at
bait; 2 workers: 19-IX-1982, mature stand of Pi-
nus clausa, SE Tract, trail 10, at bait; 3 workers:
30-XIII-1985, mature sand pine scrub habitat,
Red Hill, trail 1; 1 worker: 28-X-1982; 1 worker: 7-
VI-1984, sandhill habitat, Tract 19E; 1 dealate
queen with associated worker: 13-X-1982, in leaf
litter of Bejaria racemosa, Tract 7, road 18; 1
worker: 24-XI-1982, in dry leaf litter at base of
oak, sandhill habitat, Tract 19; 2 workers: 28-VI-
1985, in leaf litter of Carya floridana, sandhill
habitat, Tract 19E; 1 worker: 6-IV-1983, mature
Pinus clausa scrub, in pan trap below Townes
trap; 1 worker: 5-IX-1985, mature Pinus clausa
scrub, NE firelane of NE Tract; 1 worker: 14-XI-
1985, mature sand pine scrub, Red Hill, SE Tract;
1 worker: 13-VI-1985, sand pine scrub habitat; 1
worker: 17-X-1988; 1 worker: 3-X-1982, in tussock
of Andropogon; 1 worker: 6-VII-1984, mature
sand pine scrub habitat, NE firelane of NE Tract;
1 worker: 25-VII-1984, mature sand pine scrub
habitat, NE firelane of NE Tract; 1 worker: 28-VI-
1985, Tract 19E, sandhill habitat. Paratype work-
ers and queens not from holotype locality: 3 work-
ers: Highlands Co., Lake Placid, 18-IX-1987, P.
Martin, sand pine scrub habitat in former YMCA
Camp 2 mi. S. of town on Highway US 27; 1
worker: same locality, collector, habitat as previ-

June 2004

Deyrup & Cover: Odontomachus Ants of United States



E F ,F


Fig. 2. Odontomachus species. A-D: heads of males, occipital and lateral views; A: relictus, B: clarus, C: brunneus,
D: ruginodis. E-H: propodeal areas of males, lateral views; E: relictus, F: clarus, G: brunneus, H. ruginodis. At var-
ious times relictus (A, E) has been confused with clarus (B, F) and brunneus (C, G) with ruginodis (D, H). I: rugin-
odis, posterior side of petiole of worker. J-K: basalar sclerites (oval structure at upper right corer of mesopleuron)
of workers: J: clarus, K: relictus.

ous, 30-X-1987; 3 workers: same locality, habitat
as previous, 27-X-1987, J. Cronin, 1 worker: High-
lands Co., Lake Placid, 15-X-1986, M. Deyrup,
Holmes Avenue scrub site east of town; 3 workers:
Highlands Co., Sebring, 11-IX-1987, P. Martin,
former Flamingo Villas Devel., 3.7 mi. SE of Se-
bring, sand pine scrub; 2 workers: same locality,
habitat as previous, 10-XI-1987, J. Cronin; 1
dealate queen with associated worker: same local-
ity, habitat as previous, 17-IX-1990, M. Deyrup; 1
worker: Polk Co., TNC Tiger Creek Preserve, 5-X-

1989, M. Deyrup, recently burned scrub with
Quercus laevis; 1 worker: Polk Co., Lake Wales,
26-1-1988, P. Martin, Flaming Arrow Boy Scout
Camp, east of town, sand pine scrub; 1 dealate
queen: same site, habitat as previous, 24-XI-1987,
J. Cronin; 12 workers: Orange Co., Walt Disney
World, S16, T24S, R27E, 5-VII-1996, Z. Prusak,
sand pine/oak scrub zone MW-5 (unburned) #88; 2
workers: Orange Co., Walt Disney World, 22-VII-
1996, Z. Prusak, MW-5 Cons. Area, unburned
zone, sand pine scrub; 2 dealate queens, 4 work-

Florida Entomologist 87(2)

ers: Citrus Co., 12 mi. NW of Brooksville, 11-V-
2002, M. Deyrup & J. Mosley, Withlacoochee State
Forest, Sugar Mill Tract, north of 480, sandhill
with oaks; 4 workers: Citrus Co., 5 mi. W. of Inv-
erness, 14-IX-1991, M. Deyrup, sandhill habitat,
Withlacoochee State Forest; 2 workers: same lo-
cality, collector, habitat as previous, 14-XI-1991; 4
workers: Citrus Co., Withlacoochee State Forest,
14-XI-1991, M. Deyrup, 3.2 km south on forest
road that begins 7.2 km west ofjct. State Road 44
and County Road 581, open sandhill habitat,
sparse ground cover ofAristida beyrichiana, scat-
tered mature Pinus palustris and Quercus laevis;
5 workers: Citrus Co., Pine-Oak Estates, 1-IV-
1993, M. Deyrup, sandhill habitat along Rt. 488, 3
mi. NE jet. with Rt. 495. Paratype males. The fol-
lowing males are from the Archbold Biological
Station, collected by M. Deyrup in Townes flight
traps on trails through mature sand pine scrub;
dates as follows: 2: 19-VI-1985; 1: 8-VII-1985; 2:
29-VI-1985; 1: 13-VII-1985; 1: 14-VII-1985; 1: 10-
VI-1985; 1: 26-VI-1985; 2: 5-VII-1985; 4: 1-VII-
1985; 1: 9-X-1987; 1: 28-XII-1983; 1: 16-VII-1986;
1: 24-VII-1985; 1: 21-VII-1983; 1: 3-VII-1983; 1:
27-VI-1983; 2: 28-VI-1983; 1: 21-VI-1983; 1: 20-
XI-1985; 1: 8-VII-1983; 1: 23-VIII-1984. Addi-
tional paratype males: 7: Orange Co., Walt Disney
World, S16, T24S, R27E, 1-5-VII-1996, Z. Prusak,
sand pine/oak scrub zone MW-5 (unburned), Mal-
aise trap; 2: Orange Co., Walt Disney World, 8-13-
VIII-1996, Z. Prusak, MW-5 Cons. Area unburned
zone, sand pine scrub, Malaise trap.

Deposition of types

Holotype, 3 paratype dealate queens, 32
paratype workers, 12 paratype males: Museum of
Comparative Zoology, Harvard University, Cam-
bridge, Massachusetts; 6 paratype workers, 4
paratype males: The Natural History Museum,
London; 7 paratype workers, 4 paratype males:
Los Angeles County Museum, Los Angeles, Cali-
fornia; 10 paratype workers, 4 paratype males:
Florida State Collection of Arthropods, Gaines-
ville, Florida; remaining type material in the ar-
thropod collection of the Archbold Biological
Station, Lake Placid, Florida.


relictus, past participle of relinquo: left behind,
referring to the distribution of the species on relict
patches of Florida scrub and sandhill vegetation on
high sand ridges in south-central Florida.

Relationship between relictus and other species

Workers of relictus and clarus are morphologi-
cally similar, except for the striate basalar lobe
(Fig. 2K) and consistently spinose petiole of relic-
tus. Until we compared the males of the two spe-

cies, we had interpreted these differences as small
divergences between widely separated popula-
tions of a single species, especially since clarus
shows considerable variation through the South-
west. The differences between males are the only
clear evidence at present that clarus and relictus
are distinct. The extraordinarily large eyes and
ocelli of relictus (Fig. 2A) suggest that there is
some feature of relictus flight behavior that is dif-
ferent from the flight behavior ofclarus; any major
difference may be likely to confer reproductive iso-
lation. A possibly relevant feature ofrelictus flight
behavior is that male activity, as monitored by
Malaise traps, seems to be concentrated around
moonlit nights (Deyrup et al. 1985). Another major
structural difference is the longer, less declivitous
propodeum of relictus; all four species occurring in
the U.S. show conspicuous differences in the shape
and sculpture of the propodeal area (Figs. 2E-H).
It is tempting to hypothesize that relictus is
closely related to clarus, representing an eastern
offshoot of a western lineage adapted to dry habi-
tats. Some other animals, such as the Florida sand
roach (Arenivaga floridensis Caudell), the Florida
scrub-jay (Aphelocoma coerulescens) and the go-
pher tortoise (Gopherus polyphemus) seem to be
examples of western lineages in relic desert-like
habitats in Florida (Deyrup 1989). These animals
could have spread east, along with a rich fauna of
now extinct savanna-dwelling vertebrates, along
an arid corridor in southern North America dur-
ing the late Pliocene through mid-Pleistocene
(Webb 1990). In Deyrup's 1989 and 1990 papers
relictus, under the name of clarus, is specifically
mentioned as an example of such a western lin-
eage. Male relictus, however, do not support this
hypothesis. They share their large eyes and ocelli,
non-carinate scutellum and low propodeum with
the other native southeastern species, brunneus
(Figs. 2C, G). When the taxonomic status and dis-
tribution of southeastern brunneus is better un-
derstood, it may be possible to propose a new
hypothesis on the derivation of relictus.
All four of the U.S. species of Odontomachus
are combined with 17 New World species and two
Old World species in the haematodus species
group, distinguished by the reduction of the seg-
ments of the labial palps from four to three
(Brown 1976).

Habitat and distribution of relictus

This species is a subterranean nester, and
found only in areas of deep, unconsolidated, silica
sand. These areas may be covered with Florida
scrub vegetation, consisting of scattered pines,
small oaks and other small trees and shrubs.
Sometimes there are areas of bare sand, espe-
cially in the first few years following a fire. Alter-
natively, areas where relictus occurs may be
sandhill vegetation, consisting of scattered pines

June 2004

Deyrup & Cover: Odontomachus Ants of United States

above a low layer of grasses and forbs. For de-
scriptions of these habitats, see Myers (1990).
Nest entrances are not marked by a mound, but
by scattered pellets of sand. Digging into a nest
may produce a few workers, sometimes with
brood, but no large aggregation of workers.
Odontomachus relictus is known from the Lake
Wales Ridge, the southern Brooksville Ridge and
the Orlando Ridge. It has not been found in scrub
or sandhill habitats on the Atlantic Coastal Ridge,
the Northern Brooksville Ridge, or the sandy up-
lands of northern Florida. The inland south-cen-
tral sand ridges of the Florida Peninsula are over
a million years old, and are known to have many
plants and animals found nowhere else (Deyrup
1989, 1990). Some of these species appear to be
remnant populations of species that were once
more widespread, others are probably true (au-
tochthonous) endemics. Up to the discovery of the
distinct status of 0. relictus, it seemed that this
ant was an example of a series of remnant popula-
tions; now it appears that this species could just as
easily be a true endemic of south-central Florida.

The restricted range and habitat of 0. relictus
might raise questions about its conservation sta-
tus. About twenty-five years ago, its prospects
would have seemed poor. At that time there were
only two protected populations, one on the Lake
Wales Ridge (Archbold Biological Station), the
other on the southern Brooksville Ridge (Withla-
coochee State Forest). Upland areas were being
converted rapidly to housing and agriculture, and
it seemed that few scrub and sandhill areas would
remain within the range of 0. relictus. Since that
time, development and habitat destruction have
occurred at an unprecedented rate, but the estab-
lishment of ecological preserves also has been re-
markably fast, especially on the Lake Wales Ridge.
This species now appears to be adequately pro-
tected, unless it is subjected to some widespread
environmental threat that pervades natural ar-
eas. Odontomachus relictus is a good example of a
species that seemed destined for the endangered
species list, with all the trouble and expense im-
plied in such listing, but was preemptively pro-
tected by more general conservation programs.

SPECIES OF Odontomachus IN THE U.S.

The following keys distinguish between the four species known from the U.S. Brief comments on dis-
tribution, nomenclature and natural history follow the keys.

Key to Worker and Queen Odontomachus of the U.S.

1. Hairs on first gastral tergite extremely fine and dense: spaces between hairs less than one-third
as wide as the length of hairs (SE U.S., perhaps Neotropics) ....................... brunneus (Patton)
1'. Hairs on first gastral tergite sparse, spaces between hairs at least one half as wide as length of hairs ....... 2
2. Posterior face of petiole with conspicuous transverse striae (Fig. 21) (Florida, W. Indies,
perhaps elsewhere in the Neotropics) ................ .................. ruginodis M. R. Smith
2'. Posterior face of petiole smooth (as in Fig. 1C) .................................................... 3
3. Basalar lobe (oval sclerite at posterior dorsal corner of mesopleuron) conspicuously striate
(Fig. 2K) (south-central peninsular Florida) .................................. relictus, new species
3'. Basalar lobe smooth (Fig. 2J) (southwestern U.S., Mexico) ................................. clarus Roger

Key to Male Odontomachus of the U.S.

1. Each ocellus at least as wide as space between lateral ocelli and eye (Figs. 2A, C) ........................ 2
1'. Ocelli much smaller than space between lateral ocelli and eye (Figs. 2B, D) ............................. 3
2. Head and body pale orange, antennae brown ............................................... brunneus
2'. Head and body brown, antennae yellowish .................................................. relictus
3. Head and body mostly yellowish, propodeum and sometimes gaster contrasting brown;
central area of pronotum smooth and shining .......................................... ruginodis
3'. Head and body dark brown; pronotum finely striate ............................................ clarus

Notes on Species

Odontomachus brunneus. This species appar-
ently occurs throughout Florida, although there
are no records from the three westernmost coun-

ties. It is also known from southern Georgia and
Alabama. We have seen specimens from low
coastal areas in Alabama, and there is no obvious
reason why it should not occur in coastal Missis-
sippi, Louisiana and Texas, although it is not re-

Florida Entomologist 87(2)

ported from any of those states. A report of
brunneus from Cuba (Fontenla 1997) might refer
to some other species, as we have seen specimens
that would key to brunneus from the Dominican
Republic, but are probably closer to 0. insularis
Guerin. Its distribution in Central and South
America is unclear, since this species was com-
bined with 0. ruginodis in Brown's revision of the
genus (1976). Associating the various brunneus-
like forms with their males in Central and South
America and the West Indies would be an interest-
ing and useful project for local myrmecologists,
and might easily yield distributional surprises or
new species. Workers can easily be distinguished
from other U.S. species by the densely hairy gaster.
Southeastern records ofinsularis in the Formi-
cidae section of the Catalog of the Hymenoptera
(D. R. Smith 1979) refer to brunneus. Although
the catalog appeared several years after Brown's
revision, the cut-off date for changes in the Formi-
cidae section was mid-1975. Smith's treatment of
North American Odontomachus differs from
those of Creighton (1950) and M. R. Smith (1951)
in elevating to species level three subspecies of
0. haematodus (Linnaeus). This was backed by no
taxonomic references, and certainly was not in-
tended to compete with the earlier, but unavail-
able, revision by Brown.
Odontomachus brunneus occurs in both well-
drained and poorly drained habitats; nests may
be in soil or in rotten wood. This species, along
with many others, was studied by Van Pelt (1958)
at the Welaka Reserve (now Welaka State Forest)
in Putnam Co., Florida. Van Pelt found many col-
onies, which occurred in all the terrestrial habi-
tats in the area, including flatwoods, mesic forest,
swamp forest, upland scrub and sandhill. Nests
were in various microhabitats, including deep leaf
litter, fallen logs, at the bases of trees, and open or
sparsely covered sandy areas. At the Archbold Bi-
ological Station, brunneus occurs in moist habi-
tats, including flatwoods, bayheads, the edges of
seasonal ponds, and elevated tussocks or fallen
pines within seasonal ponds. It has not been
found in the more elevated upland areas of the
Station, which are occupied by relictus. This dis-
tribution gives the impression that there is some
competitive displacement based on differential
adaptation to moisture conditions, but the evi-
dence remains circumstantial. It may be relevant
that in parts of its range devoid of relictus, where
brunneus occurs in dry, upland areas individuals
never achieve the large size and dark color seen in
some specimens from wet areas. Nobody knows,
however, whether the smaller, paler individuals
represent stressed individuals in suboptimal con-
ditions, or whether they represent an adaptive
phenotypic response in a robust population.
Workers of brunneus sometimes emerge to for-
age on cloudy days, but are generally nocturnal.
The formidable jaws of brunneus are not used as

assertively as one might expect, and there is frag-
mentary evidence that brunneus is sensitive to
chemical defenses. Prey are approached tenta-
tively, and the ant recoils immediately after strik-
ing the prey (Brown 1976). There may be a delay
before the prey is picked up and carried away;
Brown (1976) suggested that these ants react to
chemical defenses, which are allowed to dissipate
before the prey is retrieved. Alex Wild, while a
student at the Archbold Biological Station, twice
observed brunneus retreating hastily when con-
fronted by aroused workers of Dorymyrmex bu-
reni (Trager) (unpublished natural history notes
on file at the Archbold Biological Station). Dory-
myrmex bureni is much smaller than 0. brun-
neus, but can release large quantities of defensive
chemicals that are pungent to the human nose.
Van Pelt (1958) reported accumulations of brun-
neus head capsules in the nests of Formica arch-
boldi Smith, and suggested the possibility that
brunneus is a regular part of the diet of F arch-
boldi. If this is the case, it is more likely that the
brunneus are subdued by chemical means than by
mandible-to-mandible combat.
During this study specimens were examined
from the following areas: FL: Alachua, Baker,
Bay, Bradford, Brevard, Broward, Citrus, Clay,
Collier, Columbia, Dade, De Soto, Dixie, Duval,
Franklin, Gadsden, Gilchrist, Glades, Hamilton,
Hernando, Highlands, Hillsborough, Indian
River, Jackson, Jefferson, Lake, Lee, Leon, Levy,
Liberty, Madison, Marion, Martin, Monroe, Nas-
sau, Okeechobee, Orange, Osceola, Palm Beach,
Pasco, Polk, Putnam, Sarasota, St. Lucie, Sumter,
Taylor, Volusia, Wakulla, Walton Counties; GA:
Clinch County; AL: Baldwin, Houston Counties.
Odontomachus clarus. This species is known
from northern Mexico and from Texas, New Mex-
ico and Arizona. It is the only Odontomachus
known from the southwestern U.S. and northern
border of Mexico, so specimens may be identified
tentatively by their source alone. The similar spe-
cies ruginodis will probably be transported to the
southwestern U.S.; ruginodis has conspicuous
striae on the posterior face of the petiole (Fig. 21).
It is also possible that the species we consider
clarus includes unrecognized cryptic species. If
this is the case, the occurrence of additional spe-
cies might be detected by finding more than one
form of male within the range of clarus. We have
seen males associated with workers of clarus
from two widely separated sites: Cochise Co., AZ,
and Jeff Davis Co., TX. In our experience, it is dif-
ficult to find workers of clarus with associated
males. Examination of specimens from light traps
or flight traps might be a convenient way to estab-
lish whether there is more than one western spe-
cies. Records of clarus from the West Indies
(Smith 1979) refer to some other species, perhaps
ruginodis. All references to clarus in Florida
(Deyrup et al. 1985; Deyrup 1989; Deyrup et al.

June 2004

Deyrup & Cover: Odontomachus Ants of United States

1989; Deyrup 1990; Sivinski et al. 1998) should be
referred to relictus, as discussed above.
The species names coninodis Wheeler and de-
sertorum Wheeler, listed in the 1979 catalog, were
synonymized under clarus by Brown (1976).
Brown reported that "coninodis," which has a
blunt petiolar spine, occurs at the higher eleva-
tions in Arizona, in isolated, low mountain
ranges, surrounded by lower areas occupied by
typical clarus with an elongate petiolar spine. The
distribution of the two forms is unlike that of a
normal pair of geographic subspecies, and Brown
characterized the short-spined forms as "depau-
perate ecotypes or ecophenotypes." As in the case
(mentioned above) of the smaller, paler brunneus
found in dry sites, it seems premature to apply
the pejorative "depauperate" to a condition that,
for all we know, could be a superb adaptive re-
In Arizona this species is found, usually in
small numbers, under rocks and grass tussocks,
in both dry and mesic sites. In western Texas it
shows a preference for more mesic sites and fine
soils; nests are usually found under rocks or logs
(Cokendolpher & Franckel990).
During this study specimens were examined
from the following sites (we provide more detailed
site information for clarus than for brunneus or
ruginodis because of evidence of geographic vari-
ability in clarus). AZ: Cochise Co. (Chiricahua
Mts: Cave Creek, Texas, and Idlewild Canyons),
Pima Co. (Tucson), Santa Cruz Co. (Patagonia
Mts., Pajarito Mts.); TX: Bosque Co. (Meridian),
Brewster Co. (Big Bend National Park: Rio
Grande Village), Denton Co. (no locality), Jeff
Davis Co. (Davis Mts.), Travis Co. (Bull Creek,
McNeil); MEXICO: Chihuahua (Riva Palacio,
Guerero, Conchos), Coahuilla (25 km E. of
Saltillo), Cuernovaca (no locality), Guanjuato
(highway 57 km 57), Hidalgo (San Miguel), Jalisco
(Guadelajara), Mexico (Pedrigales), Nuevo Leon
((Monterrey), Queretaro (3 mi. W. of Queretaro).
Odontomachus ruginodis. This species occurs
sporadically through southern and central Flor-
ida, at least as far north as Orlando, and also in
the West Indies. Its distribution in South and
Central America is unclear because it has been
confused with brunneus. Its ability to thrive in
disturbed habitats should allow it to invade main-
land Neotropical areas, if it is not already
present. It is probable that this species will be dis-
tributed by commerce to disturbed areas in the
Southwest. The conspicuous striae on the poste-
rior face of the petiole (Fig. 21) distinguish work-
ers of this species from the similar clarus and
relictus, but there are additional species with pet-
iolar striae (e.g., 0. bauri Emery) outside the U.S.
The name ruginodis was first applied by
Wheeler (1905), and Wheeler was designated as
the author of ruginodis in Deyrup et al. (1985)
and Deyrup et al. (1989). The name ruginodis,

however, was first used as a quadrinomial (Odon-
tomachus haematodus insularis ruginodis), and
therefore is not a valid name under the rules of
nomenclature. The first use of ruginodis as a tri-
nomial, or subspecies (Odontomachus haemato-
dus ruginodis), was by M. R. Smith (1937). Since
this is the first valid use of ruginodis for this spe-
cies, M. R. Smith is the author of the name. This,
along with hundreds of other tangles of nomen-
clature, was straightened out in Bolton's 1995
catalog of ants.
In Florida, this species occurs in disturbed ar-
eas, including urban and suburban habitats. It oc-
curs along the beaches in the tropical part of the
state. It has not yet been found inland in natural
habitats. In Puerto Rico it differs from another
sympatric species (perhaps 0. bauri) in its prefer-
ence for open, sunny areas, especially river bot-
toms (Smith 1937).
The defensive mandible-snapping behavior of
ruginodis was studied by Carlin and Gladstein
(1989). When a nest is attacked by other ants, the
ruginodis workers rush out, snapping at anything
that seems a threat. Enemy ants may be dismem-
bered or knocked out of the way by the mandibu-
lar strikes. If the mandibles hit a solid object, the
ruginodis may itself be flung into the air for a dis-
tance of several centimeters. This does not seem
to be an escape mechanism, as the worker, upon
landing, immediately charges back into the fray.
The nest entrance is usually guarded by a single
worker, who stands with cocked mandibles near
the entrance. If an intruder approaches within
striking distance, the mandibles snap shut, re-
sponding to signals from the antennae and long
sensory hairs at the bases of the mandibles. The
heavy apices of the mandibles do not slice into the
intruder, but knock it away a distance of about
one to fourteen centimeters. Carlin and Gladstein
call this the "bouncer defense."
During this study specimens were examined
from the following areas: FL: Brevard, Broward,
Charlotte, Collier, Dade, Glades, Hendry, High-
lands, Hillsborough, Indian River, Lee, Manatee,
Martin, Monroe, Orange, Palm Beach, Pinellas,
Orange, Polk, St. Lucie, Volusia Counties; BER-
MUDA; BAHAMAS: New Providence, San Salva-
dor, Rum Cay, North Andros Islands; PUERTO
RICO: Rio Grande.

Residual Problems

There are still some questions on the taxon-
omy and distribution of the Odontomachus spe-
cies that occur in the U.S. The distribution of
brunneus and ruginodis is unclear. Does brun-
neus occur in coastal wetlands around the Gulf of
Mexico? Is brunneus as it appears in the south-
eastern North America the same species as the
brunneus populations reported from the West In-
dies and the mainland Neotropics? Does rugino-

Florida Entomologist 87(2)

dis occur outside the West Indies and Florida?
Another kind of question deals with the morpho-
logical divergences between males of different
species. Do the kind of differences we have re-
ported relate to differences in ecology and behav-
ior of the species? Will Odontomachus males
prove useful in distinguishing species throughout
the Neotropics? The few males of insularis and
bauri that we have seen show conspicuous species
differences, but there could be species complexes
that cannot be elucidated by male morphology.


We thank Archbold Biological Station for supporting
this research, the American Museum of Natural His-
tory Southwest Research Station and Wade Sherbrooke
for hospitality during field studies of Odontomachus
clarus, and the Museum of Comparative Zoology at
Harvard University for the loan of specimens to the
Archbold Biological Station. Lloyd Davis, William
Mackay, Zachary Prusak, Paige Martin, James Cronin
and Walter Suter donated specimens of Odontomachus
for this project.


BOLTON, B. 1995. A New General Catalog of the Ants of
the World. Harvard University Press, Cambridge,
MA. 504 pp.
BROWN, W. L., JR. 1976. Contributions toward a reclas-
sification of the Formicidae. Part VI. Ponerinae, tribe
Ponerini, subtribe Odontomachiti. Section A. Intro-
duction, subtribal characters, genus Odontomachus.
Studia Entomologica 19: 67-171.
"bouncer" defense of Odontomachus ruginodis and
other odontomachine ants (Hymenoptera: Formi-
cidae). Psyche 96: 1-19.
Ants (Hymenoptera, Formicidae) of Western Texas.
Part II. Subfamilies Ecitoninae, Ponerinae, Pseudo-
mynnecinae, Dolichoderinae, and Formicinae. Mu-
seum of Texas Tech University Special Publication
30: 3-76.
CREIGHTON, W. S. 1950. The Ants of North America.
Bull. Harvard Mus. Compar. Zool. 104: 1-583.
DEYRUP, M. 1989. Arthropods endemic to Florida scrub.
Florida Scientist 52: 254-270.

DEYRUP, M. 1990. Arthropod footprints in the sands of
time. Florida Entomol. 73: 529-538.
DEYRUP, M. 2002. The exotic ant Anochetus mayri in
Florida (Hymenoptera: Formicidae). Florida Ento-
mol. 85: 658-659.
WHEELER 1989. A preliminary list of the ants of
Florida. Florida Entomol. 72: 91-101.
DEYRUP, M., J. TRAGER, AND N. CARLIN. 1985. The ge-
nus Odontomachus in the southeastern United
States (Hymenoptera: Formicidae). Entomological
News 96: 188-195.
FONTENLA, J. L. 1997. Lista preliminary de las hormi-
gas de Cuba (Hymenoptera: Formicidae). Cucujo 6:
Fast trap jaws and giant neurons in the ant Odon-
tomachus. Science 262: 561-563.
HOLLDOBLER, B., AND E. O. WILSON. 1990. The Ants.
Harvard University Press, Cambridge, MA. 732 pp.
MYERS, R. L. 1990. Scrub and high pine, pp.150-193. In
R. L. Myers and J. J. Ewel [eds.]. Ecosystems of Flor-
ida. University of Central Florida Press, Orlando.
xviii + 765 pp.
ING, AND E. PETERSSON. 1998. A natural history of
Pleotomodes needhami Green (Coleoptera: Lampyri-
dae): a firefly symbiont of ants. Coleop. Bull. 52: 23-30.
SMITH, D. R. 1979. Formicidae, pp. 1323-1467. In K. V.
Krombein, P. D. Hurd, Jr., D. R. Smith and B. D.
Burks [eds.]. Catalog of Hymenoptera in North
America North of Mexico. Smithsonian Institution
Press, Washington, D. C. 2735 pp.
SMITH, M. R. 1937. The ants of Puerto Rico. J. Agric.
Univ. Puerto Rico 20: 819-875.
SMITH, M. R. 1951. Formicidae, pp. 778-875. In C. F. W.
Muesebeck and K. V. Krombein (eds.). Hymenoptera
of America North of Mexico, Synoptic Catalog.
Smithsonian Institution Press, Washington, D.C.
1420 pp.
VAN PELT, A. 1958. The ecology of the ants of the Welaka
Reserve, Florida (Hymenoptera: Formicidae). Part II.
Annotated list. American Midland Natur. 59: 1-57.
WEBB, S. D. 1990. Historical biogeography, pp.70-100.
In R. L. Myers and J. J. Ewel [eds.]. Ecosystems of
Florida. University of Central Florida Press, Or-
lando, FL. xviii + 765 pp.
WHEELER, W. M. 1905. The Ants of the Bahamas, with
a list of the known West Indian species. Bull. Amer-
ican Mus. Natur. Hist. 21: 79-135.

June 2004

Szalanski et al.: Coptotermes formosanus Diagnostics


'Department of Entomology, University of Arkansas, Fayetteville, AR 72701

2Department of Entomology, University of Florida-Ft. Lauderdale Research and Education Center
3205 College Avenue, Ft. Lauderdale, FL 33314

'Dow AgroSciences, Indianapolis, IN 46268


Formosan subterranean termite, Coptotermes formosanus Shriaki, is a serious pest of struc-
tures in portions of United States. A 467-bp region of the mtDNA 16S rRNA gene was sub-
jected to DNA sequencing from 12 Coptotermes species, including 64 populations of C.
formosanus. Genetic diversity among species ranged from 1.8% to 7.0%, with C. formosanus
at least 3.0% divergent to the other Coptotermes taxa. No genetic variation was detected
among the C. formosanus populations for this marker making it ideal for diagnostics. Com-
parison of nucleotide sequence of mitochondrial rRNA 16S was used to design polymerase
chain reaction (PCR) primers specific for C. formosanus. The diagnostic assay consists of two
independent PCR runs of the 16S primer pair along with the C. formosanus primer set. PCR
product from samples that are not C. formosanus can be subjected to DNA sequencing and
compared with the database of termite 16S sequences on GenBank for identification. This
technique provides a non-morphological method to identify field collected termites and may
facilitate future quarantine programs for C. formosanus.

