Acquired Natural Enemies of Oxyops Vitiosa 1

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Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 1

ACQUIRED NATURAL ENEMIES OF THE WEED BIOLOGICAL CONTROL AGENT OXYOPS VITIOSA (COLEPOTERA: CURCULIONIDAE)

ROBIN M. CHRISTENSEN, PAUL D. PRATT, SHERYL L. COSTELLO, MIN B. RAYAMAJHI AND TED D. CENTER USDA/ARS, Invasive Plant Research Laboratory, 3225 College Ave., Ft. Lauderdale, FL 33314

ABSTRACT

The Australian curculionid Oxyops vitiosa Pascoe was introduced into Florida in 1997 as a biological control agent of the invasive tree Melaleuca quinquenervia (Cav.) S. T. Blake. Pop- ulations of the weevil increased rapidly and became widely distributed throughout much of the invasive tree’s adventive distribution. In this study we ask if O. vitiosa has acquired natural enemies in Florida, how these enemies circumvent the protective terpenoid laden exudates on larvae, and what influence 1 of the most common natural enemies has on O. vitiosa population densities? Surveys of O. vitiosa populations and rearing of field-collected individuals resulted in no instances of parasitoids or pathogens exploiting weevil eggs or larvae. In contrast, 44 species of predatory arthropods were commonly associated (>5 individuals when pooled across all sites and sample dates) with O. vitiosa. Eleven predatory species were observed feeding on O. vitiosa during timed surveys, including 6 pentatomid species, 2 formicids and 3 arachnids. Species with mandibulate or chelicerate mouthparts fed on adult stages whereas pentatomids, with haustellate beaks, pierced larval exoskeletons thereby by- passing the protective larval coating. Observations of predation were rare, with only 8% of timed surveys resulting in 1 or more instances of attack. Feeding by the pentatomid Podisus mucronatus Uhler accounted for 76% of all recorded predation events. Podisus mucronatus numerically responded to fourth instars but no response was observed for other life stages. Damage to M. quinquenervia plants from feeding by O. vitiosa, however, was not influenced by P. mucronatus densities, indicating that predation does not alter plant suppression.

Key Words: biological control, biotic resistance, predation, Oxyops vitiosa, Melaleuca Quin- quenervia, Podisus mucronatus

RESUMEN

El curculiónido australiano Oxyops vitiosa Pascoe fue introducido a la Florida en 1997 como un agente de control biológico para el árbol invasor, Melaleuca quinquenervia (Cav.) S. T. Blake. Poblaciones del gorgojo aumentaron rapidamente y se distribuyeron ampliamente por mucho de la distribución del árbol invasor adventivo. En este estudio, preguntamos si O. vitiosa han adquerido enemigos naturales en la Florida, como estos enemigos evitan las se- creciones de turpenoides que protejen las larvas, y que inﬂuencia tiene uno de los enemigos naturles mas comunes sobre la densidad de la población de O. vitiosa? La inspección de la po- blación de O. vitiosa y la cria de individuos recolectados en el campo resulto en no caso de parasitoides y patógenos usando los huevos o larvas de los gorgojos. En contraste, 44 especies de artrópodos depredadores fueron comúnmente asociadas (>5 individuos cuando se agrega- dos por todos los sitios y fechas de muestreo) con O. vitiosa. Se observaron once especies de depredadores alimentándose sobre O. vitiosa durante los sondeos, incluyendo 6 especies de pentatómidos, 2 formícidos y 3 arácnidos. Especies con partes bucales mandíbuladas y que- liceradas se alimentaron sobre los estadios adultos mientras que los pentatómidos, con su pico chupador, puncharon los exo-esqueletos de las larvas asi pasando el cubertura protec- tiva de las larvas. Observaciones de depredación fueron raras, con solamente 8% de los estudios que llevaron el tiempo resultaron en 1 ó más instancias de ataque. La alimentación del pentatómido Podisus mucronatus Uhler contó con 76% de los eventos de depredación re- gistrados. Podisus mucronatus respondió numericamente al los instares de cuarto estadio pero ninguna respuesta fue observada en los otros estadios de vida. El daño a las plantas de M. quinquenervia debido a la alimentación por O. vitiosa, sin embargo, no fue inﬂuenciado por la densidad de P. mucronatus, que indica que la depredación no altera la supresión de la planta.

Acquisition of novel natural enemies may in- Partland & Nicholson 2003; Norman et al. 2009; fluence the successful establishment, spread, and Paynter et al. 2010). Of the arthropods introduced impact of introduced weed biological control for control of invasive plants world wide, approx- agents in their adventive range (Goeden & Louda imately 50% suffer sufficient mortality from 1976; Semple & Forno 1987; Simberloff 1989; higher trophic levels to significantly limit sup- Cornell & Hawkins 1993; Hill & Hulley 1995; Mc- pression of target weeds (Goeden & Louda 1976).

2 Florida Entomologist 94(1) March 2011

The spider mite Tetranychus lintearius (Dufour), had acquired natural enemies in Florida, (2) how for example, was introduced into New Zealand, these enemies mitigated the defensive strategies Australia, and the United States as a biological of the herbivore, and (3) what impact the most control agent of the invasive plant Ulex europaeus abundant of these natural enemies has on O. vi- L. (Fabaceae) (Hill & Stone 1985; Hill et al. 1991). tiosa population densities? Although successfully established and widely distributed, mites in each country rarely sustained MATERIALS AND METHODS sufficient population densities to provide permanent control of the target weed (Rees & Hill 2001). Surveys for natural enemies associated with Subsequent studies demonstrated that a complex O. vitiosa were conducted at 4 locations in south of native and introduced predators suppressed T. Florida. Site 1 was located near Ft. Lauderdale, lintearius populations and limited control of the Broward Co., FL. The site was a 0.5-ha field con- invasive weed (Peterson 1993; Peterson et al. sisting of 2- to 5-m tall trees occurring at a den- 1994; Pratt et al. 2003). sity of 21,560 trees/ha. In general, M. quinquen- Considering the ecological risks (Carvalheiro ervia trees were growing in organically rich soils et al. 2008) and expense of biological control, in- typical of reclaimed ‘glades’ systems. Melaleuca creased attention in the scientific literature has quinquenervia trees at site 2 occurred under a focused on predicting susceptibility of introduced power line right of way near Weston, Broward biological control agents to natural enemies in the Co., FL. Prior to 1997 land managers cut M. quin- adventive range (Kuhlmann et al. 2006). Hill & quenervia trees near their bases, resulting in Hulley (1995), for instance, demonstrated that multi-stemmed coppices. The study area was 0.5 variation in susceptibility of introduced herbi- ha and trees were 2-5 m tall, occurring at a den- vores to parasitoids is related, in part, to evolu- sity of 2,517 trees/ha. Site 3 was located near Es- tionary strategies that render the prey less acces- tero, Collier Co., FL and consisted of an 8-ha area sible, apparent, or palatable to the attacker. of drained wetland converted to pasture. To sup- Along this continuum of use by natural enemies press M. quinquenervia growth, land managers lie those species that are highly apparent yet ex- mowed trees at 6-month intervals, resulting in perience relatively less attack due to the expres- coppices 0.5-2 m in height. These coppicing sion of chemical deterrents that render them less clumps formed a dense, nearly continuous canopy palatable or even toxic to prospective natural en- of leaves with 4,406 stumps/ha. In contrast to the emies. The introduced weevil Oxyops vitiosa Pas- previous sites, the soil type was primarily sand, coe sequesters terpenoids from leaves of its host consistent with an invaded pine flatwoods. Site 4 plant Melaleuca quinquenervia (Cav.) S. T. Blake consisted of a 1-ha area within the historically and larvae excrete these compounds through mesic flatwoods of the Picayune Forest near Belle their integument (Wheeler et al. 2002). The con- Meade, Collier Co., FL. A fire burned much of the sumption and expression of these terpenoids re- M. quinquenervia dominated areas in 1998 re- pels the red imported fire ant (Solenopsis sulting in recruitment of 129,393 trees/ha of pri- invicta Buren) and red wing blackbird (Agelaius marily small 1-2 m tall saplings, with an occa- phoeniceus L.) under controlled feeding tests sional large, mature tree interspersed (Table 1). (Wheeler et al. 2002). It remains unclear, how- Surveys were conducted monthly at each site ever, if this acquired repellency confers protection from Nov 2000 through Jun 2001 and sampling from the suite of potential novel natural enemies occurred between 10 AM and 2 PM on days without that exist in the herbivore’s adventive range. precipitation. Sampling consisted of sweeping M. Oxyops vitiosa is native to eastern Australia quinquenervia foliage, and occasionally trunks, and is a specialist herbivore of the invasive tree with a 90-cm diameter sweep net. One sample M. quinquenervia (Balciunas et al. 1994). Based consisted of 100 sweeps of the net in a sweeping on its narrow host range, the weevil was permit- motion of 180° with sweeps spaced ca. 1.0 m apart ted for release in Florida in 1997 and readily es- along a randomly selected 100-m transect. Four tablished in M. quinquenervia dominated habi- samples along separate transects were collected tats (Center et al. 2000; Pratt et al. 2003). Adult each month. The content of the net after 100 weevils feed on M. quinquenervia foliage whereas sweeps was emptied into a 4-liter sealable bag larvae consume only newly-developed leaves that and frozen at minus 20 (±1) °C until processed. are ephemerally produced in seasonal flushes at Arthropods were then separated from plant mate- branch apices (Purcell & Balciunas 1994). rial, sorted by morphological types, and pinned or Following its introduction, O. vitiosa popula- stored in 70% ethanol. tions increased rapidly and became widely dis- One limitation of our sweep sampling method tributed throughout much of the invasive tree’s included collecting arthropods that were not geographic distribution in Florida (Pratt et al. closely associated with O. vitiosa, but were tran- 2003; Balentine et al. 2009). When considering sients, merely resting on the plant foliage or dis- the large densities of these herbivores in the envi- turbed from understory vegetation while sam- ronment, we questioned (1) whether O. vitiosa pling. Additionally, this method was biased to-

Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 3

TABLE 1. RESEARCH SITE DESCRIPTION AND SUMMARY OF SURVEYS CONDUCTED FOR O. VITIOSA IN SOUTH FLORIDA.

Site GPS Coordinates Surveys conducteda Habitat Hydro-period

1 N 26.05605 W -80.25168 1, 2, 3, 4 Swale Short 2 N 26.03548 W -80.43495 1, 2, 3, 4 Swale Medium 3 N 26.42550 W -81.81033 1, 2, 3, 4, 5 Mesic flatwoods Short 4 N 26.10478 W -81.63392 1, 2, 3, 4 Mesic flatwoods Short 5 N 26.46017 W -81.70186 4 Mesic flatwoods Short 6 N 26.54698 W -81.79820 4 Wet flatwoods Short 7 N 28.47323 W -81.33632 4 Upland lake Long 8 N 25.71341 W -80.47949 4 Swale Medium 9 N 25.81208 W -80.41780 4 Swale Medium 10 N 26.16227 W -80.36269 4 Swale Long

a1 = Arthropods associated with O. vitiosa, 2 = O. vitiosa population density, 3 = O. vitiosa egg parasitism, 4 = Entomopathogens of O. vitiosa, 5 = Impacts of P. mucronatus on O. vitiosa populations. wards poor fliers. All study sites possessed interval and the nearest tree to each point was in- smaller trees that facilitated sampling but may ventoried. At all sites, O. vitiosa life stages were have biased collections to lower rather than counted along with plant resource availability. higher canopy dwelling species. Therefore, cau- Resource availability was assessed on a 5-point tion should be used when drawing inferences scale based on visual estimation of percentage of from these data due to the unknown relationships suitable foliage for feeding by O. vitiosa: 0 = no between sampled arthropods and O. vitiosa. For suitable foliage; 1 = less than 25%; 2 = 26 to 50%; this reason, a minimum of 2 observers also 3 = 51 to 75%; 4 = 76 to 100%. searched for direct predation or parasitism for 30 A partial correlation analysis was used to iden- min/survey at each site monthly. tify those predators positively associated with O. All specimens, except formicids, were submit- vitiosa (PROC CORR), after controlling for the in- ted to and deposited at the Florida State Collec- fluence of site by the PARTIAL statement (SAS tion of Arthropods (FSCA, Division of Plant In- 1999). For all tests, a P-value <0.05 was consid- dustries, Gainesville, FL) for identification and ered significant evidence for association among incorporated into their taxonomic database (Cos- predators and O. vitiosa. However, caution should tello et al. 2003). Most formicids were identified be used when interpreting these data because as- and retained by L. Davis of the Fire Ant Unit, Ag- sociation is not sufficient evidence to suggest that ricultural Research Service, USDA, Gainesville, a trophic relationship exists among the species. FL. A few formicids were identified by M. Deyrup To determine if O. vitiosa had acquired egg or of the Archbold Biological Station, Lake Placid, larval parasitoids in its adventive range, 50 eggs FL. Several dipteran specimens were identified at were collected at random from sites 1-4 at the Systematic Entomology Laboratory, Agricul- monthly intervals. Eggs were examined under a tural Research Service, USDA, Beltsville, MD. dissecting microscope (10-50X) to detect presence Population densities of O. vitiosa at sites 2 and of larval exit holes indicating larval eclosion; eggs 3 were monitored by delineating a 0.5-ha study with exit holes were discarded. The remaining site within the existing M. quinquenervia stands, eggs were left attached to leaf material and respectively. Within these plots, transects were placed in gelatin capsules. These capsules were arranged in a grid pattern with 8 transects ori- transferred into a Petri dish (10 × 1.5 cm) that ented east to west at 10-m intervals and points on was then sealed with Parafilm to retain leaf mois- each transect spaced 10 m apart. Beginning in ture. Petri dishes were placed in an environmen- Nov 2000 through Jun 2001, M. quinquenervia tal chamber at 25 (±1) °C, with a photoperiod of trees were sampled monthly at 20 randomly se- 16:8 (L:D) and a relative humidity of 65% ± 10%. lected transect points. Plants at each sampling Hatching of eggs was monitored once a week. point were selected based on the quarter method Eggs that did not hatch after 1 month were dis- of vegetation sampling (Smith 1966). The area sected. was divided into 4 quarters at each sampling To detect larval parasitoids, 50 third or fourth point based on the 4 cardinal directions. The instars of O. vitiosa were reared to the pupal nearest tree to the sample point in each quarter stage. Each larva was placed individually in 1 was examined to determine the number of O. vi- Petri dish (10 × 1.5 cm) with moistened filter pa- tiosa per plant. The ordered distance method was per and M. quinquenervia leaves. Petri dishes used to quantify weevil population densities over were sealed with Parafilm and kept in an environ- time at sites 1 and 4 (Krebs 1999). In total, 30 mental chamber under the same conditions as de- points were randomly selected at each sampling scribed earlier. Host leaves were replaced every

4 Florida Entomologist 94(1) March 2011 other day until the prepupal stage, when leaves We observed no instances of parasitoids (egg were removed for the remainder of the study. and larval) or pathogens exploiting O. vitiosa. Of Surveys for entomopathogens of O. vitiosa the 1138 O. vitiosa eggs collected from the study were conducted at 10 sites between Jun 2003 and sites (Table 1), 782 hatched and developed nor- Jan 2004 (Table 1). Late instars and adults were mally while the remaining 356 did not hatch. Of collected, packaged in an ice-cooler, transported the 1266 fourth-instars collected from study sties, to the laboratory, and examined by USDA/ARS 913 survived to become adults. Dissection of both insect pathologists at the Center for Medical, Ag- unhatched eggs and dead larvae yielded no evi- ricultural, and Veterinary Entomology, Gaines- dence that mortality was due to parasitism. Sim- ville, FL. All live individuals originating from the ilarly, no pathogens were found in the late instars same location were homogenized in 3-5 mL and adults collected from sampled sties (Table 1). of deionized water and a sample of the crude sus- These results indicate that despite the herbi- pension was examined with a phase-contrast mi- vore’s high population densities and large geo- croscope to search for pathogens, such as mi- graphic distribution (Pratt et al. 2003; Balentine crosporidia, fungal spores, or occluded viruses. et al. 2009), native parasitoids and pathogens During initial surveys, the pentatomid bug Po- have failed to exploit these lifestages of O. vitiosa. disus mucronatus Uhler was commonly associ- One explanation for the lack of O. vitiosa para- ated with O. vitiosa and observed feeding on lar- sitization may be that native parasitoids require val stages of the biological control agent at each of more than the 4 years allotted in this study to ad- the 4 study sites. Therefore, we quantified the just behaviorally and physiologically to exploit population dynamics of P. mucronatus and O. vi- the new host as well as produce sufficient densi- tiosa at site 3. Sampling was conducted as de- ties to be discovered through our sampling proto- scribed earlier except 20 transects were oriented cols. In contrast, Hill & Hulley (1995) determined east to west at 20-m intervals with 9-10 points on that 16 of the 40 established weed biological con- each transect spaced 20 m apart. Melaleuca quin- trol agents in South Africa had acquired native quenervia plants were sampled at 50 randomly parasitoids within 3 years of release. Similarly, selected transect points every 6 weeks (approxi- the biological control agent Neomusotima con- mate generation time; Purcell & Balciunas 1994) spurcatalis Warren acquired a suite of parasitoids beginning in Dec 2000 and continuing through within months of its release in Lygodium micro- Oct 2002. As before, the nearest plant to the sam- phyllum (Cav.)-dominated habitats of Florida ple point in each quadrant was examined to deter- (Kula et al. 2010). These and other examples of mine the number and life stage of each O. vitiosa rapid parasitoid acquisition by biological control and P. mucronatus individual. In addition to agents (Carvalheiro et al. 2008; Paynter et al. these data, we also noted the amount of damage 2010) suggest that the timing of our study was due to herbivory, plant resource availability (as not premature but that future parasitoid (or described earlier), and the number of dead larvae. pathogen) surveys may yield new discoveries as Herbivory damage was assessed on a 5-point the region continues to recruit exotic species scale based on visual estimation of the percentage (Klassen et al. 2002; Dobbs & Brodel 2004; of suitable foliage destroyed by O. vitiosa feeding: Childers & Rodrigues 2005). 0 = no damage; 1 = less than 25% destroyed; 2 = Surveys of O. vitiosa populations resulted in 26 to 50%; 3 = 51 to 75%; 4= 76 to 100% destroyed. the collection of 154 species of predatory arthro- Linear regression was used to test for a numerical pods, yet only 44 had an overall abundance response of predators to life stages of O. vitiosa (n greater than 5 individuals when pooled across all = 6), dead larvae, food availability, and herbivory sites and dates (Table 2). Species positively corre- damage (i.e., aggregation of predators to patches lated with O. vitiosa (all stages) included the sal- of high prey density; Schenk & Bacher 2002). ticid Eris flava (Peckham & Peckham), the crab spiders Misumenops bellulus (Banks) and Mis- RESULTS AND DISCUSSION umenops sp., and the pentatomid bug Podisus mucronatus Uhler (Table 2). Although these data Biotic resistance describes the collective influ- indicate that predators are associated with the in- ence of parasitoides, predators, pathogens, and troduced herbivore, direct observation of preda- competitors on the establishment and prolifera- tion provides conclusive evidence of these novel tion of non-indigenous species, including intro- trophic interactions. Eleven predatory species duced biological control agents (Simberloff & Von were observed feeding on O. vitiosa during timed Holle 1999). Historically, native predators, para- surveys, including 6 pentatomid species (Euthy- sitoids, and pathogens have interfered with half rhynchus floridanus (L.), P. mucronatus (Say), Po- of the published case histories involving insect in- disus jole (Stal), Podisus maculiventris (Say), Po- troductions for weed control (Goeden & Louda disus sagitta (F.), Stiretrus anchorago (F.)), 2 for- 1976). Considering this high rate of interference, micids (Pseudomyrmex gracilis (F.), Solenopsis in- we questioned if O. vitiosa had acquired natural victa) and 3 arachnids (Peucetia viridans (Hentz), enemies in its adventive range. Latrodectus mactans (F.), Latrodectus geometri- Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 5

TABLE 2. THE ABUNDANCE OF PREDACEOUS ARTHROPODS COLLECTED FROM THE INVASIVE TREE MELALEUCA QUIN- QUENERVIA AND THEIR CORRELATION WITH THE INTRODUCED BIOLOGICAL CONTROL AGENT OXYOPS VITIOSA.

Site Abundance Partial Correlation

Family Species 1234 PCC P-value

Alydidae Hyalymenus sp. A 1 12 0 0 -0.23098 0.2112 Anyphaenidae Hibana sp. 00090.04519 0.8093 Hibana sp. 4012-0.06653 0.7222 Araneidae Acacesia hamata (Hentz) 15 14 2 2 0.15511 0.4047 Eriophora ravilla (C.L. Koch) 5101-0.08230 0.6599 Neoscona sp. 27 15 2 0 -0.08867 0.6353 Neoscona arabesca (Walckenaer) 6000-0.13606 0.4655 Cixiidae Bothriocera sp. 0600-0.11674 0.5317 Clubionidae Clubiona sp. 06050.01856 0.9211 Formicidae Camponotus planatus (Roger) 75 46 2 0 0.05757 0.7584 Camponotus floridanus (Buckley) 140020.09479 0.6120 Dolichoderus pustulatus Mayr 6000-0.14389 0.4400 Paratrechina longicornis (Latreille) * 85 102 0 4 -0.14078 0.4500 Paratrechina guatemalensis (Forel) * 16206-0.08637 0.6441 Pseudomyrmex pallidus (F. Smith) * 15 11 0 0 -0.13190 0.4794 Pseudomyrmex gracilis (Fabricius) 2106-0.17915 0.3349 Solenopsis invicta Buren 8 16 16 64 0.02664 0.8869 Lycosidae Pardosa sp. 1 0 16 0 0.33110 0.0688 Pirata sp. 18 2 27 22 0.33916 0.0620 Lygaeidae Oedancala crassimana (Fabricius) 132800.33885 0.0622 Paromius longulus (Dallas) 74100.06829 0.7151 Membracidae Spissistilus festinus (Say) 00700.29864 0.1027 Mimetidae Mimetus sp. 1 0 2 16 0.07110 0.7039 Miridae Taylorilygus pallidulus (Blanchard) 22400.31276 0.0867 Miturgidae Cheiracanthium inclusum (Hentz) 96 140 10 16 -0.01003 0.9573 Oxyopidae Peucetia viridans (Hentz) 137 9 11 4 0.06787 0.7168 Pentatomidae Podisus mucronatus Uhler 6 1 98 4 0.52700 0.0023 Pisauridae Pisaurina sp. 75000.13199 0.4791 Reduviidae Zelus longipes Linnaeus 25001-0.05046 0.7875 Salticidae Eris flava (Peckham & Peckham) 4 5 16 9 0.36503 0.0435 Hentzia palmarum (Hentz) 72 45 10 55 0.00540 0.9770 Pelegrina galathea (Walckenaer) 12 5 14 0 0.18343 0.3233 Phidippus sp. 0105-0.14807 0.4267 Thiodina peurpera (Hentz) 0 19 0 0 -0.13863 0.4570 Scelionidae Trissolcus sp. 0800-0.10483 0.5746 Tetragnathidae Tetragnatha sp. 338510.11216 0.5480 Theridiidae Anelosimus studiosus (Hentz) 670300.08956 0.6318 Chrysso pulcherrima (Mello-Leitao) 10140.26530 0.1492 Theridion glaucescens (Becker) 120010.13018 0.4852 Theridion flavonotatum (Becker) 24002-0.01400 0.9404 Thomisidae Misumenops sp. 23540.30519 0.0950 Misumenops bellulus (Banks) 26 37 27 15 0.35739 0.0484 Misumenops sp. 10 0 29 2 0.39715 0.0269 Tmarus sp. 2 0 1 52 -0.24154 0.1905 cus C.L. Koch). The formicids and arachnids were counted for 76% of all recorded predation events observed feeding exclusively on adult weevils and the remaining species each represented <5% whereas the pentatomids attacked larvae of O. vi- of the events, respectively. tiosa; E. floridanus was the only species observed Ecological theory suggests that host range ex- exploiting all active stages of the introduced her- pansion is influenced in part by host phylogeny, bivore. Observing predation was rare, with only with close relatives more readily adopted than 8% of timed surveys resulting in 1 or more in- distant ones (Paynter et al. 2010). Therefore, an stances of attack. Feeding by P. mucronatus ac- alternative explanation for the lack of acquired 6 Florida Entomologist 94(1) March 2011 parasitoids and pathogens may be due to the pau- with Hawkins et al. (1997), who reported that in- city of closely related species in the biological con- sect predation is higher in late developmental trol agent’s adventive range. The Australian wee- stages due, in part, to resource concentration and vil O. vitiosa belongs to the tribe Goniopterini, handling time. Not surprisingly, a positive rela- which has no representatives in the New World tionship between P. mucronatus and larval (Alonso-Zarazaga & Lyal 1999). Similarly, inva- corpses also was observed. sion by M. quinquenervia markedly alters com- While it is clear that P. mucronatus attacks O. munity structure in ways that are likely to repel vitiosa larvae and numerically responds to the habitat specialists. Therefore, the aquisition of single most damaging stage of the herbivore, does parasitoids will likely require evolutionary rather this predation disrupt biological control of M. than ecological time scales (Hill & Hulley 1995). quinquenervia? We hypothesized that increases With the exception of E. floridanus, the exclu- in P. mucronatus densities results in concomitant sive use of adult versus larval prey observed increases in predation and ultimately decreases herein may be explained by mouthpart morpholo- in plant damage caused by O. vitiosa. Damage gies and the antipredatory activity of the viscous levels observed herein, however, were not influ- coating that covers immature stages of O. vitiosa enced by P. mucronatus densities (Table 3), indi- (Purcell & Balciunas 1994). Larvae of the intro- cating that predation does not alter plant sup- duced weevil sequester terpenoids from M. quin- pression within the sampled patch. Similarly, the quenervia leaves and excrete these compounds amount of plant resource availability for con- through their integument (Wheeler et al. 2003). sumption by O. vitiosa does not vary based on This larval coating has been shown to repel the predator loads, which suggest that predation does red imported fire ant (S. invicta) and likely con- not result in a corresponding increase in undam- fers protection against other mandibulate preda- aged plant material (Table 3). These results are tors (Wheeler et al. 2002). However, adults and supported by independent studies that also were pupae lack the coating and are susceptible to pre- conducted at site 3 and reported marked reduc- dation by a range of predator types. The larval tions in M. quinqueneriva growth and survival coating does not confer protection against pen- despite the presence of these predators (Center et tatomid species observed herein. The haustellate al. 2000; Pratt et al. 2002; Pratt et al. 2004). The mouthparts of pentatomid species pierce the lar- limited influence of P. mucronatus on O. vitiosa val integument and largely bypass the terpenoid- population growth and herbivory is likely related laden coating to access the internal contents of to low predation rates (mean = 9.5%, SE = 0.5). the larval prey. Yet, mouthpart type alone does The introduction of O. vitiosa has resulted in not facilitate exploitation of the abundant novel marked reductions in growth and survivorship of resource as other predators with haustellate the invasive tree M. quinqueneriva (Pratt et al. mouthparts (i.e., Zelus longipes L.) occurred at 2003, 2005; Rayamajhi et al. 2008; Tipping et al. the study sites but were not common or observed 2008, 2009; Balentine et al. 2009), with no direct directly feeding on O. vitiosa larvae. non-target impacts to plant species in the weevil’s Increased densities of O. vitiosa eggs, early in- adventive range (Pratt et al. 2009). The acquisi- stars, and adults did not influence patch colonization of higher trophic levels by O. vitiosa, how- tion by P. mucronatus (Table 3). A numerical re- ever, suggests that indirect effects of apparent sponse by P. mucronatus was observed, however, competition may exist as predators are subsi- on plants harboring fourth instars (Table 3), indi- dized by the introduced weevil and their result- cating a preference for larger larval stages of the ant increased population densities may exert introduced weevil. These findings are consistent asymmetrical predation on their historical prey species (Carvalheiro et al. 2008). In the absence of TABLE 3. LINEAR REGRESSION OF PODSUS MUCRONATUS pre-introduction food web analyses, it remains DENSITIES ON OXYOPS VITIOSA STAGE SPECIFIC unclear how the exploitation of O. vitiosa by na- DENSITIES, PLANT QUALITY, AND FOLIAGE tive predators affects apparent competition on AVAILABILITY. shared prey densities. The limited predation by generalists suggests that the strength of appar- Life stage df Estimate t-value Pr > t ent competition is weak but additional research is needed to quantify interactions among intro- Egg 1 -0.00097 -0.29 0.7752 duced and native prey species as mitigated by 1st instar 1 -0.00185 -0.24 0.8075 common predators. 2nd instar 1 0.01394 1.66 0.0966 3rd instar 1 0.01175 1.27 0.2025 4th instar 1 0.0251 2.88 0.004 ACKNOWLEDGMENTS Adults 1 0.00818 1.06 0.2874 Dead 1 0.13755 16.16 <.0001 We thank 2 anonymous reviewers for comments on Damage 1 -0.00265 -1.31 0.1893 earlier versions of the manuscript. We also thank Scott Wiggers, Willey Durden, Kirk Tonkel, Andrea Kral, Carl Plant foliage 1 -0.00176 -0.94 0.3462 Belnavis, Tafana Fiore, and Stacey Grassano for assis- Christensen et al.: Acquired Natural Enemies of Oxyops vitiosa 7

tance with data collection and site maintenance. Men- agents introduced to South Africa. Biol. Cont. 5: tion of trade names or commercial products in this 297-302. publication is solely for the purpose of providing specific HILL, R. L., AND STONE, C. 1985. Spider mites as con- information and does not imply recommendation or en- trol agents for weeds, pp. 443-448 In W. Helle and M. dorsement by the U.S. Department of Agriculture. This H. Sabelis [eds.], Spider Mites: Their Biology, Natu- research was supported, in part, by grants from the ral Enemies and Control, Vol. 1B. Elsevier, Amster- South Florida Water Management District, the Florida dam. Department of Environmental Protection Bureau of In- HILL, R. L., GRINDELL, J. M., WINKS, C. J., SHEAT, J. J., vasive Plant Management, and the USDA Areawide AND HAYES, L. M. 1991. Establishment of gorse spi- TAME Melaleuca Program (tame.ifas.ufl.edu). der mite as a control agent of gorse. Proc. 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PRATT, P. D., RAYAMAJHI, M. B., CENTER, T. D., TIPPING, SEMPLE, J. L., AND FORNO, I. W. 1987. Native parasi- P. W. AND WHEELER, G. S. 2009. The ecological host toids and pathogens attacking Samea multiplicalis range of an intentionally introduced herbivore: A Guenée (Lepidoptera: Pyralidae) in Queensland. J. comparison of predicted versus actual host use. Biol. Australian Entomol. Soc. 26: 365-366. Cont. 49: 146-153. SIMBERLOFF, D. 1989. Which insect introductions suc- PRATT, P. D., RAYAMAJHI, M. B., VAN, T. K., CENTER, T. D., ceed and which fail? pp. 61-75 In J. A. Drake [ed.], AND TIPPING, P. W. 2005. Herbivory alters resource al- Biological Invasions. John Wiley and Sons, New location and compensation in the invasive tree Mela- York. leuca quinquenervia. Ecol. Entomol. 30: 316-326. SIMBERLOFF, D., AND VON HOLLE, B. 1999. Positive in- PRATT, P. D., SLONE, D. H., RAYAMAJHI, M. B., VAN, T. teractions of nonindigenous species: invasional melt- K., AND CENTER, T. D. 2003. Geographic distribution down. Biol. Invasions 1: 21-32. and dispersal rate of Oxyops vitiosa (Coleoptera: SMITH, R. L. 1966. Ecology and Field Biology. Harper Curculionidae), a biological control agent of the inva- and Row, New York, N. Y. sive tree Melaleuca quinquenervia in south Florida. TIPPING, P. W., MARTIN, M. R., NIMMO, K. R., PIERCE, Environ. Entomol. 32: 397-406. R. M., SMART, M. D., WHITE, E., MADEIRA, P. T., PURCELL, M. F., AND BALCIUNAS, J. K. 1994. Life history AND CENTER, T. D. 2009. Invasion of a West Ever- and distribution of the Australian weevil Oxyops vi- glades wetland by Melaleuca quinquenervia coun- tiosa (Coleoptera: Curculionidae), a potential biologi- tered by classical biological control. Biol. Cont. 48: cal control agent for Melaleuca quinquenervia (Myrta- 73-78. ceae). Ann. Entomol. Soc. America 87(6): 867-873. TIPPING, P. W., MARTIN, M. R., PRATT, P. D., CENTER, T. RAYAMAJHI, M. B., PRATT, P. D., CENTER, T. D., TIPPING, D., AND RAYAMAJHI, M. B. 2008. Suppression of P. W., AND VAN, T. K. 2008. Aboveground biomass of growth and reproduction of an exotic invasive tree the invasive tree melaleuca (Melaleuca quinquen- by two introduced insects. Biol. Cont. 44: 235-241. ervia) before and after herbivory by adventive and WHEELER G. S., MASSEY, L. M., AND SOUTHWELL, I. A. introduced natural enemies: a temporal case study 2002. Antipredator defense of biological control in Florida. Weed Sci. 56: 451-456. agent Oxyops vitiosa is mediated by plant volatiles REES, M., AND HILL, R. L. 2001. Large-scale distur- sequestered from the host plant Melaleuca quin- bances, biological control and the dynamics of gorse quenervia. J. Chem. Ecol. 28(2): 297-315. populations. J. App. Ecol. 38: 364-377. WHEELER, G. S., MASSEY, L. M., AND SOUTHWELL, I. A. SAS. 1999. The SAS System for Windows, Version 8. 2003. Dietary inﬂuences on terpenoids sequestered SAS Institute Inc., Cary, North Carolina. by the biological control agent Oxyops vitiosa: Effect SCHENK, D., AND BACHER, S. 2002. Functional response of plant volatiles from different Melaleuca quinquen- of a generalist insect predator to one of its prey spe- ervia chemotypes and laboratory host species. J. cies in the ﬁeld. J. Anim. Ecol. 71: 524-531. Chem. Ecol. 29: 188-207.

Yang et al.: Mating Strategy of Ectropis oblique 9

COMPARATIVE MATING STRATEGIES OF MALE AND FEMALE ECTROPIS OBLIQUE (LEPIDOPTERA: GEOMETRIDAE)

YUN-QIU YANG1, XU-HUI GAO1, YAN-ZHUO ZHANG3, LONG-WA ZHANG2 AND XIAO-CHUN WAN1* 1Key Laboratory of Tea Biochemistry & Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University, Hefei, Anhui 230036, P.R. China

2Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, Anhui230036, P.R. China

3State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, P.R. China

*Corresponding author; E-mail: [email protected]

ABSTRACT

The mating strategies of male and female Ectropis oblique Prout were investigated with the aid of male antennae as an electroantennogram (EAG) detector and capillary-GC analysis. Each male was capable of mating with several females, but females that had received a spermatophore mated only once. Antennae dissected from males 0, 1, 2, 3, and 4 d post-mating and antennae from virgin males of corresponding ages displayed similar EAG responses to sex pheromone extracts from sexually active females. Pheromone extracts of mated females elicited signiﬁcantly weaker male EAG responses than the pheromone extracts of virgin females. EAG responses of males to sex pheromone extracts taken from mated females at 0, 1, 2, 3, and 4 d post-mating were consistently weak. Pheromone production in the pheromone glands of mated females was strongly suppressed and declined during each of 4 successive nights after they had mated.

Key Words: EAG, GC, mating, sex pheromone emission, spermatophore

RESUMEN

Las estrategias de apareamiento de machos y hembras de Ectropis oblique Prout fueron in- vestigadas con el uso de la antena del macho como un detector electro-antenogramatico (EAG) y de CG capilar análisis. Cada macho fue capaz de aparearse con varias hembras, pero las hembras que han recibido un espermatóforo se aparearon solamente una vez. Las antenas diseccionadas de los machos 0, 1, 2, 3, y 4 dias después de aparearse y las antenas de machos virgenes de edades corespondientes mostraron respuestas de EAG similares a los extractos de feromonas sexuales de hembras sexualmente activas. Los extractos de feromonas de hembras apareadas provocaron una respuesta del EAG de los machos signiﬁcativa- mente mas debíl que los extractos de feromonas de las hembras vírgenes. Las respuestas de EAG de machos hacia los extractos de feromonas sexuales tomados de hembras apareadas a los 0, 1, 2, 3 y 4 dias después de aparear fueron consistentemente mas debiles. La produc- ción de feromonas en las glandulas de feromonas de hembras apareadas fue fuertemente su- primida y declinó durante cada una de las noches consecutivas después de que las hembras se aparearon.