Key Words: Coptotermes formosanus, invasive species, PCR, genetic variation, molecular di-
agnostics, termite.


La termita subterraneo de Formosa, Coptotermes formosanus Shriaki, es una plaga seria de
las estructuras en algunas parties de los Estados Unidos. Una region de 467 bp del gen 16S
rARN de la ADN mitocondrial fue sujetada de la secuenciaci6n de AND de 12 species de
Coptotermes, incluyendo 64 poblaciones de C. formosanus. La diversidad gen6tica entire las
species fue de 1.8% hasta 7.0%, con la C. formosanus por lo menos 3.0% divergentes de los
otros species de Coptotermes. Ningun variaci6n gen6tica fue detectada entire las poblacio-
nes de C. formosanus para este marcador haciendole ideal para un diagn6stico. Una compa-
rici6n de la secuencia del nucle6tido del gen 16S rARN de la ADN mitocondrial fue usada
para disenar unos cebadores (primers) especificos de la reacci6n en cadena por la polimerasa
(PCR) para C. formosanus. El ensayo diagn6stico consiste de dos pruebas independientes de
PCR del par 16S del cebador junto con el grupo de cebadores de C. formosanus. El product
de PCR de las muestras que no son C. formosanus puede ser sujetado de la secuenciaci6n de
ADN y comparados con el base de datos de las secuencias de 16S de termitas en el GenBank
para la identificaci6n. Esta t6cnica provea un m6todo no morfol6gico para identificar las ter-
mitas recolectadas en el campo y puede facilitar los futures programs de cuarentena para
C. formosanus.

The Formosan subterranean termite (FST)
Coptotermes formosanus Shriaki (Isoptera: Rhi-
notermitidae), is a major economic pest world-
wide and has become a serious pest to the United
States and its territories. Native to China, FST
has been introduced into Japan, Guam, Sri
Lanka, South Africa, Hawaii and the continental
United States (Mori 1987; Su & Tamashiro 1987;
Wang & Grace 1999). Coptotermes formosanus
was first recorded in continental United States at
Charleston, SC in 1957 (Chambers et al. 1988).
Numerous well-established colonies were discov-

ered in Florida in 1980, 1982, and 1984 (Oi et al.
1992) with many additional finds since (Su &
Scheffrahn 2000). Introductions to San Diego, CA
(Atkinson et al. 1993; Haagsma et al. 1995), the
Gulf Coast states, and southeastern US also have
been documented (Spink 1967; LaFage 1987; Su
& Tamashiro 1987; Howell et al. 1987; Appel &
Sponsler 1989; Oi et al. 1992; Su & Scheffrahn
1998; Hawthorne et al. 2000; Howell et al. 2000,
Scheffrahn et al. 2001; Hu et al. 2001; Jenkins et
al. 2002). Since 2002, C. formosanus has been con-
sidered a quarantine pest in Mississippi (Missis-

Florida Entomologist 87(2)

sippi Department of Agriculture and Commerce,
Rule 40). In the city of New Orleans, the control
and repair costs due to FST are estimated at $300
million annually (Lax & Osbrink 2003) and an-
nual damage to the entire United States is esti-
mated to exceed $1 billion. It is considered the
single most economically important insect pest in
the state of Hawaii (Su & Tamashiro 1987).
The inability to quickly discriminate what
Coptotermes species one is dealing with could lead
to difficulties in evaluating the source of the infes-
tation. While introductions of FST have gained
recent notoriety, less is reported or known about
other potentially damaging Coptotermes species.
Exotic introductions of Coptotermes havilandi to
Florida (Su et al. 1997; Su et al. 2000; Scheffrahn
& Su 2000) have been detected. Although not es-
tablished, C. havilandi Holmgren, which recently
has been synonymized as C. gestroi (Wasmann) by
Kirton & Brown (2003), has been recovered in
shipping crates in Tennessee imported from East
Asia (RHS, unpublished data). Coptotermes from
South America, Central America, and the Carib-
bean also pose potential problems for the US.
There are three described endemic species of Cop-
totermes in the Americas including C. crassus
Snyder, C. testaceus L., and C. niger Snyder, and
possibly others which have not been identified.
More recent genetic surveys have uncovered old
world Coptotermes species, C. sojesti, introduced
to the West Indies (Scheffrahn et al. in press).
Mistakes have been made in both the naming
and correct identification of some Coptotermes.
For example, the inconsistencies in the pest status
of C. havilandi in different regions of its geo-
graphic range have been due to misidentification
and taxonomic confusion between C. travians
(Haviland), C. havilandi, and C. gestroi (Kirton &
Brown 2003). An examination of the type series of
C. travians indicates the species has been misi-
dentified in Peninsular Malaysian literature (Tho
1992) as C. havilandi and also has been referred to
as C. borneensis Oshima (Kirton & Brown 2003).
Correct identification is critical for pest in-
sects. Identification of termite workers is possible
at the generic level only, and finding an alate,
which can be identified, in a collection is seasonal
and can be rare. We have developed a molecular
diagnostic method capable of differentiating FST
from other Coptotermes species regardless of the
caste encountered or locality obtained. It has
been demonstrated that both nucleotide sequenc-
ing and restriction enzyme digestion of poly-
merase chain reaction (PCR) amplicons can be
used to differentiate between various termite spe-
cies (Austin et al. 2002; Austin et al. 2004; Sza-
lanski et al. 2003; Clement et al. 2001; Jenkins et
al. 2002; Uva et al. 2004). DNA sequence differ-
ences reported between Coptotermes species from
numerous disjunctive populations from around
the world in a small portion of the mtDNA were

exploited to design species-specific PCR primers
and to develop a DNA-based assay that can dis-
criminate FST from other Coptotermes species.


Coptotermes termites were collected from var-
ious locations in North America, South America,
the Caribbean, Australia, Africa, and Asia (Table
1). Morphological identification of specimens
used in this study was achieved by using Snyder
(1922), Emerson (1925, 1928), Hill (1942), Schef-
frahn et al. (1990), Scheffrahn & Su (1994), Tho
(1992), and Su et al. (1997). Voucher specimens,
preserved in 100% ethanol, are maintained at the
Arthropod Museum, Department of Entomology,
University of Arkansas, Fayetteville, AR, USA.
Alcohol preserved specimens were allowed to
dry on filter paper, and DNA was extracted from
individual worker or soldier heads with the Pure-
gene DNA isolation kit D-5000A (Gentra, Minne-
apolis, MN). Extracted DNA was resuspended in
50 pl of Tris:EDTA and stored at -20C. Polymerase
chain reaction was conducted with the primers
(Kambhampati & Smith 1995) and LR-N-13398
al., 1994). These PCR primers amplify an approx-
imately 428-bp region of the mtDNA 16S rRNA
gene. PCR reactions were conducted with 1 pl of
the extracted DNA per Szalanski et al. (2000),
and a profile consisting of 35 cycles of 94C for 45
s, 46C for 45 s and 72C for 45 s. Amplified DNA
from individual termites was purified and concen-
trated by Microcon-PCR Filter Units (Millipore,
Bedford, MA). Samples were sent to The Univer-
sity of Arkansas Medical School DNA Sequencing
Facility (Little Rock, AR) for direct sequencing in
both directions with an ABI Prism 377 DNA se-
quencer (Foster City, CA). GenBank accession
numbers for the Coptotermes termites subjected
to DNA sequencing in this study are AY558898 to
AY558914. DNA sequences were aligned by the
PILEUP command of GCG (Accelrys, San Diego,
CA), and the distance matrix option of PAUP*
4.0b10 (Swofford 2001) was used to calculate ge-
netic diversity according to the Kimura 2-param-
eter model (Kimura 1980) of sequence evolution.


The 428-bp region of the mtDNA 16S rRNA
gene was subjected to DNA sequencing from 12
described species of Coptotermes, including 64
populations of C. formosanus (Table 1). Within
the genus, genetic diversity ranged from 1.8% be-
tween C. testaceus and C. crassus to 7.0% between
C. acinaciformis and C. vastator. No genetic vari-
ation was observed between the two C. testaceus
samples, and up to 0.7% variation was observed
among the C. gestroi samples. No genetic varia-

June 2004

Szalanski et al.: Coptotermes formosanus Diagnostics


Species Location Country N

C. acinaciformis
C. caruinatus
C. crassus
C. formosanus

C. heimi
C. gestroi

C. lacteus
C. intermedius
C. michaelseni
C. sjostedti
C. testaceus

C. vasator

Belize City
San Diego, CA
Jacksonville, FL
Wilton Manors, FL
Stone Mt., GA
Oahu, HI
Maui, HI
Lake Charles, LA
New Orleans, LA
Baton Rouge, LA
St. Rose, LA
Stennis Space Ctr, MS
Spindale, NC
Forest City, NC
Rutherfordton, NC
Rockport, TX
Rockwall, TX
Galveston, TX
Garland, TX
Grapevine, TX
Lewisville, TX
San Antonio, TX
Beaumont, TX
La Porte, TX
Hong Kong

Nagasaki Prefecture

Monroe, FL
Miami, FL

Hong Kong

Oahu, HI

"Number sequenced.

tion was observed in C. formosanus, and C. formo-
sanus was most similar to the C. intermedius
sample from Togo Africa, with 3.0% DNA se-
quence divergence. Phylogenetically, C. formosa-
nus forms a distinct clade among non-Australian
Coptotermes (C. vastator, C. testaceus, C. crassus,
C. sjostedti, C. intermedius, C. gestroi, C. heimi,
and C. carvinatus) (Scheffrahn et al. in press).
Formosan subterranean termite 16S DNA se-
quences along with sequences from other Copto-

terms, Reticulitermes (Szalanski et al. 2003) and
Heterotermes (Szalanski et al. 2004) were aligned
and examined for mismatches that reflected ei-
ther substitutions or deletions. The mismatches
were exploited to design primers that were
unique to FST (Table 2). Two primers, one from
GAGGCACAA-3') were designed. Based on the
sequence, the expected sizes of the amplicon is

Grand Turk

Florida Entomologist 87(2)



10 20 30

40 50 60


70 80 90

100 110

130 140 150



151 bp. Proper sized PCR products were obtained
with conspecific DNA, whereas no product was
obtained with template from the other species,
i.e., no false positives were observed with known
DNA. Each FST primer paired with a common
primer will only amplify Formosan subterranean
termite DNA.
The FST species-specific primers were tested
for optimal annealing performance in a 470-59C
temperature gradient with 2C intervals. The op-
timal annealing temperature for the FST specific

primers was 57C. PCR reactions were the same
as the 16S conditions with the exception of the
PCR profile which consists of 30 cycles of 94C for
45 s, 57C for 45 s and 72C for 45 s. This anneal-
ing temperature, however, is too high for the 16S
universal primers, preventing multiplex PCR
with both primer pairs. To resolve this, both PCR
reactions are conducted individually and 10 pl of
each PCR reaction are loaded onto a single well of
a 2% agarose gel (Fig. 1). The FST specific primer
set successfully amplified for 52 individual FST


June 2004

Szalanski et al.: Coptotermes formosanus Diagnostics



ZIN C! 0! C jl (y (






Fig. 1. Two percent agarose gel of 428-bp mtDNA 16S amplicon and 151-bp C. formosanus diagnostic amplicon
for 4 Coptotermes spp.

from all 23 FST populations, and did not yield a
PCR product for the other Coptotermes, Reticuli-
termes and Heterotermes. PCR product from sam-
ples that are not C. formosanus can be subjected
to DNA sequencing and compared with the data-
base of termite 16S sequences on GenBank for
identification by a BLAST search (http://www.
ncbi.nlm.nih.gov/BLAST/). This technique pro-
vides a method to identify field collected termites
and facilitates the screening of the monitoring for
this species and for the introduction of other inva-
sive Coptotermes termites.
In the context of determining the species for a
large number of samples collected in connection
with distribution or competition studies, simplify-
ing the identification of the worker caste is advan-
tageous. FST has no genetic polymorphism across
its geographic distribution for the 16S marker,
whereas genetic variation has been observed in
the mtDNA COII gene (ALS unpublished data,
Jenkins et al. 2002). This lack of mtDNA 16S in-
traspecific variation makes this marker ideal for
molecular diagnostics.
These primers provide a convenient way to
identify individual termites without resorting to
more time consuming restriction fragment-length
polymorphism analysis, or extensive morphologi-
cal data which may result in overlap due to clinal
variations in size as observed in many insects in-
cluding termites. The approach is equally applica-
ble to other castes, such as soldiers and alates,
but given their obvious taxonomic importance
should be constrained to either morphologically

ambiguous samples, or when the more diagnostic
castes are unavailable. This should be an impor-
tant new tool for substantiating the identity of
FST before the onset of regulatory procedures
(i.e., quarantine).

We thank R. Gold, R. Davis, M. Merchant, G. Hend-
erson, K. Grace, J. Nixon, L. Yudin, J. Lopez, K.L. Mosg,
J. Chapman, S. Cabellero, J. Chase, B. McCullock, 0.
Miyashita, E. Phillips, P. Ban, M. Weinberg, J. Stotts,
N.Y. Su, E. Vargo, P. Fitzgerald, M.K. Rust T. Myles, D.
Muravanda, J. Darlington, and J. Woodrow for collect-
ing termite samples. Research was supported in part by
the University of Arkansas, Arkansas Agricultural Ex-
periment Station, and by the University of Florida Re-
search Foundation.

NERES, AND A. KENCE. 2002. A comparative genetic
analysis of the subterranean termite genus Reticuli-
termes (Isoptera: Rhinotermitidae). Ann. Entomol.
Soc. Amer. 95: 753-760.
FOSTER. 2004. Genetic variation and geographical
distribution of the subterranean termite genus Reti-
culitermes in Texas. Southwest Entomol. (in press).
APPEL, A. G., AND R. C. SPONSLER. 1989. Formosan ter-
mites now in Alabama. Highlights 36: 34.
ATKINSON, T. H., M. K. RUST, AND J. L. SMITH. 1993. The
Formosan subterranean termite, Coptotermes formo-
sanus Shiraki (Isoptera: Rhinotermitidae), estab-
lished in California. Pan-Pacific Entomol. 69: 111-113.




Florida Entomologist 87(2)

1988. Distribution and habitats of the Formosan
subterranean termite (Isoptera: Rhinotermitidae) in
South Carolina. J. Econ. Entomol. 81: 1611-1619.
Biosystematics of Reticulitermes termites in Europe:
morphological, chemical and molecular data. In-
sectes Sociaux 48: 202-215.
EMERSON, A. E. 1925. The termites from Kartabo, Bar-
tica District, Guyana. Zoologica 6: 291-459.
EMERSON, A. E. 1928. Termites of the Belgian Congo
and the Cameroon. Bull. Amer. Mus. Nat. Hist. 57:
AND D. A. REIERSON. 1995. Formosan subterranean
termite established in California. Calif. Agricul. 49:
C. BRIDGES. 2000. The termite (Isoptera) fauna of
South Carolina. Source: J. Agricul. Urban Entomol.
17: 219-229.
HILL, G. F. 1942. Termites (Isoptera) from the Austra-
lian Region. CSIRO, Melbourne 479 pp.
1987. The geographical distribution of the termite
genera Reticulitermes, Coptotermes, and Incisiter-
mes in Texas. Southwest. Entomol. 12: 119-125.
HOWELL, H. N., R. E. GOLD, AND G. J. GLENN. 2000.
Coptotermes distribution in Texas (Isoptera: Rhino-
termitidae). Sociobiol. 37: 687-697.
Hu, X. P., F. M. OI, AND T. G. SHELTON. 2001. Formosan
Subterranean Termites. ANR-1035. http://www.
DNA technology, Interstate commerce, and the
likely origin of Formosan subterranean termite
(Isoptera: Rhinotermitidae) infestation in Atlanta,
Georgia. J. Econ. Entomol. 95: 381-389.
KAMBHAMPATI, S., AND P. T. SMITH. 1995. PCR primers
for the amplification of four insect mitochondrial
gene fragments. Ins. Molec. Biol. 4: 233-236.
KIRTON, L. G., AND V. K. BROWN. 2003. The taxonomic
status of pest species of Coptotermes in Southeast
Asia: Resolving the paradox in the pest status of the
termites, Coptotermes gestroi, C. havilandi and C.
travians (Isoptera: Rhinotermitidae). Sociobiol. 42:
KIMURA, M. 1980. A simple method for estimating evo-
lutionary rate of base substitutions through compar-
ative study of nucleotide sequences. J. Molec. Evol.
16: 111-120.
LAFAGE, J. P. 1987. Practical considerations of the For-
mosan subterranean termite in Louisiana: a 30-
year-old problem, pp. 37-42. In M. Tamashiro and N.
Y. Su [eds.], Biology and Control of the Formosan
Subterranean Termite. Research and Extension se-
ries 083. College of Tropical Agriculture and Human
Resources, University of Hawaii, Honolulu.
LAX, A. R., AND W. L. OSBRINK. 2003. United States De-
partment ofAgriculture-Agriculture Research Ser-
vice research on targeted management of the
Formosan subterranean termite Coptotermes formo-
sanus Shiraki (Isoptera: Rhinotermitidae). Pest
Manage. Sci. 59: 788-800.
MORI, H. 1987. The Formosan subterranean termite in
Japan: distribution, damage, and current and poten-

tial control measures, pp 23-26. In M. Tamashiro
and N.-Y. Su [eds.], Biology and Control of the For-
mosan Subterranean Termite. Research Extension
Series 083. University of Hawaii, Honolulu.
FRAHN. 1992. The Formosan Subterranean Termite.
ENY-216, Florida Cooperative Extension Service,
IFAS, University of Florida. 6 pp.
tive, introduced, and structure-infesting termites of
the Turks and Caicos Islands, B.W.I. (Isoptera: Kal-
otermitidae, Rhinotermitidae, Termitidae) Florida
Entomol. 73: 622-627
SCHEFFRAHN, R. H., AND N.-Y. SU. 1994. Keys to soldier
and winged adult termites (Isoptera) of Florida.
Florida Entomol. 77: 460-474.
SCHEFFRAHN, R. H., AND N.-Y. SU. 2000. Current distri-
bution of the Formosan subterranean termite and
Coptotermes havilandi in Florida UF/IFAS. http://
FORSCHLER. 2001. New termite records (Isoptera:
Kalotermitidae, Rhinotermitidae) from Georgia. J.
Entomol. Sci 36: 109-113.
AUSTIN, AND J. NIXON. Establishment of the African
termite, Coptotermes sjostedti (Isoptera: Rhinoter-
mitidae), on the island of Guadeloupe, French West
Indies. Ann. Entomol. Soc. Amer (in press).
AND P. FLOOK. 1994. Evolution, weighting, and phy-
logenetic utility of mitochondrial gene sequences and
a compilation of conserved polymerase chain reaction
primers. Ann. Entomol. Soc. Amer. 87: 651-701.
SNYDER, T. E. 1922. New termites from Hawaii, Central
and South America and the Antilles. Proceedings of
the U.S. National Museum 61: 1-32.
SPINK, W. T. 1967. The Formosan subterranean termite
in Louisiana. Louisiana State Univ. Circ. 89, 12 pp.
SU, N.-Y., AND M. TAMASHIRO. 1987. An overview of the
Formosan subterranean termite in the world, pp. 3-
15. In M. Tamashiro & N.-Y. Su [eds.], Biology and
Control of the Formosan Subterranean Termite. Col-
lege of Trop. Agr. Human Resources, Univ. of Ha-
waii, Honolulu.
SU, N.-Y., AND R. H. SCHEFFRAHN. 1998. A review of
subterranean termite control practices and pros-
pects for integrated pest management programs. In-
tegrated Pest Management Reviews 3: 1-13.
SU, N.-Y., AND R. H. SCHEFFRAHN. 2000. Termites as
pest of buildings, pp 437-453. In T. Abe, D. E. Bignell
and M. Higashi [eds.], Termites: evolution, sociality,
symbiosis, ecology. Kluwer Academic Publishers,
Dordrecht, Netherlands.
A new introduction of a subterranean termite, Cop-
totermes havilandi Holmgren (Isoptera: Rhinoter-
mitidae) in Miami, Florida. Florida Entomol. 80:
Su, N.-Y., P. M. BAN, AND R. H. SCHEFFRAHN. 2000.
Control of Coptermes havilandi (Isoptera: Rhinoter-
miteidae) with hexafulumoron baits and a sensor in-
corporated into a monitoring-baiting program. J.
Econ. Entomol. 93: 415-421.
Swofford, D. L. 2001. PAUP*: Phylogenetic analysis us-
ing parsimony (*and other methods), ver. 4.0b10.
Sinauer, Sunderland, MA.

June 2004

Szalanski et al.: Coptotermes formosanus Diagnostics

FRITZ. 2000. Population genetics and phylogenetics
of the endangered American burying beetle, Nicro-
phorus americanus (Coleoptera: Silphidae). Ann.
Entomol. Soc. Amer. 93: 589-594.
Identification of Reticulitermes spp. (Isoptera: Rhi-
notermatidae) from South Central United States by
PCR-RFLP. J. Econ. Entomol. 96: 1514-1519.
KRECEK, AND N.-Y. SU. 2004. Molecular phylogeny
and biogeography of Heterotermes (Isoptera: Rhino-
termitidae) in the West Indies. Ann. Entomol. Soc.
Amer. 97: (in press).

THO, Y. P., AND L. G. KIRTON. 1992. Termites of Penin-
sular Malaysia. Malayan Forest Records No. 36. L.
G. Kirton [ed.]. Forest Research Institute Malaysia,
Kuala Lumpur. 224 pp.
QUINTANA, AND A. G. BAGNERES. 2004. Origin of a
new Reticulitermes termite (Isoptera, Rhinotermiti-
dae) inferred from mitochondrial DNA data. Molecu-
lar Phylogenetics and Evolution 30: 344-353.
WANG, J. S., AND J. K. GRACE. 1999. Current status of
Coptotermes Wasmann (Isoptera: Rhinotermitidae)
in China, Japan, Australia and the American Pa-
cific. Sociobiol. 33: 295-305.

Florida Entomologist 87(2)


'Department of Entomology, University of Arkansas, Fayetteville, AR 72701 (e-mail: aszalan@uark.edu)

2Department of Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK 74078

Sequencing of a portion of the mitochondrial DNA 16S gene was undertaken to determine
genetic variation and distribution of Reticulitermes in Oklahoma. From 16 Oklahoma coun-
ties, 43 R. flavipes, four R. hageni, one R. virginicus, and seven R. tibialis samples were col-
lected, identified and subjected to DNA sequencing. No genetic variation was observed in R.
virginicus, while two haplotypes were observed in R. hageni, four in R. tibialis, and 10 for R.
flavipes. Among the 10 R. flavipes haplotypes, nine nucleotides were variable and genetic
variation ranged from 0.2 to 1.4%. Phylogenetic analysis revealed several minor relation-
ships within R. tibialis and R. flavipes; however, there was no apparent geographical asso-
ciation to the haplotypes. The high amount of genetic variation, but a lack of geographically
distinct haplotypes in R. flavipes, indicate that this termite species has been distributed ran-
domly in Oklahoma by humans due to its association with structures.

Key Words: 16S, DNA sequence, genetic variation, population genetics, Reticulitermes, termite.

Se llevo a cabo la secuenciaci6n del una porci6n del gen 16S de ADN mitocondrial para de-
terminar la variaci6n gen6tica y distribuci6n de Reticulitermes en Oklahoma. De los 16 con-
dados de Oklahoma, 43 R. flavipes, cuatro R. hageni, un R. virginicus, y siete R. tibialis
muestras fueron recolectadas, identificadas y sujetas a la secuenciaci6n de ADN. Ningun va-
riaci6n gen6tica fue observada en R. virginicus, mientras que dos haplotipos fueron obser-
vados en R. hageni, cuatro en R. tibialis, y 10 en R. flavipes. Entre los 10 haplotipos de R.
flavipes, nueve nucle6tidos varian y la variaci6n gen6tica fue de 0.2 hasta 1.4%. Un analysis
filogen6tico revel6 una relacion menor entire R. tibialis y R. flavipes; sin embargo, no habia
una asociaci6n geografica aparente entire los haplotipos. La cantidad mas alta de variaci6n
gen6tica, junta con la falta de haplotipos distintos geograficos en R. flavipes, indica que esta
especie de termita ha sida distribuida al azar en Oklahoma por humans debido a su asocia-
ci6n con estructuras.

Subterranean termites in the genus Reticuli-
termes Holmgren belong to the Isopteran family
Rhinotermitidae and contain some of the most de-
structive and damaging termite species with re-
spect to their wood feeding preference. The four
principal subterranean termite species in the
United States are the eastern subterranean ter-
mite Reticulitermes flavipes (Kollar), the arid sub-
terranean termite R. tibialis Banks, and the
dark-southern subterranean termite R. virgini-
cus (Banks). Ninety percent of the termite control
business in the United States involves these four
Reticulitermes species plus Coptotermes formosa-
nus (Shiraki) (Forschler & Lewis 1997). In Okla-
homa, subterranean termites (Isoptera: Rhino-
termitidae) are found throughout the state and
cause millions of dollars in structural damage ev-
ery year. The probability that termites will attack
a wooden structure within 10 to 20 years after
construction is greater than 70% in Oklahoma
(Criswell & Pinkston 2001). While the total eco-
nomic impact of Reticulitermes spp. in Oklahoma

is uncertain, anecdotal accounts of their presence
and destructive activities within urban areas
have been documented (Affeltranger et al. 1987;
Anonymous 2001a).
Recently, Brown et al. (2004) conducted a
study involving species identification and distri-
bution, and wood consumption rates of termites
collected from over 200 sites in Oklahoma using
in-ground and surface-ground boards. The most
abundant naturally occurring termite species
found were in the genus Reticulitermes. Reticuli-
termes flavipes, the light-southern subterranean
termite R. hageni Banks, R. tibialis, and R. vir-
ginicus are known from Oklahoma (Weesner
1965). Presently, the K.C. Emerson Entomology
Museum at Oklahoma State University, Stillwa-
ter, Oklahoma has 25 Reticulitermes specimens of
which only 13 have been identified to species (all
Reticulitermes flavipes). Reticulitermes spp. have
been reported from less than 25 counties in Okla-
homa (out of 75 total) (Anonymous 1999; Anony-
mous 2000; Anonymous 2001b; Anonymous 2002).

June 2004

Austin et al.: Reticulitermes Genetics

Oklahoma probably has a similar complement of
Reticulitermes species as found in states with
which it has contiguous borders and where sur-
veys have or will be conducted given that they are
in the same geographical area with similar cli-
mates and habitats with no geographical barriers
(Howell et al. 1987; Wang & Powell 2001; Messen-
ger et al. 2002; Austin et al. 2004).
Correctly identifying termites is important be-
cause different control methods and strategies
may be used depending on the target species.
Identifying termite workers to species is difficult,
and identifying soldiers is sometimes inaccurate
because precise measurements are required and
overlap may occur between species (Scheffrahn &
Su 1994). Difficulties can arise in species determi-
nation from individual collection sites because
colonies consist mostly of the worker caste while
soldiers are less abundant. Alates are found less
frequently in collections given their seasonal oc-
currences and unpredictable swarming. Soldiers
represent only 1-3% of the total population of
Reticulitermes colonies and are morphologically
variable; use of this caste alone for identification
can result in equivocal species determinations.
Subtle clinal variations imposed by geographic
boundaries can influence morphology making cor-
rect species determinations difficult.
In contrast, molecular genetic methods are
able to differentiate species regardless of caste
(Szalanski et al. 2003). Also, genetic information
obtained from existing collections can be an inte-
gral component to phylogenetic studies as a
whole, reflecting potential changes in species dis-
tributions over time. The extent of genetic varia-
tion and subsequent gene flow in Reticulitermes
spp. from Oklahoma has never been studied. Pre-
vious genetic studies have focused on Reticuliter-
mes spp. from the southeastern United States
and Western Europe (Jenkins et al. 1998; Jenkins
et al. 2001; Marini & Mantovani 2002; Uva et al.
2003). Recently, Austin et al. (2004) conducted the
first comprehensive genetic survey of Reticuliter-
mes in Texas and found 13 haplotypes ofR. flaui-
pes, seven R. tibialis haplotypes, and one
haplotype each for R. virginicus and R. hageni.
Identification to the species level of specimens
from existing collections with molecular techniques
as outlined in this study may add significant infor-
mation on species distribution and gene flow.
Genetic variation and gene flow information may
elucidate existing patterns of spread, possible
hybridization, and general speciation ofReticuliter-
mes spp. in Oklahoma. In this study, we
investigated the extent of genetic variation within
and among Oklahoma Reticulitermes, evaluated
the utility of genetic markers used for identifying
species, expanded the known geographical distri-
bution of these taxa within Oklahoma, and deter-
mined ifReticulitermes distributions are influenced
by human activity.


Termites were collected from several locations
in Oklahoma (Table 1) and preserved in 100%
ethanol. In addition to our own collecting efforts,
we solicited the assistance of Pest Management
Professionals (PMPs) throughout the state to de-
termine the predominant species found in in-
fested structures. PMPs were provided with
collection kits and samples were mailed to our
laboratory. Fifty-five samples, representing vari-
ous geographic zones were used for molecular
analysis. When available, Reticulitermes alates or
soldiers were also morphologically identified to
species with keys by Krishna & Weesner (1969),
Scheffrahn & Su (1994), Hostettler et al. (1995),
Donovan et al. (2000). For samples consisting
only of workers, species identification was con-
ducted by using mtDNA 16S sequences (Szalan-
ski et al. 2003). Voucher specimens preserved in
100% ethanol are maintained at the Arthropod
Museum, Department of Entomology, University
of Arkansas, Fayetteville, AR.
Alcohol-preserved specimens were allowed to
dry on filter paper, and DNA was extracted from
whole individual termites with the Puregene
DNA isolation kit D-5000A (Gentra, Minneapolis,
MN) per Austin et al. (2002). Extracted DNA was
resuspended in 50 pl of Tris:EDTA and stored at
-20C. Polymerase chain reaction was conducted
with primers LR-J-13007 (5'-TTACGCTGTTATC-
CCTAA-3') (Kambhampati & Smith 1995) and
3') (Simon et al. 1994). These PCR primers am-
plify an approximately 428 bp region of the
mtDNA 16S rRNA gene. PCR reactions were con-
ducted with 1 pl of extracted DNA (Szalanski et
al. 2000) and a profile consisting of 35 cycles of
94C for 45 s, 46C for 45 s and 72C for 60 s. Am-
plified DNA from individual termites was purified
and concentrated with minicolumns (Wizard
PCRpreps, Promega Corp., Madison, WI) accord-
ing to the manufacturer's instructions. Samples
were sent to the University of Arkansas DNA Se-
quencing Facility (Fayetteville, AR) for direct se-
quencing in both directions. GenBank accession
numbers were AY538739 to AY538744 for the ter-
mite haplotypes new to this study and not present
in Austin et al. (2004). DNA sequences were
aligned by the PILEUP command of GCG (Accel-
rys, San Diego, CA). Mitochondrial DNA haplo-
types were aligned by using MacClade v4
(Sinauer Associates, Sunderland, MA).
The distance matrix option of PAUP* 4.0b10
(Swofford 2001) was used to calculate genetic dis-
tances according to the Kimura 2-parameter
model of sequence evolution (Kimura 1980). Mito-
chondrial 16S sequences from the Formosan sub-
terranean termite, Coptotermes formosanus
Shiraki (GenBank AY558910), and the desert
subterranean termite Heterotermes aureus (Sny-

Florida Entomologist 87(2)


Species City County Haplotype N

R. flauipes Stillwater

R. hageni

R. virginicus
R. tibialis

Coptotermes formosanus
Heterotermes aureus


Oklahoma City
Oklahoma City

Oklahoma City

Oklahoma City

Monkey Island
Fort Towson
Fort Towson
Baton Rouge, LA
Santa Rita, AZ

der) (GenBank AY380299) were added to the Reti-
culitermes DNA sequences to act as outgroup
taxa. The DNA sequences were aligned by the
PILEUP program in GCG (Genetics Computer
Group, Madison, WI) and adjusted manually.
Maximum parsimony analysis on the alignments
were conducted with PAUP* 4.0b10 (Swofford

2001). Gaps were treated as missing data. The re-
liability of trees was tested with a bootstrap test
(Felsenstein 1985). Parsimony bootstrap analysis
included 1,000 resamplings and used the Branch
and Bound algorithm of PAUP*. Because there
are few published accounts of the occurrence of
Reticulitermes spp. in Oklahoma, we compiled all




June 2004

Austin et al.: Reticulitermes Genetics

available data from existing sources and noted
them on our distribution map (Fig. 1).


DNA sequencing of the 16S rDNA amplicon re-
vealed an average size of 428 bp. The average
base frequencies were A = 0.41, C = 0.23, G = 0.13,
and T = 0.23. The aligned DNA data matrix, in-
cluding the outgroup taxa, resulted in a total of
433 characters. Of these characters, 86 (20%)
were variable and 46 (11%) were phylogenetically
informative. From the DNA sequence analysis of
Reticulitermes spp. samples from 16 Oklahoma
counties, a total of 43 R. flavipes, four R. hageni,
one R. virginicus, and seven R. tibialis were iden-
tified based on species diagnostic nucleotide sites
(Szalanski et al. 2003) (Table 1, Fig. 1). Morpho-
logical identifications yielded the same species
identification as the DNA sequences. An addi-
tional 10 counties, were included from published
anecdotal accounts, bringing the total number of
reported counties in Oklahoma to 24 (Fig. 1).
No genetic variation was observed in R. vir-
ginicus, while two unique haplotypes were found
in R. hageni, four in R. tibialis and 10 in R. flavi-
pes (Table 1). Pairwise Tajima-Nei distances
(Tajima & Nei 1984) among Reticulitermes taxa
ranged from 5.7% between R. flavipes and R.
hageni, to 8.3% between R. flavipes and R. tibia-
lis. A total of nine nucleotide sites varied among
the 10 R. flavipes haplotypes (Table 2) and genetic
variation among the R. flavipes haplotypes
ranged from 0.2 to 1.4% (Table 3). The most com-

mon haplotypes were L and P with 10 and 14 rep-
resentatives, respectively. Within R. tibialis, a
total of three sites varied among the four haplo-
types and variation ranged from 0.2 to 0.7%.
Within R. hageni, one nucleotide site was variable
between the two haplotypes.
Bootstrap analysis of the aligned Reticulitermes
spp. and the outgroup taxa resulted in a consensus
tree with several distinct branches (Fig. 2). These
distinct clades included R. flavipes, R. hageni and
R. virginicus; and R. tibialis. Within R. flavipes,
haplotypes Q and F formed a distinct clade. For R.
tibialis, haplotypes T8 and T5 formed a distinct
clade relative to the two other haplotypes. There
was no genetic structure observed among the R.
hageni haplotypes in the present study.


This study updates the geographic distribution
of, and genetically classifies, the genus Reticuli-
termes in Oklahoma. However, it does not repre-
sent a comprehensive survey of Reticulitermes
spp. in Oklahoma. Rather, it documents new oc-
currences ofReticulitermes spp. in Oklahoma over
a large geographic area. In the present study, ge-
netic divergence values were similar to genetic di-
vergence detected in a study of Reticulitermes in
Texas (Austin et al. 2004). In terms of population
structure, a weak relationship was observed be-
tween R. flavipes haplotypes Q and F. Haplotype F
is distributed throughout the central and eastern
portions of the state, while haplotype Q was only
observed in Ottawa County, which is located in

Fig. 1. Distribution of Reticulitermes spp. and haplotypes in Oklahoma. Counties designated with an asterisk
represent reported cases of Reticulitermes spp. but were not used in our genetic analysis and are merely included
to update the current distribution of the genus in Oklahoma.

Florida Entomologist 87(2)


Haplotype 130 131 158 162 168 179 236 271 274

F G G C *
G G G C C*
H G C *
J G G C *
L G G C *
O T C G A C *
P G C C*

the northeast corner of Oklahoma. For R. tibialis
haplotypes T5 and T8 formed a distinct clade, and
were collected from two non adjacent counties. In
general, there was no population structure for
Reticulitermes spp. based on genetic haplotypes.
Likely reasons for this could be attributed to an-
thropogenic origins, a lack of nestmate agonism
(mixing with non-nestmates imposed by foraging
traffic in complex colonies with multiple reproduc-
tive centers) (Bulmer & Traniello 2002), or from
mixing between different colonies (Clement
1986). In fact, for R. flavipes, six of the 10 ob-
served haplotypes (E, F, G, H, J, and L) are shared
with Texas. More thorough sampling including
termite specimens from more counties are desir-
able to reveal any existing genetic patterns.
Both R. virginicus (Jenks, OK) and R. hageni
(Grove, OK) were found only in the eastern part of
the state where two-thirds of Oklahoma's forest
ecosystem consisting of over two million ha of
Oklahoma's timberlands (Lewis 2001). This dis-
tribution of R. virginicus and R. hageni in Okla-
homa was also observed by Brown et al. (2004).
These species are generally found in areas of min-
imal human disturbance, which may account for
their respective occurrences in this study and pre-
vious studies. For example, in Arkansas, R. vir-
ginicus and R. hageni are more prevalent in
undisturbed habitats (JWA, unpublished). Simi-
larly, the abundance of these species in eastern

Oklahoma may indicate central and western
Oklahoma represent an east to west transition
zone which delimits the westernmost occurrence
of Reticulitermes species not commonly known
from western U.S. states. Interestingly, Okla-
homa R. tibialis from Ardmore and Stillwater
share haplotypes with R. tibialis from Texas (T2
and T5, respectively). Also, two of the three R.
hageni samples are identical to the only southern
subterranean termite haplotype observed in
Texas (Austin et al. 2004).
Competition between ecologically similar ter-
mite species can lead to coexistence through re-
source partitioning (Houseman et al. 2001).
Given Reticulitermes ability to hybridize (Cle-
ment 1979) and fuse colonies, the opportunity to
observe greater genetic diversity is probable, par-
ticularly in sympatric zones, where otherwise
strong species isolation mechanisms (behavioral,
chemical, or temporal) are inadequate to prevent
hybridized mating (Austin et al. 2002). Because
colony structure and the spatial organization of
foraging Reticulitermes spp. is less understood,
population studies such as this are important in
understanding the complex ecology of subterra-
nean termites and Reticulitermes spp. in general.
By expanding our genetic investigations of Reti-
culitermes spp. from additional geographic zones,
the ecological interactions of this genus can be
better understood.


Hap Q F P H J O E G N

F 0.002 -
P 0.007 0.005 -
H 0.005 0.002 0.002 -
J 0.007 0.005 0.005 0.002 -
O 0.009 0.007 0.007 0.005 0.005 -
E 0.009 0.007 0.007 0.005 0.007 0.009 -
G 0.012 0.009 0.005 0.007 0.005 0.009 0.012 -
N 0.007 0.009 0.009 0.007 0.009 0.007 0.012 0.014

June 2004

Austin et al.: Reticulitermes Genetics

54 R flavipes hap Q

R flavipes hap F

R flavipes hap P

R flavipes hap H

81 R flavipes hap J

R flavipes hap O

R flavipes hap E

R flavipes hap G

R flavipes hap N
100 71 R tibialis hap T7

3 R tibialis hap T5

R tibialis hap T2

R tibialis hap T8

63 R hageni hap H2
52 R hageni hap H1

R virginicus hap V1

H aureus

C formosanus
Fig. 2. Single most parsimonious tree during a branch and bound search with PAUP*. Bootstrap values for 1,000
replicates are listed above the branches supported at >50%. Tree length = 121, CI = 0.777.
ACKNOWLEDGMENTS Messenger (Dow Agrosciences, Indianapolis, IN) and
Paul Baker (University of Arizona) for their contribu-
We thank contributors who provided specimens, par- tion of samples. This research was supported in part by
ticularly the PMPs of Oklahoma. Special thanks are the University of Arkansas, Arkansas Agricultural Ex-
given to Martyn Hafley (FMC, Philadelphia, PA), Matt periment Station.

Florida Entomologist 87(2)


WERK. 1987. Damage and mortality to pecan (Carya
illinoensis (Wangenh.) K. Koch) seedlings by subter-
ranean termites in an Oklahoma forest nursery.
Annu. Rep. North Nut Grow. Assoc. Hamden, CT. 78:
ANONYMOUS. 1999. Insect-Arthropod Diagnostics Report
ANONYMOUS. 2000. Insect-Arthropod Diagnostics Report
ANONYMOUS. 200 a. Termidor Testimonial: Termites not
OK in Oklahoma. Aventis TC-TERM-002PR-500-10/
ANONYMOUS. 2001b. Insect-Arthropod Diagnostics Re-
port http://entoplp.okstate.edu/Pddl/2001ireport.htm
ANONYMOUS. 2002. Insect-Arthropod Diagnostics Report
2002. A comparative genetic analysis of the subterra-
nean termite genus Reticulitermes (Isoptera: Rhino-
termitidae). Ann. Entomol. Soc. Amer. 95: 753-760.
FOSTER. 2004. Genetic variation and geographical
distribution of the subterranean termite genus Reti-
culitermes in Texas. Southwestern Entomologist (in
BULMER, M. S., AND J. F. A. TRANIELLO. 2002. Foraging
range expansion and colony genetic organization in
the subterranean Termite Reticulitermes flavipes
(Isoptera: Rhinotermitidae). Environ. Entomol. 31:
BROWN, K. S., B. M. KARD, AND M. P. DOSS. 2004. 2002
Oklahoma termite survey (Isoptera). J. Kansas En-
tomol. Soc. 77: 1-9.
CLEMENT, J-L. 1979. Hybridization experimental entire
Reticulitermes santonensis FEYTAUD et Reticuliter-
mes lucifugus ROSSI. Annales de Sciences Naturel-
les, Zoologie, Paris 13e s6rie. Vol. 1, pp. 251-260.
CLEMENT, J-L. 1986. Open and closed societies in Reti-
culitermes termites (Isoptera: Rhinotermitidae):
Geographic and seasonal variations. Sociobiol. 11:
CRISWELL, J., AND K. PINKSTON. 2001. Choosing a Ter-
mite Control Service. Oklahoma State University
Extension publication F-7308. 4 pp.
LETON. 2000. Morphological phylogenetics of ter-
mites (Isoptera). Biol. J. Linnean Soc. 70: 467-513.
FELSENSTEIN, J. 1985. Confidence limits on phyloge-
nies: An approach using the bootstrap. Evolution
FORSCHLER, B. T., AND V. LEWIS. 1997. Why termites
can dodge your treatment. Pest Control 65: 42-53.
Resource Partitioning in two sympatric species of sub-
terranean termites, Reticulitermes flavipes and Reti-
culitermes hageni (Isoptera: Rhinotermitidae).
Environ. Entomol. 30: 673-685.
FRAHN. 1995. Intracolony morphometric variation
and labral shape in Florida Reticulitermes (Isoptera:
Rhinotermitidae) soldiers: Significance for identifi-
cation. Florida Entomol. 78: 119-129.
MAN. 1987. The geographical distribution ofReticuli-

terms, Incisitermes and Coptotermes in Texas.
Southwest Entomol. 12: 119-125.
chal genetic structure of Reticulitermes (Isoptera:
Rhinotermitidae) populations. Sociobiol. 33: 239-263.
SCHLER. 2001. Phylogenetic analysis of two mitochon-
drial genes and one nuclear intron region illuminate
European subterranean termite (Isoptera: Rhinoter-
mitidae) gene flow, taxonomy, and introduction dy-
namics. Molec. Phylog. Evol. 20: 286-293.
KAMBHAMPATI, S., AND P. T. SMITH. 1995. PCR primers
for the amplification of four insect mitochondrial
gene fragments. Ins. Molec. Biol. 4: 233-236.
KIMURA, M. 1980. A simple method for estimating evo-
lutionary rate of base substitutions through compar-
ative study of nucleotide sequences. J. Molec. Evol.
16: 111-120.
KRISHNA, K., AND F. M. WEESNER. 1969. Biology of Ter-
mites. Vol. 1. Academic Press, New York, NY.
LEWIS, D. K. 2001. Oklahoma's forest ecosystems: Their
current condition and potential contribution. Proc.
Oklahoma Acad. Sc. 81: 31-40.
MARINI, M., AND B. MANTOVANI. 2002. Molecular Rela-
tionships among European samples of Reticuliter-
mes (Isoptera, Rhinotermitidae). Mol. Phylogenet.
Evol. 22: 454-459.
2002. Current distribution of the Formosan subter-
ranean termite and other termite species (Isoptera:
Rhinotermitidae, Kalotermitidae) in Louisiana.
Florida Entomol. 85: 580-587.
SCHEFFRAHN, R. H., AND N.-Y. SU. 1994. Keys to soldier
and winged adult termites (Isoptera) of Florida.
Florida Entomol. 77: 460-474.
AND P. FLOOK. 1994. Evolution, weighting, and phy-
logenetic utility of mitochondrial gene sequences and
a compilation of conserved polymerase chain reaction
primers. Ann. Entomol. Soc. Amer. 87: 651-701.
SWOFFORD, D. L. 2001. PAUP*: Phylogenetic analysis
using parsimony (*and other methods), ver. 4.0b10.
Sinauer, Sunderland, Massachusetts.
FRITZ. 2000. Population genetics and phylogenetics
of the endangered American burying beetle, Nicro-
phorus americanus (Coleoptera: Silphidae). Ann.
Entomol. Soc. Amer. 93: 589-594.
Identification of Reticulitermes spp. (Isoptera: Rhi-
notermatidae) from south central United States by
PCR-RFLP. J. Econ. Entomol. 96: 1514-1519.
TAJIMA, F., AND M. NEI. 1984. Estimation of evolution-
ary distance between nucleotide sequences. Mol.
Biol. Evol. 1: 269-285.
QUITANA, AND A. G. BAGNERES. 2003. Origin of a new
Reticulitermes termite (Isoptera, Rhinotermitidae)
inferred from mitochondrial DNA data. Mol. Phylo-
gen. Evol. (In press).
WANG, C., AND J. POWELL. 2001. Survey of termites in
the Delta Experimental Forest of Mississippi. Flor-
ida Entomol. 84: 222-226.
WEESNER, F. M. 1965. The termites of the United
States-A Handbook. 71 pp. The National Pest Con-
trol Association, Elizabeth, NJ.

June 2004

Halbert: Greenidea (Rhynchota: Aphididae) in the United States


Florida State Collection ofArthropods, Florida Department of Agriculture and Consumer Services
Division of Plant Industry, P.O. Box 147100, Gainesville, Florida 32614-7100, USA


Two species of the Asian genus Greenidea have been introduced into the United States,
Greenidea ficicola Takahashi and Greenidea psidii van der Goot. Synonymy confusion be-
tween Greenidea formosana (Maki) and G. psidii is resolved in favor of G. psidii. Both species
colonize Ficus spp., and G. psidii colonizes a few other plants, mostly in Myrtaceae. The two
species can be distinguished by the ornamentation on the siphunculi on the apterous forms,
and usually also by the arrangement of rhinaria on antennal segment III of the alate forms.

Key Words: Rhynchota, Aphididae, Greenidea ficicola, Greenidea psidii, Greenidea formo-
sana, Ficus


Dos species del g6nero asidtico Greenidea han sido introducidas a los Estados Unidos,
Greenidea ficicola Takahashi y Greenidea psidii van der Goot. Se resuelve la confusion en la
sin6nomia entire Greenidea formosana (Maki) y G. psidii en favor de G. psidii. Ambas espe-
cies colonizan Ficus spp., y G. psidii coloniza otras pocas plants, mayormente las de la fa-
milia Myrtaceae. Se puede distinguir las dos species en la ornamentaci6n de los sifunculi
en las formas apteras, y usualmente tambi6n por el arreglo de las rinarias sobre el segment
III de las antenas de las formas aladas.

The Asian genus Greenidea (Rhynchota: Aphi-
didae) belongs to the subfamily Greenideinae
(Greenideini). Most species of aphids in this sub-
family, including those in the genus Greenidea,
have long siphunculi with correspondingly long
setae. Until recently, no species in the subfamily
were found in the Western Hemisphere except
Brasilaphis bondari Mordvilko (Cervaphidini)
(Ghosh 1982) native to Brazil, and a fossil species
in Dominican amber (Wegierek 2001). Two spe-
cies, Greenidea psidii van der Goot and Greenidea
ficicola Takahashi, now have been found in the
USA. Both species have the potential to become
pests of certain ornamental plants.

Greenidea (Trichosiphum) psidii van der Goot 1916
(= Greenidea (Trichosiphum) formosana (Maki) 1917),
new synonymy (? = Greenidea (Trichosiphum) guang-
zhouensis Chang 1979 (Remaudiere & Remaudiere
1997)) (= Greenidea (Trichosiphum) formosana subsp.
heeri D. N. Raychaudhuri, M. R. Ghosh, M. Banerjee &
A. K. Ghosh 1973 (Remaudiere & Remaudiere 1997))
(= Trichosiphum formosanum Maki 1918)

Greenidea psidii was reported (under G. formo-
sana) in Hawaii in 1993 (Beardsley 1993). In
1998, G. psidii appeared in California (Gill 1998).
Outside of the United States, G. psidii is reported
from Bangladesh, China, India, Japan, Java, Loo-
choo Islands (Ryukyus), Nepal, Philippines,
Sumatra, and Taiwan (Blackman & Eastop 1994,

2000). I have a specimen from Brisbane, Austra-
lia that also appears to be this species.
Reported hosts of G. psidii include Psidium
guajava L. and other Myrtaceae (Callistemon,
Eucalyptus, Eugenia, Melaleuca, Metrosideros,
Rhodomyrtus, Syzygium, and Tristania). It also
infests Ficus (Moraceae), Engelhardtia (Juglan-
daceae), Scurrula (Loranthaceae), Lagerstroemia
(Lythraceae) and Nesua ferrea (Clusiaceae)
(Beardsley 1993; Blackman & Eastop 1994, 2000;
Gill 1998; Noordam 1994).
There is some nomenclatural confusion about
G. psidii. The most recent catalogue (Remaudiere
& Remaudiere 1997) lists this species as G. formo-
sana; however, the most recent revision of the ge-
nus (Noordam 1994), lists it as Greenidea psidii
van der Goot 1917. The Maki description (G. for-
mosana) was listed as having been published on
October 8, 1917 in honor of the sixtieth birthday of
Mr. Yasushi Nawa. In the 1917 journal version of
the van der Goot paper (G. psidii), it says he fin-
ished his work in 1915 and added corrections in
January 1916. It seems likely that the book was
published early in 1917, but there is no month
listed for the publication date, so according to the
International Code of Zoological Nomenclature,
the date is assumed to be 31 December 1917
(ICZN 2000, Article 21.3.2). However, both the
California Academy of Sciences and the library of
the Netherlands Entomological Society have cop-
ies of an "Extrait," or separate, that lacks a title

Florida Entomologist 87(2)

page but appears to have been distributed in 1916.
In both cases, "1916" has been written on the book.
According to the International Code of Zoological
Nomenclature, "Before 2000, an author who dis-
tributed separates in advance of the specified date
of publication of the work in which the material is
published thereby advanced the date of publica-
tion" (ICZN 2000, Article 21.8). Thus, the species
should be Greenidea psidii van der Goot 1916.

Greenidea ficicola Takahashi 1921 (= Greenidea neofici-
cola A. K. Ghosh, M. R. Ghosh & D. N. Raychaudhuri
(Remaudiere & Remaudibre 1997))

Greenidea ficicola (Figs. 1 and 2) was first sus-
pected in the Western Hemisphere when a single
damaged alate form was collected in a suction trap

sample collected 22-27 XI 2002 from Kendall, Flor-
ida, near Miami (Florida State Collection of Ar-
thropods (FSCA) #E2002-5901). No other
specimens were found until colonies were located
on Ficus aurea Nutt. on 18 II 2003 (FSCA# E2003-
569). There have been several subsequent finds in
the Miami area, including more trap collections.
The newly established aphid also has been found
in Naples, in southwest Florida (FSCA# E2004-
810, 849). In addition to Florida, G. ficicola is re-
ported from Australia, Bangladesh, Burundi (re-
cent introduction), China, India, Indonesia, Japan,
Malaysia, Nepal, Pakistan, Philippines, eastern
Russia, and Taiwan (Blackman & Eastop 2000).
Greenidea ficicola seems to be restricted to
Ficus spp. throughout most of its range; however,
in India, there are reports of infestations on Psid-

Fig. 1. Adult apterous Greenidea ficicola Takahashi.

June 2004

Halbert: Greenidea (Rhynchota: Aphididae) in the United States

I d

Fig. 2. Adult alate Greenidea ficicola Takahashi.

ium guajava. Noordam (1994) had a collection
from Streblus elongatus (Miq.) Corner (Mora-
ceae). The record ofG. ficicola from litchi reported

in Blackman & Eastop (2000) is probably spuri-
ous (Victor F. Eastop, pers. comm., 12 March
2003). In Florida, we have confirmed field coloni-


Florida Entomologist 87(2)

zation on F aurea Nutt., Ficus rigo (Bailey) Cor-
ner, and Ficus microcarpa L. f. We were able to
rear colonies for several weeks on Ficus benjam-
ina L. and Ficus carica L. in the laboratory.
Because both species colonize Ficus spp., it is
important to be able to differentiate between
them. A short key is provided:
la. Apterae with reticulations covering most of the
length of the siphunculi (Fig. 3); alatae with 17-21
rhinaria on antennal segment III, in a line and not
crowded or touching each other (Fig. 4) . G. ficicola
lb. Apterae with reticulations only at the base of the si-
phunculi; siphunculi ornamented with irregularly
spaced spinules (Fig. 5); alatae with 20-31 rhinaria,
some crowded and not in line with the others, often
touching (Fig. 6) ..................... G.psidii

Both of the newly established species of
Greenidea have the potential to become pests of
certain species of ornamental plants. Both colo-
nize Ficus, a genus that includes popular land-
scape and interiorscape plants, and G. psidii

Fig. 4. Antennal segment III of alate Greenidea fici-
cola Takahashi.




Fig. 3. Siphunculus of apterous Greenidea ficicola

Fig. 5. Siphunculus of apterous Greenidea psidii van
der Goot (= G. formosana (Maki)).

June 2004


Halbert: Greenidea (Rhynchota: Aphididae) in the United States


I thank Rosser Garrison, Raymond Gill and John So-
_ rensen, California Department of Food and Agriculture,
for providing specimens of California Greenidea, Julieta
Brambila (DPI) for technical assistance, Beverly Pope
(DPI), Paul Piron (Plant Research International,
Wageningen, the Netherlands), and Larry Currie (Califor-
nia Academy of Sciences) for library assistance, Michael
Thomas (DPI) for photography and graphics assistance,
and Cal Welbourn, Greg Hodges, Michael Thomas (all
DPI), Victor Eastop (The Natural History Museum, Lon-
don), George Remaudiere (Museum National d'Histoire
Naturelle, Paris), and David Voegtlin (Illinois Natural
History Survey) for reviewing the manuscript.


Fig. 6. Antennal segment III of alate Greenidea psi-
dii van der Goot (= G. formosana (Maki)).

colonizes several species in the Myrtaceae. In the
laboratory, G. ficicola caused significant leaf drop
on F benjamin. Greenidea psidii already has
been intercepted in Florida in a shipment ofMyr-
tus communis L. cut flowers from California
(FSCA# E2003-1827). The aphids colonize the
buds and new shoots of the host plants. No holo-
cycle is known for either species (Blackman &
Eastop 2000), suggesting that freezing tempera-
tures may be limiting, but both species should do
well in the neotropics and New World subtropics
wherever suitable host plants occur. Interior-
scapes, where temperatures do not fall below
freezing, also may sustain populations.

BEARDSLEY, J. W. 1993. Greenidea formosana (Maki),
an aphid new to the Hawaiian islands (Homoptera:
Aphididae: Greenideinae). Proc. Hawaiian Entomol.
Soc. 32: 157-158.
BLACKMAN, R. L., AND V. F. EASTOP. 1994. Aphids on the
world's trees. CAB International in association with
The Natural History Museum, Wallingford. 987 pp.,
16 plates.
BLACKMAN, R. L., AND V. F. EASTOP. 2000. Aphids on the
World's Crops, Second Edition. John Wiley & Sons,
Ltd., New York. 414 pp., 51 plates.
GHOSH, A. K. 1982. Cervaphidini (Homoptera: Aphi-
doidea) of the world. Oriental Insects 15: 77-97.
GILL, R. J. 1998. New state records: an aphid. California
Plant Pest and Disease Report 17: 9.
ICZN. 2000. International code of zoological nomencla-
ture, Fourth Edition. Tipografia la Garangola, Pa-
dova. 306 pp.
MAKI, M. 1917. Three new species of Trichosiphum in
Formosa, pp. 9-20. In Kikufiro Nagano [ed.], A Col-
lection of Essays for Mr. Yasushi Nawa: Written in
Commemoration of his Sixtieth Birthday, October 8,
1917. 186 pp.
NOORDAM, D. 1994. Greenideinae from Java (Homop-
tera: Aphididae). National Natuurhistorische Mu-
seum, Leiden, The Netherlands. 284 pp.
of the World's Aphididae. INRA, Paris. 475 pp.
TAKAHASHI, R. 1921. Greenidea ficicola n. sp. Aphididae
of Formosa. Part I. Agricultural Experiment Station
Government of Formosa, pp. 66-67, plate vii.
VAN DER GOOT, P. 1916. Zur kenntnis der Blattlause
Java's. Extrait des 'Contributions a la faune des Indes
N6derlandaises, Vol. I, Fasc III'. Gifu, Japan. 301 pp.
VAN DER GOOT, P. 1917. Zur kenntnis der Blattlause
Java's. Contributions a la fauna des Indes N6derlan-
daises, Vol. I, Fasc III. pp. 1-301.
WEGIEREK, P. 2001. Quisqueyaaphis heiei gen. and sp.
new (Hemiptera: Aphididae) new species of aphid from
Dominican amber. Annales-Zoologici 51: 409-415.

Florida Entomologist 87(2)

June 2004


'Department of Entomology and Nematology, P.O. Box 110620, Building 970,Natural Area Drive
University of Florida, Gainesville, FL 32611-0620, USA

2United States Department of Agriculture, Agriculture Research Service
2727 Woodlawn Drive, Honolulu, HI 96822-1842, USA

We describe all immature stages, particularly the previously undescribed instars, of Fopius
arisanus (Sonan) (Hymenoptera: Braconidae), an egg-pupal parasitoid of tephritid fruit
flies. This is essential for quality control in mass rearing programs and for physiological
studies of host-parasite interactions. Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)
eggs were parasitized for 24 h and serial collections of hosts were made every 24 h until
adults emerged. Immature wasps were dissected from hosts and their mouthhooks and body
dimensions measured. Scatter plots of the above measurements and scanning electron mi-
croscopy indicated that there are three instars. This contrasts with the four instars previ-
ously reported. There appears to be no true fourth instar because the stage immediately
following the second instar is indistinguishable from that preceding the prepupal stage.