Male insects typically mate many times during nomenon was demonstrated by bioassay or chem- their lifetime, while females display diverse mat- ical analysis in many studies (Webster & Cardé ing strategies (Arnqvist & Nilsson 2000). In some 1984; Coffelt & Vick 1987; Ahn et al. 2002). In species, females need to mate once or only a few contrast to the numerous studies on changes in times to produce an optimal number of viable female reproductive behavior after mating, few eggs; in many other species females mate fre- studies have focused on male response to dimin- quently to maximize reproductive potential (Rad- ished pheromone production of mated females. wan & Rysinska´ 1999). No matter what strate- Recently, the electroantennogram (EAG) has gies females use, they tend to discontinue sex been widely used in studies on semiochemical in- pheromone production after mating, either tem- volvement in sex pheromones (Park et al. 2001; porarily or permanently; and this avoids prob- Gökçe et al. 2007). An EAG response proﬁle is lems associated with excessive male sexual ha- thought to represent the sensitivity and relative rassment (Giebultowicz et al. 1991). This phe- abundance of olfactory receptor neurons on the

10 Florida Entomologist 94(1) March 2011

antennae that are tuned to the compounds tested. tained by counting the spermatophores present in The EAG response amplitudes are thought to rep- her bursa copulatrix. The experiment was re- resent the quantity of semiochemicals (Pouzat & peated 40 times. The number of times a male E. ob- Ibeas 1989). Thus, the EAG may be used as a tool lique mated was determined by keeping a record of to investigate the sensitivity of males to variable the number of females mated during successive sc- amounts of sex pheromone. otophases. The experiment was also repeated 40 Ectropis oblique Prout (Lepidoptera: times. The longevity of each male and each female Geometridae) is an important tea bush pest in was recorded in order to determine whether mat- Southeast China. Population outbreaks can com- ing affected longevity of either gender. pletely defoliate leaves on the bushes (Hu et al. 1994). The sex pheromone components of the fe- Extraction of Sex Pheromones male E. oblique were identified as (Z,Z,Z)-3,6,9- octadecatriene and 6,7-epoxy-(Z,Z)-3,9-octadeca- Active sex pheromones were extracted from diene (Yao et al. 1991). Most efforts to control E. the glands of the virgin females. The terminal sec- oblique populations are focused on the develop- tion of the abdomen, which included the phero- ment of methodologies to disrupt reproduction. mone gland, was excised from the virgin female Therefore, understanding the behavior of E. ob- moth 6 h after the onset of the second scotophase, lique is necessary. In this study, the mating fre- when the virgin female had been calling for 1 h. quency and longevity of E. oblique males and fe- Experimental procedures were performed under males was investigated. At the same time, the dif- a red light to facilitate observation without dis- ferent mating strategies of the two genders were turbing the insects. Each excised abdominal tip examined using male antennae as EAG detectors. was immersed in 10 µL of redistilled hexane for 4- To verify the results of the electrophysiological 6 h at room temperature. Then, the tip was re- analysis, changes in sex pheromone titers pro- moved and the extract without any purification duced by mated and virgin females were also in- was submitted for EAG or GC analysis. The pro- vestigated using capillary-GC analysis. cedure for extracting sex pheromones from either mated or virgin females was the same. MATERIALS AND METHODS Electroantennographic Analysis of the Effect of Mating Insect Culture on Pheromone Production and on Male Responsiveness at Various Days after Mating The E. oblique insects were obtained from Qian- shan County (31.5N°, 116.3E°), Anhui Province, Electroantennograms (EAG) were obtained China, and maintained for many generations in the with an EAG apparatus (Syntech Co., 79199 laboratory. Larvae were reared on tea leaves. Adults Kirchzarten, Germany). The antennae of either and larvae were maintained in controlled condi- mated or virgin males were excised at the bases tions at 22 ± 3°C, 60-70% relative humidity, and a and a few distal segments were cut off to facilitate photoperiod of 14L:10D, with scotophase and photo- conductivity. The antennae were then attached to phase reversed from a natural light cycle to permit the electrodes of the EAG probe with Spectra 360 scotophase observations during normal working Electrode Gel (Parker Laboratories Inc., Orange, hours. Pupae were sexed based on the morphology New Jersey). Antennal preparations were ex- of the 8th-10th abdominal segments, and main- posed to a stream of humidified and charcoal-fil- tained in moist sand for eclosion. Adults were kept tered air emitted at 4 mL s-1after having flowed individually in 240-mL plastic jars and fed a 10% through a 35-cm long glass tube (inner diameter, honey solution soaked in cotton. 8 mm; outer diameter, 10 mm). To facilitate inser- tion of the Pasteur pipette used to administer the Effect of Mating Frequency on Longevity of Females pheromone test stimulus, a 3-mm hole was bored and Males 5 cm from the outlet of the glass tube. Ten µL of the extract (test stimulus) was applied to a piece In preliminary observations of behavior, both of filter paper (1 × 5 cm1 × 5 cm). The filter paper females and males copulated during the first sc- was placed in a Pasteur pipette after the solvent otophase after emergence, and were sexually ac- (hexane) had been allowed to evaporate for 5 min. tive at the second scotophase. Therefore, a 1-d-old Each test stimulus was delivered within a 0.5 s female (0 d after emergence) and a 2-d-old male pulse of 4 mL s-1 of air with a stimulus controller were paired. Copulation occurred in the sc- (type CS-55) to transport the volatiles to the an- otophase lasted approximately 6 h. After mating, tenna. The EAG signal was amplified 10× through the female insects did not resume calling during an intelligent data acquisition controller (type the same scotophase (Yang et al. 2008). Thus, each IDAC-2) and viewed on an oscilloscope. A period female in this experiment was replaced daily with of at least 30 s was allowed between 2 successive another 1-d- old virgin female. When the female stimuli for the recovery of antennal responsive- died, the number of times it had mated was ascer- ness. Redistilled hexane (10 µL) was used as a

Yang et al.: Mating Strategy of Ectropis oblique 11 control stimulus in every test. The absolute EAG perature was programmed at 50°C for 2 min, then amplitude (mV) minus the solvent response was 15°C min-1 to 250°C and held for 5 min. The tem- used for data analysis. peratures of the injector and detector were 200°C First, the influence of the male’s mating status and 250°C, respectively. Nitrogen with a flow ve- on the male’s EAG responses to active sex phero- locity of 40 mL min-1 was used as the carrier gas. mones was studied. To obtain mated males and fe- To quantify the pheromones in the female gland, males, the insects were paired in the first sc- only the amount of epo3,Z6,Z9-19:H, the major otophase and allowed to mate. Mating pairs were sex pheromone component of E. oblique, was de- then removed. After mating, the mated females termined. Each treatment was repeated 6-8 were used for the next experiment. With the same times. procedure as described above, the antennae of the mated male were dissected at 0, 24, 48, 72, and 96 Statistical Analysis h after mating for use as EAG test detectors. Sex pheromone extracts from sexually active females The data were analyzed by one-way ANOVA, were used as stimuli. The EAG responses of an- followed by a LSD multiple comparison test at P < tennae dissected from virgin males at each of 0.05 (SPSS 11.0 for Windows, 2002; SPSS Inc., these times post-mating were compared to the Chicago, IL). EAG responses of antennae obtained from mated males at corresponding times post-mating. Each treatment was repeated 6-8 times. RESULTS Secondly, the influence of the female’s mating status on the EAG responses of a male was stud- Mating Frequencies of Females and Males ied with sex pheromone extract of a mated female as the stimulus to elicit an EAG response from a The results of mating frequency are shown in 2-d-old virgin male. As described above, the sex Table 1. When 40 females were paired individu- pheromone of the mated females was extracted at ally with 2-d-old virgin males on successive days 0, 24, 48, 72, and 96 h after mating. The EAG re- until they died, only 1 spermatophore was de- sponses of antennae of 2-d-old virgin males ex- tected in the abdomens of 31 females (77.5%), posed to pheromone extract obtained from virgin while no spermatophore was detected in the abdo- females at each of the above times post-mating mens of the rest of the females (9, 22.5%). Thus were compared to the EAG responses to phero- any female that had received a spermatophore in mone extract obtained from mated females at cor- 1 mating did not copulate again. When males responding times post mating. Each treatment were repeatedly offered 2-d-old virgin females, was also repeated 6-8 times. the numbers of males that mated various times during their lifespan and the corresponding per- centages were as follows: 0 matings (10; 25.0%); 1 Pheromone Titer Analysis mating (9;22.5%); 2 matings (8; 20.0%), 3 matings To assess the effect of mating on pheromone (7; 17.5%), 4 matings (5; 12.5%); 5 matings (0; 0%) production, the sex pheromone titers of the mated and 6 matings (1; 2.5%). and virgin females were analyzed by the procedure described above. Thus sex pheromone ex- The Effect of Mating on the Longevity of Adults tract was analyzed in a gas chromatograph (Agi- lent 6890) equipped with a capillary column (DB- Males lived significantly longer than females 5, 60m × 0.5mm i.d × 0.25 µm film). The oven tem- (Table 1), whether mated or unmated (P < 0.05).

TABLE 1. MATING FREQUENCY OF ECTROPIS OBLIQUE AND ITS EFFECT ON FEMALE AND MALE LONGEVITY.

No. of Female Female No. of Male Male females mating rates Longevity males mating rates longevity Mating time observed (%) (days) observed (%) (days)

0 9 22.5 *11.36 ± 2.67 a 10 25.0 14.2 ± 0.85 b 1st 31 77.5 10.07 ± 2.87 a 9 22.5 19.6 ± 1.82 c 2nd 0 0 ** 8 20.0 16.0 ± 1.47 b 3rd 0 0 — 7 17.5 15.5 ± 2.50 b 4th 0 0 — 5 12.5 15.5 ± 1.50 b 5th 0 0 — 0 0 — 6th 0 0 — 1 2.5 15

*Values are mean ± SE. Different letters indicate signiﬁcant difference (P < 0.05) by LSD test. ** not tested.

12 Florida Entomologist 94(1) March 2011

In addition, the males that mated only once lived significantly longer than unmated males or males that had mated more than once (P < 0.05). How- ever males that had mated 2 times did not live significantly longer than males that had mated either 3 or 4 times. The life spans of mated and unmated females did not differ significantly.

Electroantennographic Analysis of the Effect of Mating on Pheromone Production and on Male Responsiveness at Various Days after Mating

The effects of mating and lapsed time after mating of males on their EAG responses to sex pheromone extracts from sexually active females are shown in Fig. 1. No significant (P > 0.05) dif- Fig. 2. Influence of differential mating status of fe- ferences in EAG responses to sex pheromone ex- males on male EAG responses to their pheromones. Sex tracts from sexually active females were observed pheromone extracts of mated females (solid bar) and between the mated and virgin males. The anten- virgin females (open bar) obtained 0, 1, 2, 3, and 4 d nae of males that had been amputated 0, 1, 2, 3, post-mating were used as stimuli. Data were presented and 4 d post-mating exhibited the same magni- as mean values ± SE (n = 6-8) and analyzed by one-way ANOVA, followed by an LSD multiple range test (P < tude of the EAG responses to sex pheromone ex- 0.05). Significant differences among various dosages of tracts from sexually active females as those of an- the same stimulant are indicated with different letters. tennae of the corresponding virgin males. The male EAG responses to female sex pheromone extracts from virgin females and mated females (Fig. 2) differed profoundly (P < 0.05) with gence (Fig. 3). Maximal pheromone titers were the former evoking much stronger responses than present in the glands during the second and third the latter. Moreover the male EAG responses to scotophase after emergence. Thereafter the pher- sex pheromones extracted from the females at 0, omone titer decreased gradually. When the fe- 1, 2, 3, and 4 d post mating remained at very low males mated on the first night after eclosion, the levels (data not shown). sex pheromone titers decreased strongly and significantly compared with the titers of virgin fe- Pheromone Titer Analysis males. Pheromone production in mated females remained suppressed during each of 4 successive The virgin females began to produce sex pher- nights after they had mated. These results are omones during the first scotophase after emer- consistent with the weak male antennographic responses coinciding with the male response to mated female sex pheromones.

Fig. 1. Influence of the mating status of males on their EAG response to female sex pheromone extracts from sexually active females. Antennae of unmated males (solid bar) and virgin males (open bar) amputated Fig. 3. Influence of mating on the production of 6,7- 0, 1, 2, 3, and 4 d post-mating were used as EAG detec- epoxy-(Z,Z)-3,9-octadecadiene, the major female sex tors. Data were presented as mean values ± SE (n = 6- pheromone. The female sex pheromone was extracted 8) and analyzed by one-way ANOVA, followed by an with hexane from pheromone glands of mated females LSD multiple range test (P < 0.05). Significant differ- (solid bar) and virgin females (open bar). Pheromone ences among various dosages of the same stimulant are glands of mated females were extracted 0, 1, 2, 3, and 4 indicated with different letters. d post-mating.

Yang et al.: Mating Strategy of Ectropis oblique 13

DISCUSSION show any increase up to 4 d after mating, it may be deduced that females may mate only once. This Our data suggest that the females in our labo- deduction is in accordance with the observation ratory colony of E. oblique are monandrous. Two that each female actually mates only 1 time. Even different views exist concerning female mating though a few females were observed to copulate strategies. Polyandry presents a variety of bene- twice, but no more than 1 spermatophore was fits to females, including full fertilization of their ever detected in a bursa copulatrix, possibly be- egg complement, increased genetic diversity of cause the first copulation was an unsuccessful offspring, receipt of non-sperm nutrients, and re- mating. duced chances of fertilization by sperm that are Mating-induced termination of sex pheromone genetically defective due to age. Conversely, poly- production has been investigated in several moth andry may decrease female fitness due to the eco- species (Raina et al. 1994; Ando et al. 1996). The logical cost of mating, including energy costs, and inactivation of pheromone production after copu- risks of physical injury and sexually transmitted lation, which reduces the ability of females to pathogens and parasites (Arnqvist & Nilsson elicit a sexual response in males, may be due to 2000). We observed that E. oblique females after the secretion of pheromonostatic peptide (Kingan having mated fended off males and did not accept et al. 1995), or the presence of viable sperm in the a second mating partner. Mated females began spermatheca (Giebultowicz et al. 1991). The laying eggs during the first scotophase and laid mechanisms involved in pheromone suppression nearly all of their eggs before the fourth day (data after copulation in E. oblique are currently un- not shown). The life spans of females ranged from known. Thus, it is suggested that further studies about ½ to ⅔ of the lengths of the life spans of must be conducted on this area. males. The lifespan of the female in the wild is The present results show that mating status likely to be even shorter than in the laboratory. did not appear to have a significant effect on the Thus, a single mating is sufficient to fertilize responses of male antennae to sex pheromone ex- nearly all eggs and minimize the above men- tracts of sexually active females. This suggests tioned risks associated with multiple matings. that a past mating does not cause the male to be Males, on the other hand, are potentially po- unresponsive to the sex pheromone, and such a lygynous. Male polygyny is to be expected because male can be expected to continue to seek females most evolutionary theories contend that the con- for additional matings. This deduction is also in tributions and consequences of mating are much accordance with direct observations of multiple greater for females than for males (Thornhill & matings by males. The same mating system de- Alcock 1983). scribed for E. oblique has been observed in some This study revealed that multiple matings re- other species (Royer & McNeil 1993; Foster& Ay- duced the longevity of males but not of females. ers 1996; Svensson et al. 1998). This result is in agreement with the results of studies on several other species (Proshold et al. ACKNOWLEDGMENT 1982; Svensson et al. 1998). It is thought that al- location of nutritional reserves for egg develop- The authors express gratitude to Professor M. Z. ment and maturation after mating may be re- FAN for assistance during this study. sponsible for causing the lifespan of mated females to be shorter than that of virgin females. In most species, there is a causal relationship REFERENCES CITED between male calling behavior and female pheromone emission. Only when the female pheromone AHN, S. J., CHOI, M. Y., AND BOO, K. S. 2002. Mating ef- gland becomes exposed to emit pheromone, may fect on sex pheromone production of the Oriental to- the male display calling behavior. Permanent or bacco budworm, Helicoverpa assulta. J. Asia-Pacific even temporary reductions in the emissions of sex Entomol. 5: 43-48. pheromones caused loss of attraction and sexual ANDO, T., KASUGA, K., YAJIMA, Y., KATAOKA, H., AND SUZUKI, A. 1996. Termination of sex pheromone pro- receptivity in males (Kingan et al. 1995). The duction in mated females of the silkworm moth. present study showed that E. oblique male EAG Arch. Insect. Biochem. Physiol. 31: 207-218. responses to mated female pheromone gland ex- ARNQVIST, G., AND NILSSON, T. 2000. The evolution of tracts were significantly diminished, which could polyandry: Multiple mating and female fitness in in- be the result of reduced pheromone release. The sects. Anim. Behav. 60: 145-164. results support the hypothesis that mating con- COFFELT, J. A., AND VICK, K. W. 1987. Sex pheromone of siderably suppressed pheromone production in fe- Ephestia cautella (Walker) (Lepidoptera, Pyralidae): males. Indeed according to our capillary-GC anal- Influence of mating on pheromone titer and release ysis, pheromone titers in pheromone gland ex- rate. J. Stored Prod. Res. 23: 119 -123. FOSTER, S. P., AND AYERS, R. H. 1996. Multiple mating tracts did not increase at all up to 4 d after mating. and its effects in the light brown apple moth, Epiph- Because the male EAG response to pheromone yas postvittana (Walker). J. Insect Physiol. 42: 657- gland extract of already mated females did not 667. 14 Florida Entomologist 94(1) March 2011

GIEBULTOWICZ, J. M., RAINA, A. K., UEBEL, E. C., AND RADWAN, J., AND RYSINSKA´ , M. 1999. Effect of mating RIDGWAY, R. L. 1991. Two-step regulation of sex- frequency on female fitness in Caloglyphus berlesei pheromone decline in mated gypsy moth females. (Astigmata: Acaridae). Exp. Appl. Acarol. 23: 399- Arch. Insect Biochem. Physiol. 16: 95-105. 409. GÖKÇE, A., STELINSKI, L. L., GUT, L. J., AND WHALON, RAINA, A. K., KINGAN, T. G., AND GIEBULTOWICZ, J. M. M. E. 2007. Comparative behavioral and EAG re- 1994. Mating-induced loss of sex pheromone and sponses of female obliquebanded and redbanded lea- sexual receptivity in insects with emphasis on Heli- froller moths (Lepidoptera: Tortricidae) to their sex coverpa zea and Lymantria dispar. Arch. Insect. Bio- pheromone components. European J. Entomol 104: chem. Physiol 25: 317-327. 187-194. ROYER, L., AND MCNEIL, J. N. 1993. Male investment in HU, C., ZHU, J., YE, G., AND HONG, J. 1994. Ectropis ob- the European corn borer, Ostrinia nubilalis (Lepi- lique Prout, a serious geometrid pest of tea bush in doptera: Pyralidae): impact on female longevity and East China. Scientific and Technical Publishers, reproductive performance. Funct. Ecol. 7: 209-215. Shanghai. pp. 226-228. SVENSSON, M. G. E., MARLING, E., AND LÖFQVIST, J. KINGAN, T. G., BODNAR, W. M., RAINA, A. K., SHABAN- 1998. Mating behavior and reproductive potential in OWITZ, J., AND HUNT, D. F. 1995. The loss of female the turnip moth, Agrotis segetum (Lepidoptera: Noc- sex pheromone after mating in the corn earworm tuidae). J. Insect Behav. 11: 343-359. moth, Helicoverpa zea: Identification of a male pher- THORNHILL, R., AND ALCOCK, J. 1983. The Evolution of omonostatic peptide. Proc. Natl. Acad. Sci 92: 5082- Insect Mating Systems. Harvard University Press, 5086. Cambridge. PARK, K. C., ZHU, J. W., HARRIS, J., OCHIENG, S. A., AND WEBSTER, R. P., AND CARDÉ, R. T. 1984. The effects of BAKER, T. C. 2001. Electroantennogram responses of mating, exogenous juvenile hormone and a juvenile a parasitic wasp, Microplitis croceipes, to host-relat- hormone analog on pheromone titer, calling and ovi- ed volatile and anthropogenic compounds. Physiol. position in the omnivorous tea leafroller moth Entomol. 26: 69-77. (Platynota stultana). J. Insect. Physiol. 30: 113-118. POUZAT, J., AND IBEAS, D. N. 1989. Electrophysiological YANG, Y. Q., WAN, X. C., ZHENG, G. Y., AND GAO, X. H. investigations of sex pheromone reception and re- 2008. Preliminary study on the habit of sex behavior lease in Bruchidius atrolineatus. Physiol. Entomol. of Ectropis obliqua Prout. Chinese Agric. Sci. Bull. 14: 319-324. 24: 339-342 (in Chinese). PROSHOLD, F. I., KARPENKO, C. P., AND GRANAM, C. K. YAO, E. Y., LI, Z. M., LOU, Z. Q., AND SHANG, Z. Z. 1991. 1982. Egg production and oviposition in the tobacco Report on structural elucidation of sex pheromone budworm: Effect of age at mating. Ann. Entomol. components of a tea pest (Ectropis obliqua Prout). Soc. America 75: 51-55. Prog. Nat. Sci. 1: 566-569.

Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas 15

DISTRIBUTION OF PSEUDACTEON CURVATUS AND PSEUDACTEON TRICUSPIS (DIPTERA: PHORIDAE) IN ARKANSAS

JAKE M. FARNUM AND KELLY M. LOFTIN Department of Entomology, University of Arkansas, 319 Agriculture Bldg., Fayetteville, AR 72701

ABSTRACT

From 1995 to 2009, four Pseudacteon species were released in the U.S. with 3 species, P. curvatus, P. tricuspis, and P. obtusus released in Arkansas. To determine Pseudacteon establishment and expansion, sticky traps were used to monitor phorid ﬂy species at and near 10 release sites, in counties bordering neighboring states, and along regional transects. Pseu- dacteon ﬂies were captured in 16 Arkansas counties: Ashley, Chicot, Clark, Desha, Drew, Hempstead, Howard, Little River, Montgomery, Nevada, Perry, Phillips, Pike, Polk, Sevier, and Union. Pseudacteon curvatus was found in areas far from release sites, suggesting dispersal from neighboring states. The range of P. tricuspis evidently also expanded from its initial release sites in southern Arkansas.

Key Words: biological control, phorid ﬂy, Solenopsis invicta, parasitoid

RESUMEN

De 1995 hasta el 2009, cuatro especies de Pseudacteon fueron liberadas en los Estados Uni- dos con 3 de ellas, P. curvatus, P. tricuspis y P. obtusus, liberadas en el estado de Arkansas. Para determinar el establecimiento y expansión de Pseudacteon, se usaron trampas pegajo- sas para monitorear las especies de fóridos en y alrededor de los 10 sitios donde fueron liberadas, en condados fronterizos con los estados vecinos, y a lo largo de las lineas regionales. Las moscas Pseudacteon fueron capturadas en los siguientes 16 condados de Arkansas: As- hley, Chicot, Clark, Desha, Drew, Hempstead, Howard, Little River, Montgomery, Nevada, Perry, Phillips, Pike, Polk, Sevier y Union. Pseudacteon curvatus fue encontrada en áreas le- jos de los sitios donde fue liberada, que indica que se disperso de los estados vecinos. Eviden- temente, el rango de P. tricuspis tambien se expandió de los sitios donde fue liberada inicialamente en el sur de Arkansas.

Pseudacteon phorid flies parasitize and kill et al. 1997; Porter 2000; Folgarait et al. 2002, their fire ant host (Solenopsis invicta Buren, S. 2005). richteri Forel and their hybrid). Due to their high Pseudacteon flies use semiochemicals to locate host specificity (Folgarait et al. 2002; Gilbert & their ant hosts (Orr et al. 1997; Vander Meer & Morrison 1997; Morrison & Gilbert 1999; Porter Porter 2002; Morrison & King 2004; Chen & Fad- 1998a, 2000; Porter & Alonso 1999; Porter & Gil- amiro 2007) and then hover over the ants before a bert 2004; Vazquez et al. 2004), several Pseudac- rapid aerial attack (Morrison & Porter 2005). teon spp. have been introduced from South Amer- Within a period of an hour a female fly can make ica as classical biological control agents against up to 120 oviposition attempts (Morrison et al. imported fire ants. While internal development of 1997), before either tiring or being captured and the fly larvae eventually decapitates and kills in- killed by the ants (Porter 1998b). A single egg laid dividual ants (Porter et al. 1995a; Consoli et al. in the thorax develops through 3 instars before 2001), perhaps the most important effect is the decapitating the ant’s head, which is then used as disruption of the foraging behavior of the ants a pupal case (Pesquero et al. 1995; Porter et al. (Feener & Brown 1992; Orr et al. 1995; Porter et 1995a). al. 1995b; Mehdiabadi & Gilbert 2002), which The first release of P. tricuspis occurred in leads to a decrease in food uptake and a decline in Texas in 1995 (Gilbert 1996), and was unsuccess- colony health (Folgarait & Gilbert 1999). Four ful due to unfavorable conditions (Vazquez et al. Pseudacteon species were released in the U.S. 2006). The first successful release of P. tricuspis in from 1995 to 2009: P. curvatus Borgmeier, P. lito- northern Florida was in 1997 (Porter et al. 1999). ralis Borgmeier, P. obtusus Borgmeier, and P. tri- Pseudacteon spp. have been released in 11 south- cuspis Borgmeier. Each species fills a different ern states: Alabama, Arkansas, Florida, Georgia, niche, in terms of diurnal activity (Pesquero et al. Louisiana, Mississippi, North Carolina, Okla- 1996), seasonal occurrence (Fowler et al. 1995; homa, South Carolina, Tennessee, and Texas Folgarait et al. 2003) and preferred size of host (Porter et al. 1999; Graham et al. 2003; Williams (Campiolo et al. 1994); all complementing traits & deShazo 2004; Parkman et al. 2005; Thead et for control of S. invicta and S. richteri (Morrison al. 2005; Henne et al. 2007; Weeks & Callcott

16 Florida Entomologist 94(1) March 2011

2008). Pseudacteon curvatus, P. obtusus, and P. with Fluon®, to prevent ants from climbing up tricuspis have been released in Arkansas and in the trap or out of the Petri dish. adjacent states except Missouri (Clemons et al. One trap was placed per location by first locat- 2003; Weeks & Callcott 2008). ing a mound of substantial size and activity. The Dispersal rates for Pseudacteon spp. are vari- mound was then disturbed, by kicking it over cre- able and the majority of flies disperse only a few ating a flat surface on which the Petri dish was hundred meters (Morrison et al. 1999). However, placed. As ants climbed into the Petri dish, a few some flies in each generation are known to travel ants were crushed by hand to induce alarm pher- 2 to 4 km or more (Porter et al. 2004) and popula- omone release and the trap was placed in the cen- tions of flies expanding on average 74 km over a ter of the Petri dish. A brightly colored pin flag (91 period of 3.5 years (Porter 2010). cm long) was positioned alongside the trap. Glo- Monitoring of Pseudacteon spp. is achieved bal positioning system (GPS) coordinates were re- with a variety of methods: actively through direct corded for each location, and a corresponding collections of flies at disturbed ant colonies with number written on the lid of the trap. either manual or electrical stimulation (Barr & Traps were retrieved 20 to 24 h after place- Calixto 2005; Morrison & Porter 2005), or pas- ment. At time of retrieval, an 8 oz expanded poly- sively by trapping with a sticky trap (Puckett et styrene foam cup (Dart® No. 8J8) was placed over al. 2007). Until the current study, monitoring of the trap, and the lid was snapped in place. The these species in Arkansas had been concentrated cup prevented damage and contamination to the at and near release sites to determine Pseudac- sticky portion of the trap. teon establishment. The objective of this study was to determine the distribution of Pseudacteon Sampling at Release Locations spp. in Arkansas through wider-scale monitoring. Fourteen releases of Pseudacteon spp. were MATERIALS AND METHODS made in Arkansas from 1998 to 2009 (Table 1). Sampling along transects in the cardinal direc- Trap Design tions from the release sites began in 2002 (Pike Co.) and 2004 (Miller Co.). In 2009, the Miller, To determine presence or absence, passive Perry, Pike, and Sevier County release sites were trapping was used based on a modified version of revaluated for this study to confirm establishment a PTS (pizza tri-stand) sticky trap (Puckett et al. of Pseudacteon spp. A 1.6-km interval was used be- 2007). The modified Puckett trap (Fig. 1) con- tween traps along each transect placed in Pike, sisted of a pizza tri-stand (Polyking® No. 20431) Miller, and Sevier Counties, and at 0.8-km inter- covered in Tanglefoot®, glued to the flat side of an vals in Perry County. Transects were determined inverted portion cup (Dart® No. 100PC), which by locating roads and highways on aerial maps was hot glued to the underside of a plastic cup lid that radiated out from the release site in north, (Dart® No. 8JL). This device was placed in the south, east, and west directions. One modified center of the bottom half of a plastic Petri dish Puckett trap was placed at each sampling location. (150 by 15 mm). The surfaces of the portion cup and the inner lip of the Petri dish were coated Sampling Bordering Counties

Sampling was intended to monitor spread of established populations of Pseudacteon spp. in bordering counties/parishes of neighboring states of Louisiana (Henne et al. 2007), Mississippi (Thead et al. 2005), and Tennessee (Graham et al. 2003; Parkman et al. 2005; Weeks & Callcott 2008), and sampling packages were sent to Uni- versity of Arkansas Cooperative Extension Ser- vice County Agents in imported fire ant infested counties in eastern and southern Arkansas in the early summer of 2009. Each package contained four modified Puckett traps, latex gloves, and an information sheet. Instructions were to place the traps during the summer months, as previously described, at 4 locations within the county. De- ployed traps were returned from 4 bordering counties: Columbia, Lafayette, Phillips, and St. Francis. Geographic coordinates were recorded on the instruction sheet with the number of the cor- Fig. 1. A modified Puckett trap. responding trap.

Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas 17

TABLE 1. HISTORY OF PSEUDACTEON SPP. RELEASES IN ARKANSAS, 1998-2009.

Date County Species Number Released

1998, Jul-Aug 1 Drew P. tricuspis 1,350 2002, May2 Pike P. tricuspis 3,000 2002, Oct2 Bradley P. tricuspis 1,200 2003, Sep2 Bradley P. tricuspis 1,500 2004, May2 Miller P. tricuspis 2,580 2005, May2 Sevier P. tricuspis 4,300 2005, Oct2 Clark P. curvatus 8,5003 2006, Sep2 Perry P. curvatus 15,8164 2007, Sep2 Perry P. tricuspis 1,900 2008, Jun2 Jefferson P. tricuspis 120 2008, Oct2 Polk P. obtusus 3,2005 2009, May2 Grant P. curvatus 25,2966 2009, Jun2 Jefferson P. tricuspis 120 2009, Sep2 Garland P. tricuspis 360

1Release made by Lynne Thompson, University of Arkansas Monticello. 2Release made by Kelly Loftin, University of Arkansas Cooperative Extension Service. 3Number released based on 35.3 g S. invicta workers, 800/gram and 30% parasitism. 4Number released based on 65.9 g S. invicta workers, 800/gram and 30% parasitism. 5Number released based on 23.7 g S. invicta workers, 450/gram and 30% parasitism. 6Number released based on 105.4 g S. invicta workers, 800/gram and 30% parasitism.

Sampling Along Regional Transects Phorid Fly Identiﬁcations

Due to possible expansion of Pseudacteon spp. Upon retrieval, modified Puckett traps were from neighboring states, sampling transects were taken to the University of Arkansas laboratory identified and mapped in 3 regions: western, (Fayetteville, AR) and GPS coordinates logged southeastern, and southwestern Arkansas into Google Earth™. Traps were examined under (Fig. 2). Transects began at the state line of the a dissecting microscope for presence of Pseudac- adjacent state and traveled inward, with modi- teon spp. The sticky portion of the trap was fied Puckett traps placed at 3-mile intervals and sprayed with liquid degreaser (Goo Gone™) if fly GPS coordinates recorded for each trap. Sampling removal was necessary. Species were identified was conducted for 36 h along transects from each based on the morphology of the female ovipositor region between 19 Sep and 3 Oct 2009. (Porter & Pesquero 2001). Voucher specimens of P. curvatus, P. tricuspis, and P. obtusus were obtained from laboratory specimens and the phorid fly rearing facility in Gainesville, FL for identification of male and female flies. Male Pseudacteon spp. are currently difficult to identify through use of keys (Morrison et al. 1997; Porter & Pesquero 2001).

RESULTS AND DISCUSSION

Release Locations

Traps placed along transects radiating from the phorid fly release sites yielded surprising results. Of the 20 traps retrieved from Sevier County, no Pseudacteon spp. were caught. Traps from Miller, Perry, and Pike Counties captured Pseudacteon spp. Along the Miller County release site transects, P. curvatus was captured at 3 locations north of Texarkana, in the southeastern Fig. 2. Regional transects in Arkansas (SE1, SE2, part of Little River County (Fig. 3). However, this SE3, SW1, SW2, SW3, SW4, W1, W2, W3, W4) (ESRI was unexpected because the phorid fly species re- Inc. 2009). leased in Miller County in 2004 was P. tricuspis 18 Florida Entomologist 94(1) March 2011

vatus. Phillips County is the one of 2 counties in Arkansas (Crittenden Co. the other) currently known to have only S. richteri and no record of S. invicta (Robert Vander Meer, USDA-ARS, personal communication). Sampling locations in Phillips Co. were located on the western levee of the Mississippi River, northwest of Friars Point, MS (Fig. 3D), adjacent to counties in Mississippi and Tennessee with known S. richteri populations (Streett et al. 2006; Oliver et al. 2009). The high number of P. curvatus found on 1 trap (~172) implies sufficient colonies of S. richteri to support P. curvatus populations.