Key Words: Braconid wasp, tephritid fruit fly host, egg-pupal parasitoid, biological control

Nosotros describimos todas los estadios inmaduros, particularmente los estadios no descri-
tos anteriormente, de Fopius arisanus (Sonan) (Hymenoptera: Braconidae), un parasitoide
del huevo-pupal de las moscas de la frutas de la familiar Tephritidae. Esto es esencial para el
control de cualidad en los programs de cria masiva y para studios fisiol6gicos de la inte-
racci6n entire hospedero y parasitoide. Los huevos de Bactrocera dorsalis (Hendel) (Diptera:
Tephritidae) fueron parasitados por 24 horas y hicieron colecciones en series de los hospede-
ros cada 24 horas hasta que los adults emergieron. Las avispas inmaduras fueron disecta-
das de sus hospederos y los ganchos bocales y las dimensions de cuerpo fueron medidos. Las
diagramas de dispersion de las medidas mencionadas y imagenes tomados por el microsco-
pio electr6nico (SEM) indicaron que habian tres estadios. Esto es contrario de los cuatro es-
tadios reportados anteriormente. Parece que no hay un cuatro estadio verdadero por que el
estadio siguiente inmediatamente al segundo estadio es indistinguible del que precede al es-
tadio prepupal.

Fopius arisanus (Sonan) is a parasitoid of
many tephritid fruit fly species (Diptera: Te-
phritidae) including the Oriental fruit fly, Bactro-
cera dorsalis (Hendel) and the Mediterranean
fruit fly, Ceratitis capitata (Wiedemann) (Vargas
& Ramadan 2000). It is one of the most effective
biological control agents of tephritids in Hawaii
(Harris & Okamoto 1991) and also parasitizes
some New World tephritids such as Anastrepha
suspense (Loew) in the laboratory (Lawrence et
al. 2000).
Previous reports based on mouthhook dimen-
sions indicated that F arisanus has four instars
(Ibrahm et al. 1992). However, there were no il-
lustrations of the larval morphology to facilitate
the identification of each instar. Other reports
have provided diagrams of the egg, first and
fourth instars (Palacio et al. 1992), and the pupa
that were useful for their identification, but gave
no diagrams of the second or third instars. The
goal of this study was to confirm the morphologies

of the first and last instars and to describe the
previously undescribed intermediate instars of
F arisanus.


Rearing of Parasites

Bactrocera dorsalis eggs were inserted into
holes punched into the rind of Carica papaya L.
and given to adult wasp females aged 10-25 d at a
ratio of 20:1 at 75-80C and 40-50% R.H. under
constant light for 24 h. Twenty-four hour sequen-
tial collections of hosts were made for 21 d when
adult wasps began to emerge. The experiment
was duplicated.

Light Microscopy

Fopius arisanus eggs, early instars [1-7 days
post parasitism (dpp)], and the heads of late in-

Rocha et al.: Immature Stages of Fopius arisanus

stars (9-14 dpp) were dissected from hosts and
placed in fluoromount-G or TE buffer (10 mM
Tris, 1 mM EDTA). Other instars (7-9 dpp) were
cleared in cellosolve [ethylene glycol monoethyl
ether (Carbide and Carbon Chemicals, New
York)] for 10 min and mounted with euparal (Bar-
bosa 1974).
Mouthhooks of F arisanus (10 individuals x 2
per time point) and body lengths were measured.
The means and standard errors of all measure-
ments were calculated and plotted against one
another. The resulting number of aggregations in-
dicated the number of instars according to Dyar's
(1890) rule that the "width of the head of a larva
in its successive stages follow a regular geometri-
cal progression." Although the rule is applied pri-
marily to lepidopteran larval head capsules, we
found that these measurements provided a reli-
able indicator of instars when used in combina-
tion with sequential dissections and other
morphological factors.

Scanning Electron Microscopy (SEM)

All larvae were placed in Trump's fixative (1%
glutaraldehyde and 4% formaldehyde in phosphate
buffer) overnight, washed with 0.1 M cacodylate
buffer (3 x 10 min), then fixed in 1% osmium tetrox-
ide for three days. After 3 x 10 min washes in deion-
ized water, the samples were dehydrated in a
graded series of ethanol, then incubated in hexam-
ethyldisilazane (HMDS) for 2 x 15 min and air
dried (Nation 1983). The larvae were then sputter
coated with gold and observed on a Hitachi S-570
scanning electron microscope at 20 kV.


Fopius arisanus eggs measured 300 (SE) 11.0
pm (range 250-350 pm) long and 55 (SE) 3.0 pm
(range 50-75 pm) wide. The egg stage lasted 1-2 d
and eggs were observed 0-2 dpp. Scatter plots of
mouthhook widths vs. mouthhook lengths (Fig.
la) and body lengths vs. mouthhook widths (Fig.
Ib) show two distinct aggregations of points, the
first occurring between 2-8 dpp and the second be-
tween 9-14 dpp. The mouthhook, cephalic, and
overall morphologies of these two groups corre-
spond to those previously described as first (Fig.
2) and last (fourth) instars, respectively (Ibrahm
et al. 1992; Palacio et al. 1992). An instar with
overall body size and morphology, differing from
the first and last instars, occurred between 7-9
dpp and had no sclerotized mouthhooks (Fig. 3a).
This time period coincides with the gap between
the two mouthhook size aggregations of the first
and last instar and no doubt represents the sec-
ond instar. Further analysis of the integument of
this putative second instar (Fig. 3b) indicated
that the integument is distinct from that of the
subsequent (last) instar (Figs. 4 and 5).

a 0.09
E 0.08
6 0.06
5 0.05
0 0.03
| 0.01
o 0

b [~.07
E 0.06
5 0.04
5 0.03
r 0.02

M 0

0 0.02 0.04 0.06 0.0
Mouthhook Width (mm)


2 4
Body Length (mm)

Fig. 1. Scatter plots of(a) mouthhook widths vs mouth-
hook lengths and (b) body lengths vs mouthhook widths
to show distinct aggregations representative of larval
instars ofFopius arisanus based on Dyar's (1890) rule.

Larval lengths from the tip of the head capsule
to the tip of the last abdominal segment were
0.848 0.06 mm (range 0.250-1.84 mm) for first
instar, 2.56 0.14 mm (range 1.50-3.22 mm) for
second instar, and 3.35 0.10 mm (range 2.99-
3.91 mm) for third instar.
The duration of the first, second, and third sta-
dia were eight, two, and six days, respectively.
Mouthhook dimensions of the first instar (Fig. 2)
were 16 1.0 pm x 24 1.0 pm, second instars had
no sclerotized mouthhooks, and third instars had

Fig. 2. Light micrographs of first instar (3 dpp)
Fopius arisanus to show sclerotized head capsule (hc),
sclerotized mouthhooks (mhk), and posterior tuft of se-
tae (tuf). Inset = enlargement of mouthhooks.



e ~rI 4L*




Florida Entomologist 87(2)

Fig. 3. Scanning electron micrograph of a second instar Fopius arisanus (8 dpp) to show (a) cephalic region lack-
ing sclerotized mouthhooks and (b) lack of spines on the integument. bc = buccal cavity; bp = buccal papilla.

Fig. 4. Scanning electron micrographs of early third instar Fopius arisanus (9 dpp) to show cephalic region (a)
and spines that cover integument (b). bss = basiconic sensillum; lbm = labrum; lbp = labial palp; mhk = mouthhook;
mth = mouth; mxp = maxillary palp; skg = silk gland; spn = spines.

June 2004

Rocha et al.: Immature Stages of Fopius arisanus

Fig. 5. Scanning electron micrographs of a late third instar (13 dpp) Fopius arisanus. (a) cephalic region to show
mouthparts. (b) spines that cover integument. bss = basiconic sensillum; cos = coeloconic sensillum; Ibm = labrum; lbp
= labial palp; mhk = mouthhook; mth = mouth; mxp = maxillary palp; skg = silk gland; spn = spines.

mouthhooks 63 3.0 pm x 42 2.0 pm (3.94x that
of first instars.) The distribution of the sensory
papillae surrounding the mouthparts of early and
late third instars is similar (Figs. 4 and 5).


Based on our direct observations of sequen-
tially dissected samples and morphology of F
arisanus larvae, we believe that this parasitoid
has three instars and a prepupal stage (Fig. 6) be-
cause antennal elongation was evident at 13 dpp.
While the third instar may have molted to a
fourth instar of similar morphology, there was no
distinct aggregation of mouthhook dimensions to
suggest an increase in size that is normally ex-
pected following a larval molt. Although Ibrahm
et al. (1992) and Palacio et al. (1992) reported a
fourth instar, our SEM evidence indicates that
the mouthhooks and cephalic region of the early
third instar which occurred immediately after (9-
11 dpp) the second instar (7-9 dpp) are similar in
morphology and dimension to those of the stage
(late third instar) immediately preceding the
prepupal stage (13-14 dpp).

Size and duration of parasitoid instars vary
with the size, age, and quality of the host in which
they are reared (Lawrence et al. 1976; Lawrence
1990). In addition we have observed size differ-
ences with different methods of fixation and
mounting (P. O. Lawrence, pers. obs.). Conse-
quently, we focused on sclerotized structures such
as larval mouthhooks and head capsules because
they are reliable characters for identification.
Nevertheless, measurements of soft tissues such
as body length, in relation to those of sclerotized
structures may prove useful for identification.
Our larval body measurements vary greatly from
those reported by Palacio et al. (1992) and Ibrahm
et al. (1992), even though the host species are the
same (B. dorsalis). This further underscores the
unreliability of soft tissue measurements for
identifying larval instars of parasitoids in general
and F arisanus in particular.
Evaluation of the integuments of the early and
late third instars (according to our definition) as
well as the pharate pupa, revealed no clear mor-
phological differences. There were no distinctions
between mouthhook sizes, integument, antenna
and labial sclerites, or distribution of cephalic

Florida Entomologist 87(2)

Fig. 6. Scanning electron micrograph of a prepupal
Fopius arisanus (13 dpp) to show differentiation of an-
tennae (ant). lbp = labial palp; mth = mouth; mhk =
mouthhook; mxp = maxillary palp; spn = spines.

sensilla between the early and late third instars.
Only direct observation of the molting of second
or third instars can definitively distinguish the
third from a presumed fourth instar. However,
our goal was to establish identification criteria
that are useful during dissections for quality con-
trol in mass rearing facilities. We believe that
standardized sequential sampling along with
these morphologies that are also visible under the
light microscope will suffice.


We thank Donna Williams from the Microbiology
and Cell Science Department at the University of Flor-
ida for her assistance with SEM and L. Matos and Y.

Hashimoto for reviewing the manuscript. Also, financial
support from the National Science Foundation (IBN
9986076) to P. O. Lawrence is gratefully acknowledged.
This article results from research conducted under
CRIS project ENY 03507 and is a Florida Agriculture
Journal Series No. R-09948.


BARBOSA, P. 1974. Manual of Basic Techniques in Insect
Histology. Autumn Publishers. Amherst, Massachu-
setts. p. 89.
DYAR, H. G. 1890. The number of molts of lepidopterous
larvae. Psyche 5: 420-422.
HARRIS, E. J., AND R. Y. OKAMOTO. 1991. A method for
rearing Biosteres arisanus (Hymenoptera: Braconi-
dae) in the laboratory. J. Econ. Entomol. 84: 417-422.
life cycle of Biosteres arisanus, with reference to
adult reproductive capacity on eggs of oriental fruit-
fly. Malaysia Appl. Biol. 21: 63-69.
ANY. 1976. Effect of host age on development of Bio-
steres (=Opius) longicaudatus, a parasitoid of the
Caribbean fruit fly, Anastrepha suspense. The Flor-
ida Entomol. 59: 33-39.
LAWRENCE, P. O. 1990. The biochemical and physiologi-
cal effects of insect hosts on the development and
ecology of their insect parasites: an overview. Arch.
Insect Biochem. Physiol. 13: 217-228.
2000. Development and reproductive biology of the
egg-pupal parasite, Fopius arisanus in Anastrepha
suspense, a new tephritid host. pp. 739-748. In K. H.
Tan [ed.], Area-Wide Control of Fruit Flies and
Other Insect Pests. Penerbit Universiti Sains Malay-
sia. 780 pp.
NATION, J. L. 1983. A new method using hexamethyld-
isilazane for preparation of soft insect tissues for
scanning electron microscopy. Stain Technology 58:
Identification of the immatures and male adults of
the opiine parasitoids (Biosteres spp.) of the Oriental
fruit fly, Bactrocera dorsalis (Hendel). The Philip-
pine Entomol. 8: 1124-1146.
VARGAS, R. I., AND M. M. RAMADAN. 2000. Comparisons
of demographic parameters: six parasitoids (Hy-
menoptera: Braconidae) and their fruit fly (Diptera:
Tephritidae) hosts, pp. 733-737. In K. H. Tan [ed.],
Area-Wide Control of Fruit Flies and Other Insect
Pests. Penerbit Universiti Sains Malaysia. 780 pp.

June 2004

Cilek & Schaediger: Black Fly Infestation and El Niio


'John A. Mulrennan, Sr., Public Health Entomology Research and Education Center
College of Engineering Sciences, Technology and Agriculture, Florida A & M University
4000 Frankford Avenue, Panama City, FL 32405

2Pasco County Mosquito Control District, 2308 Marathon Road, Odessa, FL 33556


A severe infestation of adult host-seeking black flies (Diptera: Simuliidae) occurred in west
central Florida during 1998. Collections from stationary suction traps in Pasco County re-
vealed the presence of large numbers of Simulium slossonae Dyar and Shannon. This spe-
cies peaked in traps during March (avg >40 per trap) with a lesser secondary peak in October
(avg =5 per trap). Moreover, during March, some suction traps had collected as many as
2,000 black flies for the month. It was believed that the spring outbreak of S. slossonae was
the result of above average precipitation associated with an El Niho event. Precipitation pro-
duced by this weather system during the winter of 1997/1998 provided a continuous source
of rain-swollen ditches, streams, and creeks for rapid larval and adult production the follow-
ing spring. Conversely, 1999 resulted in rainfall deficits of 1.5 cm to nearly 7.0 cm below nor-
mal. During that year, adult black fly populations were almost nonexistent (<3 black flies
collected per trap month) compared with collections obtained the previous year.

Key Words: Black fly, aquatic arthropods, El Niho, stream ecology


Una infestaci6n several de adults de la mosca negra (Diptera: Simuliidae) en busqueda de
hospederos ocurri6 en el oeste central de la Florida durante el 1998. La recolecci6n de moscas
en trampas de succi6n estacionarias en el condado de Pasco revel6 la presencia de un alto nu-
mero de Simulium slossonae Dyar y Shannon. El numero mas alto de esta especie recolec-
tados en las trampas fue durante el marzo (promedio de >40 por trampa) con el segundo
numero mas alto en octubre (promedio = 5 por trampa). Ademas, durante el marzo, algunos
de las trampas de succi6n recollectaron hasta 2,000 moscas negras por el mes. Se cree que
la erupci6n de la poblaci6n de S. slossonae en la primavera fue debido a la precipitaci6n mas
alta que el promedio asociada con el event de El Niho. La precipitaci6n producida por esta
sistema de tiempo durante el invierno de 1997/1998 provey6 un fuente continue de zanjas
llenas por la lluvia, quebradas, y arroyos para la producci6n rapida de las larvas y los adults
en la primavera siguiente. Al contrario, el 1999 result en un deficit de lluvia de 1.5 cm hasta
casi 7.0 cm menos del normal. Durante aquel ano, la poblaci6n de adults de la mosca negra
fue casi no existente (<3 moscas negras recolectadas por trampa por mes) comparada con la
recolecciones obtenidas el ano anterior.

Adult host-seeking black flies (Simulium spp.)
can often be severe biting pests of humans. The ir-
ritation associated with these bites can be consid-
erable and can often make life miserable in areas
where black fly populations are in great abun-
dance. Moreover, bites may become itchy and
swollen for a number of days. In sensitized indi-
viduals reaction to black fly saliva injected at the
feeding site may cause a syndrome known as
"black fly fever" that consists of headaches, fever,
nausea, and/or inflammation of nymph nodes
(Harwood & James 1979).
Larval habitats for black flies primarily con-
sist of swift running water, with shallow moun-
tain torrents being favored places (Harwood &

James 1979). In Florida, these habitats are not
present. Stone & Snoddy (1969) reported that
some species prefer slow flowing streams and
swamp rivers. These habitats are ubiquitous
throughout the State. In 1998, a severe outbreak
of adult Simulium slossonae Dyar and Shannon
occurred in west central Florida (particularly
Pasco County). Several reports of chicken mortal-
ity caused by adult black fly feeding had been re-
ported in the State during the first three months
of that year (Butler & Hogsette 1998). Although
this species is primarily a bird feeder, large
swarms were often attracted to people causing
considerable annoyance (Butler & Hogsette 1998).
Because this was an unusual event, the authors

Florida Entomologist 87(2)

wanted to document seasonal occurrence and
abundance of this species in Pasco County. In ad-
dition, we discuss the climatic events that led up
to that outbreak.


Black flies were collected in stationary suction
traps, primarily used for mosquito population
surveillance, by Pasco County Mosquito Control
District (PCMCD) personnel from 1997 through
1999. This trap is similar to that described by
Bidlingmayer (1971). Collection data were ob-
tained from daily catches from 35 traps placed
throughout the District (covering 855 km2). In
1998, larval samples from submerged vegetation
were periodically obtained from rain-swollen
streams by PCMCD staff to determine production
sites for emerging adults. Adult and larval sam-
ples were sent to Peter Adler, Department of En-
tomology, Clemson University for identification.
U.S. National Oceanic and Atmospheric Ad-
ministration monthly total precipitation data-
bases (including monthly normal levels) for 1997-
1999 were obtained from their data monitoring
station at Tampa International Airport (NOAA
1998b, 1999,2000).




a 30

n 25

2 15--



0W W


Suction trap collections from 1997, revealed
that adult S. slossonae were present in Pasco
County from May through November at an aver-
age of <2 black flies per trap month (Fig. 1). In
1998, collections of this species started to increase
greatly with a primary peak in March and a slight
secondary peak in October. During March, some
suction traps had collected nearly 2,000 black flies.
Suction traps located along the Anclote River,
Pithlachascotee River, and stream systems in the
Starkey Management Area, consistently collected
the greatest number of adult S. slossonae. These
watersheds were probably the primary source of
black fly infestation in the County and fit the de-
scription by Stone & Snoddy (1969) as slow mov-
ing southern swamp rivers/creeks favorable for
larval development of this species. Indeed, sub-
merged leaves and branches examined from those
watersheds revealed several hundred attached
S. slossonae larvae.
Simulium slossonae has previously been re-
ported to occur widely in Florida with immature
and adult specimens collected throughout the year
(Stone & Snoddy 1969; Pinkovsky & Butler 1978;
Butler & Hogsette 1998). But black fly populations

1997 1998 1999

Fig 1. Monthly mean adult Simulium slossonae obtained from stationary suction traps, Pasco County, FL, 1997-

June 2004

Cilek & Schaediger: Black Fly Infestation and El Niio









Fig. 2. Monthly total precipitation (cm), and associated deviation from normal, as reported by U. S. National Oce-
anic and Atmospheric Administration from Tampa International Airport weather data monitoring site, 1997-1999.

reported for the State of Florida had never before
increased to the pestiferous levels experienced in
1998. The outbreak of S. slossonae during that
year appeared to have resulted from above aver-
age rainfall during October through December,
1997, and again February, 1998 (Fig. 2). Rainfall
was reported to be 8 to 10 times above normal lev-
els in several counties (including Pasco) often
swelling stream and river systems to overflow in
early 1998 (Morris 1998). Indeed, the National Cli-
matic Data Center (NCDC) reported that the ex-
treme rainfall experienced in central Florida
during the latter part of 1997 and beginning of
1998 was associated with El Niio. This event pro-
duced 125% to nearly 300% that of normal precip-
itation levels (NCDC 1998a). According to NCDC,
November 1997 to March 1998 had been the wet-
test reported since records were started in 1895.
During April-June, 1998, adult S. slossonae pop-
ulations had declined considerably (Fig. 1). This pe-
riod was the driest interval on record (NCDC
1998a). Obviously the precipitation deficits had a
limiting effect on larval production (and subsequent
adult emergence) through decreased aquatic habi-
tat. During August, rainfall in west central Florida
returned to normal or slightly below normal levels
(Fig. 2) where, in October, a small peak of adult S.
slossonae was recorded in traps from Pasco County.

In 1999, adult black fly populations were al-
most nonexistent (<3 black flies collected per trap
month) (Fig. 1). Larval habitats were not as abun-
dant as the previous year with precipitation levels
1.5 cm to nearly 7.0 cm below normal (Fig. 2).
From our observations, and the data from west
central Florida, we found that when above average
precipitation events occur in the form of an El Niio
weather system, they can trigger a quick build up
of adult pestiferous S. slossonae populations. Ap-
parently this species can rapidly exploit rain-swol-
len watershed habitats as larval production areas
thereby producing enormous populations of host-
seeking adults. Indeed, observations during the
first half of 2003, revealed that S. slossonae again
had risen to pest population levels in Pasco County
(J.F.S., unpubl.) by exploiting rain swollen streams
produced from another El Niho system during the
winter of 2002-2003 (NOAA 2003a, b).


Appreciation is extended to Jim Robinson, Director,
and the staff of Pasco County Mosquito Control District
for their willingness to assist in the collection and colla-
tion of black fly data. Voucher specimens were deposited
in the Entomology Collection of Florida A & M Univer-
sity. We thank A. I. Watson, U.S. National Weather Ser-

Florida Entomologist 87(2)

vice, Tallahassee, FL for his help with obtaining central
Florida weather-related databases.


BIDLINGMAYER, W. L. 1971. Mosquito flight paths in re-
lation to the environment. I. Illumination levels, ori-
entation and resting areas. Ann. Entomol. Soc.
Amer. 61: 1121-1131.
BUTLER, J. F., AND J. A. HOGSETTE. 1998. Black flies,
Simulium spp. Department of Entomology, Univer-
sity of Florida. Featured Creature, EDIS Publication
#EENY-30. http://creatures.ifas.ufl.edu/livestock/bfly.
htm (accessed February 22, 2004).
HARWOOD, R. F., AND M. T. JAMES. 1979. Entomology in
Human and Animal Health (7 ed.). Macmillian Publ.
Co., NY.
MORRIS, C. 1998. Black flies hit Florida. Florida Mos-
quito Control Association. Buzz Words. January/Feb-
ruary. pg. 1.
NCDC. 1998a. Climate of 1998-Florida wild fires and
climate extremes. National Climatic Data Center,
National Oceanic and Atmospheric Admin.
Asheville, NC. 10 pp.

NOAA. 1998b. Climatological Data, Annual Summary
1997. National Oceanic and Atmospheric Admin.
101: 1-21.
NOAA. 1999. Climatological Data, Annual Summary
1998. National Oceanic and Atmospheric Admin.
102: 1-22.
NOAA. 2000. Climatological Data, Annual Summary
1999. National Oceanic and Atmospheric Admin.
103: 1-24.
NOAA. 2003a. Climate Diagnostics Bulletin. Climate Pre-
diction Center, National Oceanic and Atmospheric Ad-
min. http://www.cpc.ncep.noaa.gov/products/analysis
monitoring/CDB_archive.html (accessed August 8,
NOAA. 2003b. NOAA says El Nino to influence U.S.
weather. National Oceanic and Atmospheric Admin.
(accessed August 8, 2003)
PINKOVSKY, D. D., AND J. F. BUTLER. 1978. Black flies of
Florida I. Geographic and seasonal distribution.
Florida Entomol. 61: 257-267.
STONE, A., AND E. L. SNODDY. 1969. The black flies of Al-
abama (Diptera: Simuliidae). Auburn Univ. Agric.
Exper. Sta. Bull. 390.

June 2004

Cervantes Peredo: Cholula minute n.sp. from Jamaica


Institute de Ecologia, A.C., Apartado Postal 63 CP 91000, Xalapa, Veracruz, Mexico
E-mail: cervantl@ecologia.edu.mx


A new species of Cholula (Myodochini) from Jamaica is described. This represents the first
record of this genus for the Caribbean. Cholula minute can be differentiated from other spe-
cies of the genus mainly by its size. It is one of the smallest species described to date, being
similar in size only to C. parvus, but C. minute is unicolorous, while C. parvus has a mixture
of black, brown and white coloration.

Key Words: Cholula, Lygaeidae, Rhyparochromidae, Jamaica


Se describe una nueva especie de Cholula (Myodochini) de Jamaica. Esta represent el
primer registro de este g6nero para el Caribe. Cholula minute puede diferenciarse de otras
species del g6nero principalmente debido a su tamaio. Es una de las species mas pe-
queias descritas hasta ahora, es similar en tamaio a C. parvus, pero C. minute es de un solo
color, mientras que C. parvus es de una coloraci6n mezclada de negro, pardo y blanco.

Translation provided by author.

This paper describes a new species of Cholula
in order to make the name available for a review
of West Indian lygaeids that is in preparation by
J. A. Slater and R. Baranowski. The genus Cho-
lula includes 12 species of Neotropical distribu-
tion. None of the species has been recorded
previously from the Caribbean; six species are re-
ported from Mexico (C. bracteicola Cervantes &
Pacheco, C. irrorandus (Distant), C. lactifera
Brailovsky, C. lympha Brailovsky, C. maculatus
(Distant), and C. scapha Brailovsky), five from
Guatemala (C. bicolor Distant, C. irrorandus, C.
paruus (Distant), C. variegata Distant, and C. vi-
gens (Distant)), three from Panama (C. discoloria
Distant, C. firmus (Distant), and C. vigens), and
one from Honduras (C. parvus) (Brailovsky 1981;
Cervantes & Pacheco 2003; Distant 1882-1893).

Cholula minute Cervantes new species
(Fig. 1)

Labium reaching anterior third of abdominal
sternite III. Head and anterior pronotal lobe dark
ochraceous; posterior pronotal lobe, scutellum,
clavus and corium pale ochraceous, with ochra-
ceous punctures. Ventral surface covered with sil-
very hairs.
Head and anterior pronotal lobe covered with
tiny decumbent silvery hairs; eyes and ocelli red-
dish brown, ocelli located very close to anterior
margin of pronotum; antennae pale brown, with
joints pale yellow; rostrum pale yellow with tip of

segment IV brown. Pronotal collar and lateral
margins of posterior pronotal lobe yellow. Prono-
tum and scutellum very densely punctuate. Ace-
tabulae creamy yellow; coxae ochraceous; femora,
tibiae and tarsi pale yellow, fore femur slightly
darker. Pro-, meso-, and metapleura pale ochra-
ceous with posterior margins pale yellow. Clavus
with three complete rows of punctures and one in-
complete row. Corium with two rows of punctures
parallel to claval suture; rest of corium with
sparse punctures; membrane translucent. Ab-
dominal venter pale ochraceous.
Head slightly declivent, wider than long. Width
across eyes greater than width across anterior an-
gles of pronotum. Tylus longer than juga. Lateral
pronotal margins sinuate. Disk of scutellum
slightly elevated. Fore femur ventrally with double
ranked spines. Evaporative area occupying less
than half of metapleuron; peritreme auriculate.


Measurements in mm: Body length 3.6; head
length 0.57; width across eyes 0.95; interocular
distance 0.62; interocellar distance 0.32; postocu-
lar distance 0.02; antennal segments: I 0.22, II
0.45, III 0.37, IV 0.7; rostral segments: I 0.46, II
0.52, III 0.3, IV 0.3; pronotal length 0.87, width
across humeral angles 1.32, width across anterior
margin 0.72; scutellar length 0.68, width 0.68;
hind leg: femur length 0.88, tibia length 0.96,
tarsi length: I 0.2, II 0.07, III 0.16.

Florida Entomologist 87(2)

Fig. 1. Adult of Cholula minute new species. The bar at the right indicates the actual size of the male. The female
is slightly larger.

Male (Holotype) distance 0.02; antennal segments: I 0.2, II 0.4, III
0.35, IV 0.65; rostral segments: I 0.45, II 0.48, III
Measurements in mm: Body length 3.4; head 0.35, IV 0.3; pronotal length 0.8, width across hu-
length 0.5; width across eyes 0.85; interocular dis- meral angles 1.12, width across anterior margin
tance 0.55; interocellar distance 0.32; postocular 0.65; scutellar length 0.68, width 0.68; hind leg:

June 2004

Cervantes Peredo: Cholula minute n.sp. from Jamaica

femur length 0.82, tibia length 0.92, tarsi length:
I 0.12, II 0.07, III 0.18.
Types. Holotype, 16, JAMAICA: Manchester
Parish, Mandeville, 24-VIII-1969, R.E. Woodruff,
blacklight trap (Florida State Collection of Arthro-
pods). Paratype. 1 same locality as holotype; 23-
VIII-1960, J. Howard Frank, blacklight trap (R.
Baranowski Collection, University of Florida).


This species is similar in coloration to Cholula
lactifera and C. bracteicola, but both species are
much larger than C. minute sp. nov. In C. bracte-
icola the rostrum reaches abdominal sternite V,
and in C. minute it reaches only to anterior third of
abdominal sternite III. In C. lactifera antennal seg-
ment III is pale ochraceous; in C. bracteicola the
distal fourth of this segment is dark ochraceous,
while in C. minute all antennal segments are pale
brown. The hemelytral membrane is transparent
in C. minute and in C. bracteicola, while in C. lac-
tifera it has a milky appearance. Cholula minute is
one of the smallest species described to date, being
similar in size only to C. paruus, but C. minute is
unicolorous, while C. paruus has a mixture of
black, brown, and white coloration.
Recent sampling in Mexico has shown that
several species of Cholula are arboreal, and are

associated with figs, so probably C. minute is also
associated with figs in the Caribbean. Cholula
minute, as well as other species in the genus, is
attracted to light.