Regional Transects

A total of 176 modified Puckett traps were Fig. 3. Pseudacteon curvatus release sites in (A) placed along transects in western, southeastern, Clark Co., (B) Grant Co., (C) Perry Co., and 2009 cap- and southwestern Arkansas (Fig. 2). In the west- ture locations including (D) Phillips Co., Arkansas. ern region of transects, 28 traps contained P. cur- Pseudacteon tricuspis release sites in (E) Bradley Co., vatus, and 2 contained P. tricuspis (Fig. 3). Of the (F) Drew Co., (G) Garland Co., (H) Jefferson Co., (I) traps that captured P. tricuspis, one was located Miller Co., (J) Perry Co., (K) Pike Co., (L) Sevier Co., Ar- kansas and 2009 capture locations (ESRI Inc. 2009). 29.5 km west of the P. tricuspis release in Pike County, and the other was 46 km northwest of the release site. All 4 transects in this region included captures of P. curvatus. On the 2 northerly routes (Fig. 3, I on map). While possible, it is unlikely (W1 and W2), traps with P. curvatus were found that these captures resulted from cross contami- at regular intervals with the most easterly cap- nation of the 2 species at the rearing facility. Sev- ture 54 km from the Arkansas/Oklahoma state eral factors are in place to prevent cross contami- line. The remaining 2 transects (W3 and W4) also nation including parasitization of the ants in sep- had captures. Transect (W3) picked up P. curva- arate rooms and lack of available ants of pre- tus 13 km from the Arkansas/Oklahoma border. ferred host size. Furthermore, voucher sampling The 3 traps from the most southerly transect is conducted to insure parasitization by the cor- (W4) that contained P. curvatus were located near rect species (Amy Croft, Florida Department of the northern section of the Miller County release Agriculture and Consumer Services, personal site transect. This directional pattern of recover- communication). A plausible explanation could be ies supports the hypothesis of immigration from the expansion of P. curvatus from released and es- confirmed P. curvatus populations in Le Flore tablished populations in the bordering states of County, Oklahoma (Weeks & Callcott 2008) ap- Louisiana, Oklahoma, and Texas (Weeks & proximately 24 km northwest of Mena, Arkansas. Callcott 2008; Anne-Marie Callcott, USDA- Prevailing winds generally do not correlate with APHIS, personal communication). Similar results dispersion patterns (Morrison et al. 2000), as flies were observed along the northeast transect of the move close to the ground where wind is reduced. Pike County release site; capture of a Pseudac- Pseudacteon curvatus but not P. tricuspis was teon fly (P. curvatus) different from the species trapped along the southeastern region transects. originally released (P. tricuspis). Due to the prox- P. curvatus was found at 1 location in Union imity (5 km) with the Clark County release site County, north of El Dorado, and on 12 traps along (Fig. 3A), it is possible that this capture was from each of the other transects (SE2 and SE3) to the movement of P. curvatus from Clark County, al- east (Fig. 3). From the 2 easterly transects (SE2, though revaluation of Clark County was not con- SE3), traps with P. curvatus were found 91 km ducted in 2009. In Perry County, capture of P. north of the Arkansas/Louisiana border and 65 curvatus was recovered at the initial release site km west of the Arkansas/Mississippi border. This and at locations to the south (0.5 km) and east distribution in Arkansas is expected based on col- (2.2 km). lections of P. curvatus in bordering counties along the western side of Mississippi (Adams, Bolivar, Bordering Counties Claiborne, Coahoma, Desoto, Jefferson, Tunica, Warren, Washington, Wilkinson), and along Loui- Puckett traps from Columbia, Lafayette, and siana’s northern side (Claiborne, East Carroll, St. Francis Counties were devoid of Pseudacteon Morehouse, Union, West Carroll) (Anne-Marie flies, although all traps returned from Phillips Callcott, USDA-APHIS, personal communica- County on the Mississippi border captured P. cur- tion). Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas 19

Pseudacteon tricuspis was captured at 2 loca- necessary. With this modification, trapped fire tions along a transect (SW3) of the southwestern ants rather than fire ant midden serve as the region of Arkansas (Fig. 3). One of the traps was Pseudacteon fly attractant. located in the northern part of Nevada County, in The results suggest establishment and expan- the city of Prescott, and the other was 18 km to sion of P. curvatus from the release site in Perry the southeast. Their proximity to the Pike County County, and P. tricuspis from the release site in release site (Fig. 3K), 20 km northwest, may sug- Pike County. While limited, the current range of gest their origin, although P. curvatus appeared P. tricuspis in Arkansas appeared to be along a in no samples taken from the Pike County release narrow 86-km band stretching from northwest site transect. Weather conditions may have Pike County to south central Nevada County, and played a role in the lack of Pseudacteon spp. 25 km west of the release site. The current distri- present on the remaining 55 traps in this region. bution of P. curvatus in Arkansas suggested natu- Temperatures for the region on 3 Oct 2009 ranged ral movement from surrounding states. Despite from a low of 8°C to a high of 26°C. Temperatures extensive sampling across southern Arkansas, below 20°C inhibit activity of Pseudacteon flies many areas remained unsampled. Additional (Morrison et al. 1999; Wuellner et al. 2002). On 3 trapping would provide a better understanding of Oct 2009 temperatures were above 20°C for a 7-h the distribution of Pseudacteon spp. in Arkansas. period midday (11:00 AM to 6:00 PM CST) and below 20°C for the entire day of 4 Oct 2009, when ACKNOWLEDGMENTS the traps were retrieved. However, 2 traps located on the transect southeast of Prescott, AR collected We thank Anne-Marie Callcott of the USDA APHIS P. tricuspis. Weather data suggests similar condi- lab in Gulfport, MS for approving Pseudacteon spp. for tions for the Prescott, AR area, although slightly release, Amy Bass, Amy Croft, and Deborah Roberts of warmer temperatures (17°C) were recorded for an the Florida Department of Agriculture and Consumer additional 3.5 h on the morning of 4 Oct 2009. Services for assistance in supplying phorid flies, Ed Brown, Jerry Clemons, Randy Forst, Rex Herring, Mike Of the traps that collected Pseudacteon spp., McCarter, Shawn Payne, Doug Petty, Amy Simpson, no traps contained both species. Perhaps because Rebecca Thomas, Shaun Rhodes, Carla Vaught, Joe Ves- P. tricuspis is reliant on larger ants, P. curvatus is tal, and Danny Walker of the University of Arkansas able to establish more readily where P. curvatus Cooperative Extension Service for assistance placing and P. tricuspis overlap, and thus were not de- phorid fly traps, Michael Hamilton and Robert Goodson tected if present in low densities (Gilbert et al. for collecting imported fire ants for identification, and 2008). Another factor for the lack of both species Ricky Corder of the University of Arkansas Cooperative in the trap may be due to the modification of the Extension Service for assistance. Puckett trap. The 2 differences in the traps design were the attractant used and the placement of the REFERENCES CITED trap. Traps were placed on a disturbed mound with live ants whereas Puckett traps were placed BARR, C. L., AND CALIXTO, A. 2005. Electrical stimula- in an open area with midden (Puckett et al. 2007). tion of Solenopsis invicta (Hymenoptera: Formi- The Puckett trap as originally designed captured cidae) to enhance phorid fly, Pseudacteon tricuspis (Diptera: Phoridae), detection. Southwest. Entomol. more P. tricuspis than P. curvatus, although sea- 3: 165-168. sonal fluctuations could be a variable (Puckett et CAMPIOLO, S., PESQUERO, M. A., AND FOWLER, H. G. al. 2007). 1994. Size selective oviposition by phorid (Diptera: Phoridae) parasitoids on workers of the fire ant, So- CONCLUSION lenopsis saevissima (Hymenoptera: Formicidae). Etiologia. 4: 85-86. Passive Pseudacteon trapping with the modi- CHEN, L., AND FADAMIRO, H. Y. 2007. Behavioral and fied Puckett trap provided advantages over direct electroantennogram responses of phorid fly Pseu- collection from disturbed mounds. Because it is dacteon tricuspis (Diptera: Phoridae) to red imported fire ant Solenopsis invicta odor and trail phero- deployed quickly, multiple traps can be placed mone. J. Insect Behav. 20: 267-287. over a large area which allows continuous and si- CLEMONS, J., MCCARTER, M., GAVIN, J., PETTY, D., LOF- multaneous sampling (Puckett et al. 2007). The TIN, K., SHANKLIN, D., AND HOPKINS, J. 2003. Phorid manpower needed to achieve similar coverage us- fly releases in Arkansas: county agents perspective, ing observational sampling is cost prohibitive. pp. 40-41 In L. Greenberg, and C. Lerner [eds.], Proc. The addition of the protective cup to the original Imported Fire Ant Conference, Palm Springs, CA. design allowed longer storage time between col- CONSOLI, F. L., WUELLNER, C. T., VINSON, S. B., AND lection and examination of the trap, protection of GILBERT, L. E. 2001. Immature development of Pseu- the sticky portion, and reduced contamination. dacteon tricuspis (Diptera: Phoridae), an endopara- sitoid of the red Imported fire ants (Hymenoptera: The modification based on disturbed mounds and Formicidae). Ann. Entomol. Soc. America 94: 97-109. fluon-coated petri dishes rather than fire ant mid- ESRI, INC. 2009. ArcGIS 9.3.1. ESRI, Inc., Redlands, CA. den was advantageous in that maintenance of a FEENER, D. H., AND BROWN, B. V. 1992. Reduced forag- fire ant colony for collection of midden is no longer ing of Solenopsis geminata (Hymenoptera: Formi- 20 Florida Entomologist 94(1) March 2011

cidae) in the presence of parasitic Pseudacteon spp. opment of Pseudacteon flies (Diptera: Phoridae), (Diptera: Phoridae). Ann. Entomol. Soc. America 85: parasitoids of Solenopsis fire ants (Hymenoptera: 80-84. Formicidae). Environ. Entomol. 26: 716-724. FOLGARAIT, P. J., AND GILBERT, L. E. 1999. Phorid par- MORRISON, L. W., KAWAZOE, E. A., GUERRA, R., AND asitoids affect foraging under different availability GILBERT, L. E. 1999. Phenology and dispersal in of food in Argentina. Ecol. Entomol. 24: 1-11. Pseudacteon flies (Diptera: Phoridae), parasitoids of FOLGARAIT, P. J., BRUZZONE, O. A., PATROCK, R. J. W., Solenopsis fire ants (Hymenoptera: Formicidae). AND GILBERT, L. E. 2002. Developmental rates and Ann. Entomol. Soc. America 92: 198-207. host specificity for Pseudacteon parasitoids (Diptera: MORRISON, L. W., KAWAZOE, E. A., GUERRA, R. AND GIL- Phoridae) of fire ants (Hymenoptera: Formicidae) in BERT, L. E. 2000. Ecological interactions of Pseudac- Argentina. J. Econ. 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Environ. Entomol. 34: 308-316. L. E. 1995. Flies suppress fire ants. Nature. 373: FOWLER, H. G., PESQUERO, M. A., CAMPIOLO, S., AND 292-293. PORTER, S. D. 1995. Seasonal activity of species of ORR, M. R., SEIKE, S. H., AND GILBERT, L. E. 1997. For- fire ants in Brazil. Cientifica. 23: 367-371. aging ecology and patterns of diversification in dipter- GILBERT, L. E. 1996. Prospects of controlling fire ants an parasitoids of fire ants in South Brazil, genus with parasitoid flies the perspective from research Pseudacteon (Phoridae). Ecol. Entomol. 22: 305-314. based at Brackenridge Field Laboratory, pp. 77-92 PARKMAN, P., VAIL, K., RASHID, T., OLIVER, J., PEREIRA, In W. E. Cohen [ed.], Texas Quail Short Course II, R., PORTER, S. D., SHIRES, M., HAUN, G., POWELL, S., Texas Agricultural Extension Service, Texas A&M THEAD, L., AND VOGT, J. T. 2005. Establishment and University, Kingsville, TX. spread of Pseudacteon curvatus in Tennessee, pp. GILBERT, L. E., AND MORRISON, L. W. 1997. Patterns of 111-112 In D. J. Meloche [ed.], Proc. Imported Fire host specificity in Pseudacteon parasitoid flies Ant Conference, Gulfport, MS. (Diptera: Phoridae) that attack Solenopsis fire ants PESQUERO, M. A., PORTER, S. D., FOWLER, H. G., AND (Hymenoptera: Formicidae). Environ. Entomol 26: CAMPIOLO, S. 1995. Rearing of Pseudacteon spp. 1149-1154. (Diptera: Phoridae), parasitoids of fire ants (Sole- GILBERT, L. E., BARR, C. L., CALIXTO, A. A., COOK, J. L., nopsis spp.) (Hymenoptera: Formicidae). J. Appl. DREES, B. M., LEBRUN, E. G., PATROCK, R. J. W., Entomol. 119: 677-678. PLOWES, R. M., PORTER, S. D., AND PUCKETT, R. T. PESQUERO, M. A., CAMPIOLO, S., FOWLER, H. G., AND 2008. Introducing phorid fly parasitoids of red im- PORTER, S. D. 1996. Diurnal patterns of oviposition- ported fire ant workers from South America to Tex- al activity in two Pseudacteon fly parasitoids of fire as: outcomes vary by region and by Pseudacteon spe- ants (Hymenoptera: Formicidae). Florida Entomol. cies released. Southwest. Entomol. 33: 15-29. 79: 455-456. GRAHAM, L. C., PORTER, S. D., PEREIRA, R. M., DOR- PORTER, S. D. 1998a. Host specific attraction of Pseudac- OUGH, H. D., AND KELLEY, A. T. 2003. Field releases teon flies (Diptera: Phoridae) to fire ant colonies in of the decapitating phorid fly Pseudacteon curvatus Brazil. Florida Entomol. 81: 423-429. (Diptera: Phoridae) for control of imported fire ants PORTER, S. D. 1998b. Biology and behavior of Pseudac- (Hymenoptera: Formicidae) in Alabama, Florida, teon decapitating flies (Diptera: Phoridae) that par- and Tennessee. Florida Entomol. 86: 334-339. asitize Solenopsis fire ants (Hymenoptera: Formi- HENNE, D. C., JOHNSON, S. J., AND PORTER, S. D. 2007. cidae). Florida Entomol. 81: 292-309. Status of the fire ant decapitating fly Pseudacteon PORTER, S. D. 2000. Host specificity and risk assessment tricuspis (Diptera: Phoridae) in Louisiana. Florida of releasing the decapitating phorid fly Pseudacteon Entomol. 90: 565-569. curvatus as a classical biocontrol agent for imported MEHDIABADI, N. J., AND GILBERT, L. E. 2002. Colony fire ants. Biol. Control. 19: 35-47. level impacts of parasitoid flies on fire ants. Proc. Bi- PORTER, S. D. 2010. Distribution of the Formosa strain ol. Sci. 269: 1695-1699. of the fire ant decapitating fly Pseudacteon curvatus MORRISON, L. W., AND GILBERT, L. E. 1999. Host speci- (Diptera: Phoridae) three and a half years after re- ficity in two additional Pseudacteon spp. (Diptera: leases in North Florida. Florida Entomol. 93: 107- Phoridae), parasitoids of Solenopsis fire ants (Hy- 112. menoptera: Formicidae). Florida Entomol. 82: 404-409. PORTER, S. D., AND ALONSO, L. E. 1999. Host specificity MORRISON, L. W., AND KING, J. R. 2004. Host location of fire ant decapitating flies (Diptera: Phoridae) in behavior in a parasitoid of imported fire ants. J. In- laboratory oviposition tests. J. Econ. Entomol. 92: sect Behav. 17: 367-383. 110-114. MORRISON, L. W., AND PORTER, S. D. 2005. Phenology PORTER, S. D., AND GILBERT, L. E. 2004. Assessing host and parasitism rates in introduced populations of specificity and field release potential of fire ant de- Pseudacteon tricuspis, a parasitoid of Solenopsis in- capitating flies (Phoridae: Pseudacteon), pp. 152-176 victa. Biocontrol. 50: 127-141. In R. G. Van Driesche and R. Reardon [eds.], Assess- MORRISON, L. W., DALLAGLIO-HOLVORCEM, C. G., AND ing Host Ranges for Parasitoids and Predators Used GILBERT, L. E. 1997. Oviposition behavior and devel- for Classical Biological Control: A Guide to Best Farnum & Loftin: Distribution of P. curvatus and P. tricuspis in Arkansas 21

Practice. FHTET-2004-03, USDA Forest Service, Formicidae) populations in Mississippi. Florida En- Morgantown, WV. tomol. 89: 91-92. PORTER, S. D., AND PESQUERO, M. A. 2001. Illustrated THEAD, L. G., VOGT, J. T., AND STREETT, D. A. 2005. Dis- key to Pseudacteon decapitating flies (Diptera: persal of the fire ant decapitating fly, Pseudacteon Phoridae) that attack Solenopsis saevissima com- curvatus (Diptera: Phoridae) in Northeast Mississip- plex fire ants in South America. Florida Entomol. 84: pi. Florida Entomol. 88: 214-216. 691-699. VANDER MEER, R. K. AND PORTER, S. D. 2002. Fire ant, PORTER, S. D., PESQUERO, M. A., CAMPIOLO, S., AND Solenopsis invicta, worker alarm pheromones at- FOWLER, H. G. 1995a. Growth and development of tract Pseudacteon phorid flies, pp. 77-80 In Proc. Im- Pseudacteon phorid fly maggots (Diptera: Phoridae) ported Fire Ant Conference, Athens, GA. in the heads of Solenopsis fire ant workers (Hy- VAZQUEZ, R. J., PORTER, S. D, AND BRIANO, J. A. 2004. menoptera: Formicidae). Environ. Entomol. 24: 475- Host specificity of a biotype of the fire ant decapitat- 479. ing fly Pseudacteon curvatus (Diptera: Phoridae) PORTER, S. D., VANDER MEER, R. K., PESQUERO, M. A., from northern Argentina. Environ. Entomol. 33: CAMPIOLO, S., AND FOWLER, H. G. 1995b. Solenopsis 1436-1441. (Hymenoptera: Formicidae) fire ant reactions to at- VAZQUEZ, R. J., PORTER, S. D., AND BRIANO, J. A. 2006. tacks of Pseudacteon flies (Diptera: Phoridae) in Field release and establishment of the decapitating southeastern Brazil. Ann. Entomol. Soc. America 88: fly Pseudacteon curvatus on red imported fire ants in 570-575. Florida. Biocontrol. 51: 207-216. PORTER, S. D., NOGUEIRA DE SÁ, L. A., FLANDERS, K., WEEKS, R. D., AND CALLCOTT, A.-M. A. 2008. USDA- AND THOMPSON, L. 1999. Field releases of the decap- APHIS and consortium efforts to establish phorid itating fly, Pseudacteon tricuspis, p. 102 In L. Reeves flies (Pseudacteon spp.) in imported fire ant (Sole- [ed.], Proc. Imported Fire Ant Conference, Charles- nopsis spp.) populations in the U.S. and Puerto Rico, ton, SC. pp. 87-89 In Proc. Imported Fire Ant Conference, PORTER, S. D., NOGUEIRA DE SÁ, L. A., AND MORRISON, Charleston, SC. L. W. 2004. Establishment and dispersal of the fire WILLIAMS, D. F., AND DESHAZO, R. D. 2004. Biological ant decapitating fly Pseudacteon tricuspis in North control of fire ants: an update on new techniques. Florida. Biol. Control. 29: 179-188. Ann. Allergy Asthma Immunol. 93: 15-22. PUCKETT, R. T., CALIXTO, A., BARR, C. L., AND HARRIS, WUELLNER, C. T., DALL’AGLIO-HOLVEROCEM, C. G., M. 2007. Sticky traps for monitoring Pseudacteon BENSON, W. W., AND GILBERT, L. E. 2002. Phorid fly parasitoids of Solenopsis fire ants. Environ. Ento- (Diptera: Phoridae) oviposition behavior and fire ant mol. 36: 584-588. (Hymenoptera: Formicidae) reaction to attack dif- STREETT, D. A., FREELAND, T. B., AND VANDER MEER, R. fers according to phorid species. Ann. Entomol. Soc. K. 2006. Survey of imported fire ant (Hymenoptera: America 93: 257-266.

22 Florida Entomologist 94(1) March 2011

MINI-ASPIRATOR: A NEW DEVICE FOR COLLECTION AND TRANSFER OF SMALL ARTHROPODS TO PLANTS

MAHMUT DOGRAMACI˘ 1*, JIANJUN CHEN2, STEVEN P. ARTHURS1 CINDY L. MCKENZIE3, FABIELI IRIZARRY,1 KATHERINE HOUBEN1, MARY BRENNAN1 AND LANCE OSBORNE1 1University of Florida, Department of Entomology and Nematology, Mid-Florida Research and Education Center, Apopka, FL 32703, USA

2University of Florida, Department of Environmental Horticulture, Mid-Florida Research and Education Center, Apopka, FL 32703, USA

3U.S. Horticultural Research Laboratory, ARS-USDA, Fort Pierce, FL 34945, USA

ABSTRACT

The process of collecting and/or infesting plants with a designated number of small arthropods in biological experiments is tedious and laborious. We developed a modified mini-aspirator, powered with a vacuum pump and fitted with a specially adapted (removable) collection vial to reduce the handling effort. The efficiency of the mini-aspirator was tested with the chilli thrips, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), a predatory mite, Amblyseius (= Neoseiulus) cucumeris (Oudemans) (Acari: Phytoseiidae), and the insidious flower bug, Orius insidiosus (Say) (Heteroptera: Anthocoridae). Using the mini-aspirator, operators collected 10 A. cucumeris mites and 10 S. dorsalis thrips and transferred them onto pepper plants in 43 s and 37 s, respectively, compared with 639 and 229 s, respectively, using a camel’s hair brush as a conventional method. The use of the mini-aspirator for collecting A. cucumeris predatory mites and S. dorsalis thrips and infesting pepper plants with them represents a 15-fold and 6-fold time saving, respectively. Collection of 10 O. insidiosus flower bugs took 20 s with the mini-aspirator compared with 30 s when an unmodified aspirator was used. Proportionally, the amount of time saved with the mini-aspirator for the handling of O. insidiosus flower bugs was minimal compared with the timesavings when handling S. dorsalis thrips and the A. cucumeris predatory mites with the mini-aspirator. Additionally, the mini-aspirator can be fitted with a battery-powered Mini-Vac, which makes it portable for field applications, such as in sampling field populations when screening for pesticide resistant individuals.

Key Words: mini-aspirator, Neoseiulus cucumeris, Scirtothrips dorsalis, Orius insidiosus, manual infestation

RESUMEN

El proceso de recolectar y/o infestar plantas con un cierto número de artrópodos pequeños en experimentos biológicos es tedioso y laboroso. Desarrollamos una mini-aspiradora modificada, que funciona mediante una bomba de succión especialmente montada con un frasco de recolección especialmente adaptado (desmontable) para reducir el esfuerzo de manejo. La eficiencia de la mini-aspiradora fue probada con el trips de chile, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), un ácaro depredador, Amblyseius (=Neoseiulus) cucumeris (Oudemans) (Acari: Phytoseiidae), y el chinche pirata diminuto, Orius insidiosus (Say) (He- teroptera: Anthocoridae). Usando la mini-aspiradora, los operadores recolectaron y transfe- rieron 10 ácaros depredadores y trips de chile a plantas de chile en 43 y 37 segundos, comparado con 638 y 229 segundos usando el metodo convencional con una brocha de pintar. El uso de la mini-aspiradora para mover e infestar plantas con ácaros depredadores y trips representa un ahorro de 15 y 6 veces, respectivamente. La recolección de 10 chinches piratas diminutos tomó 20 segundos con la mini-aspiradora comparada con 30 segundos cuando se uso una aspiradora no modificada. El tiempo ahorrado proporcionalmente fue mínimo comparado con la recolección de trips de chile y los ácaros depredadores con la mini-aspiradora. Además, la mini-aspiradora puede ser mantada con una mini-bomba de succionar de bate- ría, que la hace portable para aplicaciones en el campo, como en la evaluación de resistencia de plaguicidas en poblaciones de campo.

Manual collection and infestation of small arthropods. Thrips and predatory mites are exam- thropods (<2 mm) can be labor intensive and cum- ples of small arthropods used by scientists in nu- bersome and result in injury to the handled ar- merous experiments (Mound & Palmer 1981;

Dogramaci˘ et al.: Mini-aspirator for Handling Small Arthropods 23

Chiu et al. 1991; Tatara & Furuhushi 1992; Ts- of the mini-aspirator with a paintbrush and com- chuchiya et al. 1995; Bournier 1999; Seal et al. mercial aspirator for collecting and releasing the 2006; Arthurs et al. 2009). Scirtothrips dorsalis chilli thrips, Scirtothrips dorsalis Hood and 2 of Hood, chilli thrips, is one of the smallest thrips its natural enemies, a predatory mite, Neoseiulus species with adults ranging from 1.5-2.0 mm. The cucumeris (Oudemans), and the flower bug, Orius S. dorsalis adult moves rapidly, and may jump or insidiosus (Say). fly when disturbed. Immature stages of S. dorsalis, especially first instars, are very small (<1 mm) MATERIALS AND METHODS and have fragile easily injured bodies. Thus, an important requirement for studying small arthro- Mini-aspirator pods is to have a reliable method for collecting and releasing designated numbers of individuals The mini-aspirator was built from clear 6.35- without harm. mm diam vinyl tubing fitted with a 1-mL filtered Three methods are used for infesting plants pipette tip (VWR International, West Chester, with thrips. One is manual infestation of arthro- PA). The intake tubing opening was reduced by pods with a soft-bristled camel’s hair brush using an adaptor to attach a 200-µL pipette tip, (Cloyd & Sadof 1998). This method has been which facilitated the collection of individual used widely but is labor-intensive and time-con- small arthropods (Figs. 1A and 1B). The modi- suming. For instance, Cloyd et al. (2001) re- fied mini-aspirator was powered by an electrical ported that infesting 50 plants each with 10 laboratory vacuum pump (Rocker vacuum pump, adult western flower thrips (WFT), Frankliniella Rocker Scientific Co., Ltd., Kaohsiung, Taiwan) occidentals (Pergande), required a technician 3.5 (Fig. 1A). To collect S. dorsalis, an infested leaf h. In addition, the process of mechanical transfer was placed under a stereomicroscope (Fig. 1C) involved a risk of injury to the specimen. The and the desired number of thrips was captured second method involves placement of plants in a in the collection vial for transfer onto plants. The location where thrips are known to occur in or- collection vial was removed from the mini- aspi- der to allow a natural population of the pest to rator and attached to a plant with a hair clip to build-up on the test plants. Although this allow the voluntary dispersal of the thrips onto method is less cumbersome than manual infes- the plant (Fig. 1D). To make the mini-aspirator tation, the number of thrips transferred onto portable, we integrated the mini-aspirator with test plants cannot be accurately regulated, a Mini-Vac (MV Instrument, Glendale, CA which introduces variation among test plants. 91205; http://www.mini-vac.com/index02.html) The third method is the use of a commercial (Fig. 1E). However, to compensate for the re- mouth operated aspirator purchased from Bio- duced suction power of the Mini-Vac, the pipette Quip, Rancho Dominquez, CA. An improvement tip filter was replaced with fine woven nylon fab- to the mouth-operated aspirator was reported by ric. The assembly of the pipette tip, collection Cloyd et al. (2001), who developed a “Small In- vial, filter and vacuum tube is shown in Fig. 2. sect Aspirator” for collecting WFT. The latter in- Such integration of the “mini-aspirator” with the volved the use of an aspirator from BioQuip at- battery powered Mini-Vac made the system por- tached to a battery operated ‘Mini-Vac’ (MV In- table for field use. strument, Glendale, CA 91205; http://www.mini- vac.com/index02.html). Use of the small insect Arthropods aspirator is attended with operational difficul- ties similar to those encountered with the use of Scirtothrips dorsalis specimens were obtained regular aspirators, in particular the collection of from a colony that originated from rose plants in extraneous materials. In our studies, we found Winter Park, FL. The colony was maintained on that the wide suction tubes of the aspirators col- cotton plants, Gossypium hirsutum, ‘Deltapine lect too many extraneous materials along with 493 Conventional’. The health of the colony was thrips (e.g., other organisms and plant debris) maintained by periodically introgressing thrips and sometimes causes physical damage to from naturally infested plants. Commercially thrips. Additionally, in our experience the design available thrips predators (a predatory mite, N. of the collection vial does not allow the thrips to cucumeris (Oudemans), and the insidious flower be released easily following collection. bug, Orius insidiosus (Say)) were obtained from In order to overcome some of the design limita- Koppert Biological Systems, Berkel en Rodenrijs, tions of previous small arthropod handling de- The Netherlands. vices, we developed a “mini-aspirator” that can be powered either with a laboratory vacuum pump Plant Material or with a small portable vacuum pump and fitted with a specially adapted and removable collection The infestation methods were tested on sweet vial that allows rapid transfer of the collected ar- pepper plants, Capsicum annum L. Pepper seeds thropods onto plants. We compared the efficiency were germinated on moist filter papers inside

24 Florida Entomologist 94(1) March 2011

Fig. 1. Novel minute arthropod infestation apparatus and its use. A. The complete system in use; B. The mini- aspirator small arthropod collector; C. Collecting S. dorsalis from a leaf; D. Mini-aspirator along with collected arthropods attached to a plant to allow dispersal; E. The portable mini-aspirator consisting of clear 6.35 mm diam vinyl tubing ﬁtted with a 1-mL pipette tip and nylon cloth ﬁlter connected with an adaptor to a 200-µL pipette tip; with suction provided by a Mini-Vac.

Petri dishes. Germinated seeds were transferred Collection and Infestation of Small Arthropods with the to seedling trays. Seedlings each with 4-6 fully ex- Conventional Camel’s Hair Brush panded leaves were planted into 15 cm diam pots. Pepper plants at >10 leaf stage were used for the Collecting chilli thrips and infesting plants arthropod infestation experiments. with them. Either 10 or 20 chilli thrips were cap-

Dogramaci˘ et al.: Mini-aspirator for Handling Small Arthropods 25

the mini-aspirator as described above. The mini-aspirator along with the collected S. dorsalis was attached to a pepper plant leaf with a hair clip in a manner that allowed thrips to distribute themselves on plant leaves (Fig. 1D). The time required for collection and release (the latter being the time required to attach the mini-aspirator along with the collected S. dorsalis to a pepper plant leaf) S. dorsalis was recorded. This was repeated 15 times each for groups of 10 and 20 S. dorsalis. Collecting N. cucumeris and releasing them onto plants. The entire contents of the package with N. cucumeris predatory mites were emptied into a Petri dish (15 cm diam). To separate N. cucumeris mites from the packaging material, a filter paper was placed in the Petri dish and the Fig. 2. Assembly of the pipette tip, collection vial, fil- closed Petri dish was agitated gently several ter and vacuum line of the mini-aspirator. times. Under a stereomicroscope, either 10 or 20 N. cucumeris mites on the filter paper were collected with the mini-aspirator. The collection tured with a moistened camel’s hair brush. Each tube with the N. cucumeris mites was attached of these thrips was then placed onto a pepper to a pepper leaf as described above (Fig. 1D). The plant leaf. The number of seconds needed to cap- seconds required for collection and release (the ture and transfer the above designated number of latter being the time required to attach the col- thrips was recorded. This was repeated 8 times lector containing the collected N. cucumeris to a each for groups of 10 or 20 chilli thrips. pepper plant leaf were recorded. This was re- Sorting and releasing N. cucumeris. The entire peated 15 times each for groups of 10 and 20 N. content of the package containing N. cucumeris cucumeris. and substrate was emptied into a Petri dish (15 Collecting adult O. insidiosus flower bugs and cm diam) lined with a filter paper. To separate N. releasing them onto plants. The mini-aspirator cucumeris predatory mites from the substrate, was modified by enlarging the opening of the pi- the closed Petri dish was agitated gently several pette tip to accommodate the bugs. The bugs were times. Under a stereomicroscope each predatory collected from among the vermiculate particles mite was collected individually from the filter pa- scattered on a board. Orius insidiosus in groups of per and placed onto a pepper plant with a camel’s either 10 or 20 were collected with the mini-aspi- hair brush. Groups of either 10 or 20 N. cucumeris rator. The collection tube containing the bugs was were placed on a plant. The number of seconds re- then attached to a leaf with a hair clip as de- quired to collect and release either 10 or 20 pred- scribed previously. The number of seconds re- atory mites was recorded. This was repeated 8 quired to collect and release the bugs was re- times each for groups of 10 and 20 mites. corded. This was repeated 15 times each for Collecting and releasing O. insidiosus flower groups of 10 and 20 O. insidiosus. bugs on pepper plants. Orius insidiosus flower bug adults in groups of either 10 or 20 were col- Statistical Analysis lected from a purchased colony with a BioQuip aspirator. The contents of the package (vermiculite The efficiency (time) of the mini-aspirator was substrate and bugs) were emptied onto a board compared with the conventional method for col- and bugs that crawled on the board were cap- lecting and releasing each of the three arthro- tured with the BioQuip aspirator. Each collection pods. The study was repeated 8 and 15 times for vial containing the O. insidiosus was placed at the the conventional and new method, respectively. base of a pepper plant that had been infested with Data were analyzed by ANOVA procedure (PROC thrips to allow the bugs to exit the vial and dis- GLM) and means were separated by Fisher’s pro- tribute onto the plant. The number of seconds re- tected LSD test for all the experiments (SAS In- quired to collect and to release either 10 or 20 stitute 1997). bugs was recorded. This was repeated 8 times each for groups of 10 or 20 O. insidiosus. RESULTS

Use of the Mini-aspirator to Collect Small Arthropods The collection of 10 adult thrips from cotton and Transfer Them onto Plants leaves and their release onto a pepper plant using the mini-aspirator took 37 s compared with Collecting S. dorsalis thrips and infesting plants 229 s with the camel’s hair brush method. This with them. Ten or 20 S. dorsalis were collected with represented a 6-fold reduction in infestation

26 Florida Entomologist 94(1) March 2011

= 0.0001 and F1,21 = 379.00; P = 0.0001), respectively. No predatory mite was found to be damaged by the mini-aspirator. The time required to collect and release adult O. insidiosus with a BioQuip aspirator was less than time needed for collecting and releasing by the paintbrush method. However, collecting and releasing 10 and 20 O. insidiosus with the commercial aspirator required 31 and 58 s, but only 20 and 32 s with the mini-aspirator (Figs. 3 and Fig. 3. Comparison of seconds needed for collecting 4). These time differences between the 2 methods and infesting 10 O. insidiosus, S. dorsalis and N. cucum- of collection and release were also signiﬁcantly eris onto plants with conventional methods (camel’s different (F1,21 = 43.19; P= 0.0001) (F1,21 = 33.52; P hair brush or commercial aspirator) and the developed = 0.0001), respectively. mini-aspirator.

DISCUSSION time when the mini-aspirator method was used The mini-aspirator reduces the time required (Fig. 3). The difference between the 2 methods to collect and release a designated number of

was highly signiﬁcant (F1,21 = 325.66; P < 0.0001). small arthropods to the plants. The camel’s hair The time required to place 20 thrips on a plant brush method in addition to being very slow can with the mini-aspirator was 42 s, compared with cause injury, especially to soft-bodied small ar- 461 s by using the camel’s hair brush method, thropods. An operator using the mini-aspirator which represented an 11-fold reduction in infes- can collect and transfer small arthropods using tation time when the mini-aspirator was used controlled air intake velocity, which minimizes in-

(F1,21 = 974.68; P < 0.0001) (Fig. 4). With the new jury to the collected arthropods. To avoid injury to method, the amount of time to collect and re- thrips, the vacuum was adjusted to the minimum lease per thrips decreased as number of in- sufficient to collect thrips. Although injury to creased. Such a reduction was not observed with thrips was not investigated in detail, thrips col- the paintbrush method. Some thrips adults were lected with the mini-aspirator were checked un- also observed to be injured by the commercial as- der a stereomicroscope and no serious thrips in- pirator (Fig. 5-A). We also observed some dam- jury was observed. aged thrips and reduced thrips activity when The mini-aspirator is different from the com- they were handled with a commercial aspirator monly available commercial aspirators, which (Fig. 5-B). employs larger diameter removable glass or The collection and release of 10 and 20 pred- plastic collecting vials. Initially, we used the atory mites required 639 and 1154 s with a commercial aspirators but experienced difficul- moistened camel’s hair brush compared with 43 ties in transferring the designated numbers of and 90 s, respectively, with the mini-aspirator arthropods. The commercial aspirator available (Figs. 3 and 4). The use of the mini-aspirator to us has a 4-mm diam collection tube that could saved 15-fold and 13-fold more time in collect- not be adjusted for the selective collection of in- ing and releasing 10 and 20 predatory mites, re- dividual thrips; and this is a significant limita- spectively, compared with the use of the paint- tion when working with mixed colonies of in- brush method. The difference between the two sects. Another advantage of the mini-aspirator is methods was highly significant (F1,21 = 284.86; P that unlike the traditional collection vials, the smaller removable and disposable collection tubes can easily be attached to small leaves or plant stems without disturbing the insects inside the tube. The mini-aspirator developed in this study can be powered with a Mini-Vac, which makes the technique portable for field applications. The technique may be used to quickly census wild populations for laboratory testing or for use in insecticide efficacy trials. The mini-aspirator could also be adapted for quick pesticide resistance or efficacy trials in the field (Rueda and Shelton Fig. 4. Comparison of seconds needed for collecting and infesting 20 O. insidiosus, S. dorsalis and N. cucum- 2003). The inside of the collection tube of the eris onto plants with conventional methods (camel’s mini-aspirator could be treated with pesticides of hair brush or commercial aspirator) and the developed interest or a treated leaf disk could be placed in mini-aspirator. the mini-aspirator before collecting small arthro- Dogramaci˘ et al.: Mini-aspirator for Handling Small Arthropods 27

Fig. 5. Illustration of an injured S. dorsalis when collected with a regular made aspirator. A, Injury to abdomen of S. dorsalis. B, Injury to wing of S. dorsalis.

pods, and then collected arthropods would be held CLOYD, R. A., WARNOCK, D. F., AND HOLMES, K. 2001. for a fixed exposure period to quantify pesticide Technique for collecting thrips for use in insecticide efficacy. This kind of monitoring would be helpful efficacy trials. Hort. Sci. 36: 925-926. to confirm pest susceptibility to pesticides before CLOYD, R. A., AND SADOF, C. S. 1998. Flower quality, their wide area applications. flower numbers, and Western flower thrips density on transversal daisy treated with granular insecticides. Hort. Tech. 8: 567-570. ACKNOWLEDGMENTS MOUND, L. A., AND PALMER, J. M. 1981. Identification, distribution and host plants of the pest species of We are grateful to Kenneth E. Savage, Russell D. Scirtothrips (Thysanoptera: Thripidae). Bull. Ento- Caldwell, and Younes Belmourd for help during this mol. Res. 71: 467-479. study. The study was supported by the USDA Tropical RUEDA, A., AND SHELTON, A. M. 2003. Development and and Subtropical Agricultural Research (T-STAR) Pro- evaluation of a thrips insecticide bioassay system for gram, American Floral Endowment and the USDA-ARS monitoring resistance in Thrips tabaci. Pest. Man- Floriculture and Nursery Research Initiative. age. Sci. 59: 553-558. SAS INSTITUTE. (1997) SAS User’s Guide. SAS Institute REFERENCES CITED Cary, North Carolina. SEAL, D. R., CIOMPERLIK, M. A., RICHARDS, M. L., AND ARTHURS, S., MCKENZIE C. L., CHEN, J., DOGRAMACI˘ , KLASSEN, W. 2006. Distribution of chilli thrips, Scir- M., BRENNAN, M., HOUBEN, K., AND OSBORNE, L. tothrips dorsalis (Thysanoptera: Thripidae), in pep- 2009. Evaluation of Neoseiulus cucumeris and Ambl- per fields and pepper plants on St. Vincent. Florida yseius cucumeris (Acari: Phytoseiidae) as Biological Entomol. 89: 311-320. Control Agents of Chilli Thrips, Scirtothrips dorsalis TATARA, A., AND FURUHASHI, K. 1992. Analytical study (Thysanoptera: Thripidae) on Pepper. Biol. Cont. 49: on damage to Satsuma mandarin fruit by Scirto- 91-96 thrips dorsalis Hood (Thysanoptera: Thripidae), BOURNIER, J. P. 1999. Two Thysanoptera, new cotton with particular reference to pest density. Japanese J. pests in Cote d’Ivorie. Annales de la Societe Ento- Appl. Entomol. 36(4): 217-223. mologique de France 34: 275-281. TSCHUCHIYA, M., MAUI, S., AND KUBOYAMA, N. 1995. CHIU, H. T., SHEN, S. M., AND WU, M. Y. 1991. Occur- Color attraction of yellow tea thrips (Scirtothrips rence and damage of thrips in citrus orchards south- dorsalis Hood). Japanese J. App. Entomol. Zool. 39: ern Taiwan. Chinese J. Entomol. 11: 310-316. 299-303.

28 Florida Entomologist 94(1) March 2011

TAXONOMY OF KOREAN LESTEVA WITH A DESCRIPTION OF A NEW SPECIES (COLEOPTERA: STAPHYLINIDAE: OMALIINAE)

TAE-KYU KIM AND KEE-JEONG AHN Department of Biology, Chungnam National University, Daejeon 305-764, Republic of Korea

ABSTRACT

A taxonomic study of the genus Lesteva Latreille in Korea is presented. Four species including a new species, Lesteva coreana sp. nov., are recognized. Three species, L. cordicollis Motschulsky, L. distincta Watanabe and L. miyabi Watanabe, are new to the Korean fauna, and L. plagiata Sharp previously recorded from Korea is a misidentiﬁcation of L. miyabi. A key and a comparison of morphological features of Korean Lesteva species with illustrations of the diagnostic features are provided.

Key Words: Staphylinidae, Omaliinae, Lesteva, new species, Korea

RESUMEN

Se presenta un estudio taxonómico del género Lesteva Latreille en Corea. Se reconocen cuatro especies incluyendo una nueva especie, Lesteva coreana sp. nov. Tres especies, L. cordicollis Motschulsky, L. distincta Watanabe y L. miyabi Watanabe son nuevas para la fauna de Corea y se determinó que L. plagiata Sharp, anteriormente registrada en Corea, fue basado sobre una identiﬁcación equivocada de L. miyabi. Se proveen una clave y una comparisión de las características morfológicas de las especies de Lesteva en Corea con ilustraciones de las características diagnósticas.

The genus Lesteva Latreille (tribe Anthoph- quadrate; eyes convex, large, with pubescence be- agini Thomson) is composed of 104 species dis- tween facets; ocelli distinct; temple round; vertex tributed in the Holarctic and Oriental regions with 2 longitudinal depressions; gular sutures (Watanabe 1990, 2004, 2005; Herman 2001; separated, divergent posteriorly; mandibles sub- Smetana 2004; Li 2005; Sharvrin et al. 2007). In triangular, curved inwardly with distinct internal East Asia, 19 and 15 species of the genus are re- teeth, mola distinct; maxillary palpomere 4 as ported in Japan and in China, respectively (Wa- wide and about 4.0 times as long as palpomere 3; tanabe 1990, 2004; Smetana 2004; Li 2005). antenna extending to near middle of elytra. Lesteva plagiata Sharp recorded by Cho et al. Pronotum convex, widest at anterior third or (2002) in Korea is a misidentification of L. miyabi fourth, more narrowed posteriorly than anteri- Watanabe. orly; mesoventrite with longitudinal carina along Members of Lesteva occur in montane riparian midline and several foveae on each side; elytra areas and are often found in moss or wet litter, flat, broader than pronotum, expanded posteri- sometimes in caves. Adults and larvae are preda- orly; legs long and slender, protarsus thin in both tors (Steel 1970; Newton et al. 2001). We have sexes. Abdomen broad, flat and abruptly nar- studied 20 specimens of L. cordicollis Motschul- rowed posteriorly, tergites IV-V with a pair of sky, 28 specimens of L. coreana sp. nov., 60 spec- wing folding patches. imens of L. distincta Watanabe, and 125 specimens of L. miyabi. Lesteva cordicollis Motschulsky, 1860 In this paper, we report 4 Lesteva species (L. cordicollis, L. coreana sp. nov., L. distincta, and (Figs. 1, 5, 9, 13, 17, 19, 21-22) L. miyabi) from Korea. A key, habitus photographs, and the illustrations of diagnostic fea- Lesteva cordicollis Motschulsky, 1860: 549; Sharvrin, tures are provided. All specimens are deposited in 2001: 191. the Chungnam National University Insect Collec- Description. Body (Fig. 1) length 3.6-4.0 mm tion (CNUIC), Daejeon, Korea. (head to abdominal end), covered with fine Genus Lesteva Latreille, 1797 punctures and pubescence, brown to dark brown and glossy; head and pronotum black, Lesteva Latreille, 1797: 75. mouthparts, antennae and legs light brown. Tevales Casey, 1894: 398. Synonymized by Steel, 1952: 9. Head about 1.4 times as wide as long; eye about 3.3 times as long as temple; antennae (Fig. 5) Diagnosis. Body ovoid and flattened, densely pubescent, reaching middle of elytra, 4th an- pubescent, covered with punctures. Head sub- tennomere 2.1 times as long as wide, 8th anten-

Kim & Ahn: Korean Lesteva Species 29

tures and pubescence, brown to dark brown and glossy; mouthparts, antennae and legs light brown. Head about 1.4 times as wide as long; eye about 3.1 times as long as temple; antennae (Fig. 6) pubescent, reaching middle of elytra, 4th antennomere 2.6 times as long as wide, 8th antennomere 2.4 times as long as wide. Pronotum slightly convexed with fine punctures, widest near anterior fourth with obscure U-depression near middle, 1.3 times as wide as long, about 1.2 times as wide and 1.3 times as long as head; scutellum (Fig. 10) subtriangular, prescutoscutellar suture arcuate, scutellar process narrow triangular; elytra bicolor, humeral region with indistinct yel- Figs. 1-4. Habitus. 1. Lesteva cordicollis, length 3.8 low patch and fine punctures, posterior margin mm; 2. L. coreana sp. nov., length 3.4 mm; 3. L. dis- truncated, 1.1 times as wide as long, 1.4 times as tincta, length 4.0 mm; 4. L. miyabi, length 3.9 mm. wide and 1.7 times as long as pronotum (Figs. 2 and 14); apex of metaventral process round; external surface of metatibia with 3-4 long golden nomere 1.8 times as long as wide. Pronotum setae. Abdominal segments III-VIII with micros- slightly convexed with fine punctures, widest culpture. Median lobe of aedeagus narrowed api- near anterior fourth with ambiguous U-depres- cally, lateral margin weakly arcuated; basal re- sion near middle, 1.3 times as wide as long, gion of parameres broad, narrowed apically, api- about 1.2 times as wide and 1.4 times as long as cal third constricted, slightly longer than median head; scutellum (Fig. 9) subtriangular, prescu- lobe, four setae present with two at apex (Figs. 23- toscutellar suture gently curved, scutellar pro- 24). cess broad subtriangular; elytra bicolor, hu- Type Series: Holotype, : ‘KOREA: Jeonbuk meral region with large yellow patch and fine Prov.: Muju-gun, Anseong-myeon, Mt. Deokyu- punctures, posterior margin truncated, 1.1 san, Chilyeon-fall, 27 V 2005, TK Kim, ex under times as wide as long, 1.5 times as wide and 1.7 stone near stream; Holotype, Lesteva coreana times as long as pronotum (Figs. 1, 13); apex of Kim and Ahn, Desig. T.-K. Kim and K.-J. Ahn metaventral process round (Fig. 17); external 2010.’ Deposited in CNUIC, Daejeon. Paratypes, surface of metatibia with 3-4 long golden setae same data as holotype (10 1 , CNUIC), (Fig. 19). Abdominal segments III-VIII with mi- Paratype, Lesteva coreana Kim and Ahn, Desig. crosculpture. Median lobe of aedeagus elongate, T.-K. Kim and K.-J. Ahn 2010. Other materials: parallel-sided, apical process triangular, apical Mt. Deokyusan, Chilyeon-fall, 22-23 V 1998, HJ middle area elevated, internal sac backbone- Kim, ex near stream (1 , CNUIC); same data as shaped; parameres slender, slightly longer than holotype (2 , CNUIC); Chungnam Prov.: Dae- median lobe, four setae present with two at jeon, Mt. Gyeryongsan, Keumsubong, 21 V 2000, apex (Figs. 21-22). SJ Park, ex near stream (5 5 , CNUIC); Yu- Materials Examined. KOREA: Gangwon seong-gu, Sutong-gol, 5 IX 1998, SJ Baek (1 , Prov.: Chuncheon-si, Nam-myeon, Mt. Bongh- CNUIC); Sutong-gol, 9 V 1998, KR You, HJ Lim, wasan (N37°46’1.2” E 127°35’59.0” 186m) 17 IX HJ Kim, ex near stream (1 , CNUIC). 2008, TK Kim ex under stone near stream (14, Distribution. Korea (South). CNUIC); Chungnam Prov.: Daejeon, Yuseong- Remarks. The species is similar to L. cordicol- gu, Sutong-gol, 9 V 1998, KR You, HJ Lim, HJ lis but can be distinguished by the shape and Kim, ex near stream (2, CNUIC); Jeonbuk structures of antennomeres, scutellum, and me- Prov.: Muju-gun, Anseong-myeon, Mt. Deokyu- dian lobe of aedeagus (Table 1). san, Chilyeon-fall, 27 V 2005, TK Kim, ex under stone near stream (81, CNUIC); Jinan-gun, Lesteva distincta Watanabe, 1990 Jeongcheon-myeon, Mt. Unjangsan, V 19 1998, (Figs. 3, 7, 11, 15, 18, 25-26) YB Cho (2, CNUIC); Distribution. Korea (South), Russia (East Si- Lesteva distincta Watanabe, 1990: 178; Herman, 2001: beria). 315; Smetana, 2004: 247.

Lesteva coreana Kim and Ahn sp. nov. Description. Body (Fig. 3) length 3.5-4.1 mm (Figs. 2, 6, 10, 14, 23-24) (head to abdominal end), covered with coarse punctures and pubescence, reddish brown to Description. Body (Fig. 2) length 3.1-3.5 mm black and glossy; mouthparts, antennae and legs (head to abdominal end), covered with ﬁne punc- brown. Head about 1.2 times as wide as long; eye 30 Florida Entomologist 94(1) March 2011

Figs. 5-16. 5-8. Antenna, ventral aspect. 5. Lesteva cordicollis; 6. L. coreana sp. nov.; 7. L. distincta; 8. L. miyabi. 9-12. Scutellum, dorsal aspect. 9. L. cordicollis; 10. L. coreana sp. nov.; 11. L. distincta; 12. L. miyabi. 13-16. elytron, ventral aspect. 13. L. cordicollis; 14. L. coreana sp. nov.; 15. L. distincta; 16. L. miyabi. Scales = 0.1 mm (Figs. 9-12); 0.3 mm (Figs. 5-8, 13-16). Kim & Ahn: Korean Lesteva Species 31

about 1.7 times as long as temple; antennae (Fig. 7) pubescent, reaching middle of elytra, 4th antennomere 2.1 times as long as wide, 8th antennomere 1.9 times as long as wide. Pronotum much convexed with coarse punctures, about 1.2 times as wide as long, 1.2 times as wide and 1.3 times as wide as head, widest near anterior fourth with distinct U-depression near middle; scutellum (Fig. 11) subtriangular, prescutoscutellar suture arcuate, scutellar process broad pentagonal; elytra bicolor, humeral region with red- 1.7 1.9 1.8 12 Fig. pentagonal notched 20) 10-14 dark setae (Fig. present dish brown patch and somewhat coarse punc- L. miyabi L. tures, posterior margin round, 1.1 times as long as wide, about 1.8 times as wide and 1.9 times as long as pronotum (Figs. 3 and 15); apex of metaventral process notched (Fig. 18); external surface of metatibia with 10-14 long dark brownish setae. Abdominal segments III and VIII with microsculpture. Median lobe of aedeagus broad, basal two third parallel-sided, apical third narrowed suddenly, apical process triangular, midline area elevated with longitudinal carina, lateral margin rolled dorsally; parameres robust, 1.7 2.1 1.9 11 Fig. 18) (Fig. notched present symmetrical, as long as median lobe, lateral mar- L. distincta L. gin rolled ventrally, apical region coiling ventrally and inwardly, four setae present (Figs. 25 and 26). Materials Examined. KOREA: Gangwon

Prov.: Chuncheon-si, Sabuk-myeon, Jiam-ri, 15 IV 2001, SI Lee (1, CNUIC); Chungbuk Prov.: Danyang-gun, Danyang-eup, Mt. Sobaeksan, . Cheondong-area, 8-9 V 1999, US Hwang, HJ Kim, sp. nov. sp. sifting (2, CNUIC); Yeongdong-gun, Sangchon- myeon, Mulhan-ri, Mt. Minjujisan, Mulhan- SPECIES

3.1 2.6 2.4 10 Fig. narrow triangular round pentagonal present on III-VIIIabsent VIII present on III and VIII present on III and straight (Figs. 23-24) straight (Figs. 25-26) coiling ventrally (Figs. 27-28) coiling ventrally (Figs. stream (N36°3’15” E127°52’31”), 16 VI 2006, TK Kim, ex under stone near stream (1, CNUIC); Mt. Manloi, 30 V 1998, HJ Lim, sifting (1, ESTEVA

L CNUIC); Chungnam Prov.: Daejeon-si, Yu- seong-gu, Gung-dong, Chungnam National Uni- versity (N36°22’38.7” E127°20’43.5”), 18 IV 2007, OREAN HW Kim, ex near pond (32, CNUIC); Chung- K nam National University (N36°22’38.7” OF E127°20’43.5”), 7 V 2007, YH Kim, ex near pond (74, CNUIC); Chungnam National University (N36°22’38.7” E127°20’43.5”), 14 V 2007, HW

FEATURES Kim, ex near pond (12 8 , CNUIC); Yuseong-gu,

3.3 9 Fig. broad triangular 17) round (Fig. 19)3-4 golden setae (Fig. 3-4 golden setae 10-14 dark setae present on III-VIII absent straight (Figs. 21-22) straight (Figs. Deokmyeong-dong, Sutonggol, 9 V 1998, KR You,

L. cordicollisL. coreana L. HJ Lim, HJ Kim, ex near stream (1 , CNUIC); Buyeo-gun, Naesan-myeon, Mt. Wolmyeongsan, Geumgisa, 3 V-1 VI 2000, US Hwang, HJ Kim, FIT (1, CNUIC); Geumgisa, 1 VI 2000, US Hwang, HJ Kim, sifting (1, CNUIC); Jeonbuk MORPHOLOGICAL Prov.: Buan-gun, Byeonsan-myeon, Mt. Nae- OF byeonsan, Jikso-fall, 30 V 2001, YB Cho, sifting (12, CNUIC); Jinan-gun, Jeongcheon-myeon, Mt. Unjangsan, V 19 1998, YB Cho (2, CNUIC); Jeonnam Prov.: Gurye-gun, Mt. Jirisan, Toji- myeon, Piagol, 24 V 2000, HJ Kim, ex near stream COMPARISON (11, CNUIC); Piagol, 24-27 V 2000, KJ Ahn, SJ Park, US Hwang, FIT (1, CNUIC); Jindo- 1. A gun, Uisin-myeon, Sacheon-ri, Mt. Cheomchilsan

ABLE (N34°27’53.7” E126°18’42.6” 115m), 23 II 2007 Ratio of eye to temple length Antennomere 4 (length/width ratio) Antennomere 8 (length/width ratio) Scutellum 2.1 Scutellar process 1.8 process Metaventral Long setae of metatibia Abdominal microsculpture Aedeagus with carina Apical region of paramere T 32 Florida Entomologist 94(1) March 2011

Figs. 17-20. 17-18. Metaventrite, ventral aspect. 17. Lesteva cordicollis; 18. L. distincta. 19-20. Metatibia (pubescence omitted), anterior aspect. 19. L. cordicollis; 20. L. miyabi. Scales = 0.3 mm.

TK Kim, sifting, leaf litter (1, CNUIC); Yeongg- mostly convexed with coarse punctures, widest wang-gun, Hongnong-eup, Sangha-ri (N35°23’24.9” near anterior third with distinct U-depression E126°25’57.9”), 2 V 2007, KJ Ahn, TK Kim, YH near middle, about 1.2 times as wide as long, Kim, ex near stream (2, CNUIC); Hadong-gun, about 1.1 times as wide and 1.2 times as long as Hwagye-myeon, Ssanggyesa, 25 V 2000, HJ Kim, ex head; scutellum (Fig. 12) subtriangular, prescu- near stream (1, CNUIC); Gyeongbuk Prov.: toscutellar suture round, scutellar process broad Cheongsong-gun, Budong-myeon, Mt. Juwangsan, pentagonal; elytra unicolor with coarse punc- 29 VI 1987, YB Cho, ex under moss (1, CNUIC); tures, posterior margin round, 1.04 times as long Gyeongnam Prov.: Geoje-si, Yeoncho-myeon, Mt. as wide, 1.8 times as wide and 2.0 times as long as Aengsan (N34°56’17.3” E128°36’6.6” 85m), 21 I pronotum (Figs. 4 and 16); apex of metaventral 2009, DH Lee, JH Song, ex under stone near mount process notched; external surface of metatibia stream (21, CNUIC). with 10-14 long dark brownish setae (Fig. 20). Ab- Distribution. Korea (South), Japan. dominal segments III and VIII with microsculpture. Median lobe of aedeagus broad, narrowed Lesteva miyabi Watanabe, 1990 apically with longitudinal carina, lateral margin (Figs. 4, 8, 12, 16, 20, 27-28) almost straight; parameres robust, symmetrical, as long as median lobe, lateral margin rolled ven- Lesteva miyabi Watanabe, 1990: 175; Herman, 2001: trally, apical region coiling ventrally and in- 324; Smetana, 2004: 247. wardly, four setae present (Figs. 27 and 28). Materials Examined. KOREA: Jeju Prov.: Lesteva plagiata: Cho et al., 2002: 36. Misidentiﬁcation. Jeju-si, Arail-dong, Gwaneumsa, 26 V 2003, SJ Park, ex near stream (109, CNUIC); Jeju-si, Description. Body (Fig. 4) length 3.8-4.5 mm Bonggae-dong, Muljang-oreum, 23 V 1998, YB (head to abdominal end), covered with coarse Cho (35, CNUIC); Jeju-si, Nohyeong-dong, punctures and pubescence, dark brown to black Cheonwangsa (N33°24’25.4” E126°29’42.7” 395 and glossy; mouthparts, antennae and legs brown m), 8 XI 2006, TK Kim, ex under stone near to reddish brown. Head about 1.3 times as wide as stream (1, CNUIC); Jeju-si, Orai-dong, Eorimok long; eye about 1.7 times as long as temple; anten- (N33°23’26.0” E126°29’41.1” 1000 m), 31 V 2007, nae (Fig. 8) pubescent, reaching middle of elytra, TK Kim, ex under stone near stream (2, 4th antennomere 1.9 times as long as wide, 8th CNUIC); Seoguipo-si, Hawon-dong, Seoguipo antennomere 1.8 times as long as wide. Pronotum Natural Recreation Forest (N33°18’54.2” Kim & Ahn: Korean Lesteva Species 33

Figs. 21-28. Aedeagus. 21-22. Lesteva cordicollis. 21. dorsal aspect; 22. lateral aspect. 23-24. L. coreana sp. nov. 23. dorsal aspect; 24. lateral aspect. 25-26. L. distincta. 25. dorsal aspect; 26. lateral aspect. 27-28. L. miyabi. 27. dorsal aspect; 28. lateral aspect. Scales = 0.3 mm.

E126°27’56.0” 735 m), 30 V 2007, TK Kim, sifting, Goepyeongi-oreum, 23 V 2006, SJ Park, DH Lee, ﬂood debris (1, CNUIC); Seoguipo Natural Rec- SI Lee, YH Kim, leaf litter (1, CNUIC); Goepyeo- reation Forest (N33°18’36” E126°28’9.2” 665 m), ngi-oreum (N33°25’2.7” E126°38’32.6” 530 m), 8 IX 31 V 2007, DH Lee, YH Kim, sifting, leaf litter 2006, DH Lee, ex leaf litter (15, CNUIC); (1, CNUIC); Bukjeju-gun, Aewol-eup, 1100-goji, Goepyeongi-oreum (N33°25’ 1.8” E126°38’32.2” 539 28 v 2003, CW Shin, ex near stream (2, CNUIC); m), 8 IX 2006, TK Kim, ex wet grit near pond (1, 1100-goji (N33°21’40.6” E126°27’44.6” 1097 m), CNUIC); Namjeju-gun, Namwon-eup, Dongsu- 12 × 2006, TK Kim, sifting, wet leaf litter (12, bridge (N33°22’8.4” E126°37’30.7” 640 m), 8 XI CNUIC); 1100-goji (N33°21’37.5” E126°27’45.8” 2006, TK Kim, ex under stone near stream (3, 1110 m), 31 V 2007, TK Kim, sifting, leaf litter CNUIC); Dongsu-bridge, 1 III 2007, TK Kim, ex un- (96, CNUIC); Bukjeju-gun, Jocheon-eup, der stone near stream (21, CNUIC); Dongsu- 34 Florida Entomologist 94(1) March 2011 bridge (N33°22’8.5” E126°37’30.5” 635 m), 29 V Distribution. Korea (South), Japan. 2007, TK Kim, ex under stone near stream Remarks. Cho et al. (2002) reported L. pla- (1812, CNUIC); Namjeju-gun, Namwon-eup, giata in Korea. However, we have determined Goepyeongi-oreum, 28 V 2003, SJ Park, CW Shin, that this was a misidentiﬁcation of L. miyabi, MJ Jeon, sifting (1611, CNUIC); Goepyeongi- based on our examination of their voucher speci- oreum, 28 V-27 VI 2003, YB Cho, SJ Park, CW Shin, men (1: Mt. Hallasan, 900 m alt., Jejudo Is., 17. FIT (1, CNUIC); Mt. Hallasan, 900 m alt., Jejudo VII 1994, G. Sh. Lafer leg). The species was col- Is., 17 VII 1994, G. Sh. Lafer leg (1, CNUIC). lected only in Jeju-do island.

KEY TO THE KOREAN SPECIES OF THE GENUS LESTEVA LATREILLE

1. Pronotum slightly convexed with ﬁne punctures; prosternal process without carina; posterior margin of elytra truncated with ﬁne punctures; apex of metaventral process round (Fig. 17); metatibia without long dark brownish setae (3-4 long golden setae present) (Fig. 19)...... 2 — Pronotum distinctly convexed with coarse punctures; prosternal process with short, sinuous longitudinal carina; posterior margin of elytra round with coarse punctures; apex of metaventral process notched (Fig. 18); metatibia with 10-14 long dark brownish setae (Fig. 20) ...... 3 2. Fourth antennomere 2.1 times as long as wide, 8th antennomere 1.8 times as long as wide (Fig. 5); scutellar process broad (Fig. 9); median lobe of aedeagus elongate, in basal three fourth parallel-sided, and in apical fourth abruptly narrowed in dorsal view (Figs. 21 and 22)...... L. cordicollis — Fourth antennomere 2.6 times as long as wide, 8th antennomere 2.4 times as long as wide (Fig. 6); scutellar process narrow (Fig. 10); median lobe of aedeagus narrowed apically, lateral margin weakly arcuated in dorsal view (Figs. 23 and 24) ...... L. coreana sp. nov. 3. Pronotum widest at anterior fourth; elytra bicolor with reddish patch around humeral region, moderately broad and long (Fig. 3) ...... L. distincta — Pronotum widest at anterior third; elytra unicolor, broad and long (Fig. 4)...... L. miyabi

ACKNOWLEDGMENTS tles. Vol. 1. Archostemata, Myxophaga, Adephaga, Polyphaga: Staphyliniformia. CRC Press, Boca Ra- We thank Dr. Watanabe (Tokyo University of Agri- ton, Florida. culture, Japan) for the loan of specimens. This research SHARP, D. 1889. The Staphylinidae of Japan. Ann. Mag. was supported by the project on survey and excavation Nat. Hist. (6)3: 463-476. of Korean indigenous species of the National Institute of SHAVRIN, A. V. 2001. New and little-known species of Biological Resources (NIBR) under the Ministry of En- Omaliinae from the Baikal-Transbaikal area (Co- vironment, Korea. leoptera: Staphylinidae). Zoosyst. Rossica 9: 189-193. SHAVRIN, A. V., SHILENKOV, V. G., AND ANISTSCHENKO, REFERENCES CITED A. V. 2007. Two new species and additional records of Lesteva Latreille, 1797 from the mountains of CASEY, T. L. 1894. Coleopterological notices. V. Ann. South Siberia (Coleoptera: Staphylinidae: Omalii- New York Acad. Sci. 7: 281-606. nae: Anthophagini). Zootaxa 1427: 37-47. CHO, Y. B., LAFER, G. S., PAIK, J. C., AND PARK, J. K. SMETANA, A. 2004. Staphylinidae, subfamily Omalii- 2002. Contribution to the staphylinid fauna (Co- nae, pp. 237-268 In I. Löbl and A. Smetana [eds.], leoptera, Staphylinidae) of Korea. Korean J. Soil Catalogue of Palaearctic Coleoptera. Volume 2, Hy- Zool. 7(1-2): 35-44. drophiloidea - Histeroidea - Staphylinoidea. Applo HERMAN, L. H. 2001. Catalog of the Staphylinidae (In- Books, Steustrup. secta: Coleoptera). 1758 to the End of the Second STEEL, W. O. 1952. Notes on the Omaliinae (Col., Sta- Millennium. I. Introduction, History, Biographical phylinidae). 4. On the genera Lesteva Latr., Paraleste- Sketches, and Omaliine Group. Bull. American Mus. va Casey and Tevales Casey, with a key to the British Nat. Hist. 265: 309-333. species of Lesteva. Entomol. Mon. Mag. 88: 8-9. LATREILLE, P. A. 1797. Précis des Caractères STEEL, W. O. 1970. The larvae of the genera of the Oma- Génériques des Insectes, Disposés dans un Ordre liinae (Coleoptera: Staphylinidae) with particular Naturel. xiv + 201 + 7 pp. Brive: F. Bourdeaux. reference to the British fauna. Trans. R. Entomol. LI, X.-J., LI, L.-Z., AND ZHAO, M.-J. 2005. A new species Soc. London 122(1): 1-47. of the genus Lesteva (Coleoptera: Staphylinidae: WATANABE, Y. 1990. A taxonomic study on the subfami- Omaliinae) from China. Entomotaxonomia 27(2): ly Omaliinae from Japan (Coleoptera, Staphylin- 111-113. idae). Mem. Tokyo Univ. Agric. 31: 149-179. MOTSCHULSKY, V. 1860. Énumération des nouvelles es- WATANABE, Y. 2004. Two new species of the genus Lesteva pèces de coléoptères rapportées de ses voyages. 3e (Coleoptera, Staphylinidae) from the Island of Dôgo of partie. Bull. Soc. Imper. Nat. Moscou 33(2): 539-588. the Oki Islands, West Japan. Elytra 32(1): 71-77. NEWTON, A. F., THAYER, M. K., ASHE, J. S., AND CHAN- WATANABE, Y. 2005. A new species of the genus Lesteva DLER, D. S. 2001. 22. Staphylinidae, pp. 272-342 In (Coleoptera, Staphylinidae) from Taiwan. Elytra R. H. Arnett and M. C. Thomas [eds.], American Bee- 33(1): 30-33.

Goyal et al: Corn-infesting Ulidiidae of Florida 35

DISTRIBUTION OF PICTURE-WINGED FLIES (DIPTERA: ULIDIIDAE) INFESTING CORN IN FLORIDA

GAURAV GOYAL1, GREGG S. NUESSLY1, DAKSHINA R. SEAL2, JOHN L. CAPINERA3, GARY J. STECK4 AND KENNETH J. BOOTE5 1Everglades Research and Education Center, University of Florida (UF), Institute of Food and Agricultural Sciences (IFAS), 3200 E. Palm Beach Rd., Belle Glade, FL 33430

2Tropical Research and Education Center, UF, IFAS, 18905 S.W. 280 St., Homestead, FL 33031

3Department of Entomology and Nematology, UF, IFAS, P.O. Box 110620, Gainesville, FL 32611

4Division of Plant Industry, Florida Department of Agriculture and Consumer Services, P.O. Box 147100, Gainesville, FL 32614

5Department of Agronomy, UF, IFAS, P.O. Box 110500, Gainesville, FL 32611

ABSTRACT

The picture-winged fly Euxesta stigmatias Loew (Diptera: Ulidiidae) has been a serious pest of sweet corn (Zea mays L.) in Florida since 1930. Several other species in the family are known to infest corn grown in the Caribbean, Central America, and South America. Surveys were conducted throughout Florida to evaluate species richness and distribution of corn-infesting Ulidiidae. Adults were sampled with sweep nets and reared from fly larvae-infested corn ears collected from representative corn fields in 16 and 27 counties in 2007 and 2008, respectively. Four Ulidiidae species were found in corn fields using both sampling techniques. Euxesta eluta Loew and Chaetopsis massyla (Walker) were found throughout the state on field and sweet corn. Euxesta stigmatias was only found in Martin, Miami-Dade, Okeechobee, Palm Beach, and St. Lucie Counties on field and sweet corn. Euxesta annonae (F.) was found in sweet corn in Miami-Dade, Okeechobee, and Palm Beach Counties, but field corn was not sampled in these counties. Euxesta eluta, E. stigmatias, and C. massyla were collected from corn throughout the corn reproductive stage. Raising adults from fly larvae- infested ears provided the best method for assessing rates of ear infestation and species richness. Sweep netting did not provide reliable information on the presence or species composition of ulidiid species infestation. We report for the first time E. annonae and E. eluta as pests of corn in Florida and the USA.

Key Words: Euxesta annonae, Euxesta eluta, Euxesta stigmatias, Chaetopsis massyla, maíze

RESUMEN

La mosca de alas pintadas, Euxesta stigmatias Loew (Diptera: Ulidiidae), ha sido una plaga seria de maíz dulce (Zea mays L.) en la Florida desde 1930. Varias especies de la familia Uli- diidae son conocidas de infestar maíz sembrado en el Caribe y el Centroamérica y Sudamé- rica. Se realizaron sondeos por todo la Florida para evaluar la diversidad de especies y distribución de moscas de la familia Ulidiidae que infestan maiz. Se muestrearon los adultos con redes de recolección y criandolos de mazorcas infestadas con larvas de moscas de campos representativos de maiz en 16 y 27 condados en 2007 y 2008, respectivamente. Se encontraron Euxesta eluta Loew y Chaetopsis massyla (Walker) por todo el estado en maíz de campo y maíz dulce. Euxesta stigmatias fue encontrada solamente en los condados de Martin, Miami-Dade, Okeechobee, Palm Beach y St. Lucie sobre maíz de campo y maíz dulce. Euxesta annonae (F.) fue encontrada en maíz dulce en los condados de Miami-Dade, Okee- chobee y Palm Beach, pero no se muestrearon maíz de campo en estos condados. Se recolectaron Euxesta eluta, E. stigmatias y C. massyla durante toda la etapa reproductiva del maíz. Euxesta annonae fue criada de mazorcas solamente de 8 a 21 dias de edad, pero los campos con mazorcas de ≤ 8-dias no fueron muestreados en los condados donde esta especie fue encontrada. El criar los adultos de mazorcas infestadas con larvas de moscas fue el mejor metodo para evaluar la taza de infestacion de las mazorcas y la diversidad de especies. Las recolecciones con redes no dieron un estimado conﬁable para identiﬁcar infestaciones de especies de ulidiidos. Reportamos por primera vez E. annonae y E. eluta como plagas de maíz en la Florida y EEUU.

36 Florida Entomologist 94(1) March 2011

There are 671 species of Ulidiidae worldwide, MATERIALS AND METHODS but less than 10 species in 2 genera are known to damage corn (Allen & Foote 1992; Anonymous Corn grown throughout Florida was sampled 2008c; Goyal et al. 2010; Van Zwaluwenburg for ulidiid species. Extension personnel and re- 1917). Van Zwaluwenburg (1917) first reported searchers from all 67 Florida counties provided the pest nature of Euxesta stigmatias Loew information on corn types and growing season (Diptera: Ulidiidae) (Figs. 1 g, 1h) in Puerto Rico needed to select representative fields. Corn fields where it damaged up to 100% of untreated corn. It were visited with the assistance of extension was first discovered damaging corn in Miami, agents. One to 2 corn fields were sampled for Uli- Florida in 1938 (Barber 1939) and had moved diidae in each of 16 counties from Jul through Oct north into central Florida by 1951 (Hayslip 1951). 2007 (Table 2). One to 4 corn fields were sampled This species has become a serious pest of Florida for Ulidiidae in each of 27 counties during Feb sweet corn (Zea mays L.) requiring multiple in- through Jun 2008 (Table 3), including 10 counties secticide applications during the ear stage to visited in 2007. maintain a marketable crop (Mossler 2008; Adult ulidiids can be elusive and difficult to re- Nuessly & Hentz 2004; Seal 1996, 2001; Seal & liably observe and collect. They frequently avoid Jansson 1994). Sweet corn is an important crop in direct sunlight and walk or fly away from the di- Florida with 22.8% of the total USA fresh market rect line of sight of workers approaching them. sweet corn production (Anonymous 2009). Eux- They are more easily collected from the tassels esta stigmatias also has been reported infesting and upper leaves of corn plants in the hour just sweet corn in Georgia (Daly & Buntin 2005), after sunrise and just before sunset, but it was Texas (Walter & Wene 1951), California (Fisher not possible to sample all fields at these times. 1996), Guatemala (Painter 1955) and Brazil Adults can also be killed after ovipositing on a (Franca & Vecchia 1986). The insect deposits its plant host before they are sampled, particularly eggs primarily on silks (styles) in the tips of ears. within crops that are frequently treated with in- The larvae feed on silks, kernels, and cobs. Bailey secticides, such as sweet corn. Therefore, fields (1940) observed disruption of pollination due to were sampled for both adults and immatures to larval feeding on silks. Larvae enter through the determine whether the plants served as develop- soft pericarp of milk stage kernels to completely mental hosts for ulidiid species and to determine consume the developing embryo and endosperm the feasibility of using adult collection records for (Seal & Jansson 1989). App (1938) observed lar- determining ear infestation. Preference was val feeding on cobs followed by mold development given to sampling corn that was between the silk- resulting in significant reduction in market value. ing and dough stages because both the adult and Several other ulidiid species are known maize immature stages of flies can best be collected dur- pests in the Caribbean and in the Americas south ing the first 3 weeks of corn reproduction. Neither of Texas (Arce de Hamity 1986; Barbosa et al. adults nor immatures in ears were found in fields 1986; Chittenden 1911; Diaz 1982; Evans & Zam- sampled before silking in Lake County (sweet brano 1991; Gossard 1919; Painter 1955; Wyck- corn) in 2007 and in Jefferson (field corn) and huys & O’Neil 2007), but only 1 other species is Walton (sweet corn) Counties in 2008, therefore; currently recognized as a pest in the USA. Cha- data from these 3 fields were not included in the etopsis massyla (Walker) (Figs. 1 a, b) was recently results. Sweet corn fields were preferred over determined to be a primary pest of sweet corn in field corn for sampling because the flies cause less Florida (Goyal et al. 2010). Evidence suggesting damage in field corn than in sweet corn (Scully et the possibility of additional picture-winged species al. 2000). Corn type (i.e., field, sweet, Bt-en- attacking corn in Florida include a picture of Eux- hanced, and standard corn) and variety, number esta eluta Loew (Diptera: Ulidiidae) (Figs. 1 e, f) on of days before or after first silk, and locations of the cover of Hayslip’s (1951) paper entitled “Corn the field were recorded. Visual observations were silk fly control on sweet corn” misidentified as E. taken for the presence of ulidiid adults. stigmatias. Examination of the Ulidiidae collection Flies were collected from corn fields with a at the Division of Plant Industry in Gainesville, sweep net (37.5 cm diameter). The sample size Florida revealed that E. eluta and E. annonae (F.) was adjusted depending on the estimated field (Diptera: Ulidiidae) (Figs. 1 c, d) have been col- size. In fields ≤4 ha, 3 pairs of corn rows were selected in several Florida counties since at least lected for sampling: 1 pair from each side of the 1948, but these specimens were not labeled as be- field and 1 pair in the middle of the field. In fields ing collected or reared from corn. These later 2 spe- >4 ha, 9 pairs of rows were selected for sampling: cies are recognized pests of corn in South America 1 on each side of the field, 1 in the middle of the (Diaz 1982; Frías-L 1978). Therefore, it is possible field, and 6 pairs of rows randomly selected from that additional Ulidiidae species may be feeding on between the field margins. Sweep net sampling corn in Florida. The objective of this study was to for flies was done while walking the length of the evaluate species richness and distribution of corn- field swinging the net 100 times between 2 rows infesting ulidiids throughout Florida. in each pair of selected rows. Flies were preserved

Goyal et al: Corn-infesting Ulidiidae of Florida 37

Fig. 1. Chaetopsis massyla male (a) and female (b); Euxesta annonae male (c) and female (d); E. eluta male (e) and female (f); E. stigmatias male; and (g) and female (h).