I thank Harry Brailovsky (Universidad Nacional
Autonoma de Mexico) for his comments on the manu-
script. I especially thank James A. Slater for lending me
the holotype and for encouraging me to describe this
species and continue studying lygaeids. I also thank Ri-
chard M. Baranowski for lending me the paratype spec-
imen. Financial support to visit R. Baranowski's
collection and publish this paper was provided by a
CONACyT grant (34238 V).


BRAILOVSKY, H. 1981. Hemiptera-Heteroptera de M6x-
ico XXI. Notas acerca de Cholula Distant y de-
scripci6n de nuevas species (Lygaeidae: Rhyparo-
chrominae: Myodochini). Folia Entomol6gica Mexi-
cana 47: 51-68.
ogy and description of a new species of Cholula (Rhy-
parochromidae: Myodochini) associated with a fig in
Mexico. J. New York Entomol. Soc. 111(1): 41-47.
DISTANT, W. L. 1882-1893. Biologia Centrali-Ameri-
cana. Heteroptera I. London: 210-215 and 400-404.

Florida Entomologist 87(2)

June 2004


'Entomology and Nematology Department, University of Florida, Gainesville, FL 32611-0630

2Marie Selby Botanical Gardens, 811 South Palm Avenue, Sarasota, FL 34236

3Myakka River State Park, 13207 SR 72, Sarasota, FL 34241

4Archbold Biological Station, P.O. Box 2057, Lake Placid, FL 33862

5Florida State Collection of Arthropods, 1911 SW 34th St., Gainesville, FL 32608-1268

6Department of Biological Sciences, Illinois State University, Normal, IL 61790-4120

7Fort Lauderdale Research and Education Center, University of Florida, 3205 College Ave.
Ft. Lauderdale, FL 33314-7799


Twenty four epiphytic bromeliads belonging to four species (Tillandsia fasciculata Swartz,
T recurvata (L.), T setacea Swartz, and T utriculata L.) were collected in Sarasota County,
Florida, in October-November 1997. Macroscopic invertebrate animals were extracted from
each by washing in water, filtering, and preserving in 75% ethanol. Plant sizes were mea-
sured in several ways, and their substrate was identified. Invertebrates were sorted,
counted, and identified as far as possible to the species level. Two species (T fasciculata, T
utriculata) that impound water in their leaf axils housed aquatic dipteran larvae and pupae
(Psychodidae, Culicidae, Ceratopogonidae, Chironomidae, Muscidae, and Aulacigastridae)
representing 7 species in 6 genera. Only T utriculata had a clear relationship between plant
size and number of invertebrates, which was steeper when only aquatic insect larvae were
counted. Plants of all four species housed terrestrial invertebrates, representing minimally
an additional 82 species in 75 genera and 63 families, very few of which are known to have
an obligate relationship with bromeliads, but showing that these plants support a diverse in-
vertebrate fauna. The presence of ant nests in some bromeliads complicated analysis. Such
a list of terrestrial invertebrates, identified to the species level, has not before been compiled
for bromeliads in Florida. Some collaborating taxonomists obtained specimens of species
that they could not identify, including probably undescribed species.

Key Words: Bromeliads, phytotelmata, insects, Tillandsia utriculata, bromeliad inhabitants


Se colectaron 24 bromeliaceas epifitas que pertenecen a cuatro species (Tillandsia fascicu-
lata Swartz, T recurvata (L.), T setacea Swartz, y T utriculata L.) en el Condado de Sara-
sota, Florida, durante octubre-noviembre de 1997. Se extrayeron los animals invertebrados
macrosc6picos de cada plant lavandola en agua y filtrando, seguido por preservaci6n de los
especimenes en etanol de 75%. Se midieron los tamaios de las plants por various
parametros, y se identifico su sustrato. Los invertebrados se ordenaron, contaron e identifi-
caron tanto possible al nivel de especie. Las dos species (T fasciculata, T utriculata) que em-
balsan agua entire sus axilas de hojas alojaron larvas y pupas acuaticas de moscas
(Psychodidae, Culicidae, Ceratopogonidae, Chironomidae, Muscidae y Aulacigastridae) rep-
resentando 7 species en 6 g6neros. Solo T utriculata tuvo una relaci6n clara entire tamaio
de plant y cantidad de invertebrados, la cual fue mas fuerte cuando se contaron solamente
las larvas de insects acuaticos. Plantas de las cuatro species alojaron invertebrados ter-
restres, representando un minimo de 82 species adicionales en 75 g6neros y 63 families,
muy pocas de las cuales se conocen tener una relaci6n obligada con bromeliaceas, pero de-
muestran que estas plants sostienen una diverse fauna de invertebrados. La presencia de
nidos de hormigas en algunas bromeliaceas complic6 el andlisis. Tal lista de invertebrados
terrestres, identificados al nivel de especie, no ha sido recopilado anteriormente para bro-

Frank et al.: Invertebrates from Florida Tillandsia Bromeliads

meliaceas en Florida. En este proyecto, various tax6nomos obtuvaron especimenes no-identi-
ficados, incluyendo species probablemente no-descritas.
Translation provided by the authors.

Bromeliads (Bromeliaceae) are a family of at
least 2500 species of monocotyledonous plants, al-
most restricted to the Neotropical region, but in-
cluding all of Mexico and southernmost USA. The
complex architecture of some species traps water
in leaf axils (forming phytotelmata) and harbors
many species of invertebrate animals. There are
thus three types of associations of invertebrates
with these plants: (a) those that feed on the
plants, (b) organisms aquatic at least in their im-
mature stages, and (c) those terrestrial organisms
for which bromeliads provide concealment, hu-
midity, or prey (Frank 1983). Within all three
groups are specialists, associated only with bro-
meliads, as well as generalists that occupy simi-
lar (non-bromeliad) habitats.
Four approaches have been followed in at-
tempts to unravel the mysteries of bromeliad
fauna. They may be termed (i) brief reports of new
discoveries, (ii) in-depth studies (behavioral or eco-
logical or taxonomic) of selected taxa, (iii) whole-
fauna inventories, and (iv) broad-scale hypothesis
tests. Major difficulties with the last two ap-
proaches are the need to involve teams of special-
ist taxonomists, and of distinguishing transient
species from those that have some kind of obligate
or at least usual relationship with bromeliads.
In Florida, an inventory of the macroscopic
aquatic invertebrate fauna in bromeliad tanks
(phytotelmata) is contained in an unpublished
Ph.D. dissertation (Fish 1976). A little of the con-
tent of that work was reviewed in Frank (1983).
An introduction to, and a bibliography of, studies
of the fauna and microflora of bromeliad phytotel-
mata, in Florida and abroad, are WWW-published
(Frank 1996 a, b). A complete illustrated key to
all developmental stages of all aquatic inverte-
brates in bromeliad phytotelmata in Florida can-
not now be prepared because some species are yet
undescribed (unknown to science). In contrast,
there are only 16 native species of bromeliads in
Florida, identifiable by color photographs online
as part of Frank & Thomas (1996) or (for the more
botanically adept) by a key in Wunderlin (1998).
In Florida, there has been one inventory of the
entire invertebrate fauna in the bromeliad
Tillandsia utriculata L. (Sidoti 2000), but most of
its identifications reached only the level of order.
There are works on some insects that feed on and
harm bromeliads. Detection of a moth, whose lar-
vae destroy pods of the bromeliad Tillandsia fas-
ciculata Swartz, led to a publication about larvae
of several moths that occasionally are collected
from native bromeliads in Florida (Heppner &
Frank 1998). Detection of a Mexican weevil,
Metamasius callizona (Chevrolat), in Florida led

to several publications about bromeliad-eating
weevils, reviewed and augmented by Frank
(1999) and expanded in two webpages (Larson &
Frank 2000; Larson et al. 2001) and two websites
(Frank & Thomas 1996; Larson 2000). Notable
studies in other countries are by Picado (1913) in
Costa Rica, and Beutelspacher (1971a, b) and
Palacios-Vargas (1981, 1982) in Mexico.
The Marie Selby Botanical Gardens (MSBG),
Sarasota, Florida, have an "Intern Program." Un-
der this program, students interested in plant
ecology and other aspects of botany are brought
from elsewhere to conduct a short-term (few
months) research project in one of these subjects.
Margaret Lowman (Research Director, MSBG)
and Sheeba Sreenivasan (an intern from Trinidad
and Tobago) in 1997 designed a project that was
to be a quantitative examination of the inverte-
brate fauna associated with native bromeliads in
Sarasota County. One of us (JHF) was asked to
visit MSBG to explain to Sheeba how to extract
invertebrates from bromeliads and preserve them
for examination, and also to receive her in his lab-
oratory and provide her with literature that
would help her to make preliminary identifica-
tions of the invertebrate specimens. These were
limited to insects, arachnids, myriapods, mol-
luscs, annelids, and the larger crustaceans. Fur-
ther development depended upon specialist
taxonomists to take the preliminary identifica-
tions as far as possible to the species level.
In November 1997, Sheeba visited JHF's labo-
ratory at the University of Florida, and used a
leaf-area-area meter for about 3 days to measure
the leaf-areas of the bromeliads she had collected.
To help complete the project, he sorted Sheeba's
specimens, some to family, but others only to the
level of order. He provided genus- or species-level
identification of the immature mosquitoes and a
few other dipteran larvae with which he was fa-
miliar and, much later, drafted a manuscript for
review by the other contributors. All the remain-
ing specimens had to be sent to specialist taxono-
mists for reliable identification, and the contacts
were made and specimens shipped before the end
of 1997. Fortunately, taxonomists of the Florida
State Collection ofArthropods (FSCA) were recep-
tive to providing help. Here is an account of these
invertebrates. This account recognizes the essen-
tiality of the contributions of several taxonomists,
who were offered co-authorship (some declined).


Twenty four bromeliads were collected from
sites in Sarasota County, principally from old-

Florida Entomologist 87(2)

growth hammocks in the Myakka River State
Park, and secondarily two sites in Sarasota. They
were removed from various substrates including
living trees, dead trees (snags), and a gate (Table
1). While it was being collected, each plant was
kept as upright as possible to prevent spillage.
The plant was then placed into a polyethylene bag
and briefly sprayed with insect repellent before
fastening the bag.
Invertebrates were extracted from bromeliads
by a variant of the method of Frank et al. (1976).
Each plant was cleaned with a jet of water from a
hose, with the washings directed into a bucket.
The plant was repeatedly submerged and shaken
in the bucket before being returned to its bag. The
water in the bucket was then filtered with a tea-
strainer (mesh size 500 pm) and the residue ex-
amined for invertebrates with a dissection micro-
scope. Collected invertebrates were preserved in
vials containing 75% ethanol for subsequent iden-
The collected bromeliads included whole spec-
imens of the epiphytic species Tillandsia utricu-
lata, T fasciculata, T setacea, and T recurvata.
These include all the most widespread of Florida's
16 native species except T usneoides (L.). Only
the first two of these impound water in leaf axils,
and they do this only when they have reached a
certain minimal size (exceeded by the specimens).
The volumetric capacity of the water-impounding
leaf axils of T utriculata has been related to
length of longest leaf and to age (in one habitat)
by Frank & Curtis (1981), so length of longest leaf
was one of the measurements made (Volumetric
capacity in ml = 0.003251 x leaf length in cm27799).
Other measurements were made by dismantling
each plant, leaf by leaf, starting from the outer-
most and working inward. This was done on a
white background to facilitate detection of any re-
maining invertebrates. A component part was
considered to be either a leaf or an infructescence
(the fruiting phase of an inflorescence). Each ele-
ment was further designated as live or dead. Each
bromeliad's live and dead leaves were counted.
All components were refrigerated until leaf and
infructescence areas were measured with an area
meter (LI-COR Portable Area Meter, model LI
3000, LI-COR Inc., Lincoln, NE, USA). They were
then oven-dried for 48 hr before weighing. Table
1, which lists the measurements, is thus a habitat
description rather than results.


Table 2 lists the invertebrates collected to the
level of family (with number of specimens) for
each of the 24 bromeliads sampled. Identification
of the invertebrates to the species level, where
possible, is given below. Comments are made
where deemed appropriate. Three vials contain-
ing Mollusca and four with Thysanoptera were

mislaid somewhere in the Florida State Collec-
tion of Arthropods; details of their contents would
not substantially change the conclusions.


None was identified. The three missing vials (6
specimens) may be located in FSCA. No mollusc
has a known, obligate relationship to bromeliads.
However, H. E. Luther (pers. comm.) has observed
snails eating bromeliad trichome caps in the field
and greenhouse. Assume minimally one family,
one genus, and one species.

Isopoda (identified by G. B. Edwards)
Oniscidae: genus and species unidentified (17
Rhyscotidae: genus and species unidentified (9

Diplopoda (identified by G. B. Edwards)
Chilognatha: family and genus unidentified,
species 1 (12 specimens), species 2 (1 specimen).
Pselaphognatha: Polyxenidae: Polyxenus fas-
ciculatus (Say) (5 specimens).
These were collected from all three Tillandsia
species in MRSP They have no known relation-
ship to bromeliads. All of the Chilognatha were
immature, so could not be identified reliably.

Chilopoda (identified by G. B. Edwards)
Lithobiidae: ?Neolithobius sp. (1).
This immature specimen was collected from
T utriculata in MRSP It has no known relation-
ship to bromeliads.

Araneae (identified by G. B. Edwards)
Segestriidae:Ariadna bicolor (Hentz) (2).
Theridiidae: Phoroncidia americana (Emer-
ton) (1 immature), ?genus (1 immature).
Mysmenidae: Mysmenopsis cymbia Levi (10).
Linyphiidae: Ceraticelus ?phylax Ivie & Bar-
rows (1 female).
Tetragnathidae: Dolichognatha pentagon
(Hentz) (1), ?D. pentagon (1 immature, dam-
aged), ?Tetragnatha sp. (2 immatures).
Lycosidae: ?genus (2 immatures).
Pisauridae: ?Dolomedes sp. (1 immature).
Agelenidae: ?Agelenopsis sp. (1 immature).
Hahniidae: Hahnia okefinokensis Chamberlin
& Ivie (1).
Dictynidae: Emblyna capens Chamberlin (1),
Emblyna sp. (2 immature), Lathys delicatula
Gertsch & Mulaik (2).
Anyphaenidae: Lupettiana mordax (O.P Cam-
bridge) (1).
Liocranidae: Scotinella pintura (Ivie & Bar-
rows) (3), Scotinella sp. (1 immature).

June 2004

Frank et al.: Invertebrates from Florida Tillandsia Bromeliads


No. live No. dead Longest Live leaf Dead leaf Live infr. Deadinfr. Dry wt
Code Substrate leaves leaves leaf (cm) area (cm2) area (cm2) area (cm2) area (cm2) (g)

Tillandsia fasciculata
8 Cephalanthus 64 4 58.4 2621.99 60.18 0 0 57.5
9 Cephalanthus 65 20 62.0 2794.81 316.20 0 0 71.5
10 Cephalanthus 51 11 46.4 898.86 66.24 0 0 21.5
15 snag 55 13 44.5 2002.08 199.07 0 0 33.0
17 Ulmus 33 3 44.5 881.81 44.55 0 0 10.5
18 fallen branch 55 2 22.8 350.46 37.23 0 0 6.5
19 rooted in soil 58 10 29.2 475.15 27.79 0 0 11.5
20 Quercus 76 13 55.4 5006.87 371.90 0 0 189.5
Tillandsia utriculata
1 snag 41 6 32.0 531.62 38.44 0 0 8.0
2 snag 56 29 46.5 1793.91 237.25 0 0 32.0
3 snag 55 22 33.0 824.92 63.65 0 0 11.5
4 snag 62 11 17.9 164.69 14.34 0 0 2.5
16 Quercus 60 12 91.1 9285.81 909.66 0 0 147.0
24 Quercus 59 11 93.3 7350.48 0 0 0 152.5
Tillandsia recurvata
21 Sabal 1140 71 10.1 672.1 48.60 0 0 10.5
22 wooden gate 80 1 9.3 51.69 2.37 6.02 0 1.0
23 Quercus 537 58 13.5 305.72 82.25 34.36 23.00 5.0
Tillandsia setacea
5 snag 196 82 19.0 151.37 62.86 0 0 3.5
6 snag 349 24 18.6 236.57 17.15 0 0 3.0
7 snag 158 22 27.2 210.06 27.90 28.20 5.58 8.0
11 Cephalanthus 419 63 32.1 556.39 60.18 67.68 7.97 16.5
12 Cephalanthus 338 29 28.9 437.23 62.32 18.87 16.13 9.5
13 Cephalanthus 81 34 23.5 185.37 36.46 26.92 0 3.0
14 Quercus 379 32 29.8 298.02 18.81 4.93 5.33 8.5

Clubionidae: Clubiona pygmaea Banks (3), Clu-
biona sp. (1 immature), Elaver except (L. Koch)
Gnaphosidae: Litopyllus cubanus Bryant (1),
Sergiolus sp. (1 immature).
Sparassidae: Pseudosparianthis cubana Banks
(1 immature).
Thomisidae: Bassaniana floridana (Banks) (1),
Bassaniana sp. (4 immatures).
Salticidae:Anasaitis canosa (Walckenauer) (2).
The spiders seem to represent 21 species, in 21
genera and 17 families. For only one spider (Pele-
grina tillandsia [Kaston]) is a bromeliad Ti!. .....
sia usneoides) in the southern USA known to be
the preferred habitat. In the Neotropical region,
however, other spiders typically inhabit bromeli-
ads and even are semi-aquatic in bromeliad leaf

Pseudoscorpiones (identified by G. B. Edwards)

Chthoniidae: genus and species unidentified (2

One specimen was from T fasciculata and the
other from T setacea. They were unidentifiable
because immature. Pseudoscorpions have no
known obligate relationship to bromeliads.

Acari (identified by W. C. Welbourn)

Liodidae: Liodes sp. 1 (16), Liodes sp. 2 (13), Li-
odes sp. 3 (3).
Ascidae: Lasioseius sp. (2).
Haplozetidae: genus and species unidentified
Oripodidae: genus and species unidentified (1).
Uropodidae: Uropoda sp. (2).
Oppiidae: genus and species unidentified (1).
Orbataloid: genus and species unidentified (1).
Histiostomatidae: Hormosianoetus sp. (37).
None of these 10 species in 8 genera and 8 fam-
ilies is known to have any obligate relationship
with bromeliads. There is a pressing need for more
basic taxonomic work on Floridian Acari other
than those of economic importance; only then will
specimens be identifiable to the species level.

Florida Entomologist 87(2)

June 2004


CD/PL Fauna to level of family, and number of specimens Sum

Tillandsia fasciculata
08/ M CRUSTACEA: Isopoda: Rhyscotidae (2), ARACHNIDA: Araneae: Theridiidae (1), Mys- 19
menidae (10), Hahniidae (1), Liocranidae (1), INSECTA: Homoptera: Aphididae (1), Lepi-
doptera: Tineidae (larvae, 1), Diptera: ?Muscidae (larva 1), Hymenoptera: Formicidae (1)
09/ M ARACHNIDA: Araneae: Tetragnathidae (1), Dictynidae (1), Liocranidae (1), Clubionidae 18
(2), Salticidae (1), INSECTA: Isoptera: Kalotermitidae (4), Blattodea: Blatellidae (2), Pso-
coptera: Lepidopsocidae (nymphs 2), family indet. (nymphs 2), Diptera: ?Muscidae (larva 1),
Hymenoptera: Formicidae (1)
10/ M CRUSTACEA: Isopoda: Rhyscotidae (6), ARACHNIDA: Araneae: Linyphiidae (1), Age- 21
lenidae (1), Clubionidae (2), Acari: Uropodidae (1), INSECTA: Orthoptera: Gryllidae (3),
Psocoptera: Lepidopsocidae (1), Archipsocidae (1), Peripsocidae (2), family indet. (1), Co-
leoptera: Tenebrionidae (1), Hymenoptera: Formicidae (1)
15/ M DIPLOPODA: Pselaphognatha: Polyxenidae (1), ARACHNIDA: Araneae: Segestriidae (1), 65
Pisauridae (1), Acari: Histiostomatidae (19), INSECTA: Thysanoptera (1), Psocoptera: Lep-
idopsocidae (1), Orthoptera: Gryllidae (2), Coleoptera (larvae 3, of 3 families), Diptera: Psy-
chodidae (larvae 32), Culicidae (larva 1), Ceratopogonidae (larvae 3)
17/ M CRUSTACEA: Isopoda: Rhyscotidae (1), DIPLOPODA: Chilognatha: family indet. (1), 17
ARACHNIDA: Araneae: Clubionidae (2), Pseudoscorpionida: Chthoniidae (1), INSECTA:
Blattodea: Blatellidae (4, and one egg case), Lepidoptera: family indet. (larvae 5), Co-
leoptera: Carabidae (larva 1), Diptera: Ceratopogonidae (larva 1)
18/ M ARACHNIDA: Araneae: Anyphaenidae (1), Thomisidae (1), Salticidae (1), Acari (?1), IN- 7
SECTA: Orthoptera: Gryllidae (1), Coleoptera: Scirtidae (1), Hymenoptera: Formicidae (1)
19/ M CRUSTACEA: Isopoda: Oniscidae (3), DIPLOPODA: Pselaphognatha: Polyxenidae (3), Pse- 144
laphognatha: family indet. (3), ARACHNIDA: Araneae: Lycosidae (2), INSECTA: Blattodea
(egg case 1), Homoptera: Ortheziidae (1), Coleoptera: Brentidae (1), Hymenoptera: Formi-
cidae (128 plus brood), Ichneumonidae (2)
20/ M MOLLUSCA (3), CRUSTACEA: Isopoda: Oniscidae (10), ARACHNIDA: Araneae: Dic- 36
tynidae (1), Sparassidae (1), INSECTA: Blattodea: Blattidae (1), Orthoptera: Gryllidae (1),
Lepidoptera: family indet. (larva 1), Diptera: Psychodidae (larvae 2), Ceratopogonidae (lar-
vae 8, pupae 2) Chironomidae (larvae 4), Aulacigastridae: (larva 1, pupa 1)
Tillandsia utriculata
01/ M ARACHNIDA: Araneae: Dictynidae (1), Acari: Liodidae (1) 2
02/ M CRUSTACEA: Isopoda: Oniscidae (2), DIPLOPODA: Chilognatha: family indet. (5), 39
ARACHNIDA: Araneae: Tetragnathidae (1), Liocranidae (2), Clubionidae (1), Acari: Ascidae
(2), INSECTA: Blattodea: Blatellidae (1), Psocoptera: Pseudocaeciliidae (1), Liposcelidae
(1), family indet. (nymph 1), Diptera: Ceratopogonidae (larva 1), Psychodidae (larvae 16),
Hymenoptera: Formicidae (5)
03/ M DIPLOPODA: Pselaphognatha: Polyxenidae (1), ARACHNIDA: Araneae: Clubionidae (1), 16
Thomisidae (1), INSECTA: Psocoptera: Caeciliusidae (1), Archipsocidae (1), Liposcelidae (1),
Lepidopsocidae (nymph 1), family indet. (nymph 1), Blattodea: Blattidae (2), Homoptera:
Coccidae (1), Coleoptera: Curculionidae (3), Diptera, Ceratopogonidae (larvae 2)
04/ M DIPLOPODA: Chilognatha: family indet. (2), ARACHNIDA: Araneae: Clubionidae (1), Th- 10
omisidae (2), Acari: Liodidae (1), INSECTA: Blattodea: Blatellidae (2), Coleoptera: Curcu-
lionidae (1), Diptera: Chironomidae (1)
16/ M CHILOPODA: Lithobiidae (1), DIPLOPODA: Chilognatha: family indet. (1), ARACHNIDA: 226
Acari: Histiostomatidae (18), INSECTA: Blattodea: Blattidae (1), Psocoptera: family indet.
(1 nymph), Thysanoptera (2), Lepidoptera: family indet. (larva 1), Diptera: Psychodidae:
(larvae 108, pupa 1), Culicidae: (larvae 28), Ceratopogonidae (larvae 47), Chironomidae (lar-
vae 3), Aulacigastridae: (5), Cecidomyiidae (adults 2, pupa 1), Hymenoptera: Formicidae (6)
24/S ARACHNIDA: Araneae: Dictynidae (1), Gnaphosidae (1), INSECTA: Collembola: Entomobry- 135
idae (22), Psocoptera: Trogiidae (2), family indet. (nymph 1), Coleoptera: Coccinellidae (1),
Diptera: Psychodidae (larvae 31), Hymenoptera: Formicidae (76, of 2 spp., each with brood)
Tillandsia recurvata
21/S INSECTA: Psocoptera: Lepidopsocidae (1), Coleoptera: Elateridae (1), Hymenoptera: For- 4
micidae (2)
22/S No animals were collected 0

Frank et al.: Invertebrates from Florida Tillandsia Bromeliads


23/S MOLLUSCA (1), ARACHNIDA: Araneae: Gnaphosidae (1), INSECTA: Hemiptera: Miridae 6
(nymphs 2), Psocoptera: Trogiidae (1), Hymenoptera: Formicidae (1)
Tillandsia setacea
05/ M Diplopoda: Pselaphognatha: Polyxenidae (1), ARACHNIDA: Acari: Liodidae (4), Haploz- 11
etidae (1), Oripodidae (1), INSECTA: Psocoptera: Peripsocidae (1), Archipsocidae (1
nymph), Lepidopsocidae (1 nymph), Orthoptera: Gryllidae (1)
06/ M ARACHNIDA: Araneae: Clubionidae (2), Thomisidae (1), Acari: Liodidae (9), INSECTA: 19
Psocoptera: Peripsocidae (2), Homoptera: Aphididae (4), Hymenoptera: Aphelinidae (1)
07/ M MOLLUSCA (2), ARACHNIDA: Araneae: Segestriidae (2), INSECTA: Blattodea: Blatel- 60
lidae (2), Lepidoptera: family indet. (larvae 2), Hymenoptera: Formicidae (52)
11/ M ARACHNIDA: Araneae: Dictynidae (1), Clubionidae (1), Acari: Liodidae (2), INSECTA: 6
Thysanoptera (1), Coleoptera: Scirtidae (1)
12/ M ARACHNIDA: Araneae: Theridiidae (1), Tetragnathidae (2), Thomisidae (1), Acari: Lio- 18
didae (2), Oppiidae (1), 'orbataloid' (1), Pseudoscorpionida: Chthoniidae (1), INSECTA: Col-
lembola: Hypogastruridae (5), Thysanoptera (2), Psocoptera: Peripsocidae (nymph 1),
Hymenoptera: Formicidae (1)
13/ M CRUSTACEA: Isopoda: Oniscidae (2), ARACHNIDA: Araneae: Clubionidae (1), Acari: Lio- 18
didae (13), INSECTA: Psocoptera: Liposcelidae (1), Diptera: Cecidomyiidae (1)
14/ M ARACHNIDA: Araneae: Clubionidae (1), Acari: Uropodidae (1), INSECTA: Lepidoptera: 4
Gelechiidae (larva in flower) (1), Hymenoptera: Formicidae (1)

Notes: CD = code number (collection no.)/PL = place (M = Myakka River State Park, S = Sarasota). SUM = total number of in-
vertebrate animals of the groups sampled. Presence of immature stages suggests that development was taking place in the brome-
liad unless the individuals fell from the tree above.

Collembola (identified by R. J. Snider)

Entomobryidae: Seira steinmetzi Wray (22).
Hypogastruridae:Xenylla sp. (5).
The specimens of Xenylla represent an unde-
scribed species and were retained by R. J. Snider.

Orthoptera (identified by T J. Walker)

Gryllidae: Cycloptilum trigonipalpum (Rehn &
All 8 specimens of Orthoptera were identified
as of this species or were unidentifiable because
immature, but probably belong to this species.
Seven of them were collected from T fasciculata
and one from T setacea, all within MRSP.

Blattodea (identified by M. C. Thomas)

Blattellidae: Cariblatta sp. prob. lutea (Saus-
sure & Zehnter) (12).
Blattidae: Eurycotis floridana (Walker) (4).
Neither species has any known obligate rela-
tionship with bromeliads.

Isoptera (identified by R. H. Scheffrahn)

Kalotermitidae: Crytotermes ?cauifrons Banks
No termite is known to have an obligate rela-
tionship with bromeliads. Most likely these had
fallen from a dead tree limb.

Psocoptera (identified by E. L. Mockford)

Trogiidae: Cerobasis guestfalica (Kolbe) (4).
Lepidopsocidae: Echmepteryx (Thylacopsis)
madagascariensis (Kolbe) (3), Echmepteryx sp. (1),
Nepticulomima sp. (1), unidentified nymphs (4).
Liposcelidae: Liposcelis ornata Mockford (1),
Liposcelis sp. (2).
Archipsocidae: Archipsocus sp. (2), unidenti-
fied nymphs (1).
Peripsocidae: Peripsocus sp. (5).
Caeciliusidae: Valenzuela indicator (Mockford)
(= Caecilius indicator Mockford) (1).
Pseudocaeciliidae: Pseudocaecilius citricola
(Ashmead) (1).
Epipsocidae: Mesepipsocus niger (New) (3), un-
identified nymphs (4).
None of these 12 species in 9 genera and 8 fam-
ilies is known to have an obligate relationship to
bromeliads. As in so many other groups, it is dif-
ficult to identify nymphs reliably to the species
level. The surprise among these specimens was
the finding of specimens of C. guestfalica from
two Tillandsia specimens (T recurvata, T utricu-
lata) from the city of Sarasota; it is an adventive


None was identified. The four missing vials (6
specimens) may be located in FSCA. Assume min-
imally one family, one genus, and one species.