38 Florida Entomologist 94(1) March 2011 in 70% ethyl alcohol for later identification and culated by box fans. Corn ears collected on Mar 6, counting with a dissecting microscope. Identified 2007 were placed collectively in 3.78 L Ziploc bags Ulidiidae specimens housed at the Division of and then transferred to plastic containers with Plant Industry, Gainesville, FL and keys of Eux- mesh tops. Pupae were removed from the bags and esta (Ahlmark & Steck unpublished, Curran plastic containers, and placed on moistened filter 1928, 1934, 1935) and Chaetopsis (G. Steyskal un- paper (Whatman® 3, Whatman International Ltd., published) were used to confirm identifications. Maidstone, England) in covered Petri dishes for Corn ears were examined for the presence of fly adult emergence. The dishes were sealed with Para- larvae in the same fields sampled with sweep nets. film® (Pechiney Plastic Packaging, Chicago, IL) to Ears found to contain larvae were collected and held reduce moisture loss. Adults that emerged were pre- for adult emergence to confirm species infestation. served in 70% ethyl alcohol for later identification The number of ears sampled per field was adjusted and counting as above. depending on the number of planted rows in each field. Fifty-six ears were examined in fields with Statistical Analysis <90 rows and 88 ears were examined in fields with >90 rows. In a field with <90 rows, 10 groups of 4 The results were tested by analysis of variance plants each were randomly selected for ear inspec- to examine the effects of sample technique, corn tion. In a field with 90 rows, ears were examined in type (field and sweet), corn ear age day and every tenth row starting from the first row and con- month of sampling (1-7, 8-14, 15-21 d) and sample tinuing to the other side of the field (total of 10 year on the mean numbers of each species col- rows). In a field with >90 rows, 6 rows were sampled lected (Proc GLM, Version 9.0; SAS Institute from each side of the field (each sampled row sepa- 2008). Year was used as a random variable in the rated by 10 rows), and 6 additional rows were ran- model. The mean number of flies sweep netted per domly selected and sampled in the middle of the pair of rows used in the data analysis was calcu- field. One ear on each of 4 plants in the middle of lated for each field by dividing the total number of each selected row was examined for fly larvae (40 flies caught in sweep nets by the number of pairs and 72 ears per field for <90 and >90 rows, respec- of rows sampled in that field. The mean number of tively). An additional 4 plants in each corner of the flies per infested ear was calculated for each field field were examined for larvae-infested ears (16 by dividing the total number of flies reared from ears per field). The top third of each infested ear was infested corn ears by the number of infested ears removed with a knife and placed individually in a in each field. Different numbers of ears and plant Ziploc® bag (1.83 L, S.C. Johnson & Son, Inc., Ra- rows were sampled in each field and more fields cine, WI). Two paper towels were added to each bag were sampled in 2008 than in 2007; therefore the to reduce moisture accumulation. Bags were stored results were presented as least square means in portable coolers in the field and during transpor- rather than arithmetic means of flies caught per tation back to the laboratory. row and reared per corn ear. Infested ears kept in the Ziploc bags were held in an air conditioned room maintained at 26.0 ± 1°C RESULTS and L14:D10 h photoperiod to collect pupae for adult identification. To reduce the accumulation of The mean number of ulidiid adults caught in moisture and associated fungus growth, bags with sweep nets was significantly affected by fly spe- corn were left partially open, paper towels were cies, corn type, survey year, and the species × year changed frequently, and the air was constantly cir- interaction (Table 1). Significantly more E. eluta

TABLE 1. ANALYSIS OF VARIANCE FOR FLIES CAPTURED BY SWEEP OR REARED FROM CORN EAR ON SPECIES, CORN TYPE, AGE OF CORN AND YEAR

Sweep net Corn ears

Source dfFPFP

Species of Ulidiidae 3 8.12 <0.0001 3.73 0.0120 Corn type 1 6.77 0.0099 6.53 0.0113 Age of corn 2 1.82 0.1638 1.09 0.3368 Year 1 33.17 <0.0001 13.82 0.0003 Species × corn type 3 0.68 0.5642 1.52 0.2091 Species × age of corn 6 0.31 0.9304 0.67 0.6739 Species × year 3 5.50 0.0011 2.00 0.1145

ANOVA (Proc GLM, SAS Institute 2008); denominator df = 236.

Goyal et al: Corn-infesting Ulidiidae of Florida 39

(least squares mean ± SEM; 3.80 ± 0.63) and C. were reared from ears collected from all but Lee, massyla (3.62 ± 0.63) were caught in sweep nets Lake and St. Johns Counties. These 2 species per row than E. stigmatias (1.33 ± 0.63) and E. were reared from infested ears in 82% of corn annonae (0.14 ± 0.63). More adults were caught fields statewide. Euxesta annonae and E. stigma- per row with sweep nets in sweet corn (2.95 ± tias were reared only from corn ears collected 0.34) than in field corn (1.49 ± 0.49). Sweep net from Miami-Dade, Okeechobee, and Palm Beach counts per row were greater in 2007 (3.89 ± 0.51) Counties, amounting to only 18% of fields sam- than in 2008 (0.55 ± 0.32). pled. Adults of E. eluta and C. massyla were The mean number of adults emerged per ear reared from both field and sweet corn ears. Eux- were significantly affected by fly species, corn esta annonae and E. stigmatias emerged from type, and survey year (Table 1). Significantly sweet corn ears in fields from Miami-Dade, more E. eluta (1.41 ± 0.30) were reared from each Okeechobee, and Palm Beach Counties, but field corn ear than E. stigmatias (0.40 ± 0.30) and E. corn fields were not sampled in these counties. annonae (0.05 ± 0.30). The mean number of C. Euxesta eluta and C. massyla were each reared massyla per ear (0.82 ± 0.30) was not significantly from 50% of field corn and 92% of sweet corn different than the other species. More adults were fields. Euxesta annonae and E. stigmatias were reared from each corn ear in 2007 (1.19 ± 0.25) reared from 100% of the sweet corn fields in above than in 2008 (0.15 ± 0.15). Significantly more mentioned Counties. adults per ear were reared from sweet corn (1.02 The age of sampled corn in 2007 ranged from 4 ± 0.16) than from field corn (0.33 ± 0.24). Results to 21 d after first silk (Table 2). Chaetopsis mas- for species by county and reared from fields were syla was sweep netted in fields of all ages sam- presented separately for 2007 (Table 2) and 2008 pled. Euxesta eluta was sweep netted in fields 7- (Table 3) due to significant differences in mean 21 d after first silk. Euxesta annonae and E. stig- counts between years. matias were sweep netted from fields 8 to 21 d af- The correlation between adults caught in ter first silk, but no fields <8 d after first silk were sweep nets and those reared from ears varied by sampled in counties infested with these 2 species. species. Correlation coefficients were as follows: Euxesta eluta and C. massyla emerged from corn 0.79 (P < 0.0001) for E. stigmatias, 0.62 (P < ears collected from fields 4 to 21 d after first silk, 0.0001) for C. massyla, 0.58 for (P < 0.0001) E. an- while E. annonae and E. stigmatias from ears col- nonae, and 0.51 (P < 0.51) for E. eluta. lected 8 to 21 d after first silk (Table 4). Corn ears were not collected from fields <8 d after first silk 2007 Field Survey in counties with E. annonae and E. stigmatias.

Four Ulidiidae species were caught in sweep 2008 Field Survey nets and reared from fly larvae-infested ears in Florida corn during the first survey year (Table The same 4 species were again collected in 2). Chaetopsis massyla was collected in more sweep nets and reared from fly-larvae infested counties throughout the state than other species ears in Florida corn during the second study year and was netted in 100% of the sampled fields. (Table 3). Ulidiid adults were netted in 23 of 27 This was followed by E. eluta, which was netted in counties sampled in 2008. No adult picture- 88% of sampled fields in all counties except Lake winged flies were captured in corn in Dixie, Jack- and Lee Counties (Table 2). Euxesta annonae and son, Sumter, Taylor or Volusia Counties. Chaetop- E. stigmatias were netted from only 3 counties in sis massyla was collected from more counties central and southern Florida, i.e., Miami-Dade, than other species throughout the state and was Okeechobee, and Palm Beach Counties. As a re- netted in 66% of the fields sampled. Euxesta eluta sult of the more limited distribution, both E. an- was netted in 49% of the fields sampled. Chaetop- nonae and E. stigmatias were netted in only 18% sis massyla was the only species collected from of fields sampled. The species netted and reared corn in Alachua, Jefferson and Marion Counties, varied by corn type. Adults of E. annonae and E. while E. eluta was the lone species collected from stigmatias were netted only from sweet corn corn in Okaloosa County. Euxesta stigmatias was fields in Miami-Dade, Okeechobee, and Palm netted only in Martin, Okeechobee, Palm Beach, Beach Counties, but field corn was not sampled in and St. Lucie Counties amounting to only 11% of these counties (Table 2). Adults of E. eluta and C. the fields sampled. Euxesta annonae was not col- massyla were netted from both field and sweet lected in sweep samples in 2008. Euxesta eluta, E. corn fields throughout the state. Euxesta eluta stigmatias and C. massyla were netted from field and C. massyla were netted from 50 and 100% of and sweet corn fields (Table 3). Euxesta eluta field corn fields, respectively, while both species were netted from 27 and 59%, and C. massyla were netted from 100% of sweet corn fields. from 40 and 78% of the field and sweet corn fields The percentage of ulidiid-infested ears ranged throughout the state, respectively. Euxesta stig- from 5% in Escambia to 38% in Santa Rosa matias was caught from 100 and 67% of the field County (Table 2). Euxesta eluta and C. massyla and sweet corn fields, respectively, in Martin, 40 Florida Entomologist 94(1) March 2011 E. annonaeE. eluta E. stigmatias E. massyla C. Mean no. adults emerged per ear (per infested ear) Mean no. No. ears No. sampled (no. infested) (no. 3 , 2007. 4.22.9 56 (6)3.8 88 (14)6.94.8 0.0 (0.0) 56 (5)2.3 0.0 (0.0) 56 (3) 0.9 (8.3)3.2 0.7 (4.4) 56 (6)1.5 0.0 (0.0) 0.0 (0.0) 56 (11) 0.0 (0.0)1.8 0.0 (0.0) 56 (0) 0.2 (2.2) 1.4 (13.5) 0.0 (0.0) 56 (0) 1.6 (9.9) 0.3 (6.3)2.9 0.0 (0.0) 0.0 (0.0) 56 (4)(10.3) 1.1 0.0 (0.0) 0.6 (3.2) 0.0 (0.0) 0.0 (0.0)8.3 0.3 (3.2) 0.0 (0.0) 88 (6) 0.0 (0.0) 0.0 (0.0) 0.6 (10.3) 0.0 (0.0) 1.0 (9.3) 0.0 (0.0)2.7 0.0 (0.0) 56 (21) 0.6 (2.8) 0.3 (4.3)3.3 0.0 (0.0) 0.1 (1.3) 0.0 (0.0) 0.0 (0.0) 56 (12) 0.6 (8.3) 0.0 (0.0) 0.0 (0.0) 88 (16) 0.5 (7.3) 0.3 (4.3)(12.1) 4.5 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.8 (12.3) (13.2) 2.8 11.3 (62.3) 1.1 (3.0) 0.0 (0.0) 0.0 (0.0) 4.6 (21.3) 3 (16.3) LORIDA F IN

EARS

enhanced sweet corn. INFESTED

FROM

Mean no. adults captured per 100 sweeps Mean no. REARED

E. annonaeE. eluta E. stigmatias E. massyla C. Bacillus thuringiensis- OR

with FIELDS

sampled No. rows No. sweep net IN

2 Ear age (d) COLLECTED

date Sample 1 SPECIES

Corn type Fld 17 Oct 15-21 3 0.0 0.0 0.0 LIDIIDAE 2. U Corn type: Fld = field corn; Swt sweet Bt swt Estimated days after first silk at time of sampling. 0 = no flies were detected in sweep nets. 1 2 3 ABLE AlachuaBradfordMiami-Dade Swt SwtEscambia - 1 Swt FldAug 16 Escambia - 2 Bt swt 17 Oct 15-21 6 MarGadsden 15-21 Aug 2 15-21Aug 2 Holmes 7 Lake 8-14 Swt 3 3Lee 9 Swt 17 SepLiberty 3Marion 7 16 Oct Fld 9Okeechobee 15-21 0.0 Fld 2.8 BeachPalm Swt 14 Sep 0.0 Swt SwtSanta Rosa 0.0 4-5 13 Sep 3 18 Sep 3 Swt JohnsSt. 1.8 0.0 15-21 4 Sep 26.0 14 Nov 8-14 Suwannee 0.9 8-14 8-14Aug 3 Swt 11.5Washington 9 Swt Swt 3 8-14 11.2 5.8 0.0 13 Sep 0.0 0.0 3 13 Sep 3 15-21 Aug 3 2 0.0 15-21 0.0 3 15-21 0.0 3.7 6.3 0.0 3 0.0 11.7 3 1.1 1.7 3 0.0 0.0 0.0 56 (16) 0.0 0.0 2.3 0.9 21.3 0.0 16.5 0.0 0.4 (1.3) 0.0 0.0 3.7 17.5 (61.3) 0.0 10.7 33.5 11.5 1.0 5.6 (19.4) 0.9 4.3 5.9 (20.7) 0.0 0.0 0.0 18.3 26.0 0.0 56 (17) 11.3 56 (11) 1.7 (5.5) 0.0 (0.0) 56 (0)(19.1) 5.8 (21.2) 4.2 8.8 (29.1) 0.0 (0.0) 0.0 (0.0) 1.8 (5.9) 0.0 (0.0) 7.1 (36.3) 0.0 (0.0) 0.0 (0.0) County-ﬁeld no. T Goyal et al: Corn-infesting Ulidiidae of Florida 41 E. annonaeE. eluta E. stigmatias E. massyla C. Mean no. adults emerged per ear (per infested ear) Mean no. No. ears No. sampled (no. infested) (no. 3 * 56 (0)* 0.0 (0.0) 0.0 (0.0) 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) , 2008. 1.38.75.7 56 (8)2.2 56 (15)0.0 56 (17)1.3 0.0 (0.0) 88 (2)2.1 0.0 (0.0) 0.2 (1.4) 88 (0)0.8 0.0 (0.0) 3.3 (12.3) 88 (6) 10.2 (33.5)1.3 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 88 (4)0.0 0.0 (0.0) 0.0 (0.0) 0.1 (2.5) 88 (0)1.4 0.0 (0.0) 0.0 (0.0) 1.1 (4.3) 0.0 (0.0) 56 (0)0.7 0.4 (1.5) 0.0 (0.0) 0.0 (0.0) 0.6 (9.3) 56 (0)2.9 0.0 (0.0) 0.0 (0.0) 0.1 (2.3) 88 (18)0.0 0.05 (2.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (0)0.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 88 (9)0.0 0.0 (0.0) 0.8 (11.3) 0.0 (0.0) 0.0 (0.0) 56 (0)0.7 2.7 (13.3) 0.2 (5.3) 0.0 (0.0) 0.0 (0.0) 56 (0)1.3 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 88 (0)1.7 0.0 (0.0) 0.0 (0.0) 0.4 (3.4) 56 (10)0.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 4.4 (21.4) 0.0 (0.0) 56 (0)5.2 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (0)1.8 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 88 (0) 0.4 (2.4) 0.1 (1.4) 0.0 (0.0) 0.0 (0.0) 56 (0)1.3 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (22)0.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)0.9 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (0)4.7 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (9) 1.7 (4.4) 0.0 (0.0) 0.0 (0.0) 88 (0)0.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (8)0.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)0.0 0.0 (0.0) 0.0 (0.0) 0.4 (2.3) 88 (0) 4.1 (10.4) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (0) 3.1 (19.0) 0.0 (0.0) 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.5 (3.3) 0.0 (0.0) 0.5 (3.4) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) LORIDA F IN

EARS

enhanced sweet corn. INFESTED

FROM

Mean no. adults captured per 100 sweeps Mean no. REARED

E. annonaeE. eluta E. stigmatias E. massyla C. Bacillus thuringiensis- OR

with FIELDS

sampled No. rows No. sweep net IN

2 Ear age (d) COLLECTED

date Sample 1 SPECIES

Corn type Fld 4 Jun 2-3 9 0.0 0.0 0.0 LIDIIDAE 3. U Corn type: Fld = field corn; Swt sweet Bt swt Estimated days after first silk at time of sampling. 0 = no flies were detected in sweep nets; * fly species was observed only, not collected. 1 2 3 ABLE AlachuaBradford - 1Bradford - 2 SwtColumbia Swt Swt 23 JunDixie 4 Jun 18-21 23 Jun - 1Gilchrest Swt 10-14 7 - 2Gilchrest Swt 24 Jun 3 - 3Gilchrest Bt fld 23 Jun 8-14Hamilton - 1 23 Jun 3 Bt fld 15-21 Hamilton - 1 23 Jun 7 Swt 3Hendry 7 Fld 9 24 Jun 0.0 9Holmes - 1 24 Jun 7 0.0Holmes - 2 14 - 1Jackson 9 Swt Swt 0.0 2.3 - 2Jackson 9 Fld 0.0 26 Jun 26 Feb 0.0 3.3 Bt fld -1Jefferson 21 15-21 26 Jun 3 5 Jun Bt fld - 1Jefferson 3 0.0 21 0.0 Fld 5 Jun - 1Lafayette 0.0 1.1 2-3 0.7 Swt 0.0 9 - 2Lafayette 25 Jun 2-3 0.0 Swt 25 Jun 3 - 3Lafayette 0.0 8-14 Swt 24 Jun 14 0.0 0.0 0.0Lake - 1 9 9 Fld 14 24 Jun 0.0 1.8 0.0 Lake - 2 9 14 0.0 24 Jun 9Marion 1.3 0.0 14 0.0Martin - 1 Swt 0.0 3 0.0 0.0Martin - 2 3 Swt 0.0 4.6 6 Jun 0.0Martin - 3 3 Swt Bt fld 8.3 21 0.0 0.0 6 JunMartin - 4 0.0 9 Swt 3 Jun 11Mar 21 2.1 0.0 0.0Nassau - 1 14 Swt 11 Mar 0.0 0.0 7 0.0 Nassau - 2 Swt 11 Mar 0.0 1-2 0.0 0.0 Okaloosa - 1 Swt 9 11 Mar 1-2 0.0 0.0Okaloosa - 2 0.0 Fld 9 14 0.0 23 Jun Swt 1.0 3 15-21 0.0 23 Jun Swt 3 3 0.0 0.0 5 Jun 15-21 9 0.0 0.0 5 Jun 0.0 5 3 3 0.0 10 0.0 0.0 9 0.0 0.0 0.0 0.0 0.0 0.0 2.0 3 0.0 0.0 0.0 3 3.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 * 0.0 0.0 1.0 0.0 0.0 14.3 2.0 0.0 0.0 0.0 0.0 0.0 County-field no. T 42 Florida Entomologist 94(1) March 2011 E. annonaeE. eluta E. stigmatias E. massyla C. Mean no. adults emerged per ear (per infested ear) Mean no. , 2008. No. ears No. sampled (no. infested) (no. LORIDA F 3 IN

*** 56 (0) 56 (0) 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)* 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (6) 0.0 (0.0) 0.0 (0.0) 0.3 (3.2) 0.0 (0.0) 0.3 (2.3) 1.91.7 88 (0) 56 (16) 0.0 (0.0)0.0 0.0 (0.0)5.7 0.0 (0.0) 16.1 (56.4)0.0 0.0 (0.0) 56 (0)1.0 0.0 (0.0) 56 (0)0.0 0.0 (0.0) 56 (0)0.0 0.4 (1.4) 0.0 (0.0) 56 (0) 0.0 (0.0) 0.0 (0.0) 56 (0)0.0 0.0 (0.0) 0.0 (0.0) 56 (1)2.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0)0.0 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 88 (4) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 56 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 88 (0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.1 (2.3) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.02 (1.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) 0.0 (0.0) EARS

INFESTED

FROM

enhanced sweet corn. REARED

FIELDS

Mean no. adults captured per 100 sweeps Mean no. IN

E. annonaeE. eluta E. stigmatias E. massyla C. Bacillus thuringiensis- with COLLECTED sampled

No. rows No. sweep net 2 Ear SPECIES

age (d) date LIDIIDAE Sample ) U 1 Corn type ONTINUED 3. (C Corn type: Fld = field corn; Swt sweet Bt swt Estimated days after first silk at time of sampling. 0 = no flies were detected in sweep nets; * fly species was observed only, not collected. 1 2 3 ABLE Okeechobee Beach Palm SwtSanta Rosa - 1 Swtswt Bt Apr 19 Santa Rosa - 2 5 JunApr 10 Swt 14Santa Rosa - 3 10 15-21swt Bt - 1 Johns St. 5 Jun 5 Jun - 2 Johns St. 14 15-21swt Bt Lucie 3St. 6 Jun 9swt Bt 3Sumter 6 Jun 8-14 3 14Suwannee Bt ﬂd 3Taylor 29 May 0.0 3Volusia 14 0.0 Fld Swt 0.0Walkulla 0.0 3 6 JunWalton 4 Jun 11.3 0.0 Fld 15-21 15-21Walton 3.6 Swt 0.0 0.0 9 SwtWalton 25 Jun 2.3 25 Jun 6 Jun 15-21 20.7 3 0.0 3 Fld 14 21 * Swt 7.2 6.3 26 Jun 0.0 0.0 3 Swt 26 Jun 14 0.0 0.0 15-21 26 Jun 12.7 0.0 0.0 3 3 15-21 0.0 0.0 0.0 3 0.0 9 56 (15) 0.0 0.0 9 1.0 0.0 0.0 4.1 0.9 (3.2) 0.0 0.0 0.0 3.3 (12.3) 0.0 0.0 0.0 2.0 0.0 1.1 (4.3) 0.0 1.8 * 2.5 (9.2) 0.0 0.0 0.0 0.0 0.0 0.0 County-ﬁeld no. T Goyal et al: Corn-infesting Ulidiidae of Florida 43

TABLE 4. PERCENTAGE OF FIELDS WITH ULIDIIDAE SPECIES SWEEP NETTED OR REARED FROM CORN EARS BY EAR AGE

Sweep netted Reared from corn ears Ear age (d) Ear age (d)

Species 0 to 7 d 8 to 14 d 15 to 21 d 0 to 7 d 8 to 14 d 15 to 21 d

2007 E. annonae 0 100 100 0 100 100 E. eluta 67 100 89 67 100 78 E. stigmatias 0 100 100 0 100 100 C. massyla 100 100 100 67 100 78

2008 E. annonae 000 00100 E. eluta 36 42 64 27 32 35 E. stigmatias 33 100 100 33 33 100 C. massyla 54 68 71 18 21 41

Okeechobee, Palm Beach, and St. Lucie Counties. ing both sweet and field corn fields across the No E. annonae adults were netted in field or Florida panhandle from Escambia to Nassau sweet corn fields. Counties and south through the peninsula to Mi- Ulidiid-infested ears were found in 13 of 27 ami-Dade County. Flies were collected in sweep counties sampled (Table 3). The percentage of ul- nets or reared from corn ears from 29 out of 33 idiid infested ears ranged from 2% in Volusia to sampled counties during the 2 survey years 39% in Lake County. Only E. eluta were reared (Fig. 2). Flies were more common in the 2007 from corn ears collected from Alachua, Jefferson, compared to 2008 surveys probably due to differ- and Walton Counties. Chaetopsis massyla was the ences in sampling times. Corn fields in 2007 were only species reared from corn ears collected from largely sampled from Aug to Oct, except for Mi- Volusia County. Euxesta eluta and C. massyla ami-Dade County that was sampled in Mar. In were reared from 32% and 28% of the corn fields contrast, surveys were conducted from Feb to Jun sampled throughout the state. Euxesta eluta were in 2008. The flies may be more common in mid- reared from 20 and 38% and C. massyla from 13 summer through fall months in northern Florida. and 34% of the field and sweet corn fields, respec- While more research has been conducted on E tively throughout the state. Euxesta stigmatias stigmatias than the other species, it was found to was only reared from infested sweet corn ears in be much less common than C. massyla and E. Martin and Palm Beach Counties amounting to eluta in this survey. Euxesta eluta and C. massyla 6% of the total fields sampled. Adults of E. an- were distributed in most fields sampled through- nonae were only reared from infested sweet corn out the state in both years, while E. stigmatias ears collected from Palm Beach County amount- and E. annonae were found in only several coun- ing to approximately 2% of the total fields sam- ties of southern Florida (Martin, Miami-Dade, pled. Field corn fields were not sampled in the Okeechobee, Palm Beach, and St. Lucie). counties where E. stigmatias and E. annonae The distribution of alternate host plants and were reared from ears. differences in acceptable temperature ranges for The age of corn ears in surveyed fields ranged each species may explain some of the variation from 1 to 21 d after first silk (Table 3). More E. present in the distribution of ulidiids infesting eluta and C. massyla were caught in sweep nets corn in Florida. Euxesta eluta, E. stigmatias, and and reared from corn ears 15 to 21 d post-silking C. massyla were collected from both field and compared to 0 to 14 d post-silking. More E. stig- sweet corn, while E. annonae was collected only matias were caught in sweep nets and reared from sweet corn fields. Sweet corn is mostly grown from corn ears in fields with 15 to 21 d post-silk- in southern Florida in comparison to northern ing ears than in younger fields. In counties where Florida where field corn predominates (Anony- E. annonae was found, it was only reared from mous 2008a). However, E. stigmatias was not col- fields sampled 15 to 21 d after first silk. lected or reared from sweet corn fields in northern Florida. Frías-L (1978) in Chile found that higher DISCUSSION temperature and lower relative humidity led to greater numbers of E. annonae while the reverse The results of this 2-year study confirmed that led to greater numbers of E. eluta. several species of Ulidiidae flies were infesting Sampling with both sweep nets and collecting corn in Florida. Ulidiidae flies were found infest- infested corn ears gave a more complete picture of 44 Florida Entomologist 94(1) March 2011

Fig. 2. Distribution of Ulidiidae species infesting corn in Florida by county during the 2007-2008 surveys. Sym- bols in ﬁgure represent species collected using sweep nets and reared from corn ears in each of the sampled (shaded) counties.

fly distribution in Florida corn fields than either is currently the only method available for differ- sampling technique alone. Low correlation values entiating the species developing within ears. The indicate that sweep netting is not an efficient external physical characteristics of the immature method to estimate ulidiid species infesting corn stages of Ulidiidae infesting Florida corn are cur- ears. The relationship between sweep nets and fly rently being examined by the authors to deter- species that emerged from infested ears ac- mine the possibility of using eggs, larvae or pupae counted for >60% of the variation for E. stigma- for the identification of species of flies infesting tias and C. massyla, but <60% for E. eluta and E. corn. annonae. There were also a few locations where Euxesta eluta, E. stigmatias and C. massyla were flies were observed but not collected with sweep collected from corn throughout the reproductive nets. These were the places where flies were un- stage of corn. Adult E. annonae may be present in common (1 or 2 per site) and netting was not the fields during the first week of silking, but only fields best sampling technique for insects at low densi- ≥8 d after first silk were sampled in counties where ties. Seal et al. (1996) found that E. stigmatias this species was found. In general, there was a ten- congregated on the top of plants late in the dency for greater infestation by all 4 species as evening. Fly species in our study may have been sweet corn ears neared harvest and as field corn more active or more accessible with nets at times ears approached the dough stage. The authors also of the day other than when sampling was con- have frequently reared E. eluta, E. stigmatias, and ducted. Therefore, sweep netting can be used to C. massyla from tassels and stems of corn plants. indicate the potential for ear infestation, but the Therefore, the potential host period on this crop is identification of adults reared from infested ears longer than just the reproductive stage. Goyal et al: Corn-infesting Ulidiidae of Florida 45

This is the first report of E. annonae infesting (Cyperales: Poaceae), tomato fruit, Lycopersicon corn in Florida and the USA. This species was not esculentum L. (Solanales: Solanaceae) (Seal & common in any location but was always netted Jansson 1989), and decaying carrot roots, Daucus from fields and reared from ears along with other carota L. (Apiales: Apiaceae) (Franca & Vecchia Ulidiidae species. Euxesta annonae was the least 1986). collected species in sweep nets and it was reared Chaetopsis massyla was caught in sweep nets from corn ears collected only from the southern and reared from corn ears in the majority of sur- end of the Florida peninsula (Fig. 2). Euxesta an- veyed counties (Fig. 2). This fly was common in nonae is also reported as a pest of corn in Chile field and sweet corn fields throughout the year in (Frías-L 1978). The authors have frequently ob- southern Florida counties. The relative abun- served E. annonae on Annona spp. (Magnoliales: dance and development range across corn types Annonaceae) and Chinese long bean, Vigna un- indicates this species is a much greater threat to guiculata ssp. sesquipedalis (L.) Verdc. (Fabales: Florida corn than previously recognized. Its habit Fabaceae) in southern Florida and reared E. an- of feeding on a range of monocots may help ex- nonae adults from field collected Annona spp. plain its widespread distribution throughout fruit (Magnoliales: Annonaceae). Plants of An- Florida. Allen & Foote (1992) reported it to be a nona spp. are recorded in several southern and secondary invader of wetland monocots. Chaetop- central Florida counties (Brevard, Broward, Col- sis massyla has been reared from cattail, Typha lier, De Soto, Glades, Hendry, Highlands, Indian spp. (Typhales: Typhaceae) in California (Keiper River, Lee, Manatee, Martin, Miami-Dade, Mon- et al. 2000). Typha spp. are found in most Florida roe, Palm Beach, and St. Lucie) (Wunderlin & counties except Flagler, Gadsden, Glades, Her- Hansen 2008) where they may provide alterna- nando, and Suwannee (Wunderlin & Hansen tive food resources for this species. The authors 2008). The authors made several personal obser- have reared this species from decaying corn vations of C. massyla plant associations during stalks and from spiny amaranth, Amaranthus the course of this statewide survey. Chaetopsis spinosus L. (Caryophyllales: Amaranthaceae) massyla was frequently observed by the authors roots collected from the field at Belle Glade, Flor- on cattail plants on ditch banks and feeding on ida. sugary exudates from sugarcane plants (a com- Euxesta eluta was widely collected in this plex hybrid of Saccharum spp.) in Belle Glade study from fields sampled throughout Florida (Palm Beach County). Chaetopsis massyla adults (Fig. 2). These flies were commonly observed in were reared from sugarcane stems that were ac- fields and as many as 62 were reared from an in- tively infested with the sugarcane borer, Diatraea dividual ear. While this is the first known record saccharalis (F). (Lepidoptera: Crambidae) col- of E. eluta being a pest of corn in Florida and the lected by the authors in November 2009 from sug- USA, its image in Hayslip (1951) suggests that it arcane fields at Clewiston (Hendry County) and was present in Florida corn fields >50 yr ago, but Sebring (Highlands County), Florida. Chaetopsis incorrectly identified as E. stigmatias. The wide massyla was also successfully reared by the au- distribution of E. eluta in Florida and its discov- thors from otherwise healthy sugarcane stems ex- ery on both sweet and field corn indicates this fly posed to colonies in the laboratory in which 0.5 cm is a much greater threat to corn than E. stigma- diam holes were drilled in billets to simulate tias, which is found in a much smaller portion of emergence and frass evacuation holes produced Florida. Euxesta eluta was recognized as infesting by D. saccharalis. Other plants from which C. corn in Puerto Rico >60 yr ago (Wolcott 1948) and massyla has been reared include hairy sedge, has been recorded as an ear pest in Ecuador Carex lacustris Willd. (Cyperales: Cyperaceae) (Evans & Zambrano 1991), Chile (Frías-L 1978; (Allen & Foote 1992), Narcissus spp. (Liliales: Lil- Olalquiaga 1980), Peru (Diaz 1982), Argentina iaceae) (Blanton 1938) and onions, Allium cepa L. (Arce de Hamity 1986), and Brazil (Franca & Vec- (Asparagales: Alliaceae) (Merrill 1951). Carex chia 1986). Euxesta eluta is a pest of loquat, Erio- spp. are found throughout the state while the dis- botrya japonica (Thumb.) Lindl. (Rosales: Ro- tribution of Narcissus spp. is considered to be lim- saceae) in Alachua County, Florida (Anonymous ited to Alachua, Calhoun, Escambia and Leon 2008b). Loquat is grown as a dooryard plant and Counties (Wunderlin & Hansen 2008). is distributed in several counties throughout the Two additional Chaetopsis spp. have been re- state (Wunderlin & Hansen 2008). ported feeding on corn, but neither was found in Euxesta stigmatias was found in sweep net col- this 2-year survey of Florida corn fields. Large lections and reared from corn ears from southern populations of fly larvae that were discovered in and central Florida counties only (Fig. 2). corn stalks within tunnels likely produced by Eu- Weather differences in southern and northern ropean corn borer, Ostrinia nubilalis Hübner Florida may explain part of the variation in dis- (Lepidoptera: Pyralidae) in Ohio were reared to tribution of the species. Adult E. stigmatias have adults and identified as Chaetopsis aenea (Wiede- been reared from damaged or decayed inflores- mann) by Gossard (1919). Larvae of C. fulvifrons cences of sorghum, Sorghum bicolor (L.) Moench (Macquart) were reared from within the tunnels 46 Florida Entomologist 94(1) March 2011 of southwestern corn borer, Diatraea grandiosella dae), a secondary invader of herbaceous stems of Dyar (Lepidoptera: Crambidae), in the Texas high wetland monocots. Proc. Entomol. Soc. Washington plains (Knutson 1987). Langille (1975) reported 94: 320-328. that Chaetopsis spp. larvae were commonly asso- ANONYMOUS. 2008a. Florida Statistical Abstract. Uni- ciated with diapausing D. grandiosella within versity of Florida, Gainesville, FL. ANONYMOUS. 2008b. Florida Cooperative Agricultural corn stalks in Missouri and hypothesized that ul- Pest Survey Program. Quarterly Report No. 2-2008. idiid larvae feed on the decaying stalks or micro- Division of Plant Industry, Florida Department of bial growth within the bored stalks. Agriculture and Consumer Services, Gainesville, In conclusion, 4 species of picture-winged flies FL. were found infesting corn in Florida. Evidence ANONYMOUS. 2008c. Biosystematic Database of World presented herein is the first known documenta- Diptera. Available at http://www.sel.barc.usda.gov/ tion for E. annonae and E. eluta as pests of corn in diptera/names/Status/bdwdstat.htm (verified 10 Florida and the USA. The 4 species were not uni- Nov 2009). formly distributed throughout Florida corn grow- ANONYMOUS. 2009. USDA vegetables 2008 summary, January 2009. Available at http://usda.mannlib.cor- ing regions. Euxesta eluta and C. massyla were nell.edu/usda/current/VegeSumm/VegeSumm-01- found infesting field and sweet corn throughout 28-2009.pdf (verified 10 Nov 2009). Florida. Euxesta stigmatias was only found in- APP, B. A. 1938. Euxesta stigmatias Loew, an otitid fly festing corn in Martin, Miami-Dade, Okeechobee, infesting ear corn in Puerto Rico. J. Agric. Univ. Pu- Palm Beach, and St. Lucie Counties. Euxesta an- erto Rico 22: 181-187. nonae (F.) was found in sweet corn in Miami- ARCE DE HAMITY, M. G. 1986. Biology of Euxesta eluta Dade, Okeechobee, Palm Beach Counties, but (Diptera: Ulidiidae): Behavior in the attack and pu- field corn was not sampled in these counties. Eu- trefaction of corn ears. Acta Zool. Lilloana 38: 119- xesta eluta, E. stigmatias and C. massyla were 128. BAILEY, W. K. 1940. Experiments in controlling corn ear collected from corn throughout the corn reproduc- pests in Puerto Rico. Puerto Rico Agric. Exp. Stn. tive stage. Euxesta annonae was reared from 8-21 Circular no. 23. d old ears only, but fields with ears <8 d old were BARBER, G. W. 1939. Injury to sweet corn by Euxesta not sampled in the counties where this species stigmatias Loew in southern Florida. J. Econ. Ento- was found. The relative abundance of E. eluta and mol. 32: 879-880. C. massyla in Florida field and sweet corn indi- BARBOSA, P., SEGARRA-CARMONA, A. E., AND COLON- cates the need for more research into their biology GUASP, W. 1986. Eumecosomyia nubila (Wiede- and ecology. The discovery of E. eluta and C. mas- mann), a new otitid in Puerto Rico, with notes on the syla attacking corn ears in many of the northern- habits of the dipteran species complex of corn. J. Ag- ric. Univ. Puerto Rico 70: 155-156. most Florida counties suggests that further sur- BLANTON, F. S. 1938. Some dipterous insects reared veys of corn growing areas across the borders into from narcissus bulbs. J. Econ. Entomol. 31: 113-116. neighboring states is warranted to determine the CHITTENDEN, F. H. 1911. Some insects injurious to truck extent of corn infesting picture-winged fly infesta- crops - Notes on various truck-crop insects. USDA tions in the southern U.S. The statewide distribu- Bur. Entomol. 82: 90-90. tion of E. eluta and C. massyla in reproducing CURRAN, C. H. 1928. Insects of Porto Rico and the Virgin corn also suggests that studies should be con- Islands: Diptera or two-winged flies. New York Acad. ducted to evaluate additional food sources that Sci. Scientific Survey of Porto Rico and the Virgin Is- support these species in the absence of corn. lands 11: 1-118. CURRAN, C. H. 1934. The families and genera of North American Diptera, American Museum of Natural History, New York, NY 512 p. ACKNOWLEDGEMENTS CURRAN, C. H. 1935. New American Diptera. American We thank Harsimran K. Gill and Bijayita Thapa for Mus. Novit. 812: 1-24. assistance in rearing Ulidiidae from collected ears, and DALY, T., AND BUNTIN, G. D. 2005. Effect of Bacillus thu- Nicholas A. Larsen for help contacting extension agents ringiensis corn for lepidopteran control of nontarget and field navigation. We acknowledge Jaya Das for help arthropods. Environ. Entomol. 34: 1292-1301. with the Florida county maps. The photographic assis- DIAZ, W. 1982. Danos de Euxesta eluta y E. mazorca tance of Lyle Buss was instrumental in producing the (Dipt: Otitidae) sobre maices amilaceos en la Costa fly images used in this report. We are also thankful to Central del Peru. Rev. Peruvian Entomol. 1: 51-53. University of Florida Cooperative Extension Agents and EVANS, D. C., AND ZAMBRANO, E. 1991. Insect damage in researchers for their help in selecting corn fields and ar- maize of highland Ecuador and its significance in ranging the visits, and corn growers of Florida for allow- small farm pest management. Trop. Pest Manage. ing us to survey their fields. This research was made 37: 409-414. possible by a Hand Fellowship awarded by the Dolly FISHER, E. 1996. Two new insect pests of corn in Califor- and Homer Hand Group. nia. New Pest/Disease Advisory, 31 December 1996. State of California, Dept. Food & Agric., Div. Plant Ind. REFERENCES CITED FRANCA, F. H., AND VECCHIA, P. T. D. 1986. Damages caused by Euxesta stigmatias on carrot roots in com- ALLEN, E. J., AND FOOTE, B. A. 1992. Biology and imma- mercial seed field. Pesq. agropec. bras., Brasília 21: ture stages of Chaetopsis massyla (Diptera: Otiti- 789-791. Goyal et al: Corn-infesting Ulidiidae of Florida 47

FRÍAS-L, D. 1978. Ecological studies in Euxesta eluta SAS INSTITUTE. 2008. Proc user’s manual, version 9th and Euxesta annonae (Diptera: Otitidae). Agric. Téc- ed. SAS Institute, Cary, NC. nica (Chile) 38: 109-115. SCULLY, B. T., NUESSLY, G. S., AND BEIRIGER, R. L. GOSSARD, H. A. 1919. Insects resembling the European 2000. Resistance in maize to Euxesta stigmatias corn borer. Mon. Bull. Ohio Agric. Exp. Stn. 4: 372-379. Loew (Diptera: Otitidae). J. Entomol. Sci. 35: 432- GOYAL, G., NUESSLY, G. S., STECK, G. J., SEAL, D. R., 443. CAPINERA, J. L., AND BOOTE, K. J. 2010. New report SEAL, D. R. 1996. Insect control in sweet corn, 1995. Ar- of Chaetopsis massyla (Diptera: Ulidiidae) as a pri- thropod Manage. Tests 21: 111-111. mary pest of corn in Florida. Florida Entomol. 93: SEAL, D. R. 2001. Control of the corn silk fly using vari- 198-202. ous insecticides, 2000. Arthropod Manage. Tests 26: HAYSLIP, N. C. 1951. Corn silk fly control on sweet corn, E31. Entomol. Soc. Am. Available at http:// pp. 1-6. Univ. Florida Agric. Exp. Stn. Circ S-41, www.entsoc.org/Protected/ AMT/Amt26/INDEX.ASP Gainesville, FL. (verified 10 Nov 2009). KEIPER, J. B., SANFORD, M., JIANNINO, J., AND WALTON, SEAL, D. R., AND JANSSON, R. K. 1989. Biology and man- W. E. 2000. Invertebrates inhabiting wetland mono- agement of corn silk fly, Euxesta stigmatias Loew cots damaged by Lepidoptera. Entomol. News 111: (Diptera: Otitidae), on sweet corn in southern Flori- 348-354. da. Proc. Florida State Hort. Soc. 102: 370-373. KNUTSON, A. 1987. Dynamics and natural enemies of SEAL, D. R., AND JANSSON, R. K. 1994. Insect control in the southwestern corn borer in the Texas High sweet corn, 1991. Arthropod Manage. Tests 19: 96- Plains. PhD Dissertation, Texas A & M University, 96. College Station, TX. SEAL, D. R., JANSSON, R. K., AND K. BONDARI. 1996. LANGILLE, R. N. 1975. Observations on the overwinter- Abundance and reproduction of Euxesta stigmatias ing survival and spring development of the south- (Diptera: Otitidae) on sweet corn in different envi- western corn borer, Diatraea grandiosella Dyar. PhD ronmental conditions. Florida Entomol. 79: 413-422. Dissertation, University of Missouri, Columbia, MO. VAN ZWALUWENBURG, R. H. 1917. Report of the Ento- MERRILL, JR., L. S. 1951. Diptera reared from Michigan mologist: A new corn pest. Puerto Rico Agric. Exp. onions growing from seeds. J. Econ. Entomol. 14: Stn. 31-34. 1015-1015. WALTER, E. V., AND WENE, G. P. 1951. Tests of insecti- MOSSLER, M. A. 2008. Crop profile for sweet corn in Flor- cides to control larvae of Euxesta stigmatias and Me- ida. CIR 1233, University of Florida. Available at ht- gaselia scalaris. J. Econ. Entomol. 44: 998-999. tp://edis.ifas.ufl.edu/pi034 (verified 10 Nov 2009). WOLCOTT, G. N. 1948. The insects of Puerto Rico. J. Ag- NUESSLY, G. S., AND HENTZ, M. G. 2004. Contact and ric. Univ. Puerto Rico 32: 417-748. leaf residue activity of insecticides against the sweet WUNDERLIN, R. P., AND HANSEN, B. F. 2008. Atlas of corn pest Euxesta stigmatias Loew (Diptera: Otiti- Florida Vascular Plants. Institute for Systematic dae). J. Econ. Entomol. 97: 496-502. Botany, University of South Florida, Tampa, FL. OLALQUIAGA, G. F. 1980. Aspectos fitosanitarios de la is- WYCKHUYS, K. A. G., AND O’NEIL, R. J. 2007. Local agro- la de pascua. Rev. Chil. Entomol. 10: 101-102. ecological knowledge and its relationship to farmers’ PAINTER, R. H. 1955. Insects on corn and teosinte in pest management decision making in rural Hondu- Guatemala. J. Econ. Entomol. 48: 36-42. ras. Agric. Hum. Values 24: 307-321.

48 Florida Entomologist 94(1) March 2011

A CHECKLIST AND KEY TO SPECIES OF THE GENUS BETACIXIUS MATSUMURA (HEMIPTERA: FULGOROMORPHA: CIXIIDAE) WITH DESCRIPTIONS OF TWO NEW SPECIES FROM GUIZHOU PROVINCE, CHINA

PEI ZHANG 1, 2, 3 AND XIANG-SHENG CHEN 1, 2, * 1The Provincial Key Laboratory for Agricultural Pest Management of Mountainous Region, Guizhou University, Guiyang, Guizhou 550025, P.R. China

2Institute of Entomology, Guizhou University, Guiyang, Guizhou 550025, P.R. China

3 Xingyi Normal University for Nationalities, Xingyi, Guizhou 562400, P.R. China

*Corresponding author; E-mail: [email protected]

ABSTRACT

Two new species of Betacixius Matsumura, 1914 (Hemiptera: Fulgoromorpha: Cixiidae: Cixiini), B. bispinus Zhang and Chen sp. nov. (China: Guizhou) and B. ﬂagellihamus Zhang and Chen sp. nov. (China: Guizhou), from southwest China, are described and illustrated. A key for identifying 23 known species of Betacixius is provided.

Key Words: Hemiptera, Fulgoroidea, Cixiidae, Oriental region, Betacixius, new species, China

RESUMEN

Se describen e ilustran dos nuevas especies de Betacixius Matsumura, 1914 (Hemiptera: Fulgoromorpha: Cixiidae: Cixiini), B. bispinus Zhang y Chen sp. nov. (China: Guizhou) y B. ﬂagellihamus Zhang y Chen sp. nov. (China: Guizhou) del suroeste de China. Se provee una clave para identiﬁcar las 23 especies conocidas de Betacixius.

The cixiid planthopper genus Betacixius (Cixi- trations were scanned with Canon CanoScan inae: Cixiini) was established by Matsumura LiDE 200 and imported into Adobe Photoshop 8.0 (1914) for the type species, B. ocellatus Mat- for labeling and plate composition. Specimens ex- sumura, 1914, from Japan. To date, 21 species amined are deposited in the Institute of Entomol- with 2 subspecies have been recorded worldwide ogy, Guizhou University, Guiyang, Guizhou Prov- and all species occur in the southern region (Mat- ince, China (IEGU). sumura 1914; Schumacher 1915; Metcalf 1936; Jacobi 1944; Fennah 1956; Hori 1982; Chou et al. DESCRIPTIVE TAXONOMY 1985, 1988; Tsaur et al. 1991; Hua 2002). Betacixius Matsumura, 1914 (Figs. 1-25) During the course of studying species biodiversity of the suborder Auchenorrhyncha in south- Betacixius Matsumura 1914: 412; Chou et al. 1985: 23; west China, 2 specimens belonging to unde- Tsaur et al. 1991: 27. scribed species of the genus Betacixius were found. The purpose of this paper is to describe Type species: Betacixius ocellatus Matsumura these 2 new species and to provide an identiﬁca- 1914, by original designation. tion key to all species of Betacixius. Description. This is a redescription incorporat- ing the descriptions previously published by Chou et al. (1985) and Tsaur et al. (1991) as follows. MATERIALS AND METHODS Morphological terminology follows Löcker et Body Size and Coloration. Small to medium- al. (2006). Dry specimens were used for the de- size cixiids (4.3-7.3 mm). Body coloration varying scription and illustration. External morphology from green, brown to fulvous, mostly bearing spe- was observed under a stereoscopic microscope cial markings on anteclypeus; lateroapical parts and characters were measured with an ocular mi- of frons, otherwise unicolorous throughout. crometer. The genital segments of the examined specimens were macerated in 10% KOH and Head and Thorax. Head including eyes drawn from preparations in glycerin jelly with slightly narrower than pronotum. Vertex much the aid of a Leica MZ 12.5 stereomicroscope. Illus- wider than long in midline, widest basally or

Zhang & Chen: New Species of Betacixius from China 49

Figs. 1-13. Betacixius bispinus Zhang and Chen sp. nov. 1. Head and thorax, dorsal view; 2. Frons; 3. Forewing; 4. Male genitalia, lateral view; 5. Pygofer and genital styles, ventral view; 6. Anal segment, dorsal view; 7. Anal segment, caudal view; 8. Connective, caudal view; 9. Right genital style, ventral view; 10. Aedeagus, left side; 11. Aede- agus, right side; 12. Aedeagus, dorsal view; 13. Aedeagus, ventral view. Scale bars = 0.25 mm (Figs. 6, 7), 0.5 mm (Figs. 1, 2, 4, 5, 8-13), 1 mm (Fig. 3).

50 Florida Entomologist 94(1) March 2011 apically, lateral carinae moderately elevated, Male Genitalia. Pygofer symmetrical, U- disc shallowly hollowed on each side of median shaped, with thumb-shaped dorsolateral angles carina. Frons rounded at base, usually with in- in ventral view. Medioventral process triangular complete median carina not reaching anterior or subtriangular in ventral view, generally wider margin of vertex, lateral carinae slightly ele- at base than long in midline. Anal segment tubu- vated below level of eyes, with small median lar. Genital styles symmetrical in ventral view. ocellus and semicircular frontoclypeal suture. Aedeagus slender in lateral view. Clypeus tricarinate, convex to midline. Prono- Distribution. Oriental and Palaearctic regions. tum small, with distinct median carina, inter- Remarks. This genus may be easily distin- median carinae curving laterad, angularly guished from other genera of Cixiini by the presence rounded posteriorly. Mesonotum tricarinate, of 4-5 subapical cells and 8-9 apical cells on the convex between lateral carinae, flattened pos- forewing, vertex much wider than long in midline, teromedially. Forewings broadest at apical frons with incomplete median carina distinct near third, rounded at apex, with 4-5 subapical cells frontoclypeal suture, and chaetotaxy of hind tarsus and 8-9 apical cells, hyaline, sometimes with an 7/7. The 2 new species, B. bispinus Zhang and Chen oblique band or ocellated stripe. Hind tibia with sp. nov. (China: Guizhou) and B. flagellihamus 2-4 lateral spines and 6 apical spines. Chaetot- Zhang and Chen sp. nov. (China: Guizhou), fit into axy of hind tarsus 7/7. the genus by the presence of features as above.

WORLD CHECKLIST OF SPECIES OF BETACIXIUS MATSUMURA

B. bispinus Zhang and Chen sp. nov.; southwestern China (Guizhou). B. brunneus Matsumura (1914); China (Taiwan), Japan. B. clypealis Matsumura (1914); China (Taiwan). B. clypealis vittifrons Matsumura (1914); China (Taiwan). B. delicatus Tsaur & Hsu (1991); China (Taiwan). B. euterpe Fennah (1956); China (Guangdong). B. ﬂagellihamus Zhang and Chen sp. nov.; southwestern China (Guizhou). B. ﬂavovittatus Hori (1982); China (Taiwan). B. fuscus Tsaur & Hsu (1991); China (Taiwan). B. herbaceus Tsaur & Hsu (1991); China (Taiwan). B. kumejimae Matsumura (1914); South China, Japan (Okinawa). B. maculosus Tsaur & Hsu (1991); China (Taiwan). B. michioi Hori (1982); China (Taiwan). B. nelides Fennah (1956); China (Zhejiang, Guangdong). B. nigromarginalis Fennah (1956); China (Hubei). B. obliquus Matsumura (1914); South China, Japan (Gifu). B. obliquus pallens Matsumura (1914); Japan (Tokyo, Harima, Kumanoto) B. ocellatus Matsumura (1914); China (Taiwan), Japan. B. pallidior Jacobi (1944); South China (Fujian). B. rinkihonis Matsumura (1914); China (Taiwan), Japan. B. robustus Jacobi (1944); South China (Fujian). B. shirozui Hori (1982); China (Taiwan). B. sparsus Tsaur & Hsu (1991); China (Taiwan). B. tonkinensis Matsumura (1914); South China, Vietnam. B. transversus Jacobi (1944); South China (Fujian).

KEY TO SPECIES OF THE GENUS BETACIXIUS MATSUMURA

1. Forewings with markings (Figs. 3, 16, 24, 25) ...... 2 — Forewings without any markings ...... 9 2. Forewings with a large ocellate marking in apical half (Figs. 16 and 25) ...... 3

Zhang & Chen: New Species of Betacixius from China 51

Figs. 14-23. Betacixius ﬂagellihamus Zhang and Chen sp. nov. 14. Head and thorax, dorsal view; 15. Frons; 16. Forewing; 17. Male genitalia, lateral view; 18. Pygofer and genital styles, ventral view; 19. Anal segment, dorsal view; 20. Anal segment, caudal view; 21. Genital styles, ventral view; 22. Aedeagus, dorsal view; 23. Aedeagus, right side. Scale bars = 0.25 mm (Figs. 18-21), 0.5 mm (Figs. 14, 15, 17, 22, 23), 1 mm (Fig. 16).

52 Florida Entomologist 94(1) March 2011

— Forewings without such a marking in apical half (Figs. 3 and 24)...... 6

3. Forewings with an oblique brown band extending from clavus across middle of corium ...... B. tonkinensis

— Forewings without such a band (Figs. 16 and 25) ...... 4

4. Flagellum of aedeagus with 1 spine, hook-shaped (Figs. 22 and 23) ...... B. ﬂagellihamus sp. nov.

— Flagellum of aedeagus with 2 spines, not hook-shaped...... 5

5. Aedeagus with 2 L-shaped processes apically ...... B. maculosus

— Aedeagus with 1 nearly straight and 1 arched process apically...... B. ocellatus

6. Forewings with an oblique band extending from stigma passing through its middle part ...... 7

— Forewings without such a band (Figs. 3 and 24) ...... 12

7. Frons with median carina distinct on apical third; hind basitarsus much longer than the 2nd and 3rd segment put together...... B. michioi

— Frons and hind basitarsus without features as above...... 8

8. Forewings with apical cells of M and Cu strongly infuscate...... B. transversus

— Forewings with apical cells not infuscate...... 9

9. Forewings with apical margin black or distinctly darkened...... 10

— Forewings with apical margin fuscous or not distinctly darkened ...... 11

10. Frons with a pallid spot at centre of lateral margins, clypeus dark, mesonotum testaceous ...... B. kumejimae

— Frons without such spots; mesonotum, except scutellum, castaneous-piceous ...... B. euterpe

11. Forewings with an oblique dark band extending from clavus into centre of corium, slightly distad of level of union of claval veins...... B. obliquus

— Forewings with a spot near sutural margin of clavus near union of claval veins, no oblique dark band at this level extending into corium ...... B. pallidior

12. Forewings with a long black stripe from base, along clavus extending to Rs ...... B. fuscus

— Forewings without such a stripe (Figs. 3 and 24) ...... 13

13. Fore tibiae with black, longitudinal stripes...... 14

— Fore tibiae without such stripes ...... 15

14. Mid- and hind- tibiae with black, longitudinal stripes ...... B. delicatus

— Mid- and hind- tibiae without such stripes ...... B. sparsus

15. Forewings infuscated at base, extending along clavus to end of Cu1; mesonotum with a large, very distinct brown marking between lateral carinae ...... B. shirozui

— Forewings and mesonotum without spots as above (Figs. 3 and 24) ...... 16

16. Forewings with apical margin black or very dark (Figs. 3 and 24) ...... 17

— Forewings with apical margin not particularly dark...... 18

17. Medioventral process of pygofer in ventral view right-angled triangular, pointed at apex (Fig. 5); on ventral margin, periandrium with lobate processes, 2 broad processes basally, bending forward, directed ventrocephalad, ﬂagellum semisclerotised, with several serrated processes near apex (Figs. 10 and 12) ...... B. bispinus sp. nov.

— Medioventral process in ventral view subtriangular, rounded at apex; periandrium without lobate processes, but with serrated processes basally, ﬂagellum, without any processes ...... B. rinkihonis

18. First apical cell of forewing piceous ...... B. robustus

— A dark suffusion over all apical cells and across base of forewing ...... B. nelides Zhang & Chen: New Species of Betacixius from China 53

19. Aedeagus with a curved spine on left near apex and a short ledge in a similar position on right; flagellum arising above left margin, sides parallel for most of length, distally a short curved spine directed cephalad, and a subquadrate plate with a stout spine directed ventrad ...... B. nigromarginalis — Aedeagus and flagellum without features as above ...... 20 20. Frons without median carina ...... B. clypealis — Frons with median carina ...... 21 21. Frons with white or yellowish lateroapical parts; anteclypeus entirely black ...... B. flavovittatus — Frons and clypeus unicolorous ...... 22 22. Body pale brown; flagellum of aedeagus with 2 processes on right side ...... B. brunneus — Body green; flagellum of aedeagus with one process on each side ...... B. herbaceus Betacixius bispinus Zhang and Chen sp. nov. teromedially, carinae concolorous. Frons yellow- (Figs. 1-13, 24) ish brown except lateral carinae blackish brown. Postclypeus yellowish brown, anteclypeus black- Description. Body length (from apex of vertex ish brown. Rostrum generally yellowish brown, to tip of forewings): male 5.0-6.1mm (n = 2), fe- blackish brown near tip. Forewings pale brown, male 5.5-6.8mm (n = 7); forewing length: male hyaline; veins brown, tubercles dark brown; 4.0-4.9mm (n = 2), female 4.5-5.7mm (n = 7). stigma black; clavus with a short transversal

brown band, just distad of fork PCu+A1. Hind-tib- Coloration. General color brown. Body slightly iae yellowish brown, lateral- and apical- spines covered with powdery wax. Eyes yellowish brown yellowish brown basally, black apically; platellae to blackish brown; ocelli reddish yellow. Vertex of tarsi dark brown. Abdomen black ventrally. yellowish brown. Pronotum blackish brown except median carina yellowish brown. Mesonotum Head and Thorax. Vertex narrowing to apex as blackish brown, with yellowish brown area pos- shown in Figs. 1 and 24, wider than distance be-

Figs. 24-25. Body of adult in dorsal view. 24. Betacixius bispinus Zhang and Chen sp. nov.; 25. Betacixius flagellihamus Zhang and Chen sp. nov. 54 Florida Entomologist 94(1) March 2011 tween eyes, 2.5 times wider than long in midline; touching each other; in lateral view, ventral mar- anterior margin arched convex, with small emar- gin curving upward, outside slightly corrugated, gination at midpoint, posterior margin arched dorsal margin smooth and bent forming a right concave; median carina indistinct, lateral carinae angle approximately; incompactly connected with without branches, subapical carina absent. Me- connective, freely movable; apical margin with dian ocellus very small, located the centre of the bristles in ventral view. Aedeagus broad, short, frontoclypeal suture. Pedicel of antenna 1.6 times connected with anal segment by 2 points; each longer than wide. Frons broad, with small spots side with broad spine arising at apex of aedeagal on middle area, widest at level of lateral ocelli, shaft, curving upward; periandrium ventrally narrowing to both ends, 1.25 times longer than with lobate processes, basally 2 broad processes, wide in midline; median carina indistinct, extend- bending forward, directed ventrocephalad. Con- ing from slightly above level of lateral ocelli to nective anchor-like, relatively long, the width of median ocellus, lateral carinae distinct and aedeagal shaft 1.65 times as wide as width of con- ridged, arched convex; anterior margin arched nective plus ventral arm. Flagellum semi-sclero- concave. Clypeus with median carina distinct and tised, structure simple, generally curving left, elevated throughout, widest at level of endpoints with several serrated processes near apex. of frontoclypeal suture; lateral carinae distinct and elevated. Rostrum relatively short, reaching Type Material. Holotype: , Mayanghe Na- hind coxae, apical and subapical segments tional Natural Reserve (600m), Yanhe County, equally long. Pronotum short and narrow, collar- Guizhou Province, China, 5-12 June 2007, X.-S. like, twice as long as vertex in midline; median Chen. Paratypes: 1, 7, same data as holo- carina distinct and complete; intermedian cari- type. nae corrugated and curving into posterior margin which is concave in obtuse angle. Mesonotum 1.89 Etymology. The name is derived from the Latin times longer than pronotum and vertex com- words bi- (double) and spinus (spine), which re- bined; 3 longitudinal carinae all reaching ante- fers to the 2 spines on ventral margin of perian- rior and posterior margins, median carina indis- drium. tinct on posteromedian area, which bears trans- Distribution: Southwest China (Guizhou Prov- verse striations. Forewings 2.22 times longer ince). than wide, with sparse setae on surface, tubercles Remarks. This new species is similar in ap- along veins, claval veins without tubercles; 2 in- pearance to B. rinkihonis, but differs from the lat- distinct subapical lines of cross veins; fork Sc+RP ter in the shape of the medioventral process and distad of fork CuA1+CuA2; r-m crossvein slightly the anal segment and by having 2 spines on the distad of fork MA+MP; RP apically bifid, MA api- ventral margin of the periandrium and several cally bifid, MP apically bifid; fork PCu+A1 serrated processes near apex of flagellum. slightly basad of centre of clavus; Sc+R and M fused at superior-outside angle of basal cell; fork Betacixius flagellihamus Zhang and Chen sp. nov. (Figs. 14-23, 25) MA1+MA2 distad of fork MP1+MP 2. 2nd hindtarsus with 5 platellae; hind-tibia with 4 lateral Description. Body length (from apex of vertex spines, 6 apical spines: 2 large, 1 medium, 3 to tip of forewings): male 5.0-5.8mm (n = 13), fe- small, divided into 2 groups. male 5.4-6.2mm (n = 15); forewing length: male 4.5-5.0mm (n = 13), female 4.6-5.1mm (n = 15). Male Genitalia. In ventral view, pygofer stout, slightly concave medially, widening laterally; dor- Coloration. General color brown. Body covered sal margin caudad obliquely raised in lateral with powdery wax. Median area of eyes black, ven- view, inferior part with bristles; lateral lobes sym- tral margin yellow, other part reddish brown to metrical, medium-inferior part arched convexly blackish brown. Median ocellus pale yellow, semi- in lateral view. Medioventral process right-angled hyaline; lateral ocelli red. Vertex pale yellow. triangular in ventral view, relatively broad, 1.5 Pronotum pale yellow. Mesonotum brown. Frons times wider than long, distance between tips of 2 generally brown, bright yellow above frontoclypeal lateral lobes 3.06 times as long as width of medio- suture; postclypeus yellow to brown, with oblique ventral process; narrow triangular in lateral view, streaks; anteclypeus black. Apical segment of ros- covered basally. Anal segment short and stout as trum brown, subapical segment yellow. Forewings shown in Figs 4, 6 and 7; 2.13 times longer than pale brown, semihyaline, with ocellated marking wide in dorsal view; incompactly connected with as shown in Figs. 16 and 25; veins and tubercles pygofer, freely movable; anal style, finger-like api- brown; stigma brown to dark brown; clavus with cally, not beyond anal segment; anal opening dark brown stain on apical third, sometimes ex- nearly subelliptical in dorsal view. Genital styles tending to end of clavus; clavus suture brown. as shown in Figs. 4, 5 and 9, in ventral view, wid- Hind-tibia brown, lateral- and apical-spines brown ening to apex, apical margin truncated, with at base, black apically; membranous tooth of tarsi sharp angle outside, internal processes broad, dark yellow. Abdomen black ventrally. Zhang & Chen: New Species of Betacixius from China 55

Head and Thorax. Vertex narrowing towards touching each other; in lateral view, ventral mar- apex as shown in Figs. 14 and 25, wider than dis- gin curving upward, dorsal margin strongly bend- tance between eyes, 2.3 times wider than long in ing upward; compactly connected with connec- midline, separated into 2 hollow areolets by me- tive, immovable. Aedeagus stout, structure sim- dian carina; anterior margin generally arched ple; each side with a broad spine at apex of convex slightly, slightly concave at midpoint, pos- aedeagal shaft, right one curving dorsad, left one terior margin concave in obtuse angle; median ca- relatively straight, directed up-cephalad. Connec- rina distinct and complete, subapical carina ab- tive relatively slender, the width of aedeagal shaft sent. Median ocellus indistinct, in the centre of fr- 1.33 times as wide as width of connective plus ontoclypeal suture. Frons with well-distributed ventral arm. Flagellum strongly sclerotised, small spots, widest at level of antennae, narrow- freely movable, structure simple, with a barb- ing to both ends, length equal to width; median shaped spine at apex. carina distinct and elevated near frontoclypeal suture; lateral carinae slightly S-shaped, ele- Type Material. Holotype: , Leigongshan Na- vated; apex of frons elevated and lobate; anterior tional Natural Reserve, Leishan County, Guizhou margin semicircle concave. Median carina of Province, China, 13 May 1985, Z.-Z. Li. clypeus distinct and elevated; lateral carinae of Paratypes: 7, 9, same data as holotype; postclypeus elevated. Rostrum reaching hind- 2, 3, Guiyang, Guizhou Province, China, femora, apical and subapical segments equally June 1983, Students of Grade 79, Major Plant- long. Pronotum short and narrow, collar-shaped, protecting; 2, 1, Huaxi (1000m), Guiyang, 2.25 times longer than vertex in midline; median Guizhou Province, 20 May 2007, Q.-Z. Song; 1, carina distinct and elevated; posterior margin 2, Forest Park (1000m), Guiyang, Guizhou concave in obtuse angle. Mesonotum 1.44 times Province, China, 20 May 2007, X.-S. Chen. longer than pronotum and vertex combined; three longitudinal carinae elevated except for median Etymology. The name is derived from the Latin carina weakly elevated at base, all reaching ante- words flagell (flagellate) and hamus (hook), which rior and posterior margins. Forewings 2.2 times refers to the hook-like spine of flagellum. longer than wide; surface of Forewings with se- Distribution: Southwest China (Guizhou Prov- tae, basal part less, apical part more; veins with ince). distinct tubercles, C vein with 34 tubercles, claval Remarks. This new species is similar in ap- apically vein with tubercles; 2 indistinct subapi- pearance to B. ocellatus, but differs from the lat- cal lines of cross veins; fork Sc+RP distad of fork ter in the shape of the anal segment and the num- CuA1+CuA2; r-m crossvein distad of fork ber of spines on the flagellum. MA+MP; RP apically bifid, MA apically bifid, MP apically bifid; fork PCu+A1 slightly basad of cen- ACKNOWLEDGMENTS tre of clavus; Sc+R and M fused at superior-outside angle of basal cell; fork MA1+MA2 distad of We are grateful to Prof. Zi-Zhong Li, Ms. Qiong- Zhang Song (Institute of Entomology, Guizhou Univer- fork MP1+MP2. 2nd hindtarsus with 3 platellae; sity, China) for providing valuable specimens. We thank hind-tibia with 3 lateral spines, 6 apical spines, Prof. Dr. Shun-Chern Tsaur (Research Center for Biodi- being divided into 2 groups by a relatively wide versity, Academia Sinica, Taibei, Taiwan), Kun-Wei gap, one group with 3 equal spines, arranged Huang (National Museum of Natural Science, Tai- closely, the other group with 1 large and 2 small, chung, Taiwan) and Ms. Gail Charabin (Saskatoon Re- arranged sparsely. search Centre, Agriculture and Agri-Food Canada) for obtaining literature. This work was supported by the Male Genitalia. In ventral view, pygofer stout, National Natural Science Foundation of China (No.30560020, 31060290), the China Postdoctoral Sci- shallowly U-shaped, slightly widening from base ence Foundation (No. 2005037111), the Program for to end, ventral margin slightly concave; dorsal New Century Excellent Talents in University (NCET- margin caudad obliquely upward in lateral view; 07-0220), the Specialized Research Fund for the Doc- lateral lobes symmetrical, inferior half slightly toral Program of Higher Education (No. 20060657001), arched concave in lateral view. Medioventral pro- and the International Science and Technology Coopera- cess mastoid in ventral view, with bristles, 1.25 tion Program of Guizhou (20107005). times wider than long, distance between tips of 2 lateral lobes 2.4 times as long as width of medio- REFERENCES CITED ventral process; tongue-shaped in lateral view. Anal segment short and stout as shown in Figs CHOU, I., LU, J.-S., HUANG, J., AND WANG, S.-Z. 1985. 17, 19 and 20; in dorsal view 2 times longer than Homoptera, Fulgoroidea. Economic Insect Fauna of wide; compactly connected with pygofer, immov- China. Fasc. 36. Science Press, Beijing pp. 1-152. CHOU, I. WANG, Y.-L, HUANG, B.-K., AND YUAN, X.-Q. able; anal styles, not beyond anal segment; anal 1998. Homoptera: Fulgoroidea: Cixiidae, pp. 379-382 opening pear-like in dorsal view. Genital styles as In B.-K Huang [ed.], Insect Fauna of Fujian Prov- shown in Figs 17, 18 and 21, in ventral view, wid- ince, Volume II. Fuzhou, China, Science and Tech- ening to apex, internal processes broad, not nology Press. 56 Florida Entomologist 94(1) March 2011

FENNAH, R. G. 1956. Fulgoroidea from Southern China. LÖCKER, B., FLETCHER, M. J., LARIVIÈRE, M.-C., AND Proc. California Acad. Sci. 28: 441-527. GURR, G. M. 2006. The Australian Pentastirini HORI, Y. 1982. The Genus Betacixius Matsumura, 1914 (Hemiptera: Fulgoromorpha: Cixiidae). Zootaxa (Homoptera: Cixiidae) of Formosa, pp. 175-182 In M. 1290: 1-138. Satô, Y. Hori, Y. Arita and T. Okadome [eds.], Special MATSUMURA, S. 1914. Die Cixiinen Japans. Annota- issue to the memory of retirement of Emeritus Pro- tiones Zoologicae Japanenses, Tokyo 8: 393-434 fessor Michio Chûjô. Association of the Memorial Is- METCALF, Z. P. 1936. General Catalogue of the Homoptera sue of Emeritus Professor M. Chûjô C/O Biological Fascicle IV Fulgoroidea, Part 2, Cixiidae. pp 269. Laboratory, Nagoya Women’s University, Nagoya, SCHUMACHER, F. 1915. Der gegenwärtige Stand unserer Japan. Kenntnis von der Homopteren-Fauna der Insel For- HUA, L.-Z. 2002. Catalogue of Insects of China I. Guang- mosa unter besonderer Berücksichtigung von Sau- zhou, China: Sun Yat-Sen University Press. ter’schem Material. Mitteilungen aus dem Zoologis- JACOBI, A. 1944. Die Zikadenfauna der Provinz Fukien chen Museum in Berlin. Berlin 8: 73-134 in Südchina und ihre tiergeographischen Beziehun- TSAUR, S.-C., HSU, T.-C., AND VAN STALLE, J. 1991. Cixi- gen. Mitteilungen der Münchner Entomologischen idae of Taiwan, Part V. Cixiini except Cixius. J. Tai- Gesellschaft 34: 5-66. wan Museum 44: 1-78.

Scheffrahn & Crowe: Termites on Boats 57

SHIP-BORNE TERMITE (ISOPTERA) BORDER INTERCEPTIONS IN AUSTRALIA AND ONBOARD INFESTATIONS IN FLORIDA, 1986-2009

RUDOLF H. SCHEFFRAHN1 AND WILLIAM CROWE2 1Fort Lauderdale Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, 3205 College Avenue, Davie, Florida, 33314, U.S.A. rhsc@uﬂ.edu

2Australian Quarantine and Inspection Service, P.O. Box 222, Hamilton Central, Queensland, 4007, Australia [email protected]

ABSTRACT

Alate termite ﬂights from mature colonies infesting marine vessels is a primary mechanism for anthropogenic transoceanic establishment of invasive termite species. A taxonomic review is given of 133 recorded termite infestations onboard vessels in Australia and Florida between 1986 and 2009. The differing governmental approaches to regulating entry by foreign boats appears to reﬂect the relative frequency of exotic termite establishments in Aus- tralia and Florida.

Key Words: invasive species, biosecurity, overwater dispersal, Kalotermitidae, Rhinotermiti- dae, Termitidae

RESUMEN

El vuelo de termitas aladas de colonias maduras que infestan barcos es el mecanismo principal para el movimiento transoceánico y establecimiento de especies de termitas invasoras. Se provee una revisión taxonómica de 133 infestaciones de termitas encontradas en barcos en Australia y la Florida entre 1986 y 2009. Los diferentes enfoques gubernamentales en regular la entrada de barcos extranjeros tienden a reﬂejar la frecuencia relativa de establecimiento de termitas exóticas en Australia y la Florida.