Florida Entomologist 87(2)

Hemiptera/Homoptera (identified by S. E. Halbert)
(Ortheziidae by A. B. Hamon)
Miridae: unidentified nymphs.
Aphididae: Myzocallis sp. (4), unidentified ge-
nus (1 cast skin).
Ortheziidae: Orthezia sp. (1 nymph).
Coccidae: genus and species unidentified (1
adult male).
The Myzocallis aphids are known to feed on
oak; one of the specimens was parasitized byAph-
elinus sp. (Hymenoptera: Aphelinidae) (det. G. A.
Evans, FSCA). The only species of Orthezia re-
ported from Tillandsia in Florida is 0. tillandsiae
Morrison, but the specimen obtained (from T fas-
ciculata in MRSP) is immature and could not be
identified with certainty. The Miridae were uni-
dentifiable because they were immature.

Coleoptera (identified by M. C. Thomas)
Scirtidae: Ora sp. (1), Cyphon sp. (1).
Elateridae: Conoderus amplicollis (Gyllenhal)
Tenebrionidae: Glyptotus cribratus LeConte (1).
Brentidae:Apion sp. (1).
Curculionidae: Acalles clavatus Say (3);
Conotrachelus maritimus Blatchley (2).
Larvae of Scirtidae are aquatic. It is possible
that the two adults of Scirtidae are associated
with bromeliad phytotelmata, but both specimens
were found in T setacea, which does not form phy-
totelmata. Alternatively, their larvae may de-
velop in treeholes. Specimens mislaid include one
adult of Coccinellidae and several beetle larvae, of
which one is of Carabidae.

Lepidoptera (identified by D. H. Habeck)
Tineidae: genus and species unidentified (1
Gelechiidae: genus and species unidentified (1
Family uncertain: (8 larvae).
Lepidoptera were represented only by larvae of
"primitive" families. Their identification was un-
certain. It is not clear whether these larvae were
feeding on bromeliads, on debris in the bromeli-
ads, or on the tree canopy (or epiphytes) above.
The gelechiid larva was found clinging to a flower
(which was not a bromeliad flower) in the brome-
liad, but there is no evidence it was feeding on that
flower. There is a clear need to rear lepidopteran
larvae encountered in bromeliads, to allow identi-
fication of adults and associate larvae with adults.

Diptera (identified by J. H. Frank)
(Cecidomyiidae by R. J. Gagn6, Ceratopogonidae by
G. J. Steck)
Cecidomyiidae: Campylomyza sp. (1 adult),
Lestodiplosis laticaulis Gagne (2 adults and 1

Psychodidae: Alepia sp. (190 larvae and pu-
Culicidae: Wyeomyia mitchellii (Theobald) (25
larvae), W vanduzeei Dyar & Knab (3 larvae).
Ceratopogonidae: Forcipomyia sp. or spp. (62
larvae, 2 pupae)
Chironomidae: Monopelopia tillandsia Beck &
Beck (53 larvae), (1 damaged unidentified adult).
Aulacigastridae: Stenomicra (7 larvae).
?Muscidae: genus and species unidentified (2
larvae) (perhaps this is the Neodexiopsis sp. of
Fish [1976]).
Specimens of Cecidomyiidae were shipped in
vials of alcohol to R. A. Gagne; he found both spe-
cies interesting and retained one specimen of each.
He reports that L. laticaulis is known as a preda-
tor of Diaspis echinocacti (Bouche) (Homoptera:
Diaspididae) a scale insect on Opuntia cacti-so
its presence in T utriculata is unexpected. Larvae
of Psychodidae, Culicidae, many Ceratopogonidae,
Chironomidae, and Aulacigastridae are aquatic
and, expectedly, were found only in T fasciculata
and T utriculata. Identification of these larvae
was made by J. H. Frank (who makes no claim to
be an expert on larvae of Diptera), either by prior
experience (larvae of Wyeomyia mosquitoes), by
use of keys to larvae of Chironomidae (Epler
2001), or (for the other families) according to the
brief descriptions by Fish (1976). Although 27
years have passed since Fish (1976) reported his
collections, the "Neurosystasis" (Psychodidae) and
"Stenomicra" (Aulacigastridae) occurring in Flor-
ida bromeliads have not yet been formally de-
scribed. G. J. Steck (FSCA) questioned the name
Neurosystasis (he identified them as belonging to
Telmatoscopus) and suggested contacting Larry W.
Quate (Poway, California), a specialist in the fam-
ily. Quate requested adult specimens reared from
field-collected larvae in order to make a precise
identification and, if necessary, prepare a formal
description. Thereupon, JHF (with help from M.
M. Cutwa and G. F. O'Meara, Florida Medical En-
tomology Laboratory, Vero Beach) obtained larvae
from bromeliads in southeastern Florida and pro-
vided them to G. J. Steck who from them reared a
few adults and shipped them to Quate in 1999-
2000. Quate reported that they represent the first
Nearctic record for a member of the genusAlepia,
and he was pleased to see the associated larvae.
Tragically, Larry Quate died in January 2002. It
should be fairly easy to obtain more larval speci-
mens and rear more adults to replace those unre-
turned by his estate.

Hymenoptera (ants identified by M. A. Deyrup)

Formicidae: Camponotus floridanus (Buckley)
(2), Camponotus planatus Roger (26), Cremato-
gaster ashmeadi Mayr (64), Paratrechina longi-
cornis (Latreille) (50), Pheidole megacephala
(Fabricius) (5), Pheidole moerens Wheeler (129).

June 2004

Frank et al.: Invertebrates from Florida Tillandsia Bromeliads

Ichneumonidae: genus and species unidenti-
fied (2) (det. L. A. Stange, FSCA).
Aphelinidae: Aphelinus sp. (1) (see under
Hemiptera and Homoptera).
Ant nests with brood were detected in T utric-
ulata (C. planatus and P. longicornis), T fascicu-
lata (P. moerens), and T setacea (C. ashmeadi).
The other ant specimens doubtless were foraging
from nests elsewhere. It has long been known
that ants will nest in the dry, outer leaf axils of
bromeliads such as T fasciculata and T utricu-
lata that hold water in their inner axils. One
plant of T utriculata in Sarasota provided space
for nests of two species: C. planatus and P longi-
cornis. Paratrechina longicornis, P megacephala,
P. moerens, and C. planatus are adventive species.
Ants were identified from Tillandsia spp. in
various Neotropical countries and Florida by
Wheeler (1942). However, the Tillandsia were not
identified to species level, nor were the localities
in Florida nor dates of collection specified.
Table 2 arranges the collection data by sample
number, with invertebrates identified to the level
of family. This arrangement was designed to allow
extraction of numerical data for statistical analy-
sis. However, the Table suggested few patterns
that would yield useful analysis. To further com-
plicate the table by including species names
would have been unwieldy.
A simple analysis was made by contrasting the
content of the three smallest with three largest
plants within each species (Tables 1 and 2), a
valid statistical method. For T fasciculata the
smallest plants were nos. 17, 18, and 19 (with 17,
7, and 144 invertebrates). The three largest were
8, 9, and 20 (with 19, 18, and 36 invertebrates).
The presence of an ant nest in plant 19, with 128
adult ants was the cause of the high count in a
small plant. Even if all data for ants were omitted,
the evidence for relationship of plant size to num-
ber of invertebrates would have been negligible.
For T utriculata, the three smallest plants
were 1, 3, and 4 (with 2, 16, and 10 invertebrates),
and the three largest were 2, 16, and 24 (with 39,
226, and 135 invertebrates). In plant 24, ants ac-
counted for 76 of the invertebrates. Whether or
not we exclude data for ants, the largest plants
clearly have more invertebrates, and these were
mainly aquatic dipteran larvae (Ceratopogonidae,
Culicidae, and Psychodidae; except in plant 24). If
we exclude ants and aquatic insect larvae, the
three smallest plants had 1, 14, and 9 inverte-
brates whereas the three largest had 17, 28, and
28; again there is a relationship between plant
size and number of invertebrates, but it is shal-
lower than when including the aquatics. If we in-
clude only the aquatic invertebrates, then the 3
smallest plants had 0, 2, and 1 invertebrates,
whereas the three largest had 17, 191, and 31; the
larger plants clearly had many more, but variance
is huge. We might expect that the number of

aquatic dipteran larvae would best be associated
with volumetric capacity ofbromeliad axils (calcu-
lated from length of longest leaf). But intraplant
variance in numbers of contained invertebrates
warns us that the fitting of regressions will suffer
from high sums-of-squares errors. The presence of
ant nests adds greatly to variance.
For the three T recurvata plants sampled, the
number of invertebrates was not related to plant
size. For T setacea, the three smallest plants were
5, 6, and 13 (with 11, 19, and 18 invertebrates)
and three largest were 11, 12, and 14 (with 6, 18,
and 4 invertebrates); there was no relationship of
plant size to number of invertebrates.


The total number of invertebrates in leaf axils
of T utriculata was related to plant size, but the
number of aquatic insect larvae increased more
strongly with plant size. The numbers of inverte-
brates were not or not clearly related to plant size
in the other three bromeliad species, although
such a relationship is something that would be
expected given a very large number of samples
(because larger plants provide more habitat).
Data in Tables 1 and 2 could be the materials for
hundreds of regression analyses, should anyone
wish to do these.
This study scratches the surface of Florida's
bromeliad fauna. It reaffirms that larvae of sev-
eral aquatic Diptera (Psychodidae, Culicidae,
Ceratopogonidae, Chironomidae, Muscidae, and
Aulacigastridae), perhaps one species of scale in-
sect (Ortheziidae), and perhaps one or more spe-
cies of Lepidoptera (Tineidae and/or Gelechiidae)
have an obligate relationship with bromeliads.
The null hypothesis for all the remaining species
is that they "just happened to be there" and may
additionally be found in tree canopies or in leaf
litter on the ground. This null hypothesis cannot
now be tested for lack of studies of the canopy
fauna or the leaf litter fauna in Myakka River
State Park.
This in no way discounts the importance of
bromeliads as habitat for large numbers of inver-
tebrate species: how many other small plants
have such a diversity of invertebrates? At least 70
families with 82 genera and 90 species are repre-
sented in the few (24) samples. Further sampling
should yield very many more species (and genera
and families) at least in Coleoptera, and perhaps
some other orders, including species that just
happen to be represented in the bromeliads at the
time of sampling.
If sampling is to be repeated, this should be (a)
with very many more samples to allow more rep-
lication and thus a more useful comparison be-
tween the faunas of the four bromeliad species,
(b) with prior agreement (probably involving
funded written subcontracts for expenditure of

Florida Entomologist 87(2)

time) from numerous specialist taxonomists to
devote time to the project, (c) with the collector
charged with rearing representatives of all the
immature arthropods to the adult stage. The ad-
vantage of having more samples will be the avail-
ability of a series of adult specimens of every
species represented, except perhaps a few of the
transients. The advantage of rearing the imma-
ture arthropods will be that adult specimens will
be available for identification, and identifiers will
then have immature specimens reliably associ-
ated with the adults; thereafter, the specialists
may be able to provide identification keys to the
immature stages. The collector should be profi-
cient in invertebrate classification, and should
have the time to rear immature arthropods to the
adult stage.
Raw data used to compile Table 2, on inverte-
brates associated with the 24 plants, will be of-
fered to the "Bromeliad tank dwellers database"
on the website of the Florida Council of Bromeliad
Societies (http://www.fcbs.org). It records any an-
imal species detected in or on a bromeliad, not
just tank dwellers (the aquatic species in tanks).
This could lead to detection of other animals fre-
quently associated with bromeliads, even if it
takes tens of thousands of records. It was not easy
to obtain identifications to the species level of in-
vertebrates collected from bromeliads in Florida,
and we were only partially successful, and only
for some groups. We warn investigators who
would like to conduct similar studies in the Neo-
tropics that they will encounter severe taxonomic
problems. The effort to collect the specimens is
small compared with the effort required to iden-
tify the specimens reliably to the species level.
Identification not made to the species level is
worth rather little. Taxonomists need to be con-
vinced that the project is worth their support. In
this project, some taxonomists obtained useful
and interesting specimens, at least ofXenylla (an
undescribed species), Cerobasis guestfalica, Cam-
plomyza sp., Alepia (the first Nearctic record),
and various mites of uncertain identity.
The sampling method did not collect micro-
scopic aquatic organisms. For these, it would be
better to use a siphon or large syringe (such as an
"oven baster") to extract the water from leaf axils
of T fasciculata and T utriculata, and to decant
this water directly into Petri dishes for micro-
scopic examination. Such a method should collect
bacteria, Fungi Imperfecti, algae, rotifers, nema-
todes, platyhelminthes, annelids, ostracods and
copepods. But identification of these would have
been beyond the skills of the taxonomists in-
volved in the present study.
Future projects of this nature in Florida with
all four of these plant species are unlikely in the
near future. This is because the weevil Metama-
sius callizona was detected in Myakka River
State Park in September 1999 and, since then,

has been relentlessly destroying the park's popu-
lations of T utriculata and T fasciculata (T. M.
Cooper in Larson 2000). Similar destruction has
been detected in almost all southern Florida
counties. These two bromeliad species are right-
fully listed in the Florida Administrative Code as
endangered species. Recovery of their populations
is unlikely unless the weevil can be brought un-
der biological control.


We thank M. M. Cutwa and G. F. O'Meara (Florida
Medical Entomology Laboratory, Vero Beach) for collect-
ing psychodid larvae from bromeliad leaf axils, G. R.
Buckingham (USDA-ARS, Gainesville) for loan of a leaf-
area meter, G. A. Evans (FDACS-DPI, Gainesville) for
identification of an aphelinid specimen, L. A. Stange
(FDACS-DPI, Gainesville) for family-level identifica-
tion of two wasp specimens, D. H. Habeck (Entomology
and Nematology Dept., University of Florida, retired)
for identification of lepidopteran larvae, R. J. Gagn6
(Systematic Entomology Laboratory, USDA-ARS) for
identification of cecidomyiids, R. J. Snider (Michigan
State University) for identification of Collembola, E. E.
Baskerville and C. Gambetta (MSBG) and J. Y. Miller
(Allyn Museum of Entomology, Sarasota) for transport,
E. E. Baskerville, B. K. Holst and H. E. Luther (MSBG)
for plant identification, H. E. Luther and J. Y. Miller for
access to literature, H. E. Luther for permission to cite
his observation on snails eating bromeliad trichome
caps, R. E. Rivero (MSBG) for lending a dissecting mi-
croscope, M. A. Blanco, E. Kr6tz and D. Mondrag6n
(MSBG) and staff of MRSP for help, B. C. Larson and
R. D. Cave (UF-IFAS) for critical reviews of the manu-
script, and R. D. Cave for preparation of a Spanish ab-
stract. This is Florida Agricultural Experiment Station
journal series R-09639.


BEUTELSPACHER B., C. R. 1971a. Una bromeliacea como
ecosistema. Biologia (Mexico) 2(7): 82-87.
BEUTELSPACHER B., C. R. 1971b. Fauna de Tillandsia
caput-medusae E. Morren 1880 (Bromeliaceae).
Anales Inst. Biol., Univ. Nac. Aut6n., Mexico (Ser.
Zool.) 43: 25-30.
EPLER, J. H. 2001. Identification manual for the larval
Chironomidae (Diptera) of North and South Caro-
lina. A guide to the taxonomy of the midges of the
southeastern United States, including Florida. Spec.
Publ. SJ 2001-SP 13. North Carolina Dept. Envir.
Nat. Resources, Raleigh, NC and St. Johns Water
Management District, FL, 526 pp.
FISH, D. 1976. Structure and composition of the aquatic
invertebrate community inhabiting epiphytic bro-
meliads in south Florida and the discovery of an in-
sectivorous bromeliad. Ph.D. dissertation, Univ.
Florida, Gainesville, ix + 78 pp.
FRANK, J. H. 1983. Bromeliad phytotelmata and their
biota, especially mosquitoes, pp. 101-128. In J. H.
Frank and L. P. Lounibos. Phytotelmata: Terrestrial
plants as hosts for aquatic insect communities.
Plexus; Medford, NJ, vii + 293 pp.
FRANK, J. H. 1996a. Bromeliad biota. (http://Bromeliad-
Biota.ifas.ufl.edu/) (September 2002).

June 2004

Frank et al.: Invertebrates from Florida Tillandsia Bromeliads

FRANK, J. H. 1996b. Bibliography of bromeliad phytotel-
mata in Bromeliad Biota (http://BromeliadBiota.
ifas.ufl.edu/fitbibl.htm) (September 2002).
FRANK, J. H. 1999. Bromeliad-eating weevils. Selbyana
20: 40-48.
FRANK, J. H., AND G. A. CURTIS. 1981. Bionomics of the
bromeliad-inhabiting mosquito Wyeomyia vanduzeei
and its nursery plant Tillandsia utriculata. Florida
Entomol. 64: 491-506.
FRANK, J. H., G. A. CURTIS, AND H. T. EVANS. 1976. On
the bionomics of bromeliad-inhabiting mosquitoes. I.
Some factors influencing oviposition by Wyeomyia
vanduzeei. Mosquito News 36: 25-30.
FRANK, J. H., AND M. C. THOMAS. 1996. Weevil pests of
bromeliads in Pests ofbromeliads in Bromeliad Biota
(http://BromeliadBiota.ifas.ufl.edu/wvbrom.htm) (Sep-
tember 2002).
HEPPNER, J. B., AND J. H. FRANK. 1998. Bromeliad pod
borer. Univ. Florida, Entomol. Nematol. Dept., Fea-
tured Creatures, EENY-40 (http://Creatures.ifas.ufl.
edu/orn/bromeliad_pod_borer.htm) (September 2002).
LARSON, B. C. 2000. Save Florida's native bromeliads.
Published on WWW at (http://SaveBromeliads.ifas.
ufl.edu/) (September 2002).
LARSON, B. C., AND J. H. FRANK. 2000. Mexican brome-
liad weevil. Univ. Florida, Entomol. Nematol. Dept.,

Featured Creatures, EENY-161 (http://Creatures.
ifas.ufl.edu/orn/m_callizona.htm) (September 2002).
LARSON, B. C., J. H. FRANK, AND O. R. CREEL. 2001. Flor-
ida bromeliad weevil. Univ. Florida, Entomol. Nema-
tol. Dept., Featured Creatures, EENY-209. (http//
Creatures.ifas.ufl.edu/orn/m_mosieri.htm) (September
PALACIOS-VARGAS, J. G. 1981. Collembola asociados a
Tillandsia (Bromeliaceae) en el Derrame Lavico del
Chichinautzin, Morelos, Mexico. Southw. Entomol.
6: 87-98.
PALACIOS-VARGAS, J. G. 1982. Microartr6podos asocia-
dos a bromeliaceas. Actas VIII Congreso Latino
Americano Zool. 1: 535-545.
PICADO, C. 1913. Les bromliac6es 6piphytes consid-
er6es comme milieu biologique. Bull. Sci. France Bel-
gique 47: 215-360.
SIDOTI, B. J. 2000. Faunal inhabitants of a Florida bro-
meliad. J. Bromeliad Soc. 50: 227-233.
WHEELER, W. M. 1942. The epiphytic Bromeliaceae and
their fauna, p. 133-143. In Studies of Neotropical ant
plants and their ants. Bull. Mus. Comp. Zool. Har-
vard 90: 1-262, pl. 1-56.
WUNDERLIN, R. P. 1998. Bromeliaceae, pp. 192-195. In
R. P. Wunderlin, Guide to the Vascular Plants of
Florida. Univ. Press of Florida, Gainesville, 806 pp.

Florida Entomologist 87(2)


1USDA-ARS CMAVE, 1700 SW 23'd Drive, Gainesville, FL 32608

2Texas A&M University-Kingsville Citrus Center, P.O. Box 1150, Weslaco, TX 78599-1150

Two parasitoid species,Amitus hesperidum Silvestri and Encarsia opulenta (Silvestri), were
released in an augmentative program to control citrus blackfly, Aleurocanthus woglumi
Ashby, in the citrus growing areas of southern Texas. Releases were made with laboratory-
reared and field insectary parasitoids. Six citrus groves were closely monitored, and evalu-
ations made during and after releases suggested that both parasitoid species became rees-
tablished and exerted control over pest populations. Dissection of citrus blackfly immatures
led us to suggest that E. opulenta increased in larger numbers thanA. hesperidum, and that
a stable host-natural enemy relationship became established.

Key Words: Aleurocanthus woglumi, Amitus hesperidum, Encarsia opulenta, augmentative
biological control


Dos species de parasitoides, Amitus hesperidum Silvestri y Encarsia opulenta (Silvestri)
fueron liberadas en un program de aument6 para el control de la mosca prieta de los citri-
cos. Aleurocanthus woglumi Ashby, en areas donde siembran los citricos en el sur de Texas.
Las liberaciones fueron hechas usando parasitoides criados en el laboratorio y los del insec-
tario del campo. Un monitoreo precise de seis huertos de citricos fue hecho, y las evaluacio-
nes hechas durante y despu6s de la liberaci6n que sugerieron que ambas species de
parasitoides se re-establecieron y ejercieron un control sobre la poblaci6n de la plaga. La di-
secci6n de inmaduras de la mosca prieta de los citricos tambien sugeri6 que el E. opulenta
se aumento en numeros mas altos que elA. hesperidum, y que una relaci6n stable entire el
hospedero y el enemigo natural fue establecida.

The citrus blackfly, Aleurocanthus woglumi
Ashby, first invaded the Lower Rio Grande Valley
of Texas in 1955 on residential citrus (Smith et al.
1964), and again near Brownsville in 1971 in both
residential citrus and commercial groves (Hart et
al. 1973). An augmentative biological control pro-
gram to establish parasitoids was initiated in
1974 with release of three species,Amitus hesperi-
dum Silvestri (Hymenoptera: Platygasteridae),
Encarsia (= Prospaltella) opulenta (Silvestri), and
E. clypealis (Silvestri) (Hymenoptera: Aphelin-
idae). These releases were made from laboratory-
reared and field-collected cultures in Mexico
(Hart 1978). Evaluations undertaken from 1977-
1982 indicated a widespread distribution ofE. op-
ulenta, but fewA. hesperidum, and no E. clypealis,
suggesting local competitive displacement by E.
opulenta in groves with effective parasitoid regu-
lation (Summy et al. 1983).
Citrus blackfly population densities remained
stable under excellent biological control until the
mid 1980s (Summy et al. 1983). Following a severe
freeze in December 1983, citrus blackfly densities
surged while concomitant parasitoid densities ap-
parently remained low. Citrus blackfly popula-

tions reached damaging levels during the 1988
and 1989 seasons, especially in central valley
groves (French et al. 1990). A biological control
program of parasitoid augmentation was initiated
in June 1989, and results suggested reestablish-
ment of E. opulenta and A. hesperidum (Meagher
et al. 1991). However, another severe freeze in De-
cember 1989 halted evaluation efforts.
Although most commercial groves had no cit-
rus blackfly populations after the freeze (unpub-
lished data), populations were discovered in
citrus nurseries and on residential citrus trees in
early 1990 (French & Meagher 1992). Newly
planted and commercial groves located near resi-
dential areas soon were infested with dense,
largely unparasitized citrus blackfly populations
(unpublished data). An augmentation program
was initiated in 1992 to increase biological control
efficacy in commercial citrus groves. This was ac-
complished by releasing parasitoids into residen-
tial citrus so that "field insectaries" could be
developed. Commercial groves were then sampled
with yellow sticky traps (Harlan et al. 1979;
Summy et al. 1986) and leaf observation, so that
groves containing large citrus blackfly densities

June 2004

Meagher & French: Citrus Blackfly Biological Control

could be identified as release sites. The final step
was to transfer laboratory- and field insectary-
produced parasitoids into infested groves. This re-
port describes the seasonal progression of the
host and its natural enemies in commercial citrus
groves in southern Texas.


Groves Sampled

Grapefruit and orange groves selected for this
study were located throughout the Lower Rio
Grande Valley in both Cameron and Hidalgo
counties. They included groves near Bayview
(10 ha., 'Rio Red' grapefruit, sampled 21 July
1992-21 March 1995; 7.2 ha., 'Marrs' and 'Valen-
cia' orange, sampled 23 November 1993-21 March
1995), Donna (16.2 ha., 'Ruby Red' and 'Star
Ruby' grapefruit, 27 July 1992-22 February
1994), Mercedes (16.2 ha., 'Ruby Red' grapefruit
and'Valencia' orange, 7 December 1993-21 March
1995), Edinburg (8.3 ha., 'Ruby Red' grapefruit,
22 July 1992-12 April 1994), Mission-grapefruit
(12.1 ha., 'Rio Red' grapefruit, 15 July 1992-22
March 1994), and Mission-orange (3.4 ha.,'Marrs'
orange, 15 July 1992-22 March 1994).

Parasitoid Releases

Augmentation of both A. hesperidum and
E. opulenta was accomplished with laboratory-
produced and field-collected specimens from Flor-
ida (Florida Department of Agriculture, Division
of Plant Industry, Gainesville) and field insec-
tary-produced parasitoids from Texas. Both the
cup and paper bag methods of French et al. (1990)
and Meagher et al. (1991) were used to disperse
parasitoids. From January 1992-February 1993,
over 92,000A. hesperidum and 18,000 E. opulenta
were released throughout the citrus growing re-
gion, and selected releases of both species of par-
asitoids were made later during 1993.

Citrus Blackfly Sampling

Citrus blackfly infestation and parasitization
levels were sampled by two techniques. First, the
percentage of leaves infested with citrus blackfly
was determined by examining four branches of
eight trees. The total number of leaves and num-
ber with live citrus blackfly immature stages
were recorded. Selection of each branch was
based on directional orientation (quadrants:
southeast, southwest, northwest, northeast). Five
of the trees were 'station' trees that were sampled
each time; an additional three trees were selected
randomly each sample date. This sampling tech-
nique was not conducted in the Bayview-orange
grove. Analysis of variance (PROC GLM, LSD
mean separation test, SAS Institute 1995) was

used to examine variation among trees or among
quadrants. Parasitization was calculated by dis-
secting and examining a subsample of at least
100 fourth stage nymphs ('pupae').
Beginning in late 1993, an additional sampling
technique was developed to provide a closer ex-
amination of citrus blackfly parasitization. These
samples were taken in the grapefruit and orange
sections of the Bayview grove and in the Mer-
cedes grove. From each tree, one hundred leaves
that contained citrus blackfly pupae were col-
lected. From this collection, ten leaves containing
pupae were selected and returned to the labora-
tory for dissection. Each pupa was categorized as
live, dead, or emerged citrus blackfly; or live,
dead, or emerged parasitoid. Dead citrus blackfly
and parasitoids were represented by desiccated
remains. Emerged individuals were represented
by either the characteristic pupal exuvia split by
citrus blackfly or circular exit holes created by
parasitoids. Parasitization by species was not
identified, although exit hole numbers per pupa
at times provided species information. Generally,
two exit holes indicated the presence of A. hes-
peridum, although rarely an individual larva of
both species was found live in one host pupa. Par-
asitoid adults searching on leaves were noted
during sampling.


Citrus blackfly-infested leaves exhibited sig-
nificant inter- and intra-tree variation when each
grove was analyzed individually (P < 0.0001; P <
0.003, respectively). The northwest quadrant of
the tree always contained more infested leaves
than the southeast quadrant (Table 1). These re-
sults are in agreement with previous studies in
Florida and Texas which suggested high inter-
tree variation (Dowell & Cherry 1981), although
Gilstrap et al. (1980) found more citrus blackfly in
the northwestern quadrant of the tree. Although
our sampling described clear quadrant differ-
ences in population level, we agree with other re-
searchers that collection of leaves for population
sampling or parasitoid efficacy should be from all
tree areas, especially when pest densities are low
(Cherry & Fitzpatrick 1979; Gilstrap et al. 1980).
The Donna, Edinburg and two Mission grove
locations selected for monitoring in 1992 had ini-
tial citrus blackfly populations below 10% in-
fested leaves (Fig. 1). Leaf samples from these
groves showed maximum citrus blackfly infesta-
tions ranging from 44.5 to 82.1%, but leaf samples
taken at the end of sampling showed little to no
citrus blackfly infestation (Table 2). Parasitiza-
tion from both species ranged from 62.1-100% in
these groves, but by the end of sampling in 1994
was much reduced due to the scarcity of hosts.
Population dynamics of host and parasitiza-
tion in the Bayview grove showed low numbers of

Florida Entomologist 87(2)

June 2004


Tree quadrant

Grove df F Northwest Northeast Southwest Southeast

Bayview 3, 1271 7.2 18.2 1.5 a 15.3 1.4 b 16.0 1.4 ab 12.2 1.3 c
Donna 3, 775 30.1 19.9 1.7 a 17.9 1.7 a 13.3 1.3 b 8.4 1.1 c
Mercedes 3, 491 5.6 81.1 2.2 a 80.7 2.2 a 77.9 2.6 a 73.0 2.8 b
Edinburg 3, 759 21.0 23.1 1.9 a 20.2 1.8 b 19.9 1.7 b 12.8 1.4 c
Mission-grapefruit 3, 775 4.9 32.7 2.5 a 31.8 2.5 a 31.1+ 2.5 a 28.8 2.4 ab
Mission-orange 3, 806 3.1 19.2 1.7 a 17.9 1.6 ab 15.7 1.6 b 15.8 1.6 b

Means ( SE) within the same row followed by the same letter are not significantly different, LSD (P > 0.05).

citrus blackfly initially when sampled in summer
1992 (Fig. 2). Citrus blackfly populations rose to
over 60% leaves infested by late fall 1993, until




c 40

a 30



0 (N
a S a


increasing parasitization appeared to reduce host
populations. A closer examination of grapefruit
trees showed proportionally high levels of para-


90 Edinburg I
80 \
I CBF-infested leaves I
70 t 9
S -0- Parasitization
50 I
40 I
30 -

20 I p I

L- to C -
(N N 0

o CO -


C Li -4

- U, cc-


's o ; 0t s a0 0' ' 0 0 0 0 0 '
M .0
08P ~ C ~f

Fig. 1. Population densities of citrus blackfly (percent leaves infested) and parasitization (percent fourth stage
parasitized) due to Amitus hesperidum and Encarsia opulenta, in three Lower Rio Grande Valley, Texas citrus
groves, 1992-1994.