Natural overwater dispersal of infested flot- termitidae) or by alates (all taxa) flying onboard sam and anthropogenic dispersal by maritime during dockage, either on water or in dry dock. vessels are the 2 primary means by which ter- Hochmair & Scheffrahn (2010) showed a strong mites are transported across distant sea barriers correlation between land-borne infestations of (Scheffrahn et al. 2009). The cessation of rapid Coptotermes spp. in Florida and their distance to late Pleistocene/Holocene sea level rise at about maritime boat dockage suggesting that marine 7K years before present (ybp, Fleming et al. 1998) vessels are predominant vehicles for dissemina- predates the first known long-distance human tion of this pest genus. maritime voyages by some 3.5K ybp (Anderson et Within the last century, 6 exotic termite spe- al. 2006). Therefore, contemporary nonathropo- cies have become established in Florida (Schef- genic termite distributions were established be- frahn et al. 1988; Scheffrahn et al. 1992; Schef- tween these 2 periods. Distant termite dispersal frahn et al. 2002; Scheffrahn & Su 1995; Su et al. by flotsam can be presumed to be a very rare 1997), more than any other state or territory in event with a success rate inversely proportional America, followed by Hawaii with 5 species (or 4 to distance. Establishment of the depauparate species if Zootermopsis angusticollis Hagen is not native terrestrial faunas on distant oceanic volca- established, Woodrow et al. 1999; Yeap et al. nic islands such as Hawaii was the result of tran- 2007). Australia has a slightly greater human soceanic dispersal (Cowie & Holland 2006). Near- population than Florida, a much larger tidal shore islands like the Krakatau can be colonized coastline, and both share similar economies, cli- much more frequently by both flotsam transport mates, pleasure boating industries, and proximi- and cross-water termite dispersal flights (Gath- ties to tropical nations to the north and south, re- orne-Hardy & Jones 2000). spectively. Yet, in the last century, a single exotic Shipboard transport of termite colonies, where termite, Cr. brevis (Walker), has become estab- success is not affected by travel distance, has lished in Australia (Peters 1990). As for other sus- been suspected in recent (Gay 1967; Scheffrahn & pected exotic Cryptotermes, Gay & Watson (1982) Su 2005) and early (Scheffrahn et al. 2009) trans- determined that Cr. cynocephalus Light and Cr. oceanic termite establishments. Vessels can be domesticus (Haviland) are endemic to northeast colonized during construction (usually only Kalo- Australia. Gay (1967) reported that Cr. dudleyi

58 Florida Entomologist 94(1) March 2011

Banks, an exotic drywood termite from Southeast infestations (Fig. 1) with Co. formosanus Shiraki Asia, was already established in Darwin by 1913. being the most common onboard pest in both In a further attempt to understand the dynam- Florida (27 records) and Australia (13 records) ics and taxa involved in exotic termite establish- followed by Co. gestroi with 15 and 4 records, re- ments, we provide a summary of onboard termite spectively. Three other infestations by subterra- infestations in Florida and border interceptions nean termites were recorded including 2 by Reti- in Australia and we contrast the regulatory pro- culitermes virginicus (Banks) in Florida and 1by cedures used for boats arriving from foreign Heterotermes sp. in a boat that sailed from Flor- ports. ida to Grand Cayman Island. The second most prevalent genus, at 30% of boat infestations, was MATERIALS AND METHODS Cryptotermes (Family Kalotermidae). Australian interceptions yielding 8 infestations each of Cr. Termite specimens from Australia were found brevis (Walker) and Cr. domesticus (Haviland), 3 during interceptive inspections by WC and other infestations of Cr. dudleyi Banks, 2 of Cr. cyno- Australian Quarantine and Inspection Service cephalus Light, and 15 Cr. species undetermined. (AQIS) personnel. Florida samples were collected Only 3 infestations of Cr. brevis were recorded by or submitted to RHS by pest control profes- from Florida; however, fumigations for this spe- sionals and boat owners or operators. In both cies are so routine in Florida that samples are sel- cases, onboard specimens were collected and dom collected for identification. One pest control stored in ethanol. Identifications were made by company in Fort Lauderdale estimates that it is the authors using voucher specimens from their contracted to fumigate about 15 boats a year for respective collections. Other information sought drywood termites (read Cr. brevis, Edwards, J. K., included date of collection, given in Table 1 only in personal communication). On the other hand, 6 years, location collected (city), vessel origin (if infestations of I. minor were recorded in both known), and vessel and/or infestation type. For Australia and Florida. Alates of I. minor are species identification, samples were required to much more robust and dark (reddish pronotum contain morphologically robust winged imagos and head) than Cr. brevis and have a different and/or soldiers. Workers were identified morpho- flight season and diel periodicity. Therefore, I. mi- logically only to genus. nor flights prompt elevated identification re- quests by the Florida pest control industry. Aus- RESULTS AND DISCUSSION tralia recorded a single shipboard infestation each of I. immigrans (Snyder) and I. sp., while in During 1986-2009, 74 and 59 termite incidents Florida, a single infestation of I. snyderi (Light) onboard boats were recorded in Australia and Flor- was observed on a houseboat in Key West. The ida, respectively (Table 1). The Australian records most unexpected find of this study was a mature are comprehensive and represent all known AQIS infestation in 2009 of Rugitermes panamae (Sny- interceptions. The Florida incidents represent an der) from an itinerant yacht intercepted while informal and very incomplete sampling of the ac- visiting Bundaberg Australia. The yacht was ap- tual number of boat infestations occurring around parently infested by this “dampwood” species dur- the State. Three vessels were infested simulta- ing a voyage in 2003 to Central America. Colonies neously by 2 species and each is recorded as a sep- of the predominantly arboreal genus Nasutiter- arate incident in Table 1. Unlike Australia where mes (Termitidae) were found on boats 3 times only Cr. brevis is established, most boats in Florida during the last 25 years. Nasutitermes acajutlae are infested in their home waters where exotic spe- (Holmgren) was found twice and N. nigriceps cies abound. This phenomenon enhances the (Haldeman) once in Florida. Although not re- spread of termites in Florida from boat-to-land or corded in Table 1, N. corniger (Motschulsky) was land-to-boat by dockside dispersal flights and also found infesting 2 boats in dry dock in Dania elevates the likelihood that boats voyaging from Beach, Florida, as part of a land-borne infestation Florida could spread termites to foreign ports. Al- of this pest (Scheffrahn et al. 2002). though long, open ocean voyages are not the norm We suggest herein that the difference in the for Florida boaters, some will “island-hop” number of exotic termite species established in throughout the West Indies. One yacht, suspected Florida versus Australia is attributable, at least of acquiring Co. gestroi Wasmann (= havilandi, in part, to differing laws and regulations intended Kirton & Brown 2003) during a winter dockage in to exclude exotic pests. The U.S. Customs and the Turks and Caicos Islands (Scheffrahn & Su Border Patrol (CBP) requires that pleasure ves- 2005) was simultaneously infested with Incisiter- sels arriving in the U.S. from a foreign port must mes minor (Hagen). It was presumed that the lat- report their arrival by telephone and be directed, ter species infested this boat while under construc- with passengers and crew, to the nearest port of tion in San Diego, California. entry or nearest designated reporting location for The subterranean genus Coptotermes (Family a CBP face-to-face interview and/or vessel inspec- Rhinotermitidae) was observed in 53% of all boat tion (Anonymous 2009). Inspections focus on im-

Scheffrahn & Crowe: Termites on Boats 59

TABLE 1. TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).

Vessel location where termites found Vessel origin1 Genus Species Year Vessel comments

Islamorada Key, FL Coptotermes formosanus 1986 boat North Palm Beach, FL Tennessee Coptotermes formosanus 1995 boat Fort Lauderdale, FL Coptotermes formosanus 1995 11 m boat Jacksonville, FL Coptotermes formosanus 1997 cable ship Lighthouse Point, FL Coptotermes formosanus 1998 boat Hypoluxo, FL Coptotermes formosanus 1998 26 m boat Palm Beach, FL Coptotermes formosanus 1999 9 m boat Brunswick, Georgia Coptotermes formosanus 1999 10 m boat Palm Beach Gardens, FL Coptotermes formosanus 2000 9 m boat Hillsborough Beach, FL Coptotermes formosanus 2000 large boat Hallandale, FL Hong Kong Coptotermes formosanus 2000 23 m boat Tampa, FL Coptotermes formosanus 2001 10 m boat Pompano Beach, FL Coptotermes formosanus 2001 16 m boat Tampa, FL Coptotermes formosanus 2002 10 m boat Fort Lauderdale, FL Coptotermes formosanus 2002 26 m speed boat Fort Lauderdale, FL Coptotermes formosanus 2002 15 m cabin cruiser Holmes Beach, FL Coptotermes formosanus 2002 boat Dania Beach, FL Coptotermes formosanus 2003 11 m boat Hollywood, FL Coptotermes formosanus 2004 boat Fort Lauderdale, FL Coptotermes formosanus 2004 boat Fort Lauderdale, FL Coptotermes formosanus 2004 18 m fishing yacht Fort Lauderdale, FL Coptotermes formosanus 2005 15 m sailboat Jacksonville Beach, FL Coptotermes formosanus 2006 13 m boat Marathon Key, FL Coptotermes formosanus 2006 small boat Fort Lauderdale, FL Coptotermes formosanus 2008 15 m boat Volusia County, FL Hong Kong Coptotermes formosanus 2008 18 m cabin cruiser Panama City, FL Coptotermes formosanus 2008 9 m boat Lake Park, FL Coptotermes formosanus 2008 8 m boat Fort Pierce, FL Jamaica Coptotermes gestroi 1991 Boat Hollywood, FL Virgin Gordo, B.V.I. Coptotermes gestroi 1995 boat in dry dock Fort Lauderdale, FL Turks, Caicos Coptotermes gestroi 2001 27 m yacht Key West, FL Coptotermes gestroi 2003 15 m sailboat Key West, FL Coptotermes gestroi 2005 houseboat, nest with queen Key West, FL Coptotermes gestroi 2005 8 m motor boat Tequesta, FL Coptotermes gestroi 2005 9 m fishing boat Key Largo, FL Cuba Coptotermes gestroi 2006 Sailboat Key West, FL Coptotermes gestroi 2007 Sailboat Key West, FL Coptotermes gestroi 2007 Boat Miami Beach, FL Coptotermes gestroi 2007 12 m fishing boat transom Stock Island Key, FL Key West Coptotermes gestroi 2007 15 m cabin cruiser St. Petersburg, FL Coptotermes gestroi 2007 11 m Yacht Boca Chica Key, FL Key West Coptotermes gestroi 2007 sailboat Stock Island (Key West), FL Coptotermes gestroi 2007 Boat Franklin, Louisiana Florida Cryptotermes brevis 2000 17 m boat Marathon Key, FL Cryptotermes brevis 2005 Sailboat Cudjoe Key, FL Cryptotermes brevis 2006 Boat Grand Cayman, Cayman Is. Florida Heterotermes sp. 1995 Boat

1Unknown if blank. 2dead imagos only, no live infestation. 60 Florida Entomologist 94(1) March 2011

TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).

Vessel location where termites found Vessel origin1 Genus Species Year Vessel comments

Fort Lauderdale, FL Incisitermes minor 2000 Boat Miami, FL Los Angeles, CA Incisitermes minor 2000 26 m boat Fort Lauderdale, FL San Diego, CA Incisitermes minor 2001 27 m yacht Marathon Key, FL Taiwan Incisitermes minor 2006 Boat Dania Beach, FL western Mexico Incisitermes minor 2007 20 m boat St. Augustine, FL FL Keys Incisitermes minor 2008 Boat Key West, FL Incisitermes snyderi 2000 Houseboat Fort Lauderdale, FL St. Thomas U.S.V.I. Nasutitermes acajutlae 2002 15 m boat Jacksonville, FL Puerto Rico Nasutitermes acajutlae 2002 container on ship Fort Lauderdale, FL Nasutitermes nigriceps 1996 Sailboat Key West, FL Reticulitermes virginicus 2000 Houseboat Jacksonville Beach, FL Reticulitermes virginicus 2003 Boat Darwin, NT, AUS China Coptotermes formosanus 1994 boat (refugee) Perth, WA, AUS Hong Kong Coptotermes formosanus 2000 Boat Brisbane, Qld, AUS China Coptotermes formosanus 2002 boat, fibreglass Sydney, NSW, AUS Coptotermes formosanus 2003 Yacht Brisbane, Qld, AUS Hong Kong Coptotermes formosanus 2003 boat Brisbane, Qld, AUS China Coptotermes formosanus 2003 Yacht Brisbane, Qld, AUS USA / Japan Coptotermes formosanus 2005 9 m boat Brisbane, Qld, AUS USA Coptotermes formosanus 2005 boat (with I. minor) Townsville, Qld, AUS Hong Kong/Asia Coptotermes formosanus 2005 Boat Bundaberg, Qld, AUS Hawaii Coptotermes formosanus 2006 itinerant yacht Brisbane, Qld, AUS Japan Coptotermes formosanus 2007 Boat Newcastle, NSW, AUS China Coptotermes formosanus 2008 Boat Brisbane, Qld, AUS USA Coptotermes formosanus 2009 itinerant yacht Darwin, NT, AUS Thailand Coptotermes gestroi 1986 Yacht Darwin, NT, AUS Thailand Coptotermes gestroi 1994 Boat Bundaberg, Qld, AUS Marshall Islands Coptotermes gestroi 1996 Yacht Brisbane, Qld, AUS China Coptotermes gestroi 2003 Boat Brisbane, Qld, AUS USA Coptotermes sp. 2002 Yacht Cairns, Qld, AUS AUS Coptotermes sp. 2005 dinghy from TI to Cairns Sydney, NSW, AUS Unknown Coptotermes sp. 2006 navy boat Brisbane, Qld, AUS New Caledonia Coptotermes sp. 2008 boat (returning AUS yacht) Brisbane, Qld, AUS Taiwan Coptotermes sp. 2008 Boat Brisbane, Qld, AUS Hong Kong Coptotermes sp. 2005 yacht (flybridge in lockers) Mackay, Qld, AUS Taiwan Coptotermes sp. 2005 Boat Brisbane, Qld, AUS France Coptotermes sp. 2008 Yacht Brisbane, Qld, AUS USA Coptotermes sp. 2008 Boat Perth, WA, AUS Singapore Coptotermes travians? 2002 boat, fibreglass & wood Townsville, Qld, AUS Cryptotermes brevis 1989 Yacht Brisbane, Qld, AUS USA Cryptotermes brevis 2003 wooden yacht (with I. Minor) Brisbane, Qld, AUS USA Cryptotermes brevis 2005 Superyacht Cardwell, Qld, AUS Cryptotermes brevis 2006 Trimaran Brisbane, Qld, AUS South Africa Cryptotermes brevis 2007 Boat Brisbane, Qld, AUS USA Cryptotermes brevis 2008 Boat Airlie Beach, Qld, AUS USA Cryptotermes brevis 2009 Catalina 400 MK II

1Unknown if blank. 2dead imagos only, no live infestation. Scheffrahn & Crowe: Termites on Boats 61

TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).

Vessel location where termites found Vessel origin1 Genus Species Year Vessel comments

Mackay, Qld, AUS USA Cryptotermes brevis? 2009 28 m super yacht Bundaberg, Qld, AUS USA Cryptotermes cavifrons1 2008 Yacht Darwin, NT, AUS Indonesia Cryptotermes cynocephalus 2005 foreign fishing vessel Darwin, NT, AUS Indonesia Cryptotermes cynocephalus 2009 foreign fishing vessel Darwin, NT, AUS Indonesia Cryptotermes domesticus 1986 Yacht Darwin, NT, AUS Indonesia Cryptotermes domesticus 1987 yacht Brisbane, Qld, AUS Vanuatu Cryptotermes domesticus 1999 boat Sydney, NSW, AUS Cryptotermes domesticus 2003 yacht Broome, WA, AUS Papela, Roti, Indonesia Cryptotermes domesticus 2005 foreign fishing vessel Gove, NT, AUS Karja Sama, Indonesia Cryptotermes domesticus 2006 foreign fishing vessel Gove, NT, AUS Indonesia Cryptotermes domesticus 2006 foreign fishing vessel Broome, WA, AUS Indonesia Cryptotermes domesticus 2007 foreign fishing vessel Darwin, NT, AUS Indonesia Cryptotermes dudleyi 1994 boat Darwin, NT, AUS Philippines Cryptotermes dudleyi 2006 boat Darwin, NT, AUS Indonesia Cryptotermes dudleyi 2008 foreign fishing vessel Darwin, NT, AUS Indonesia Cryptotermes sp. 1993 boat Gove, NT, AUS Indonesia Cryptotermes sp. 1993 foreign fishing vessel Darwin, NT, AUS Vietnam Cryptotermes sp. 2001 boat Gove, NT, AUS Indonesia Cryptotermes sp. 2004 foreign fishing vessel Thursday Island, AUS Indonesia Cryptotermes sp. 2004 foreign fishing vessel Darwin, NT, AUS Indonesia Cryptotermes sp. 2004 boat Darwin, NT, AUS Indonesia Cryptotermes sp. 2005 foreign fishing vessel Broome, WA, AUS Papela Roti, Indonesia Cryptotermes sp. 2006 foreign fishing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2007 foreign fishing vessel Bundaberg, Qld, AUS USA Cryptotermes sp. 2007 boat Broome, WA, AUS Sulawesi, Indonesia Cryptotermes sp. 2005 foreign fishing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2005 foreign fishing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2005 foreign fishing vessel Darwin, NT, AUS Indonesia Cryptotermes sp. 2005 foreign fishing vessel Broome, WA, AUS Indonesia Cryptotermes sp. 2009 foreign fishing vessel Brisbane, Qld, AUS Thailand Drepanotermes2 sp. 2008 boat Bundaberg, Qld, AUS Hawaii Incisitermes immigrans 2007 boat Brisbane, Qld, AUS Incisitermes minor 2001 yacht Brisbane, Qld, AUS USA Incisitermes minor 2003 wooden yacht (with Cr. brevis) Brisbane, Qld, AUS USA Incisitermes minor 2005 boat (with Co. formosanus)

1Unknown if blank. 2dead imagos only, no live infestation. 62 Florida Entomologist 94(1) March 2011

TABLE 1. (CONTINUED) TERMITE INFESTATIONS BY GENUS AND SPECIES ONBOARD VESSELS DURING 1986-2009 (VOUCHER SPECIMENS IN THE UNIVERSITY OF FLORIDA TERMITE COLLECTION OR AQIS RECORDS).

Vessel location where termites found Vessel origin1 Genus Species Year Vessel comments

Brisbane, Qld, AUS Fiji (made in USA) Incisitermes minor 2006 super yacht Bundaberg, Qld, AUS USA Incisitermes minor 2006 trimaran Cairns, Qld, AUS USA Incisitermes minor 2009 trimaran Broome, WA, AUS Papela Roti, Indonesia Incisitermes sp. 2006 foreign ﬁshing vessel Bundaberg, Qld, AUS Central Amer. Rugitermes panamae 2009 Yacht

1Unknown if blank. 2dead imagos only, no live infestation.

Fig. 1. Frequency of termite genera collected on vessels in Australia and Florida. “Other” includes Drepanoter- mes and Rugitermes from Australia and Heterotermes from Florida. migration compliance by the passengers and must also be inspected by AQIS. The level of AQIS crew, possible illegal contraband, and agricul- inspection required will depend on the amount of tural pests in cargo. Structural and household timber present and the construction/re-fit and pests, which are usually disassociated with cargo sailing history of the vessel. The inspection can be and dwell within the vessel’s own structure, are conducted by an AQIS quarantine officer or AQIS not mandated for inspection. In contrast to Flor- entomologist with or without a licensed pest con- ida practices, passengers and crew aboard vessels trol professional and approved termite detection arriving to Australia from a foreign port must ob- method. If termites are found upon inspection, tain clearance by the Australian Customs and the vessel must be fumigated with methyl bro- Border Protection Service and the Australian mide (AQIS method T9047) or sulfuryl fluoride Quarantine and Inspection Service (AQIS). Ves- (AQIS method T9090) at the owner’s expense sels with timber in their cargo or construction (Anonymous 2010). Scheffrahn & Crowe: Termites on Boats 63

ACKNOWLEDGMENTS KIRTON, L. G., AND BROWN, V. K. 2003. The taxonomic status of pest species of Coptotermes in Southeast We thank boat owners and pest control companies in Asia: Resolving the paradox in the pest status of the Florida for submitting termite samples to RHS. termites, Coptotermes gestroi, C. havilandi and C. travians (Isoptera: Rhinotermitidae). Sociobiol. 42: 43- 63. PETERS, B. C. 1990. Infestations of Cryptotermes brevis REFERENCES CITED (Walker) (Isoptera: Kalotermitidae) in Queensland, Australia. 1. History, detection and identification. ANDERSON, A., CHAPPELL, J., GAGAN, M., AND GROVE, R. Australian Forester 53: 79-88. 2006. Prehistoric maritime migration in the Pacific SCHEFFRAHN, R. H., AND SU, N.-Y. 1995. A new subterra- islands: An hypothesis of ENSO forcing. The Ho- nean termite introduced to Florida: Heterotermes locene 16: 1-6. Froggatt (Rhinotermitidae: Heterotermitinae) estab- ANONYMOUS. 2009. Code of Federal Regulations (United lished in Miami. Florida Entomol. 78: 623-627. States) 19CFR4.2. SCHEFFRAHN, R. H., AND SU, N.-Y. 2005. Distribution of ANONYMOUS. 2010. ICON Condition C9645. Australian the termite genus Coptotermes (Isoptera: Rhinoter- Quarantine and Inspection Service. mitidae) in Florida. Florida Entomol. 88: 201-203. COWIE, R. H., AND HOLLAND, B. S. 2006. Dispersal is SCHEFFRAHN, R. H., CABRERA, B. J., KERN JR., W. H., fundamental to biogeography and the evolution of AND SU, N.-Y. 2002. Nasutitermes costalis (Isoptera: biodiversity on oceanic islands. J. Biogeography 33: Termitidae) in Florida: First record of a non-endemic 193-198. establishment by a higher termite. Florida Entomol. FLEMING, K., JOHNSTON, P., ZWARTZ, D., YOKOYAMA, Y., 85: 273-275. LAMBECK, K., AND CHAPPEL, J. l998. Refining the SCHEFFRAHN, R. H., KRECK˘ ˘ , J. RIPA, R., AND LUPPICHINI, eustatic sea-level curve since the last glacial maxi- P. 2009. Endemic origin and vast anthropogenic dis- mum using far- and intermediate-field sites. Earth persal of the West Indian drywood termite. Biol. Inva- and Planetary Science Letters 163: 327-342. sions 11: 787-799. GATHORNE-HARDY, F. J., AND JONES, D. T. 2000. The re- SCHEFFRAHN, R. H., MANGOLD, J. R., AND SU, N.-Y. 1988. colonization of the Krakatau islands by termites A survey of structure-infesting termites of peninsular (Isoptera), and their biogeographical origins. Biol. J. Florida. Florida Entomol. 71: 615-630. Linnean Soc. 71: 251-267. SU, N.-Y., SCHEFFRAHN, R. H., AND WEISSLING, T. 1997. A GAY, F. J. 1967. A world review of introduced species of new introduction of a subterranean termite, Coptoter- termites. CSIRO Bulletin Melbourne, Australia 286: mes havilandi Holmgren (Isoptera: Rhinotermitidae) 1-88. in Miami, Florida. Florida Entomol. 80: 408-411. GAY, F. J., AND WATSON, J. A. L. 1982. The genus Cryp- WOODROW, R. J., GRACE, J. K., AND YATES III, J. R. 1999. totermes in Australia (Isoptera: Kalotermitidae). Hawaii’s Termites: An Identification Guide. Honolulu Australian J. Zool. Supplementary Series 30, 88: 1- (HI): University of Hawaii. 6 p. (Household and Struc- 64. tural Pests; HSP-1). HOCHMAIR, H. H., AND SCHEFFRAHN, R. H. 2010. Spatial YEAP, B.-K., OTHMAN, A. S., LEE, V. S., AND LEE, C.-Y. association of marine dockage with land-borne infes- 2007. Genetic relationship between Coptotermes tations of invasive termites in urban South Florida. gestroi and Coptotermes vastator (Isoptera: Rhinoter- J. Econ. Entomol. 103: 1338-1346. mitidae). J. Econ. Entomol. 100: 467-474.

64 Florida Entomologist 94(1) March 2011

FIRST RECORD OF THE GENUS ADOXOMYIA (DIPTERA: STRATIOMYIDAE) WITH FOUR SPECIES FROM TURKEY

TURGAY ÜSTÜNER1 AND ABDULLAH HASBENLI·2 1Selçuk University, Faculty of Science, Department of Biology, Alaaddin Keykubat Kampüsü, 42100 Selçuklu, Konya, Turkey E-mail: [email protected]

2Gazi University, Faculty of Arts and Sciences, Department of Biology, 06500 Teknikokullar, Ankara, Turkey.

ABSTRACT

Adoxomyia aureovittata (Bigot, 1879), A. cinerascens (Loew, 1873), A. obscuripennis (Loew, 1873) and A. sarudnyi (Pleske, 1903) are recorded from Turkey for the first time. Both sexes of the first 3 species and the male of A. sarudnyi are redescribed and photographs of all species are provided. The distributions of all species are briefly discussed. The male genitalia and some other diagnostic characters of all the examined species are illustrated. An identification key to all East-Mediterranean species was constructed and is included in this report.

Key Words: Adoxomyia aureovittata, A. cinerascens, A. obscuripennis, A. sarudnyi, new records, distribution, Turkey

RESUMEN

Se registran por primera vez Adoxomyia aureovittata (Bigot, 1879), A. cinerascens (Loew, 1873), A. obscuripennis (Loew, 1873) y A. sarudnyi (Pleske, 1903) para Turquia. Se proveen redescripciones y fotos de ambos sexos de las primeras 3 especies y del macho de A. sarudnyi. Se discutan la distribución de todas las especies. Se ilustran las genitalias de los machos y algunas de las caracteristicas diagnósticas de las especies examinadas. Una clave para la identiﬁcación de todas las especies de la región Este del Mediterráneo es incluida.

The family Stratiomyidae belongs to the sub- The larvae of Adoxomyia are known only for order Brachycera in Diptera (Rozkosny 1982). some Nearctic and one Oriental species; they This large family includes more than 2650 species were found in decaying cacti and nests of pack in 375 genera composed of 12 subfamilies world- rats (Neotoma sp.) (McFadden 1967; James & Mc- wide of which 426 species in 55 genera in 7 sub- Fadden 1969). families occur in the Palaearctic region (Woodley In addition to the 4 species recorded in Turkey at 2001). So far 48 species in 14 genera and 7 sub- least 5 additional species may occur here. A. dahlii families (Beridinae, Pachygastrinae, Clitellarii- (Meigen 1830) is known from southern Europe (incl. nae, Hermetinae, Sarginae, Stratiomyinae, Nem- Ukraine), Armenia and Israel. A. ruﬁcornis (Loew otelinae) have been recorded in Turkey (Rozkosny 1873) occurs in Azerbaijan, Iran and Kyrgyzstan. A. & Nartshuk 1988; Woodley 2001; Üstüner et al. hermonensis Lidner, 1975 and A. paleastinensis 2002, 2003; Üstüner & Hasbenli 2003, 2004). Lindner, 1937 were described from Israel and A. The subfamily Clitellariinae contains 50 gen- transcaucasica Nartshuk, 2004 is based on types era worldwide, 10 genera in the Palaearctic re- from Armenia and Azerbaijan. According to a recent gion and 1 genus (Pycnomalla) in Turkey (Wood- paper by Nartshuk (2004), A. portschinskii (Pleske) ley 2001; Üstüner et al. 2002). The genus Adox- is a mere synonym of A. dahlii (Meigen). omyia (Kertész, 1907) belongs to the subfamily Clitellariinae and includes 36 described species. MATERIALS They are distributed in the Palaearctic region (16 A total of 16 specimens (12 males and 4 fe- species), the Nearctic region (13 species), the Neo- males) were collected by hand net at Antalya, Is- tropical region (4 species), the Oriental region (3 parta, and Konya in Turkey between 1999 and species) and the Afrotropical region (2 species) 2001. Some specimens of Adoxomyia were caught (Woodley 2001; Hauser 2002; Nartshuk 2004). sunbathing on stones or on the ground in dry Palearctic species of Adoxomyia are found mainly creek beds. The specimens are deposited in the in south-eastern Europe, Transcaucasus, Near collection of the Selçuk University Department of East, Central Asia and China (Rozkosny 1983; Biology in Konya, Turkey. The fact that previ- Rozkosny & Nartshuk 1988; Nartshuk 2004). ously the genus Adoxomyia had not been recorded Adoxomyia had not been recorded in Turkey be- from Turkey reﬂects the poor knowledge of the fore this report. Turkish Stratiomyidae fauna.

Üstüner & Hasbenli: First Records of Adoxomyia Species in Turkey 65

THE GENUS ADOXOMYIA Kertész 1907 were chiefly devoted to descriptions of separate additional species (Ôuchi 1938; Dusek & Rozko- The generic name Adoxomyia was proposed sny 1963; Lindner 1975; Nartshuk 2004). by Kertész (1907) and according to the catalog In Turkey, the species belonging to Adoxomyia publishad a year later (Kertész 1908) this ge- are mid-sized (6-11 mm) with a predominantly nus embraced 23 species. In 1923, Kertész tried black body. The eyes, which are contiguous in to establish a new separate genus, Euclitellaria males and widely separated in females, are cov- Kertész, but it was not accepted by Pleske ered with dense hairs. The antennae are rela- (1925) and subsequent authors. The last com- tively long, predominantly black, but can be red- prehensive key to the Palaearctic species is that orange to dark brown in some species. Scape and by Lindner (1937), who followed Pleske’s con- pedicel are of equal length. The flagellum consists cept of Adoxomyia in a broad sense. Besides the of 8 flagellomeres. No thoracic spines but two work by Rozkosny (1983), which treated the Eu- scutellar spines are present. All of 4 M-veins ropean species, some other relevant papers reach the wing margin.

KEY TO THE EAST MEDITERRANEAN SPECIES OF ADOXOMYIA The following key is based partly on Lindner (1937). The male of A. hermonensis and the female of A. palaestinensis are unknown.

1. Legs completely black ...... 2 — Legs bicoloured or mainly ...... 5 2. Antenna entirely black...... 3

— At least basal half of antennal flagellum red...... A. ruficornis (Loew 1873) 3. Scutellar spines short, slender and bare, basal 3-4 flagellomeres in female unusually broad ...... 4 — Femora - Scutellar spines longer, thickened and haired, basal 3-4 flagellomeres not as broad (Figs. 19 and 20) ...... A. obscuripennis (Loew 1873) 4. Female eyes black haired, postocular band wider than scape is long; male unknown ...... hermonensis Lindner 1975 — Female eyes white haired, postocular band as wide as scape is long ...... A. transcaucasica Nartshuk 2004

5. Legs entirely yellow ...... A. sarudnyi (Pleske 1903) — At least femora black...... 6 6. Antenna black...... 7

- Abdomen with silverish white hair patches ...... A. palaestinensis Lindner 193 8. Abdomen with golden yellow hair patches ...... A. aureovittata (Bigot 1879) — Abdomen with silverish white (rarely coppery) hair patches ...... 9 9. Male flagellum almost cylindrical, female apical flagellomere at base half as broad as scape at distal end ...... A. dahlii (Meigen 1830) — Male flagellum distinctly swollen in middle, female apical flagellomere broader, at most slightly narrower than scape at distal margin (Figs. 17 and 18) ...... A. cinerascens (Loew 1873)

Redescription of the 4 Adoxomyia species recorded in covered with appressed yellowish hairs. Face, Turkey cheeks and posteroventral part of head covered with hairs as long as scape, erect, black and 1. Adoxomyia aureovittata (Bigot 1879); partly brown. Antenna entirely black and more see Figs. 1-4, 15-16 and 22-23. slender in male than in female. Scape and pedicel black with black and erect hairs being as long as Male: Head transversely oval, in dorsal view. scape. Flagellum about four times as long as Eyes touching on frons. Hairs on eyes as long as scape and pedicel combined, the ﬁrst 5 ﬂagellom- pedicel, dense and black. Black postocular area eres with dense yellowish pubescence. Scutum swollen in lower half and narrowed in upper part, black, with semi-erect and black hairs. Scutellum

66 Florida Entomologist 94(1) March 2011

Figs. 1-14. Adoxomyia aureovittata: 1- male in dorsal view, 2- female in dorsal view, 3- male in lateral view, 4- female in lateral view; 5-8 Adoxomyia cineracens 5- male in dorsal view, 6- female in dorsal view, 7- male in lateral view, 8- female in lateral view; 9-12 Adoxomyia obscuripennis 9- male in dorsal view, 10- female in dorsal view, 11- male in lateral view, 12- female in lateral view; 13-14. Adoxomyia sarudnyi 13- male in dorsal view, 14- male in lateral view (Scale 1 mm).

and scutellar spines mainly black but tip of on tergite 4 and a transverse, golden yellow band scutellar spines brown. Scutellum covered with on tergite 5. semi-appressed sparse black hairs. Legs black and yellow. Coxa, trochanter, and femur black ex- Female: Eyes densely black haired, hairs only cept for yellow bases of femora. Tibia mainly one-fourth as long as pedicel. Black postocular black, its both ends narrowly yellowish brown. area approximately as wide as fore tibia and cov- Tarsi yellow, front tarsus darkened dorsally as ered with appressed yellowish hairs. Frons about well as tarsomeres 3-5 of mid and hind legs. Coxa, 1/3 of head width, with ﬁne longitudinal groove in trochanter, and femur with semi-erect black middle, black, densely punctate, with yellowish hairs. Tibia covered with appressed yellowish hairs. Face black, with yellowish hairs on sides hairs. Tarsi with appressed golden yellowish below antennae. Remainder of face, cheeks and hairs. Abdomen entirely black, abdominal pile posteroventral part of head covered with erect, black except for golden yellow lateral markings black hairs being as long as scape. Antenna in-

Üstüner & Hasbenli: First Records of Adoxomyia Species in Turkey 67

serted at middle of head profile, partly black. verse, pale yellow hair band on posterior margin Scape black but reddish brown at tip. Pedicel and of tergite 4. first 5 flagellomeres dark red, last 3 flagellomeres black. Scape and pedicel with erect, black hairs as Female: Hairs on eyes black, about 0.3 times as long as antenal scape. Flagellum about 4.5 times long as pedicel. Postocular area black, covered as long as both basal antennal segments com- with pale yellow and semi-appressed hairs. Frons bined, first 5 flagellomeres densely golden yellow about 0.3 of head-width, shining black and dusted. Thorax black, scutum with 2 golden yel- densely punctate, with fine longitudinal groove in low dusted longitudinal stripes which can be re- middle and with sparse pale yellow hairs. Face duced to absent in some specimens (especially black, with whitish, dense, erect hairs being as smaller ones). Scutellum black with appressed long as pedicel. Antenna long, about 1.5 times as golden yellow hairs. Tips of scutellar spines long as head in lateral view, bicolored and in male brown. Legs black and yellow, femora black ex- more slender than in female. Scape brownish or- cept for yellow base, tibiae mainly darkened, with ange on lower surface but darkened on upper sur- both ends broadly yellowish brown on fore and face. Pedicel and first 3 flagellomeres brownish mid legs. Hind tibia mainly black, with both ends orange, rest of flagellum black. Flagellomeres 2-3 broadly yellowish brown. Tarsi yellow, with dor- darkened on outer surface. Thorax black, with ap- sally darkened tarsomeres 3-5. pressed dense pale yellow hairs, but tip of post- All legs with yellowish hairs that are semi- pronotal callus brownish. Scutellum black with erect on femora and appressed on tibiae and tarsi. pale yellow hairs, scutellar spines yellow. Wing Abdomen black but lateral markings on tergites veins brown. Legs bicoloured. Coxae black, tro- 2-4 and a triangular apical spot on tergite 5 with chanters yellow. Femora and tibiae mainly black golden yellow hair patches. and partly yellow on tips. Fore and hind tarsi yel- Adoxomyia aureovittata (Bigot 1879) was de- lowish on inner surface, darkened on outer sur- scribed from an unknown locality. Apparently, face, basal 2 tarsomeres of mid tarsi yellow, other this species is distributed in the eastern part of tarsomeres darkened on outer surface. Femora the Mediterranean area. In addition to Turkey it with semi-erect pale yellow hairs. Tibiae with ap- was also found in Greece (M. Hauser, personal pressed yellowish hairs. Tarsomeres with ap- communication). pressed yellow hairs. Abdomen mainly black but Material Examined: Turkey: Konya, Hadim, with pale yellow hair patches at posterior margin between Tosmur and Gevne Village, Gevne Valley, of tergite 4. 1450-2020 m, 10 June 1999, 1 male and 1 female; Material Examined: Turkey: Antalya, Gündog- Konya, Taskent, Begreli Village, Gevne Valley, mus district, Güneycik Village, Topraktepe place, 1570 m, 10 August 2001, 1 male; all T. Üstüner elev. 200 m, 23 June 1999, 7 males, 12 July 2000, leg. 1 male; Konya, Taskent, Begreli Village, Gevne Distribution: Greece, Turkey. Valley, elev. 1585 m, 10 July 2000, 1 female; all T. Üstüner leg. 2. Adoxomyia cinerascens (Loew 1873); see Figs Distribution: Palaearctic: Iran, Israel, Kaza- 5-8, 17-18 and 24-25. khstan, Kyrgyzstan, Tajikistan (Kertész 1908; Lindner 1937, 1974, 1975; Rozkosny & Nartshuk Male: Hairs above compound eyes as long as 1988; Woodley 2001). This is the first record for pedicel, densely black. Postocular area black, cov- the fauna of Turkey. ered with pale yellowish, appressed hairs. Fron- tal triangle black with dense, erect, pale yellow- 3. Adoxomyia obscuripennis (Loew 1873); Figs 9- ish hairs being about 1.5 times as long as the 12, 19-20 and 28-29. scape. Hairs on black face erect, as long as pedicel, pale yellowish. Antenna as long as head Male: Head transverse, hemispherical, eyes in lateral view. Scape, pedicel and first 3 basal touching on frons. Hairs on eyes dense, black, as flagellomeres brownish orange, rest of flagellum long as scape. Frontal triangle black, with silver- black. Hairs on scape and pedicel erect, as long as ish white pubescence. Face slightly produced in scape, pale yellowish. Thorax including scutellum lateral view, with dense, black hairs, as long as black, with dense yellowish hairs. Scutellar scape. Cheeks and posteroventral part of head spines yellow. Wing veins brown. Legs bicoloured, with erect black hairs. Black postocular area coxae black, trochanters brownish, femora and prominent but narrower than pedicel in upper tibiae mainly black, partly yellow at tips. Fore half and somewhat swollen in lower half, about as and hind tarsi yellowish on inner surface and wide as antennal scape is long, covered with ap- darkened on outer surface, mid tarsi yellow but pressed silverish white hairs. Antenna entirely basal 2 tarsomeres darkened on outer surface. black, flagellomeres 1-3 thickened, following Femora with sparse semi-erect pale yellow hairs. flagellomeres small and slender, last flagellomere Tibiae and tarsomeres with dense appressed yel- longer than 4 preceding combined. Thorax com- low hairs. Abdomen mainly black, with trans- pletely black. Scutum covered with long erect 68 Florida Entomologist 94(1) March 2011