S -A- CBF-infested leaves
I0- PaIsitization
SI "- Parasitizatioo

..-pppg1-i -p-

- Mission Grapefruit o

|I -t CBFinfted leaves
/ II -- P....,it.n .on


-a 96 II~ k,,~

a a
g -s
'' u-

Meagher & French: Citrus Blackfly Biological Control

TEXAS, 1992-1995.

Highest Highest Final Final
Grove infested leaves (%) parasitization (%) infested leaves (%) parasitization (%)

Bayview 65.5 5.9 79.3 2.8 0.5 48.7
Donna 44.5 5.4 62.1 0.03 0.03 0
Mercedes 100.0 0 92.9 68.1+ 3.1 59.2
Edinburg 55.7 4.3 100.0 0 0
Mission-grapefruit 82.1 2.5 88.0 0 0
Mission-orange 56.8 5.2 79.0 0 0

sitization occurred during late winter 1993 and
spring 1994, with over 50 immature parasitoids
per leaf found in the 15 February, 29 March, and
19 April samples, and 65 per leaf in the 20 June
sample (Fig. 3 a, b). Adults from many of the par-
asitized pupae during this period had already
emerged. Adult A. hesperidum was the predomi-
nate species observed on leaves during early sam-
pling, and although E. opulenta adults were
present, their numbers did not increase until fall
1994. The highest citrus blackfly population den-
sity also occurred during spring and summer
1994, with a peak of 109.7 30 live immatures
found in the 10 May sample (Fig. 3a). The grower,
without our recommendation, applied an un-
known insecticide in February and May, resulting
in mortality of both citrus blackfly and parasi-
toids (20.7 6.8 dead parasitoid immatures per
leaf, 20 June sample) (Fig. 3b). Populations of cit-
rus blackfly, A. hesperidum, and E. opulenta all
declined after the 20 June sample. Samples taken
in early 1995 showed low populations levels of the
host, but active populations of both parasitoids
Overall population density of citrus blackfly
was lower in the Bayview oranges, peaking at
26.7 8.4 live immatures per leaf in late 1994
(Fig. 3c). As in the grapefruit, high mortality oc-
curred during June through August as a result of
an insecticide application. However, both citrus
blackfly and parasitoid activity increased in the
late fall. Over 10 live or emerged parasitoids per
leaf were found by the end of the study, including
both A. hesperidum, and E. opulenta (Fig. 3d).
Sampling in the Mercedes grove showed initial
citrus blackfly populations already close to 40%
infested leaves (Fig. 2). This grove was the only
one that had high levels of citrus blackfly at the
conclusion of sampling, although parasitization
was also high (Fig. 2). Intensive sampling in late
1993 through early 1994 showed medium levels of
live and dead citrus blackfly, with low levels of
parasitization (Fig. 4a). By spring, citrus blackfly
populations increased to >100 live pupae per leaf
in May. Populations remained high through sum-
mer and early fall, peaking with an average of

120.3 live and 3.8 emerged citrus blackfly pupae
per leaf in October 1994. However, parasitoid ac-
tivity was increasing by August, as exemplified by
observation of searching adult E. opulent on
leaves. Parasitization increased through fall and
into 1995, with >200 live or emerged parasitoid
immatures found in the 1 November sample (Fig.
4b). Citrus blackfly populations decreased after
the 22 November sample, and from the 19 Decem-
ber sample to the conclusion of the study, over 70
live or emerged immature parasitoids (predomi-
nately E. opulenta) per leaf were documented.


Release of these and other exotic parasitoid
species against citrus blackfly in Mexico during
the early 1950s formed the resource for future re-
leases in the Western Hemisphere. By the end of
1953, over 300 million adults (242 million of A.
hesperidum alone) were dispersed in Mexico
(Flanders 1969). The discovery of the pest in Ft.
Lauderdale, FL residential citrus during Janu-
ary, 1976 led to the release of A. hesperidum,
E. opulenta, and E. clypealis from laboratory cul-
tures in General Teran, Mexico (Hart et al. 1978).
Since then, over 250,000 parasitoids from labora-
tory colonies, field collections, and movement of
infested and parasitized citrus leaves were re-
leased in Florida from October 1979 through May
1980 (Nguyen et al. 1983). Laboratory cultures
and field collections from Florida formed the basis
for the material that was augmented into Texas
Our results suggest the successful establish-
ment of A. hesperidum and E. opulenta in the
Texas citrus agroecosystem following severe
freezes, and the reduction of citrus blackfly popu-
lations. Insecticide applications, especially within
the grapefruit trees at Bayview, limited our abil-
ity to follow natural enemy interactions. How-
ever, postbloom and summer selective pest
management chemical sprays have been shown to
have only short term influence on citrus blackfly
natural enemy populations (Fitzpatrick et al.
1978, 1979).

Florida Entomologist 87(2)

June 2004


nf m n ON 00
e. rDa ti


A I a

- Mercedes

i \


0 \
i \
I\ I
I \ I '
I l \ I I


I I '

I \ I
Q I \ I
\ /

~ I I I -- I I
i i 5 i i a i


ON ON = S t '. -
-1 -

SCBF-infested leaves

-- Parasitization

CI C, C" 0>1^1 C ") ( a
> > Q C

Z Zo
0 0 0

N O ON 00 -
(N N r^


Fig. 2. Population densities of citrus blackfly (percent leaves infested) and parasitization (percent fourth stage
parasitized) due toAmitus hesperidum and Encarsia opulenta, in two Lower Rio Grande Valley, Texas citrus groves,

. . . . . . ..I I I I I




Meagher & French: Citrus Blackfly Biological Control

5 0 CBF-live
E3 Parasitized


O CBF-ive
I CBF-dead




. 40
" 30




S 8

S 6



S 8 0g g Date' '

71f l
N -7n O. -
Dat e ad


Fig. 3. Population densities of citrus blackfly and two parasitoid species in a grapefruit (a, b) and orange (c, d)
grove, Bayview, Texas. Bars correspond to live, dead, or parasitized citrus blackfly per leaf (a, c), or live, dead, or
emerged parasitoids per leaf (b, d).

In the Mercedes grove, results suggested in-
creasing populations of E. opulenta and popula-
tion suppression of citrus blackfly. Several reports
have documented population regulation by this
parasitoid, even within groves under pest man-
agement chemical applications (Cherry & Pastor
1980; Swezey & Cano Vasquez 1991). Encarsia
opulenta has been shown to be able to competi-
tively displace populations of other Encarsia spe-
cies and A. hesperidum because of its ability to
maintain a stable interaction with its host under
low host populations due to density-dependent
searching of adult parasitoids (Summy et al.
1983, 1985). In a laboratory study, E. opulenta fe-
males showed preferences for citrus blackfly that
were previously parasitized by A. hesperidum
(Dowell et al. 1981), although A. hesperidum lar-
vae can avoid predation by E. opulenta larvae by
"hiding" in the midgut (Flanders 1969).

This report suggests that citrus blackfly popu-
lations were reduced in groves selected for parasi-
toid augmentation. Parasitoid populations in
these groves increased temporally either due to
our reestablishment program or due to the in-
crease of naturally-occurring populations that
survived the freeze in residential and commercial
citrus trees. Since we did not determine if parasit-
ism was attributed to naturally-occurring or re-
leased individuals, the role of our augmentation
program on parasitoid reestablishment cannot be
identified explicitly. Only carefully planned exper-
iments comparing parasitoid populations in "con-
trol" and "treated" groves with similar residential
and commercial citrus habitats will provide this in-
formation. This type of experimentation has not
been accomplished on a large scale in studies in-
volving citrus blackfly biological control because
of grower, citrus industry, and logistical demands.


. 15



192 Florida Entomologist 87(2) June 2004


225 CBF-live

200 CBF-dead

175 -
S 15 Parasitized

d 125

00 100 7

S 75
1 75 / De // d

25 jl

75 -c.'^
50 /


200 --- Live ^

175 Dead ,

S 150 l a Edmerged "


n 100 7 C

04 150- Z 1mre


b. 00|C|C, C)

Fig. 4. Population densities of citrus blackfly and two parasitoid species, in a citrus grove, Mercedes, Texas. Bars
correspond to live, dead, or parasitized citrus blackfly per leaf (a), or live, dead, or emerged parasitoids per leaf (b).

Meagher & French: Citrus Blackfly Biological Control


A debt of gratitude is owed to the Florida Depart-
ment of Agriculture, Division of Plant Industry (Richard
Gaskella, Director) and its entomologist Ru Nguyen,
and to Francisco Gomez Garcia, Montemorelos, N.L.,
and Jorge Alfonso Gongora Rodriguez, Director SARH,
Monterrey, Mexico, for assistance in supplying parasi-
toids. John Worley, USDA-APHIS, Daniel S. Moreno,
USDA-ARS, the Texas Agricultural Extension Service,
and Texas Department of Agriculture were helpful dur-
ing the early stages of parasitoid release. Nancy Epsky
(USDA-ARS) and John Sivinski (USDA-ARS) kindly re-
viewed an earlier version of the manuscript. Elias Her-
nandez, Jr., Robert Saldana, Hilario Perez, and
Santiago Villarreal are thanked for technical assis-
tance. This research was in part supported by Texas Cit-
rus Mutual (Ray Prewett, Executive Director, William
Watson, Assistant to the Executive Vice-President.


CHERRY, R., AND G. FITZPATRICK. 1979. Intra-tree disper-
sion of citrus blackfly. Environ. Entomol. 8: 997-999.
CHERRY, R., AND S. PASTOR, JR. 1980. Variations in pop-
ulation levels of citrus blackfly, Aleurocanthus
woglumi (Hom.: Aleyrodidae) and parasites during
an eradication program in Florida. Entomophaga 25:
DOWELL, R. V., AND R. H. CHERRY. 1981. Detection of,
and sampling procedures for, the citrus blackfly in
urban southern Florida. Res. Popul. Ecol. 23: 19-26.
Searching and ovipositional behavior ofAmitus hesperi-
dum (Hym.: Platygasteridae) and Encarsia opulenta
(Hym.: Aphelinidae) parasitoids of the citrus blackfly
(Hom.: Aleyrodidae). Entomophaga 26:233-239.
1978. Short-term effects of three insecticides on
predators and parasites of the citrus blackfly. Envi-
ron. Entomol. 7: 553-555.
1979. Effects of Florida citrus pest control practices on
the citrus blackfly (Homoptera: Aleyrodidae) and its as-
sociated natural enemies. Can. Entomol. 111: 731-734.
FLANDERS, S. E. 1969. Herbert D. Smith's observations on
citrus blackfly parasites in India and Mexico and the
correlated circumstances. Can. Entomol. 101: 467-480.
FRENCH, J. V., AND R. L. MEAGHER, JR. 1992. Citrus
blackfly: chemical control on nursery citrus. Sub-
tropical Plant Sci. 45: 7-10.
1990. Release of two parasitoid species for biological

control of citrus blackfly in south Texas. J. Rio
Grande Valley Hort. Soc. 43: 23-27.
Within-tree distribution of resident populations of cit-
rus blackfly in Texas. J. Econ. Entomol. 73: 474-476.
1979. A yellow coffee lid trap for the citrus blackfly,
Aleurocanthus woglumi Ashby Southwestern Ento-
mol. 4: 25-26.
HART, W. G. 1978. Some biological control successes in
the southern United States. Proc. Int. Soc. Citric. 3:
1973. Aerial photography with infrared color film as
a method of surveying for citrus blackfly. J. Econ. En-
tomol. 66: 190-194.
BALLERO, AND R. L. GARCIA. 1978. The introduction
and establishment of parasites of citrus blackfly,
Aleurocanthus woglumi Ashby, in Florida. Ento-
mophaga 23: 361-366.
1991. Monitoring and biological control of citrus
blackfly in South Texas. Subtrop. Plant Sci. 44: 19-24.
ulation density of the citrus .1 :i. kI. .. ........ ..
woglumi Ashby (Homoptera: Aleyrodidae), and its
parasites in urban Florida in 1979-1981. Environ.
Entomol. 12: 878-884.
SAS INSTITUTE. 1995. SAS/STAT guide for personal
computers, version 6.11 ed. SAS Institute, Cary, NC.
Biological control of the citrus blackfly in Mexico.
USDA Tech. Bull. 1311: 30 pp.
Aleurocanthus woglumi (Hom.: Aleyrodidae) and
Encarsia opulenta (Hym.: Encyrtidae): density-de-
pendent relationship between adult parasite aggre-
gation and mortality of the host. Entomophaga 30:
Correlation between flight trap response and foliar
densities of citrus blackfly, Aleurocanthus woglumi
(Homoptera: Aleyrodidae). Can. Entomol. 118: 81-83.
LERO, AND I. SAENZ. 1983. Biological control of citrus
blackfly (Homoptera: Aleyrodidae) in Texas. Envi-
ron. Entomol. 12: 782-786.
SWEZEY, S. L., AND E. CANO VASQUEZ. 1991. Biological
control of citrus blackfly (Homoptera: Aleyrodidae)
in Nicaragua. Environ. Entomol. 20: 1691-1698.

Florida Entomologist 87(2)

June 2004


Department Of Entomology & Nematology, University of Florida, Gainesville, FL 32611-0620


We reared newly hatched Phaon Crescent butterfly larvae to the adult stage on a completely
artificial diet. About 37% of first instars survived to the adult stage. Addition to the diet of
freeze-dried host plant leaves equal to 10% by weight of dry ingredients produced up to 66%
survival to the adult stage. Survival of larvae and production of adults on the artificial diet
without host plant leaves was increased to equal that of diet with host plant leaves by adding
5% glucose or 5% Beck's salt mix. Although the ovaries of females produced on host-free arti-
ficial diet appeared to be mature at emergence and contained mature-looking eggs, we never
obtained viable eggs from them. In contrast, females produced on the artificial diet containing
at least 10% by weight of freeze-dried host plant leaves laid viable eggs, and four successive
generations were reared on the artificial diet with 10% freeze-dried host plant leaves. Males
produced on the artificial diet without host plant tissue displayed abnormalities in the shape
of the testes and parts of the vas deferentia, compared to males reared on the diet with freeze-
dried host leaves or on living host plants. The role of host plant tissue in nutrition and repro-
duction of both male and female Phaon crescents remains to be determined.

Key Words: artificial diets, butterflies, Phaon crescent, Nymphalidae, insect-plant interac-
tion, Phyla nodiflora, ovary, testes


Nosotros criamos larvas de la mariposa cresciente Phaon reci6n eclosionadas hasta la esta-
dia adulta sobre una dieta artificial. Aproximadamente, 37% de las larvas de la primera es-
tadia sobrevivieron hasta la estadia adulta, pero los adults no se aparearon ni
reproducieron. La adici6n de hojas congeladas y secadas de la plant hospedera a la dieta
igual a 10% del peso seco de los ingredients produjieron una sobrevivencia de 66% hasta la
estadia adulta, y los adults se aparearon y pusieron huevos que eclosionaron. La sobrevi-
vencia de las larvae y la producci6n de adults sobre una dieta artificial sin hojas de la
plant hospedera fue aumentada para ser igual que la sobrevivencia con hojas de la plant
hospedera por aiadir 5% de glucosa o 5% de la mezcla de sal de Beck, pero los adults no se
apareararon a menos que la dieta tenia hojas de la hospedera. Los ovarios de las hembras
producidas sobre una dieta artificial sin la plant hospedera aparecieron ser maduras al
emergir y tenian huevos maduros, igual como los ovarios de las hembras criados sobre la
plant hospedera. Ningun abnormalidad en las estructuras reproductivas internal de las
hembras producidas sobre la dieta artificial fue detectada. Los machos producidos sobre la
dieta artificial sin el tejido de la plant tenian abnormalidad en las estructuras reproductivas
internal. Asi, ambos sexos criados sobre una dieta artificial evidentemente tienen algunas
abnormalidad funcionales para prevenir el apareamiento y el 6xito reproductive. El papel
del tejido de la plant hospedera en influenciar el comportamiento y la reproducci6n de los
crescientes Phaon es desconocido.

Many moths, beetles, crickets, grasshoppers,
and other insects, but only two or three butterfly
species, can be reared on artificial diets (Singh
1977). The cabbageworm butterfly (Pieris rapae
(L.)) (Webb & Shelton 1988) and the painted lady
butterfly (Vanessa cardui (L.)) have been reared
on artificial diets. Semiartificial diets that con-
tain host-plant material have been published for
rearing the Monarch butterfly. The need to study
and control pest insects probably has contributed
to the development of artificial diets for many in-
sects, but most butterflies are not pests on eco-
nomic crops and little effort has been devoted to
developing artificial rearing media for them. But-

terflies tend to be restricted to one or only a few
host plants as larvae, and possibly they are very
sensitive to the balance of nutrients and/or pres-
ence of specific feeding cues in their host plants. A
practical difficulty in working with butterflies is
that many are active only part of the year, and
their larval host plants are often seasonal.
The Phaon crescent, Phyciodes phaon (Ed-
wards), is a small butterfly present over much of
the southeastern United States. A number of fac-
tors make the Phaon crescent a suitable butterfly
for study, including year-round distribution of the
butterfly and its host plant in Florida. The adults
do not diapause, and they mate and lay eggs in

Genc & Nation: Artificial Diet for Phaon Crescent Butterfly

the laboratory. Females lay their eggs on the un-
derside of leaves of the host plant Phyla nodiflora
(L.) Greene in the family Verbenaceae (Minno &
Minno 1999; Emmel & Kenny 1997; Genc 2002).
In preliminary trials with several published diets
for rearing butterflies and moths, Genc (2002)
found that the only diet formulation that allowed
a few adults to be produced was the pinto bean
(PB) diet developed for certain moths (Guy et al.
1985). Survival on the PB diet was poor, however,
and adults produced did not mate. Our objectives
in this paper are to describe (1) diets that improve
survival of Phaon crescent larvae and adult pro-
duction, (2) diets that promote mating and repro-
duction of adults, and (3) female and male
internal reproductive systems of Phaon crescents
reared on completely artificial diet with those
reared on the host plant.


The Butterfly and Host Plant

A colony of Phyciodes phaon was started from
females collected on the University of Florida
campus. The host plant was collected from the
campus and vicinity, and maintained in contain-
ers and small outdoor plots. Leaves of the host
plant were frozen in liquid nitrogen, ground while
frozen in a mortar, and freeze-dried. The freeze-
dried host plant leaves were stored at -20C until
needed. Adults were allowed to lay eggs on the
leaves of the living host plant, and newly hatched
first instars were removed and placed on diets. A
breeding colony of the butterfly was maintained
in the laboratory on host plants, and adults were
provided with 10% honey in water.

Preparation of the Pinto Bean Diet from Components

The pinto bean (PB) diet was prepared from in-
dividual components purchased from BioServ
(One 8th Street, Frenchtown, NJ 08825, USA). We
mixed pinto bean meal (19 g), wheat germ (14 g),
torula yeast (8 g), casein (7 g), gelcarin (3 g), me-
thyl paraben (0.5 g), and sorbic acid (0.3 g) and
added the mixture to 182 ml cold water with stir-
ring by a mechanical mixer. The aqueous mixture
was heated slowly (requiring about 15 minutes)
on a hot plate to 70C with continuous stirring.
Formaldehyde (1 ml) was added and mixing was
continued for about 3 minutes without further
heating. Ascorbic acid (0.9 g) was added and mix-
ing was continued an additional 3 minutes with-
out heating, and finally tetracycline (0.01 g),
BioServ Vitamin Mix #F8095 for Lepidoptera (0.8
g), and propionic acid (0.3 ml) were added with
additional mixing for 2-3 minutes. The mixture
was poured into paper cups, allowed to cool and
gel at room temperature, and stored in a refriger-
ator until needed. When ground, freeze-dried

plant leaves were added to the pinto bean diet,
addition was made with the vitamin mixture to
minimize heat damage to the host plant material.

Diet Modifications

Diets were formulated by incorporating 1%
glucose, 5% glucose, 1% Beck's salt mixture, 5%
Beck's salt mixture, 1%, 5%, 10%, or 20% ground,
freeze dried host plant leaves into the PB diet. Di-
ets were tested by placing 25 newly hatched lar-
vae on each of 3 replicates of each diet. The
criteria for evaluating a diet were number of
adults reared and whether the adults mated and
females laid viable eggs.


The abdomen of adults was brushed with a
camel's hair brush dipped in 70% ethyl alcohol to
remove scales, and then opened ventrally from
the first to the terminal abdominal segment. The
terminology used by Dong et al. (1980) was used
to describe the internal reproductive structures.


For comparing the number of adults produced on
modifications of the PB diet, we used binary logistic
regression analyses (Harrell 2001; Hosmer & Leme-
show 2000). Chi Square tests were used to deter-
mine the statistical significance of the model
parameters and an overall Chi Square test assessed
the hypothesis of no overall treatment difference.
When a significant Chi Square value was obtained,
the means for adult production on each tested diet
were transformed from non-linear function to linear
function and least square estimates of the diet-spe-
cific probabilities, P, of survival to the adult stage
were obtained by inverting the log odds model.


Diet Modifications

Survival to the adult stage was statistically
higher on PB diet with 10% or 20% freeze-dried
host plant leaves than with only 1% or 5% leaves
(Table 1). Moreover, adults from the diets contain-
ing 10% and 20% leaves reproduced and enabled
us to maintain a colony, but adults produced with
1% or 5% leaf tissue in the PB diet did not repro-
duce. Addition of 5% glucose or 5% Beck's salt mix
to the PB diet produced adults in numbers statis-
tically equal to numbers of adults produced with
10% host plant leaves in the PB diet, but none of
the adults from glucose or salt modified diets re-
produced. Numbers of adults produced on diets
with 1% host leaves, 5% host leaves, 1% glucose,
1% Beck's salt mix, or the original PB formula
were not statistically different from each other.

Florida Entomologist 87(2)

June 2004


Mean ( SD) number of adults
PB diet + Amendment produced per replicate Percent adults

1% host plant leaves 8.0 0.0 a 32
5% host plant leaves 12.0 1.4 a 48
10% host plant leaves 16.5 0.7 b 66
20% host plant leaves 13.5 2.1 b 54
PB diet 8.5 0.7 a 34
1% glucose 10.0 1.4 a 40
5% glucose 17.5 2.1 b 70
1% Beck's salt mix 11.5 0.7 a 46
5% Beck's salt mix 16.5 0.7 b 66
Values in a column followed by the same letter are not significantly different from each other at a = 0.01 level.

Anatomy of Phaon Crescent Internal Reproductive

The structure of the internal reproductive sys-
tem of a 10-day-old female produced on the host
plant is shown in Figure 1A and the ovary of a
newly emerged female adult from the PB diet is
shown in Figure lB. Adult females produced on
both food sources appeared to have mature or
nearly mature eggs in the terminal follicles of
each ovariole at emergence, with four ovarioles in
each of two ovaries. Although a large amount of
fat body associated with the ovaries makes count-
ing individual egg chambers very difficult, one
newly emerged female was determined to have

approximately 49 egg chambers in each ovariole.
Not enough observations were made, however, to
determine an average for number of egg cham-
bers per ovary or eggs laid. The lateral oviducts
guide eggs to a common, medial oviduct leading to
the genital chamber (the bursa copulatrix).
Paired lateral accessory glands are each con-
nected to the median oviduct.
The structure of the internal reproduction or-
gans from a male produced on the host plant is
shown in Figure 2A, and those from a male pro-
duced on the PB diet is shown in Figure 2B. Fused
testes form one testicular body in males. The tes-
ticular body is round and dark reddish brown in
males produced on the host plant and on PB diet

Fig. 1. Internal reproductive structure of Phyciodes phaon female. A. The ovary was dissected from a 10-day-old
female reared on the host plant. The fat body has been almost entirely used up in production of eggs. The four ova-
rioles per ovary, and individual egg chambers can be seen. B. Ovary dissected from a newly emerged female pro-
duced on PB diet. Mature-looking eggs are present surrounded by large amounts of fat body, which is characteristic
of newly emerged females produced on host plant, PB diet with 10% leaves, or PB diet.

Genc & Nation: Artificial Diet for Phaon Crescent Butterfly

Fig. 2. Internal reproductive structures of Phyciodes phaon male. A. Internal structures dissected from a newly
emerged male produced on the living host plant. The fused testes (T) seminal vesicle (SV) leading to the ductus ejac-
ulatorius duplex (D) and accessory glands (AC). The duplex loops join to form the ductus ejaculatorius simplex lead-
ing to the aedeagus. B. The photograph shows the fused testes and related internal structures from a newly
emerged male reared on PB diet. Note larger, discolored testes (T) and atrophied seminal vesicles (SV).

with 10% freeze-dried leaves. In males produced
on the PB diet, the testicular body is not uniformly
colored as in males from the host plant. There are
differences also in the appearance of the vas defer-
entia of the males. The vas deferentia of males
produced on the host plant or PB diet with 10%
freeze-dried leaves have swollen vas deferentia
near the midlength, forming the seminal vesicles.
The seminal vesicles of males produced on the PB
diet are not swollen and appear to be atrophied.


The PB diet designed for moths clearly is not
satisfactory as a diet for the Phaon crescent. As
originally formulated, it allows only about 37% of
newly hatched larvae to become adults, and the
adults do not reproduce. Thus, a colony cannot be
maintained on the artificial diet. We improved the
diet with respect to both survival and ability of
adults to reproduce by adding freeze dried host
plant leaves equal to 10% by weight of dry ingredi-
ents of the PB formula. This improved diet pro-
duced from 66% up to 78% adults in some
experiments from first instars started on the diet,
and the adults mated and reproduced, maintain-
ing the colony. Although we also improved the PB
diet to give good production of adults by addition of
5% glucose or 5% Beck's salt mix, the adults did
not reproduce. Glucose in the PB formula may be a
feeding stimulant, and/or a readily available car-
bohydrate energy source. The original PB formula
does not include a simple carbohydrate, nor does it
include a salt mixture. Lepidopterans, most of
which are phytophagous, typically have a rela-

tively high K/Na' ratio in the hemolymph, in con-
trast to omnivorous and some carnivorous feeders
which have low K/Na ratios. Beck et al. (1968) de-
veloped a salt mixture (now sold as Beck's salt
mixture) relatively high in K and Mg2' and low in
Na' and Ca2 and showed that it improved the
growth and survival of the European corn borer,
Ostrinia nubilalis (Hiibner). Wesson's salt mix of-
ten has been used in insect diets (Singh 1977), but
it was developed for vertebrate animals, and it has
high Na /K and Ca2 /Mg2' ratios suitable for verte-
brates. Although it works for some insects, proba-
bly it is not optimal for phytophagous insects.
Dethier (1954) and Fraenkel and Blewett
(1943) emphasized that host plant selection is de-
termined by the presence or absence of nonnutri-
tive secondary plant substances that act as
feeding deterrents or stimulants. Various imbal-
ances of the nutrients in a diet can stress insects,
and reduce growth and survival (House 1965;
House 1969). The small amount of host leaves in
the artificial diet may aid digestion and assimila-
tion of nutrients, and may help balance some of
the nutrients in the PB diet formula.
Newly emerged females reared on the host
plant and on the PB diet with added host plant
leaves have mature ovaries with apparently ma-
ture eggs at the time of emergence. In this respect
they are similar to the cecropia moth Hyalophora
cecropia, the silkmoth, Bombyx mori, and the fall
armyworm Spodoptera frugiperda, all of which
develop the ovaries and eggs during some part of
the pupal stage (Tsuchida et al. 1987; Sorge et al.
2000). In contrast, the noctuid moth Heliothis
virescens (Zeng et al. 1997) and the monarch but-

Florida Entomologist 87(2)

terflyDanaus plexippus (Pan & Wyatt 1971) have
a relatively immature ovary at adult emergence.
Female Phaon crescents have four ovarioles in
each of 2 ovaries, and each ovariole contains about
49 egg follicles, with apparently mature eggs ready
to be fertilized and laid a few days after emer-
gence. Thus, a female might be able to lay about
400 eggs, and we found that one individual did lay
434 eggs. The failure of females produced on the
host-free PB diet to lay eggs may be due to a failure
to mate. Despite substantial time in observations,
we never observed mating in butterflies produced
on the PB diet, whereas observations of mating
were common in butterflies produced on PB diet
with 10% freeze-dried host leaves or those pro-
duced on living host plants. Mating is a stimulus
for oviposition and oogenesis in some insects. For
example, oviposition in the Australian field cricket,
Teleogryllus sp., and the onion fly, Delia sp., is en-
hanced as a result of mating (Chapman 1998).
Males of some lepidopterans transfer prostaglan-
dins or prostaglandins-synthesizing chemicals to
the female during mating and these stimulate ovi-
position (Stanley-Samuelson 1994).
Male Phaon crescents produced on host plants
have a mature reproductive system and mate
within 2-3 days after emergence. The male system
includes fused testes, vas deferentia, paired acces-
sory glands, and an ejaculatory structure and duct.
The enlarged regions of the vas deferentia that
serve as a sperm reservoir and seminal vesicle in
males produced on the host plant or PB diet with
10% freeze-dried host plant leaves appear to be at-
rophied in males produced on PB diet. The testes
from PB diet reared males are larger (swollen) and
light red in color, compared to those reared on liv-
ing host plant or PB diet with 10% freeze-dried
host leaves. These defects observed in the internal
reproductive system of males produced on the PB
diet, coupled with the failure to get any reproduc-
tion from sexes produced on the PB diet suggest
that these males may not produce viable sperm.
Although no apparent abnormalities were de-
tected in the internal reproductive system of fe-
males produced on PB diet, they could have
physiological defects in the reproductive system
that are not evident from simple dissections.


We thank Drs. Norman Leppla, Grazyna Zimowska,
Jeff Shapiro, and Simon Yu for helpful criticism of earlier
drafts. We thank Kathy Milne for technical assistance in
the laboratory. We thank Dr. Kenneth Portier for assis-
tance with statistical analyses. Hanife Gene received
support from the government of Turkey. Florida Agricul-
tural Experiment Station Journal Series No. R-09650.


1968. Nutrition of the European corn borer, Ostrinia

nubilalis. VI. A larval rearing medium without crude
plant fractions. Ann. Entomol. Soc. Am. 61: 459-462.
CHAPMAN, R. F. 1998. The Insects: Structure and Func-
tion. 4th Edition. Cambridge University Press, UK.
770 pp.
DETHIER, V. G. 1954. Evolution of feeding preferences in
phytophagous insects. Evolution 8:33-54.
HABECK. 1980. Morphological Studies on the beet ar-
myworm Spodoptera exigua (Hubner) (Lepidoptera:
Noctuiidae). Technical Bulletin 816, Agricultural
Experiment Stations, Institute of Food and Agricul-
tural Sciences, University of Florida, Gainesville.
EMMEL, T. C., AND B. KENNEY. 1997. Florida's Fabulous
Butterflies. Tampa, FL, World Publications. 96 pp.
FRAENKEL, G., AND M. BLEWETT. 1943. The sterol re-
quirements of several insects. J. Biochem. 37:692-695.
GENC, HANIFE. 2002. Phaon crescent, Phyciodes phaon:
Life cycle, nutritional ecology and reproduction,
Ph.D. dissertation, University of Florida, Gaines-
ville, FL 32611, USA.
GuY, R., N. C. LEPPLA, J. R. RYE, C. W. GREEN, S. L.
BARRETTE, AND K. A. HOLLIEN. 1985. Trichoplusia
ni, pp. 487-493. In P. Singh and R. F. Moore [eds.],
Handbook of Insect Rearing Vol. II, Elsevier Science
HARRELL, F. E. 2001. Regression Modeling Strategies:
with Applications to Linear Model, Logistic Regres-
sion, and Survival Analysis. Springer-Verlag, 568 pp.
HOSMER, D. W., AND S. LEMESHOW. 2000. Applied Logis-
tic Regression. Wiley & Sons, NY, 373 pp.
HOUSE, H. L. 1965. Effects of low levels of the nutrient
content of a food and of nutrient imbalance on the
feeding and the nutrition of a phytophagous larva,
Celerio euphorbiae (Linnaeus) (Lepidoptera: Sphin-
gidae). Can. Entomol. 97: 62-68.
HOUSE, H. L. 1969. Effects of different proportions of
nutrients on insects. Ent. Exp. Appl. 12: 651-669.
MINNO, M. C., AND M. MINNO. 1999. Florida Butterfly
Gardening: A Complete Guide to Attracting, Identi-
fying, and Enjoying Butterflies of the Lower South.
University Press of Florida, Gainesville. 210 pp.
PAN, M.-L., AND G. R. WYATT. 1971. Juvenile hormone
induces vitellogenin synthesis in the monarch but-
terfly. Science 174: 503-505.
SINGH, P. 1977. Artificial Diets for Insects, Mites, and
Spiders. IFI/Plenum, New York. 594 pp.
STANLEY-SAMUELSON, D. W. 1994. Prostaglandins and
related eicosanoids in insects. Advances in Insect
Physiology, 24: 115-212.
2000. Regulation of vitellogenesis in the fall army-
worm, Spodoptera frugiperda (Lepidoptera: Noctu-
idae). J. Insect Physiol. 46: 969-976.
monal control of ovarian development in the silk-
worm, Bombyx mori. Arch. Insect Biochem. Physiol.
5: 167-177.
WALDBAUER, G. P., AND S. FRIEDMAN. 1991. Self-selec-
tion of optimal diets by insects. Annu. Rev. Entomol.
36: 43-63.
WEBB, S. E., AND A. M. SHELTON. 1988. Laboratory
rearing of the imported cabbageworm. New York's
Food and Life Sciences Bulletin 122: 1-6, N.Y State
Agricultural Experiment Station, Geneva.
ZENG, E., S. SHU, AND S. B. RAMASWAMY. 1997. Vitello-
genin and egg production in the moth, Heliothis vire-
scens. Arch. Biochem. Physiol. 34: 287-300.

June 2004

Florida Entomologist 87(2)

June 2004


'Everglades Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences
3200 E. Palm Beach Rd., Belle Glade, FL 33430-4702

2Department of Entomology and Nematology, P.O. Box 110620, Gainesville, FL 32611-0620

3Department of Horticultural Sciences, P.O. Box 110690, Gainesville, FL 32611-0690

4Indian River Research and Education Center, 2199 S. Rock Rd., Ft. Pierce, FL 34945-3138


One hundred faba bean (Vicia faba L., Fabales: Fabaceae) accessions from the USDA-NSSL
Seed Repository in Prosser, WA were grown outdoors in southern Florida from December
2000 through April 2001 and October 2001 through April 2002 to both evaluate their poten-
tial as a forage crop and to initiate selections of superior genotypes. Insect herbivores and
their predators were observed for feeding associations and collected for identification
throughout the two seasons of trials. Sixty-one species of insect herbivores and nectaring
predators and parasitoids were observed feeding on or were captured on faba bean leaves,
stems, flowers, extra-floral nectaries or pods. Additionally, thirty-two species of predacious
and parasitic insects were observed eating herbivorous insects or captured while searching
for prey or hosts on faba beans plants. The most significant damage was caused by large pop-
ulations ofAphis craccivora Koch (Hemiptera: Aphidae) that fed on terminals and young leaf
and stem tissue. Six Coccinellidae species fed upon aphids and reproduced on the crop. Pods
were damaged by reproducing populations of Leptoglossus phyllopus (L.) (Hemiptera: Core-
idae) and Nezara viridula (L.) (Hemiptera: Pentatomidae).

Key Words: Aphis craccivora, Leptoglossus phyllopus, Nezara viridula, bidens mottle mo-
saic, faba bean, Vicia faba


Cien accesiones de haba (Vicia faba L., Fabales: Fabaceae) del Repositorio de Semillas de
USDA-NSSL en Prosser, WA fueron sembradas en el campo en el sur de Florida de diciembre
2000 hasta abril 2001 y de octubre 2001 hasta abril 2002 para evaluar su potential como cul-
tivo de forraje y para iniciar la selecci6n de genotipos superiores. Los insects herbivoros y
sus depredadores fueron observados para determinar las asociaciones alimentarias y reco-
lectados para identificarlos durante dos estaciones de pruebas. Sesenta y una species de in-
sectos herbivoros y depredadores que se alimentaban del nectar, parasitoides que fueron
observados alimentandose de la plant, o fueron capturados en las hojas, tallos, flores, n6c-
tar extra-floral o las vainas de haba. Ademas, treinta y dos species de insects depredadores
y parasiticos fueron observados alimentandose de insects herbivoros, o capturados mien-
tras estaban buscando press u hospederos sobre el haba. El dano mas significativo fu6 cau-
sado por la alta poblaci6n de Aphis craccivora Koch (Hemiptera: Aphidae) que se aliment6
de los terminales y del tejido tierno de las las hojas y el tallo. Seis species de Coccinellidae
se alimentaron de los afidos y se reprodujeron en el cultivo. Las vainas fueron danadas por
poblaciones de Leptoglossus phyllopus (L.) (Hemiptera: Coreidae) y de Nezara viridula (L.)
(Hemiptera: Pentatomidae) reproduci6ndose sobre el cultivo.

The faba bean, Vicia faba L., is a cold hardy, horse bean (V faba var. equina Pers.) and the pi-
grain legume originally domesticated in the Hin- geon or tick bean (V faba var. minor Beck) are
dustani region of central Asia, but now cultivated grown primarily for animal feed or as a green ma-
from tropic to sub-arctic climates (Zeven & Zhuk- nure crop. In Europe, these two later species are
ovsky 1975). This taxa has been artificially di- referred to as "field beans" (Bond et al. 1985). In
vided by seed size into three subspecies (Polhill & Florida, faba bean production is uncommon, and
van der Maesen 1985). The broad bean (V faba broad beans are only rarely seen in Florida gar-
var. major Harz) is mostly grown as a grain vege- dens (Stephens 1994). However, Florida does
table because of its large seed size, while the have significant and diverse legume based indus-

Nuessly et al.: Insects on Faba Bean in Southern Florida

tries throughout the state, which range from ex-
otic oriental vegetables such as the winged bean
(Psophocarpus tetragonolubus (L.) DC.) to forage
legumes including clovers. Large commercial in-
dustries are in place for peanuts (Arachis hy-
pogaea L.) and fresh market beans (Phaseolus
vulgaris L.), with smaller production of cowpea
(Vigna unguiculata (L.) Walp.) (Florida Agricul-
tural Statistics Service 2001). Additionally, uses
of feral legumes such asAeschynomene spp. vacil-
late from weed to cover crop to domesticated for-
age. With the rare exception of the Austrian pea
(Pisum satiuum var. arvense (L.) Poiret) used by
recreational hunters for deer browse, most le-
gumes grown in Florida are warm season crops
and frost intolerant. The faba bean is one of a few
freeze tolerant winter legumes that could be inte-
grated into Florida agriculture as either a vegeta-
ble or forage crop. It could enlarge the array of
winter vegetable crops or be inserted into a silage
cropping system that includes corn (Zea mays L.)
and sorghum (Sorghum bicolor Moench) to sup-
port the cattle and dairy industries. It has the an-
cillary benefits of nitrogen fixation and thus a
reasonably low fertility requirement.
Any assessment of a crop's potential in a new
region would be aided by the knowledge of the in-
sect fauna that would be associated with its pro-
duction. Insect and nematode pests of faba beans
were broadly reviewed by Bardner (1983) and
Cammel & Way (1983). Economically important
faba bean insect pests include aphids that cause
direct feeding damage and transmit plant viruses
(e.g., Aphis fabae Scopoli, A. craccivora Koch,
Acyrthosiphon pisum (Harris), and Megoura vi-
ciae Buckton) (Hemiptera: Aphidae), as well as
leafhoppers, thrips, moth larvae, leafmining fly
larvae, seed beetles and weevils. Many insect spe-
cies are found on warm season legumes in Flor-
ida, some of which are considered to be
commercial pests (Pernezny et al. 2004). It is rea-
sonable to assume that some of these insects
would overlap onto faba beans, but an actively
growing legume crop in the winter season could
host additional insect species not typically found
on warm season legumes. The purpose of this re-
search was to identify insects and their associa-
tion (i.e., herbivorous, predacious, parasitic) with
experimental plots of faba beans grown from Oc-
tober to April in southern Florida. Our findings
are discussed in relation to other known insect
pests of faba beans in the western hemisphere
and of Florida legumes in general.


One hundred faba bean accessions in the range
from PI 301011 through PI 577748 were acquired
from the USDA-NSSL Seed Repository in
Prosser, WA. The accessions were split planted in
two seasons at the Everglades Research and Ed-

ucation Center, Belle Glade, Palm Beach County,
Florida. Sixty-seven of these accessions were
planted on December 1, 2000 and grown through
April 30, 2001. Selections were made based on
horticultural and agronomic characters and
planted with the remaining 33 accessions in Octo-
ber 2001 and grown through April 2002. Plants
were grown outdoors in 40 above-ground, con-
crete-walled production bins, 76.2 cm deep and
2.1 m long, filled with Palm Beach soil mix (50%
compost, 25% clean sand, 25% bark, Odum's,
West Palm Beach, FL). Seeds were planted 10 to
15 cm apart in rows spaced 46 cm on center, five
rows per bin. Six seeds of each accession were
planted in a row with final plant density averag-
ing four plants per row and 20 plants per bin.
Plots were provided with a complete fertilizer
plus micro nutrients mixed with the soil at plant-
ing. Additional fertilizer (20-20-20 plus micro nu-
trients and ammonium nitrate) was applied at
label rates on a regular basis from early February
to early April in both seasons. The plants were
grown insecticide free until March of both years
when imidacloprid (Provado 1.6 Flowable, Bayer
CropScience LP, Research Triangle Park, NC)
was applied at 3 fl. oz per acre to control excessive
populations of cowpea aphids,Aphis craccivora. A
composite population of PI lines from seeds left
over from selections from the previous season was
mixed together and planted in the field on 31 Oc-
tober 2001 for observation and collection of asso-
ciated insects. Hand-held planters (Almaco,
Nevada, IA) were used to plant the seeds 10 cm
apart in 4 rows 76 cm on center and 114 m long in
a Lauderhill organic soil (i.e., euic, hyperthermic
Lithic Medisaprists) at the Everglades Research
and Education Center, Belle Glade, FL.
Plants were examined weekly for presence of
insects at various times from early morning to
early evening to survey the entire photophase.
Observations of feeding associations with faba
bean leaves, stems, flowers, and pods, as well as
predacious and parasitic activity against insect
herbivores was recorded whenever possible be-
fore specimens were collected and preserved for
identification. Insects were identified to species
where possible through the use of published sys-
tematic keys and direct comparisons with mu-
seum specimens housed at the Division of Plant
Industry in Gainesville, Florida.


Plant and Nectar Feeders

Insects found in association with faba beans
during the two seasons are divided into plant and
nectar feeders (Table 1) and predators and para-
sitoids (Table 2). Notes on feeding associations
are included for only those directly observed. In-
sects that caused visible damage to terminals,

Florida Entomologist 87(2)

June 2004

IN 2001 AND 2002.

Order Family Insect Life stage' Plant part




Acrididae Chortophaga australion Rehn & Hebard
Tettigoniidae Microcentrum rhombifolium (Saussure)
Thripidae Frankliniella bispinosa (Morgan)
Frankliniella insularis (Franklin)
Frankliniella kelliae (Sakimura)
Miridae Creontiades rubinervis (Stal)
Lygaeidae Oncopeltus cayensis Torre-Bueno
Oncopeltus fasciatus (Dallas)
Ozophora trinotata Barber
Pyrrhicoridae Dysdercus mimulus Hussey
Coreidae Acanthocephala femorata (F.)
Anasa scorbutica (F.)
Leptoglossus phyllopus (L.)
Zicca taeniola (Dallas)
Alydidae Stenocoris tipuloides (DeGeer)
Pentatomidae Acrosternum hilare (Say)
Acrosternum marginatum (Palesot de Bearvois)
Edessa bifida (Say)
Euschistus ictericus (L.)
Euschistus quatrator Raulston
Nezara viridula (L.)
Thyanta perditor (F.)
Cicadellidae Draeculocephala mollipes (Say)
Gypona sp.
Delphacidae Perkinsiella saccharicida Kirkaldy
Aphidae Acyrthosiphon pisum (Harris)
Aphis craccivora Koch
Pseudococcidae Planococcus citri (Risso)

Coleoptera Scarabaeidae


Lepidoptera Pyralidae







Anomala marginata (F.)
Euphoria sepulcralis (F.)
Trigonopeltastes delta Forster
Chauliognathus marginatus (F.)
Diabrotica balteata Leconte
Diabrotica undecimpunctata howardi Barber
Diaprepes abbreviatus (L.)
Hellula rogatalis (Hulst)
Herpetogramma phaeopteralis (Guenee)
Spoladea recurvalis (F.)
Spilosoma virginica (F.)
Feltia subterranea (F.)
Spodoptera eridania (Cramer)
Automeris io io (F.)
Lerema accius (J. E. Smith)
Hedriodiscus triuittatus (Say)
Hermetia illucens (L.)
Euxesta annonae (F.)
Xanthaciura insecta (Loew)
Liriomyza trifolii (Burgess)
Chrysis sp.
Agapostemon splendens (Lepeletier)
Halictus sp.
Xylocopa micans Lepeletier
Apis mellifera L.





Leaf and stem
Root and stem

Seedling stem


1Life stage: L, larva; N, nymph; A, adult.

Nuessly et al.: Insects on Faba Bean in Southern Florida

FLORIDA IN 2001 AND 2002.

Order Family Insect Life stage' Observed association


Forficulidae Doru taeniatum (Dohrn)
Reduviidae Repipta taurus (F.)

Sinea sp.
Zelus longipes (L.)
Podisus maculiventris (Say)
Calleida decora (F.)
Brachiacantha decora Casey
Coelophora inaequalis (F.)
Cycloneda sanguinea (L.)
Harmonia axyridis (Pallas)
Hippodamia convergens Guerin-Meneville
Olla v. nigrum (Mulsant)

Coleoptera Carabidae


Calliphoridae Phaenicia caeruleiviridis (Macquart)
Sarcophagidae Sarcodexia sp.
Tachinidae Lespesia sp. 1
Lespesia sp. 2
Nilea sp.
Winthemia sp.
Hymenoptera Mutillidae Timulla sp.
Vespidae Eumenes fraternus Say

Pachyodynerus nasidens (Latreille)

Polistes dorsalis (F.)

Polistes major Beauvois

Polistes metricus Say

Pompilidae Anoplius sp.
Sphecidae Liris sp.


General predator
General predator
General predator
General predator
General predator
General predator
Aphid predator
Aphid predator
Aphid predator
Aphid predator
Aphid predator
Aphid predator

A Lepidoptera predator/
A Lepidoptera predator/
A Lepidoptera predator/
A Lepidoptera predator/
A Lepidoptera predator/

Lepidoptera predator/

Braconidae Bracon sp.
Cotesia sp.
Ichneumonidae Coccygomimus marginellus (Brulle)
Exetastes sp.
Pterocormus sp.
Trogomorpha trogiformis (Cresson)
Chalcididae Brachymeria sp.
Conura sp.

Extra floral nectary
Extra floral nectary

'Life stage: L, larva; N, nymph; A, adult.

leaves and pods appeared to be evenly distributed cessions and both study years. Sixty-one species
across the tested accessions and none were ob- of insect herbivores and nectaring predators and
served to be more attractive than another to the parasitoids were observed feeding or captured on
insect herbivores and natural enemies. Collection faba bean leaves, stems, flowers, extra-floral nec-
records in Tables 1 and 2 are pooled across all ac- taries or pods.

Dolichopodidae Condylostylus sp.
Plagioneurus univittatus Loew
Syrphidae Allograpta oblique (Say)
Palpada vinetorum (F.)
Toxomerus sp.


Florida Entomologist 87(2)

Cowpea aphids were the most abundant in-
sects feeding on faba bean leaves in both years of
the study. Their feeding was concentrated on the
youngest leaf and stem tissue and resulted in
stunted terminal growth and distorted leaf ex-
pansion. They are known as faba bean pests
throughout the Mediterranean and some subtrop-
ical and tropical areas where they cause damage
from both direct feeding and virus transmission
(Cammell & Way 1983). The pea aphid,Acyrthosi-
phon pisum, was a late season colonizer of the
crop after initiation of pod set in February 2001,
but not in 2002. It utilized a different microhabi-
tat of the plants compared withAphis craccivora,
concentrating instead on the underside of leaves
in the more protected middle region of the canopy.
Lowe (1967) found that A. pisum first preferred
faba bean stems in the growing terminal before
moving to developing leaves. Pea aphids are
known for causing more damage from virus trans-
mission than from direct feeding damage (Cam-
mell & Way 1983). Aphis fabae is known from
Florida (Halbert & Nuessly 2001) and is an aphid
pest of faba bean in Nova Scotia, Canada
(Patriquin et al. 1988), but it was not observed
feeding on the crop in our studies.
A crippling virus, Bidens mottle mosaic, in-
fected the PI accessions tested during the middle
of the first year causing stunted terminal growth
and chlorotic, disfigured leaves and pods (Baker
et al. 2001). While the disease is known from
southern Florida on leafy vegetables, and both of
the aphids colonizing the plants in our study are
known vectors, faba beans are a new host for this
virus. No difference is colonization rates of acces-
sions were observed for either cowpea or pea
aphids. Two other aphid vectors of Bidens mottle
mosaic, Myzus persicae (Sulzer) andAphis spirae-
cola Patch (both Hemiptera: Aphidae), are known
from the area (Halbert & Nuessly 2001), but they
were not found feeding on or colonizing faba
beans during this study. Aphid transmitted vi-
ruses have also been reported on faba bean in
Guatemala (Vasquez 1988). Plants infested with
broad bean mosaic virus in Egypt serve as better
hosts of A. craccivora allowing them to produce
more progeny on infected than on non-infected
plants (El-Kady & Salem 1974).
Other piercing-sucking insects observed feed-
ing on leaves (Table 1) included the plant bug Cre-
ontiades rubinervis (Stal), the seed bug Ozophora
trinotata Barber, and the leafhoppers Draeculo-
cephala mollipes (Say) and Gypona sp. Other mir-
ids, including Lygus sp., have been reported to
produce necrotic spots on leaves that later col-
lapse to form holes (Bardner 1983). Leafhopper
feeding damage was also noted by Bardner (1983)
to produce distorted growth and stunting on faba
beans. While necrotic lesions were observed on
leaves in our plantings, it was not confirmed
whether they were the result of feeding by these

heteropterous and homopterous insects. The lyg-
aeid Oncopeltus cayensis Torre-Bueno was ob-
served probing stems and pods, while 0. fasciatus
(Dallas) was not observed feeding on any of the
plant structures. Both are known to specialize on
various milkweeds (Slater & Baranowski 1990).
Two species of leafminers were found attack-
ing faba bean leaves. The American serpentine
leafminer, Liriomyza trifolii (Burgess), is a com-
mon pest of leafy vegetables throughout Florida
(Spencer & Stegmaier 1973). Damage by this in-
sect consisted of feeding and oviposition stipples
and mines on leaves, but not pods. Another spe-
cies of dipterous leafminer produced much wider
and longer tunnels lined with a dark residue that
was quite obvious without light transmission.
This leafminer remains unidentified because re-
peated attempts to rear adults from larvae in in-
fested leaves held in plastic cups at room
temperature were unsuccessful.
Species from several orders were found chew-
ing on faba bean foliage (Table 1). The grasshop-
pers Chortophaga australion Rehn & Hebard and
Microcentrum rhombifolium (Saussure) ate large
jagged edge sections from leaves. Granulate cut-
worm, Feltia subterranea (F.), cut off seedling
faba beans at their base. Both cucumber beetle
species found in southern Florida, banded cucum-
ber beetle (Diabrotica balteata Leconte) and spot-
ted cucumber beetle (D. undecimpunctata howardi
Barber), produced irregular sized notches on the
edge and holes within the youngest fully ex-
panded leaves. These cucumber beetles have a
wide adult host feeding range andD. balteata is a
pest of leafy vegetables and sweet corn in south-
ern Florida (Nuessly & Webb 2002a, b). A single
adult Diaprepes root weevil (Diaprepres abbre-
viatus (L.)) was found feeding on the edge of a
leaf. The adults of this species have been reported
to feed on a variety of vegetables and weeds and
the larvae are pests of many crops, including cit-
rus and sugarcane (Simpson et al. 1996), which
are grown extensively throughout central and
southern Florida. Larvae of the tiger moth (Spilo-
soma virginica (F.)) and Io moth (Automeris io io
(F.)) were the only Lepidoptera observed to com-
plete development on the plants. Larvae of other
species, including the southern armyworm
(Spodoptera eridania (Cramer)), were collected
on plants, but were likely predated by wasps, bee-
tles, bees and assassin bugs (Table 2) before they
could complete development. Adults of three spe-
cies of pyralids were captured while they rested
on the plants (Table 1).
Flower and nectar feeders included thrips, bee-
tles, skippers and wasps (Table 1). The thrips
Frankliniella bispinosa (Morgan), F insularis
(Franklin), and F kelliae (Sakimura) fed on pol-
len, anthers, and other flower parts, but did not
cause any noticeable problems with pollination or
seed set. Adults of three scarab beetle species were

June 2004

Nuessly et al.: Insects on Faba Bean in Southern Florida

found feeding on pollen and nectar within faba
bean flowers.Anomala marginata (F.) and Eupho-
ria sepulcralis (F.) are common flower feeders,
with the latter species found feeding at ear tips
and armyworm feeding holes of sweet corn (Zea
mays L.) ears (Nuessly et al. 1999). Trigonopel-
tastes delta Forster is commonly found feeding on
fragrant inflorescences of many plants, including
the sable or cabbage palm (Sabal palmetto (Walt.
Lodd.)) (G.S.N., unpublished data). The soldier
beetle Chauliognathus marginatus (F.) became
very common as the seasons progressed, feeding
on nectar and pollen within flowers during late af-
ternoon and early evening. Mating pairs were fre-
quently observed. Adults of the hesperiid Lerema
accius (J.E. Smith) were observed feeding on faba
bean flowers. Various bees (Anthophoridae, Halic-
tidae and Apidae) were observed feeding at the
flowers (Table 1). While wasps are discussed be-
low, paper wasps (Vespidae), spider wasps (other
Sphecidae), and the cuckoo wasp Chrysis sp. (Ta-
bles 1 and 2) were also observed flying between
and feeding from numerous flowers during the
day. Two Chalcidoidae species were also observed
feeding from extra floral nectaries.
Pod feeders composed the largest guild of faba
bean herbivores observed in the experiments (Ta-
ble 1). The pyrrhocorid Dysdercus mimulus Hus-
sey, four species of Coreidae and seven species of
Pentatomidae fed on developing pods. Leptoglos-
sus phyllopus (L.) was the most common and de-
structive of the Coreidae that fed and reproduced
on the crop. Their nymphs were observed to feed
in small groups on pods. This species feeds on a
wide variety of cultivated crops, including cowpea
(Baranowski & Slater 1986). Pod feeding pro-
duced raised, pitted black bumps on the pod sur-
face and black spots on developing seeds. The
other coreids found on faba beans in our studies,
Acanthocephala femorata (F.), Anasa scorbutica
(F.), and Zicca taeniola (Dallas), are more com-
monly found associated with native plants and
have not been identified as pests of leguminous
plants (Baranowski & Slater 1986). Pod damage
similar to that caused by L. phyllopus also was
produced by the most commonly encountered
stink bug, Nezara viridula (L.). This insect also
reproduced on the faba beans, although few were
observed to complete development. Six other
stink bug species (Table 1) were not commonly en-
countered and were not observed to reproduce on
faba beans. Three of these six species, Acroster-
num hilare (Say) (Simmons & Yeargan 1990), A.
marginatum (Palesot de Bearvois) (Hallman et al.
1985), and Thyanta perditor (F.) (Saunders et al.
1983) are known to cause at least some damage to
soybeans or other cultivated legumes.
Dipterous species in the families Stratiomyidae,
Otitidae, and Tephritidae were captured while
they rested on bin and field planted faba beans, but
no feeding associations were noted for these flies.

These fly species are commonly found on many spe-
cies of agronomic crops and weeds throughout
southern Florida (G.S.N., unpubl. data).

Predacious and Parasitic Insects

Twenty-seven species of predator and parasi-
toid insects were collected during our studies.
Larvae of six coccinellid species (Table 2) fed on
cowpea aphids and their adults were reared from
pupae collected from stems and under leaves of
test plants. Raymond et al. (2000) found that
Aphis fabae feeding on V faba attracted the coc-
cinellid Adalia bipunctata L. in laboratory test-
ing, whereas plants without aphids or ones with
aphids recently removed did not attract the bee-
tles. Larvae of all three syrphids feed on the sug-
arcane aphids Melanaphis sacchari (Zehntner)
and Sipha flava (Forbes) (both Hemiptera:
Aphidae) in Florida sugarcane fields (Hall 1988).
Calleida decora (F.) is a red and iridescent
green predacious ground beetle commonly en-
countered on various cultivated crops throughout
the southeastern and into the mid-western
United States (Erwin et al. 1977). Larvae and
adults of this species were found on the soil and
up into the faba bean canopy. It is an important
predator of several lepidopterous pests of cotton
and soybean (Harris et al. 1985).
Solitary and social wasps (Sphecidae and
Vespidae, Table 2) were frequently observed
searching leaves that exhibited feeding damage.
These wasps normally anesthetize their prey and
then either macerate them into "meat balls" to
bring back to their nests or use them to provision
solitary mud or sub-soil nests for their progeny.
Feeding damage associated with medium through
large sized Lepidoptera larvae was not common
on our faba beans and a few late instar southern
armyworm, tiger, and io moth larvae were the
only large larvae found. Resistance to armyworm
pests in faba beans was not noted in the Clement
et al. (1994) review of plant resistance achieve-
ments in cool season food legumes. Therefore, we
believe that lepidopteran larvae in their early to
mid instars succumbed to predation rather than
to plant resistance mechanisms.
The assassin bugs Repipta taurus (F), Zelus
longipes (L.) and Sinea sp. were each observed to
feed on Lepidoptera larvae, cucumber beetle
adults and spiders on faba bean leaves. They are
generalist predators found throughout the United
States (Blatchley 1926; Reinert 1978; Altieri &
Whitcomb 1980). The earwig Doru taeniatum
(Dohrn) and velvet ant Timulla sp. were captured
on faba beans without any specific feeding associa-
tion, however, the former is known as a predator of
armyworms, aphids and other soft bodied insects
in corn (Jones 1985) and sugarcane (Hall 1988).
Several species of insects caused damage to the
crop in our studies. Aphid (Aphis craccivora) feed-

University of Florida Home Page
© 2004 - 2010 University of Florida George A. Smathers Libraries.
All rights reserved.

Acceptable Use, Copyright, and Disclaimer Statement
Last updated October 10, 2010 - - mvs