Figs. 15-21. Adoxomyia Antennae: 15- A. aureovittata male, 16- A. aureovittata female, 17- A. cineracens male, 18- A. cineracens female, 19- A. obscuripennis male, 20- A. obscuripennis female, 21- A. sarudnyi male (Scala 1 mm.): 22-23 Adoxomyia aureovittata male terminalia: 22- dorsal part of male genitalia, 23- ventral part of male genitalia; 24-25 Adoxomyia cineracens male terminalia: 24- dorsal part of male genitalia, 25- ventral part of male genitalia; 26-27 Adoxomyia sarudnyi male terminalia: 26- dorsal part of male genitalia, 27- ventral part of male genitalia; 28- 29 Adoxomyia obscuripennis male terminalia: 28- dorsal part of male genitalia, 29- ventral part of male genitalia (Scale 0.1 mm.). Üstüner & Hasbenli: First Records of Adoxomyia Species in Turkey 69 black hairs and short appressed silverish white area black, about 0.4 times as wide as length of hairs. Scutellum with very strong and thick, antennal scape, somewhat swollen in lower half, black scutellar spines. Legs black, but knees about 0.75 times as long as scape, covered with brown. Femora with semi-erect silverish white dense, silverish white hairs. Antenna relatively hairs. Tibiae with semi-appressed, dense, silver- long, about twice as long as head in lateral view. ish white hairs. Tarsomeres black on outer sur- Scape and pedicel orange but darkened on outer face, brown on inner surface, with semi-ap- surface, with strong, erect, black hairs. First 3 pressed, dense, silverish white hairs. Halteres flagellomeres orange on inner surface but dark- pale yellow. Abdomen entirely black, tergites 1-2 ened on outer surface, rest of flagellum black. with erect, moderately long, white hairs. White Thorax black, with short appressed yellowish hairs also distinct on distal half of tergite 4 and on golden hairs and long semi-appressed black hairs entire tergite 5. intermixed. Tops of postpronotal callus reddish brown. Scutellum black and scutellar spines yel- Female: Hairs above compound eyes dense and low. Wings transparent brownish, basal wing black, as long as antennal scape. Frons black, as veins bright orange and distal veins brown, wing wide as flagellum is long, with fine median tip much darker and contrasting to clear wing groove. Frontal hairs as long as antennal scape, base. Legs entirely bright yellow to orange except pale. Face black, covered below antennae with for black coxae. Fore tarsi yellowish on inner sur- long black hairs. Cheeks and posteroventral part face, darkened on outer surface. Mid and hind of head with erect, black and yellowish hairs. Pos- tarsi yellow, but last three tarsomeres darkened tocular area as wide as flagellum, black and with on outer surface. Femora with semi-erect, sparse, appressed, dense, silverish white hairs. Antenna yellowish hairs. Tibia with appressed, dense, yel- slender, about 1.1 times as long as head. Scape, lowish hairs. Tarsi with thick, adpressed, dense, pedicel and flagellomeres 4-7 shining black, three yellowish hairs. Abdomen black with posterolat- basal flagellomeres black, whitish grey dusted. eral, silverish white lateral markings on tergirtes Last flagellomere about 1.5 times as long as 4 pre- 3-4. ceding. Thorax black, covered with appressed, sil- Material Examined: Turkey: Konya, Taskent, verish white hairs. Scutellum, including strong Begreli Village, Gevne Valley, elev. 1570 m, 1 July and short scutellar spines, black. Top of post- 2001, 1 male, T. Üstüner leg. pronotal callus and postalar callus brownish. Distribution: Palaearctic: Afghanistan, Iran Legs black but knees brown, femora with semi- (Kertész 1908, 1923; Lindner 1937; Pleske 1925; erect white hairs, tibiae with semi-appressed, Rozkosny & Nartshuk 1988; Woodley 2001). dense, silverish white hairs. Tarsomeres black on outer surface, brown on inner surface, with semi- DISCUSSION appressed, dense, silverish white hairs. Halteres pale yellow. Wings transparent and partly brown- The genus Adoxomyia was previously un- ish, with brown veins. Abdomen entirely black, known from Turkey. This fact is fairly surprising with erect, moderately long, silverish white hairs because many species were recorded from adja- on sides of tergites 2-3. White hairs also devel- cent countries and are known from southern, of- oped on distal half of tergite 4 and on entire terg- ten arid parts of the Palaearctic region. Therefore ite 5. it was to be expected that at least some species of Material Examined: Turkey: Isparta, Yalvaç, this genus would be found in Turkey as well. That The Sultan Mountains, elev. 1660 m, 29 May is why in this report we constructed an actual 2001, 1 male, 2 females, T. Üstüner leg. identification key to the all East-Mediterranean Distribution: Palaearctic: Azerbaijan, Kazakh- species. Due to intense collecting efforts in many stan, Russia, Tajikistan, Uzbekistan (Kertész different localities in Turkey, 4 species of this ge- 1908, 1923; Lindner 1937; Nartshuk 2004; Pleske nus were collected. The most remarkable record is 1925; Rozkosny 1983; Rozkosny & Nartshuk Adoxomyia aureovittata which was described as 1988; Woodley 2001). Adoxomyia obscuripennis Euparyphus aureovittatus by Bigot in 1879 from (Loew 1873) is recorded from Turkey for the first an unknown locality. The record from Turkey rep- time. resents the first evidence that it is a Palaearctic species and an unpublished record from Greece 4. Adoxomyia sarudnyi (Pleske 1903); Figs 13-14, (Hauser, personal communication) confirms that 21 and 26-27. this species probably has an East- Mediterranean distribution. Male: Head transverse, hemispherical. Eyes Adoxomyia cineracens is distributed in Tran- touching on frons, dense, black eye hairs about scaspia, in the Near East (Iran, Israel) and Cen- 0.3 times as long as scape. Frontal triangle shin- tral Asia. The type locality is Kizilkum (Kazakh- ing black, with fine median groove and white pile stan). Our record thus closes the distribution gap in upper part. Face, cheeks and posteroventral between the known records from Israel and the part of head black with whitish hairs. Postocular type locality. 70 Florida Entomologist 94(1) March 2011

Our record of Adoxomyia obscuripennis repre- der palaearktischen Region. E. Schweizert’sche Ver- sents the most southern and western point of its lagsbuchhandlung, Stuttgart, 218 pp. range and the second evidence of its occurrence in LINDNER˙ , E. 1974. On the Stratiomyidae (Diptera) of the Near East. Israel J. Entomol. 9: 93-108. the western part of Asia (cf. a record from Azer- ˙ baijan in Nartshuk 2004). LINDNER, E. 1975. On some Stratiomyidae (Diptera) from the Near East. Israel J. Entomol. 10: 41-49. Adoxomyia sarudnyi was only known from Af- MCFADDEN, M. W. 1967. Soldier ﬂy larvae in America ghanistan and Iran. Our record in Turkey repre- north of Mexico. Proc. U.S. Natl. Mus. 121: 1-72. sents the most western locality of this very rare NARTSHUK, E. P. 2004. New data on Adoxomyia Bezi species. from the Caucasus and Eastern Europe (Diptera: Stratiomyidae). Zoosystematica Rossica 12: 263-266, St. Petersburg. CKNOWLEDGMENTS A ÔUCHI, Y. 1938. On some stratiomyiid ﬂies from eastern China. J. Shanghai Sci. Inst., Section III 4: 37-61. Our thanks to Dr. Martin Hauser and Prof. Dr. R. PLESKE, T. 1925. Études sur les Stratiomyiinæ de la ré- Rozkosny for critically reviewing the manuscript. gion paléarctique—III.—Revue des espéces paléarc- tiques de la sous-famille des Clitellariinae. Encyclo- pédie Entomologique, Série B (II), Diptera 1(3-4): REFERENCES CITED 105-119, 165-188. ROZKOSNY, R. 1983. A Biosystematic Study of the Euro- DUSEK, J., AND ROZKOSNY, R. 1963. Revision mitteleu- pean Stratiomyidae (Diptera), Volume 2: Clitellarii- ropäischer Arten der Familie Stratiomyidae nae, Hermetiinae, Pachygasterinae and Bibliography. (Diptera) mit besonderer Berücksichtigung der Fau- Dr. W. Junk, The Hague, Boston, London, 431 pp. na der CSSRˇ I. Acta Societatis Entomologicae Cˇech- ROZKOSNY, R., AND NARTSHUK, E. P. 1988. Family Stra- osloveniae 60(3): 202-221. tiomyidae, pp. 42-96 In A. Soós and L. Papp [eds.], HAUSER, M. 2002. A new species of Adoxomyia Kertész, Catalogue of Palaearctic Diptera. Volume 5. Atheri- 1907 (Diptera: Stratiomyidae) from Socotra, Yemen. cidae-Asilidae. Akadémiai Kiadó Budapest, 446 pp. Fauna of Arabia 19: 463-466. ÜSTÜNER, T., AND HASBENLI˙, A. 2003. First record of JAMES, M. T., AND MCFADDEN, M. W. 1969. The genus the subfamily Beridinae (Diptera: Stratiomyidae) Adoxomyia in America North of Mexico (Diptera: from Turkey. Studia Dipterologica 10(1): 186-188, Stratiomyidae). Journal of the Kansas Entomologi- Halle (Saale). cal Society 42: 260-276. ÜSTÜNER, T., HASBENLI˙, A., AND AKTÜMSEK, A. 2002. KERTÉSZ, K. 1907. Eine neuer Dipteren-Gattungsname. Contribution to subfamily 360 Clitellariinae Annalen Historico-Naturales Musei Nationalis Hun- (Diptera, Stratiomyidae) in Fauna of Turkey. J. En- garici 5(2): 499. tomol. Res. Soc. 4(1): 19-24. KERTÉSZ, K. 1908. Catalogus dipterorum hucusque de- ÜSTÜNER, T., HASBENLI˙, A., AND ROZKOSNYˇ ´ , R. 2003. scriptorum. Volumen III. Stratiomyiidae, Erinnidae, First record of Hermetia illucens (Linnaeus, 1758) Coenomyidae, Tabanidae, Pantophthalmidae, Rha- (Diptera, Stratiomyidae) from the Near East. Studia gionidae. Museum Nationale Hungaricum, Budap- Dipterologica 10(1): 181-185, Halle (Saale). estini, 366 pp. ÜSTÜNER, T., AND HASBENLI˙, A. 2004. A new species of KERTÉSZ, K. 1923. Vorarbeiten zu einer Monographie Oxycera Meigen (Diptera: Stratiomyidae) from Tur- der Notacanthen. XLV-L. Annales Historico Natu- key. Entomol. News 115(3): 163-167. rales Musei Nationalis Hungarici, 20: 85-129. WOODLEY, N. E. 2001. A World Catalog of the Stratiomy- LINDNER˙ , E. 1937. 18. Stratiomyiidae [part]. Lieferung idae (Insecta: Diptera). Backhuys Publishers, 114. Pp. 145-176. In Lindner, E. [ed.], Die Fliegen Leiden, 474 pp.

Wiesenborn: Nitrogen Contents in Riparian Arthropods 71

NITROGEN CONTENT IN RIPARIAN ARTHROPODS IS MOST DEPENDENT ON ALLOMETRY AND ORDER

WILLIAM D. WIESENBORN U.S. Bureau of Reclamation, Lower Colorado Regional Ofﬁce, P.O. Box 61470, Boulder City, NV 89006

ABSTRACT

I investigated the contributions of body mass, order, family, and trophic level to nitrogen (N) content in riparian spiders and insects collected near the Colorado River in western Arizona. Most variation (97.2%) in N mass among arthropods was associated with the allometric effects of body mass. Nitrogen mass increased exponentially as body dry-mass increased. Significant variation (20.7%) in N mass adjusted for body mass was explained by arthropod order. Ad- justed N mass was highest in Orthoptera, Hymenoptera, Araneae, and Odonata and lowest in Coleoptera. Classifying arthropods by family compared with order did not explain significantly more variation (22.1%) in N content. Herbivore, predator, and detritivore trophic-levels across orders explained little variation (4.3%) in N mass adjusted for body mass. Within orders, N content differed only among trophic levels of Diptera. Adjusted N mass was highest in predaceous flies, intermediate in detritivorous flies, and lowest in phytophagous flies. Nitro- gen content in riparian spiders and insects is most dependent on allometry and order and least dependent on trophic level. I suggest the effects of allometry and order are due to exoskeleton thickness and composition. Foraging by vertebrate predators, such as insectivorous birds, may be affected by variation in N content among riparian arthropods.

Key Words: nutrients, spiders, insects, trophic level, exoskeleton, cuticle

RESUMEN

Se investiguo las contribuciones de la masa de cuerpo, orden, familia y el nivel trófico al contenido de nitógeno (N) en arañas e insectos riparianos (que viven en la orilla del rio u otro cuerpo de agua) recolectadaos cerca del Rio Colorado en el oeste del estado de Arizona. La ma- yoría de la variación (97.2%) en la masa (N) entre los artrópodos fue asociado con los efectos alométricos de la masa de cuerpo. La masa de nitrógeno aumentó exponencialmente con el au- mento de masa-seca del cuerpo. La variación significativa (20.7%) en la masa N ajustada por la masa del cuerpo se explica según el ordén del artrópodo. La masa ajustada N fue mas alta en Orthóptera, Hymenóptera, Araneae, Odonata y mas baja en Coleoptera. Al clasificar los ar- trópodos por familia comparado con el ordén no explica la variacion mayor significativa (22.1%) en el contenido de N. Los niveles tróficos de los herbívoros, depredadores y detritívoros en todos los ordenes explica la pequeña variación (4.3%) en la masa N ajustada por la masa del cuerpo. Entre los ordenes, el contenido N varía solamente entre los niveles tróficos de Diptera. El valor ajustado de la masa de N fue mayor para las moscas depredadores, intermedio para las moscas detritívoras y menor para las moscas fitófagas. El contenido de nitrógeno en arañas e insectos riparianos es mas dependiente sobre la alometría y ordén y menos dependiente sobre el nivel trófico. Sugiero que los efectos de alometría y ordén son debidos al grosor y la com- posición del exo-esqueleto. El forraje por los depredadores vertebrados, como aves insectivoras, puede ser afectado por la variación del contenido N entre los artrópodos riparianos.

Nitrogen concentrations in organisms are de- predators by analyzing data compiled from vari- pendent on trophic level. This is most apparent ous sources. Concentrations of N in spiders and between plants and herbivores, because N com- insects were dependent on trophic level after con- prises 0.03-7% of dry mass in plants compared trolling for body length, representing allometry, with 8-14% in animals (Mattson 1980). Variation and taxonomic group, representing phylogeny in N concentration among and within plants, and (Fagan et al. 2002). Predators generally con- its effects on abundances of herbivores including tained higher %N than herbivores. Predaceous arthropods, especially agricultural pests, has arthropods may concentrate N from food similar been frequently examined (reviewed in Mattson to phytophagous arthropods. 1980; Scriber 1984). Fewer studies have consid- Variation in N concentration among spiders ered variation in N concentration among spiders and insects may affect foraging by arthropod- and insects. Bell (1990) and Studier & Sevick feeding vertebrates and the qualities of food they (1992) tabulated measurements of %N in various obtain. Diet protein has been implicated as affect- insects from different studies. Fagan et al. (2002) ing egg production (Ramsay & Houston 1997) and compared %N between arthropod herbivores and nestling growth (Johnston 1993) in insectivorous

72 Florida Entomologist 94(1) March 2011

birds. Identifying sources of variation in arthro- for Prosopis spp., which grew together. Each spe- pod N content may improve our understanding of cies was swept 10-15 min on 9 dates: 30 Apr, 14 the prey composition required to support species May, 27 May, 08 Jun, 22 Jun, 30 Jun, 21 Jul, 4 of insectivorous wildlife. Aug, and 18 Aug 2009. All plant species were in I examined variation in N content among spi- flower or fruit except for P. fremontii. Arthropods ders and insects collected from trees and shrubs swept from plants were placed into plastic bags, established to restore riparian habitat for insec- kept in a refrigerator, and killed in a freezer. Fly- tivorous vertebrates, especially birds. Variation ing insects were trapped with a Malaise trap in N mass was partitioned into various sources. I (Santee Traps, Lexington, KY) that was placed in first determined the allometric relationship be- the center of a plot supporting S. gooddingii and tween N mass and body dry-mass. After adjusting P. sericea and elevated 1 m aboveground with N mass for this relationship, N contents of arthro- fence posts. Trapped insects were collected into a pods were compared among orders and families dry plastic bottle containing a nitrogen-free, di- and among trophic levels across and within or- clorvos insecticide strip. Insects were trapped for ders. I interpreted N contents in relation to exosk- 6.1-7.3 h during 0855-1640 PDT on each of the eleton scaling and chemical composition and con- above dates except 30 Apr, 14 May, and 18 Aug cluded by applying the results to diets of insectiv- 2009. orous birds. Spiders and insects collected on each date were sorted under a microscope into morphotypes (sim- MATERIALS AND METHODS ilar-looking specimens). Representatives of each morphotype were placed into 70% ethanol for Arthropod Collections identification. I counted and split the remaining specimens of each morphotype into samples each Spiders and insects were collected next to the with an estimated maximum dry mass of 10 mg. Colorado River within Havasu National Wildlife Individual specimens with dry masses ≥10 mg Refuge in Mohave County, Arizona. Most arthro- were placed into separate samples. Arthropod pods were collected at an irrigated 43-ha riparian samples for N analyses were cleaned by vortexing restoration area (34°46’N, 114°31’W; elevation in water, transferred to filter paper with a Büch- 143 m) of planted or volunteer trees and shrubs ner funnel, dried 2 h at 80ºC, and stored in stop- 12 km southeast and across the river from Nee- pered vials. dles, California. Plots were planted during 2003- 2005 with cuttings that were taken from nearby Arthropod Identifications and Trophic Levels areas along the river and rooted in containers. The area is straddled by Topock Marsh (16 km2) Spiders and insects were identified to the low- and Beal Lake (0.9 km2), 2 impoundments con- est taxon possible, at least to family and typically taining mostly emergent cattails (Typhus sp., to genus. Vouchers of adult insects were deposited Typhaceae) and open water. Undeveloped areas of at the Bohart Museum of Entomology, University the surrounding floodplain support mostly natu- of California, Davis, and vouchers of spiders were ralized tamarisk (Tamarix ramosissima Ledeb., deposited at the California Academy of Sciences, Tamaricaceae) shrubs. The floodplain is flanked San Francisco. Arthropod taxa were classified by Sonoran desertscrub dominated by creosote into the trophic levels of herbivore, predator, and bush (Larrea tridentata (DC.) Cov., Zygophyl- detritivore based on published descriptions (Ta- laceaae). Maximum temperatures average 42.7ºC ble 1). Holometabolous insects were classified by during Jul, and minimum temperatures average larval diet. Herbivores included consumers of pol- 5.6ºC during Jan at Needles (DRI 2010). len, nectar, or honeydew (homopteran egesta). I collected arthropods from plants and trapped Predators included parasites and consumers of insects in flight. Arthropods were swept with a already-dead animals. 38-cm diameter muslin net from planted cotton- wood (Populus fremontii S. Watson, Salicaceae) Arthropod Nitrogen Estimates and Goodding’s black willow (Salix gooddingii C. Ball, Salicaceae) trees, planted narrow-leaved The mass of N in each arthropod sample was willow shrubs (Salix exigua Nutt.), volunteer estimated with the Kjeldahl method adapted honey mesquite (Prosopis glandulosa Torrey, Fa- from Isaac & Johnson (1976). Samples of dried ar- baceae) and screwbean mesquite (Prosopis pubes- thropods were weighed (±0.01 mg) with a mi- cens Benth.) trees, and volunteer arrowweed crobalance (model C-30, Cahn Instruments, Cer- shrubs (Pluchea sericea (Nutt.) Cov., Asteraceae). ritos, CA) and ground into water with a 5-mL I also swept arthropods from T. ramosissima bor- glass tissue homogenizer. Homogenized samples dering the plots. Additional arthropods on S. ex- were poured and rinsed with water, to a total vol- igua were swept from plants growing along a dirt ume of 20 mL, into 100-ml digestion tubes. I irrigation canal 2 km northwest of the restoration added 6 mL of concentrated sulfuric acid, contain- area. Plant species were swept separately except ing 4.2% selenous acid, and 3 mL of 30% hydrogen

Wiesenborn: Nitrogen Contents in Riparian Arthropods 73 % N 9.3 ± 1.2 9.2 9.0 8.6 9.1 ± 1.5 8.9 ± 1.2 10.5 ± 2.0 11.8 10.6 ± 0.9 13.0 12.1 ± 1.8 14.3 13.8 ± 0.1 12.3 ± 1.0 13.9 ± 2.7 14.6 11.0 ± 1.5 11.6 13.3 10.1 ± 2.2 11.2 ± 1.5 11.4 ± 0.0 14.6 10.1 10.6 12.5 Mean ± SD . . CONTENT

7.20 1.37 1.93 6.29 0.07 2.03 2.46 2.35 0.46 6.62 3.36 0.68 0.35 4.37 1.24 5.22 1.51 8.99 5.72 39.7 13.0 49.2 55.2 17.1 14.1 ramosissima 115.0

Mean body dry mass (mg) NITROGEN

Tamarix 3 FOR

; T, P P P P P P P P P P P P H H H H H H H H H H H H H H ANALYZED

Pluchea sericea AND

; S, RIZONA A 3-4 1 9 1-2 6 6-7 1 1-3 1 1 1 1 1 1-3 1-3 2 5 4 2 2 2-14 1 pubescens

67 28-41 19-22 11 IN sp.

P. per sample Trophic level No. specimens No. or RIVER

Misumenops 2 1 2 1 2 4 6 1 1 1 1 1 1 4 2 1 1 1 1 1 OLORADO C No. Samples No. Prosopis glandulosa THE 2

sp.; Thomisidae, NEAR E,SS S E 2 S S P S S S S F,G,PE P F,P,S 4 E,F,GG 6 T F 5 S S G,TG F,G,SG 2 F 9

Source 6 HABITAT

Habronattus 1 7 ; M, Malaise trap; P, — — — — — 6 sp. & sp. Genus RIPARIAN

Philodromus Habronattus Metaphidippus Misumenops Pachydiplax Acridinae Scudderia Largus Nysius Brochymena Thyanta Pselliopus Zelus Cicadellinae Gyponinae Opsius Typhlocybinae Oecleus Ormenis Chrysoperla Myrmelion FROM

Misumenops Salix gooddingii Metaphidippus ; G, 4,6 5,6 COLLECTED

sp.; Thomisidae, sp.; Salticidae, Myrmeliontidae Philodromidae Salticidae Thomisidae 2 families 3 families Tettigoniidae Lygaeidae Pentatomidae Reduviidae Cixiidae Flatidae Membracidae Populus fremontii ARTHROPODS ; F,

Hypsosinga Habronattus DULT 1. A Salix exigua Subfamily in Acrididae and subfamily or genus Cicadellidae. E, D, Detritivore; H, Herbivore; P, Predator. Reference for all (Borror et al. 1981) except Apioceridae (Cole 1969) and Andrenidae, Formicidae, and Tettigoniidae (Essig 1926). Salticidae, Araneidae, Adults and immatures. Immatures. 1 2 3 4 5 6 7 ABLE Homoptera Cicadellidae Araneae OdonataOrthopteraHeteroptera Libellulidae Acrididae Largidae Neuroptera Chrysopidae Order or suborder Family T

74 Florida Entomologist 94(1) March 2011 . % N 8.3 9.8 ± 1.2 6.6 ± 2.8 9.9 ± 2.0 7.8 ± 1.0 8.1 ± 4.6 9.2 ± 2.3 5.1 ± 1.5 9.8 8.8 8.5 21.2 11.4 11.7 11.5 11.6 10.9 ± 2.2 10.9 ± 1.8 11.7 14.1 16.7 13.4 14.0 Mean ± SD CONTENT

NITROGEN

FOR

3.01 4.75 6.26 52.87 42.3 0.39 1.31 2.37 1.68 15.0 13.8 7.66 1.01 1.74 0.76 7.42 5.57 2.71 33.5 7.23 4.54 28.8 10.6 Mean body dry mass (mg) ANALYZED Tamarix ramosissima .

3 ; T, AND

P P P P P P P P P P P P P D D D H H H H H H H RIZONA A IN

Pluchea sericea ; S, RIVER

6 2-4 2-8 1 1 4-5 2-6 2 1 2-3 1-2 7-9 2 6-16 1 3 9 1 1 1 6 1 17-113 P. pubescens per sample Trophic level OLORADO No. specimens No. C THE

Misumenops sp. 1 3 3 1 1 1 2 2 1 1 1 1 1 1 1 1 1 NEAR

13 13 No. Samples No. Prosopis glandulosa or 2 HABITAT

P F,P F,S M M M F,GF,GF,GM 2 M 2 M 1 F S E,SE S E 4 M M M E G Source RIPARIAN

Habronattus sp.; Thomisidae, 1 FROM

sp. Genus Algarobius Chilocorus Hippodamia Apiocera Proctacanthus Asyndetus Homoneura Minettia Eumacronychia Apatolestes Tabanus Zaira Acinia Perdita Formica Agapostemon Dieunomia Lasioglossum Bembix Cerceris Tachysphex Myzinum Polistes COLLECTED

Misumenops Salix gooddingii ; M, Malaise trap; P, Metaphidippus sp. & ; G, ARTHROPODS

DULT Coccinellidae Asilidae Dolichopodidae Lauxaniidae Sarcophagidae Tabanidae Tachinidae Tephritidae Formicidae Halictidae Sphecidae Tiphiidae Vespidae ) A Populus fremontii ONTINUED 1. (C Subfamily in Acrididae and subfamily or genus Cicadellidae. E, Salix exigua ; F, D, Detritivore; H, Herbivore; P, Predator. Reference for all (Borror et al. 1981) except Apioceridae (Cole 1969) and Andrenidae, Formicidae, and Tettigoniidae (Essig 1926). Salticidae, Habronattus sp.; Thomisidae, Araneidae, Hypsosinga sp.; Salticidae, Adults and immatures. Immatures. 1 2 3 4 5 6 7 ABLE Order or suborder Family Diptera Apioceridae Hymenoptera Andrenidae Coleoptera Bruchidae T Wiesenborn: Nitrogen Contents in Riparian Arthropods 75 peroxide and heated samples 1 h at 400ºC with a explained more variation in adjusted log(mg N) block digestor (model 2040, Tecator, Herndon, with the general linear test approach (Neter et al. VA). After cooling, water was added to 60 mL. 1996). This approach tests if mean square error in The ammonia concentration formed in the clear, an analysis of variance decreases signiﬁcantly digested samples was measured by colorimetry, when the model becomes more complex. Samples against standards prepared from dried ammo- containing more than 1 family (3 samples of Ara- nium-sulfate, with a segmented flow analyzer neae, or spiders) were classiﬁed only to order. (model FS-4, OI Analytical, College Station, TX). Arthropod N-contents adjusted for body mass Salicylate, hypochlorite, and sodium nitroprus- were compared among trophic levels across and side were used as the indicator. I converted am- within orders or suborders. I compared N masses monia concentration to mg N. among trophic levels across orders or suborders I adjusted estimates of mg N in arthropod with analysis of variance. Separate analyses were samples with chitin samples containing known N performed within Heteroptera, Diptera, and Hy- masses. Chitin is a nitrogenous polysaccharide menoptera, the 3 orders or suborders with 2 or

(C8H13NO5)n abundant in arthropod exoskeleton, more trophic levels each containing more than 1 or cuticle (Neville 1975), that typically comprises sample. Analyses within orders or suborders 25-40% of exoskeleton dry-mass in insects (Rich- weighted adjusted values of log(mg N) by 1/s2 in ards 1978). Various masses (2, 4, 8, 16, 32, 64 mg) each trophic level to correct for uneven variances of powdered chitin (Tokyo Chemical Industry) among trophic levels (Neter et al. 1996). containing 6.89% N were weighed, placed in 20 mL water, digested, and measured for ammonia RESULTS within each batch (n = 4) of arthropod samples. I increased estimates of mg N in arthropod sam- Collected Arthropods ples in each batch to correct for the batch’s mean I collected 121 samples of spiders and insects underestimate of %N (5.76, 6.23, 6.44, 6.08%) in containing 1,490 specimens in 9 orders or subor- chitin samples. I calculated %N in arthropod ders, 33 families, and 43 subfamilies or genera samples as 100(mg N/mg dry mass). Two arthro- (Table 1). All of the arthropods collected were pod samples of Acinia and Chrysoperla with un- adults except for 8 samples in 3 taxa (families, usually low N concentrations (<0.9%) were ex- subfamilies, or genera) with adults and imma- cluded as outliers. Dry mass and mg N of each ar- tures and 6 samples in 1 taxon with only imma- thropod sample were divided by the number of tures. Body dry-masses of adult arthropods specimens in the sample to estimate dry mass ranged from 0.35 mg in Typhlocybinae leafhop- and N mass per specimen. pers (Cicadellidae) to 115 mg in the fork-tailed bush katydid Scudderia furcata Brunner (Tet- Statistical Analysis tigoniidae). Two orders or suborders (Orthoptera and Ho- Body masses of arthropods, transformed moptera) of collected spiders and insects were log(mg) to normalize residuals, were compared only herbivorous, 3 orders (Araneae, Odonata, among trophic levels with analysis of variance and Neuroptera) were only predaceous, and 4 or- (SYSTAT version 12, San Jose, CA). Nitrogen ders or suborders (Heteroptera, Coleoptera, masses in spiders and insects were analyzed se- Diptera, and Hymenoptera) included both trophic quentially. I first determined the relationship be- levels. All Coleoptera were predaceous except for tween N mass and body dry mass by regressing 1 sample. The only detritivores collected were log(mg N) against log(mg body mass) for each ar- ﬂies (Diptera). Across orders or suborders, herbi- thropod sample. I verified that the relationship vores included 42 samples in 22 taxa, predators was allometric (exponential) by testing with an included 62 samples in 24 taxa, and 17 samples in approximate t test the null hypothesis that the re- 3 taxa were detritivores (Table 1). Trophic levels gression coefficient b = 1 (Neter et al. 1996). 1 contained arthropods with different body dry- Transformed N masses were adjusted for their al- masses (F = 25.5; df = 2, 118; P < 0.001). Preda- lometric relationship with transformed body tors were largest (back-transformed mean = 6.37 mass by adding the residuals from the regression mg) followed by herbivores (4.03 mg) and detriti- to the overall mean of transformed N mass (Sokal vores (0.55 mg). & Rohlf 1981). Adjusted, transformed N masses were com- Allometric Nitrogen Contents pared among arthropod orders with analysis of variance. Hemiptera were split into suborders Nitrogen mass in riparian spiders and insects Heteroptera and Homoptera, because the diges- was allometrically related to body dry mass tive systems of most homopterans have ﬁlter (Fig. 1). Transformed N mass per specimen in ar-

chambers that concentrate nitrogenous com- thropod samples was positively related (F = 4, 066; pounds (Borror et al. 1981). I tested if classifying df = 1, 119; P < 0.001) to transformed body dry- arthropods by family instead of order or suborder mass per specimen by: 76 Florida Entomologist 94(1) March 2011

Fig. 1. Mean N mass vs. mean body dry-mass in riparian arthropods from the lower Colorado River classiﬁed by family. Abbreviations are orders or suborders (in Hemiptera): A, Araneae; C, Coleoptera, D, Diptera; He, Het- eroptera; Ho, Homoptera; Hy, Hymenoptera; N, Neuroptera; Od, Odonata; Or, Orthoptera. Single point labeled Ara- neae represents mixed samples of Araneidae, Salticidae, and Thomisidae. Axes are log scales. Line ﬁt to transformed data by linear regression weighted by sample size.

log mg N = -1.006 + 1.039(log mg dry mass) Nitrogen Content in Arthropod Orders Back-transforming this equation produced: Nitrogen mass adjusted for body mass in ripar- mg N = 0.0986(mg dry mass)1.039 ian arthropods (Fig. 2) differed (F = 3.64; df = 8, 112; P < 0.001) among orders or suborders. These The exponent (1.039 ± 0.016 SD) differs from taxonomic levels explained 20.7% of variation in unity (t* = 2.43; df = 119; P = 0.008), verifying adjusted N mass. Orthoptera (mean 14.0% N), that the relationship is exponential rather than Hymenoptera (12.4% N), Araneae (11.9% N), and linear. This allometric relationship explained Odonata (12.3% N) contained the highest ad- 97.2% of variation in N mass. Percentage of N in justed N contents, and Coleoptera (8.2% N) con- riparian arthropods (Table 1) increased as body tained the lowest adjusted N content. Orthoptera mass increased. were mostly immature slant-faced grasshoppers

Wiesenborn: Nitrogen Contents in Riparian Arthropods 77

pended on classification (Fig. 2). Across orders or suborders, N mass did not vary (F = 0.62; df = 2, 118; P = 0.54) among trophic levels. Trophic levels explained 1.0% of variation in N mass after accounting for body mass. Back-transformed means of adjusted N mass (and mean % N) were 0.413 mg (11.1% N) in herbivores, 0.397 mg (10.9% N) in predators, and 0.380 mg (9.44% N) in detritivores, the smallest arthropods collected. Within orders or suborders, N mass varied among trophic levels in Diptera (F = 4.60; df = 2, 35; P = 0.017) but not in Heteroptera (F = 0.62; df = 1, 12; P = 0.45) or Hymenoptera (F = 0.13; df = 1, 11; P = 0.91). Adjusted N contents in flies (Fig. 2) were lower in herbivores (mean 5.1% N) compared with predators (10.9% N) or detritivores (9.4% N). All phytophagous flies collected were 2 samples of the fruit fly (Tephritidae) Acinia picturata (Snow), swept from P. fremontii. Adjusted N concentrations in predaceous or parasitic flies (Apio- ceridae, Asilidae, Sarcophagidae, Tabanidae, and Tachinidae) and detritivorous flies (Dolichopo- didae and Lauxaniidae) were similar. Fig. 2. Nitrogen mass allometrically adjusted for body mass in riparian arthropods from the lower Colo- DISCUSSION rado River classiﬁed by order or suborder (in Hemi- ptera). Letters are means (± SE) and trophic levels: D, detritivores; H, herbivores; P, predators. Adjacent num- Allometric Nitrogen Contents bers are sample sizes. Y-axis is log scale. The allometric relationship between N mass and body mass in riparian arthropods resembles a similar relationship between exoskeleton mass (Acridinae) along with the sole katydid S. furcata. and body mass in terrestrial arthropods. Ander- Hymenoptera included ants (Formicidae), 2 fami- son et al. (1979) dissected the exoskeletons from 3 lies of bees (Andrenidae and Halictidae), and 3 species of immature and adult spiders, weighing families of wasps (Sphecidae, Tiphiidae, and between 25 mg and 1.2 g, and determined exosk- Vespidae). Spider samples contained several fam- eleton dry-mass and body wet-mass were posi- ilies (Table 1). The only odonate collected was the tively related by: dragonfly Pachydiplax longipennis Burmeister. Coleoptera included 1 sample of the herbivorous g exoskeleton = 0.078(g body mass)1.135 seed beetle (Bruchidae) Algarobius prosopis Le- Conte, collected from Prosopis spp., and 6 sam- Body mass in spiders explained 94.1% (their r- ples containing 2 species of predaceous ladybird value squared) of variation in exoskeleton mass. beetles (Coccinellidae), Chilocorus cacti L. and Anderson et al. attributed this allometric rela- the widespread Hippodamia convergens Guerin- tionship to scaling. The exoskeleton of terrestrial Meneville. Insects in other orders, including the 2 arthropods must increase in thickness as body Hemiptera suborders, contained intermediate N weight increases to support the organism and concentrations (Fig. 2). withstand the stresses of bending and twisting Classifying arthropods by family instead of or- (Prange 1977; Anderson et al. 1979). der or suborder did not explain more variation in Allometric relationships between N mass and N mass adjusted for body mass. Error variance of body mass, and between exoskeleton mass and adjusted N mass did not decrease (F = 1.45; df = body mass, may be primarily due to exoskeleton 26, 86; P = 0.10) when arthropods were classiﬁed N. Trim (1941) estimated N concentrations of by family compared with order or suborder. Clas- 11.8% in abdominal cuticles of 2 Orthoptera spe- sifying arthropods by family instead of order or cies, approximating the mean concentration suborder explained 22.1%, a 1.4% improvement, (10.7%) in riparian arthropods. A large proportion of variation in adjusted N mass. of N in terrestrial arthropods likely resides within the exoskeleton due to its greater density Nitrogen Content in Trophic Levels compared with internal tissues and hemolymph. The allometric relationship between exoskeleton Differences in N content among the trophic mass and body mass may have produced the sim- levels of herbivore, predator, and detritivore de- ilar relationship between N mass and body mass. 78 Florida Entomologist 94(1) March 2011

A linear increase in N mass in internal tissues as Nitrogen Contents in Trophic Levels body mass increases would dampen the exponential increase in cuticular N mass. The lower expo- I did not detect an overall difference in N con- nent relating N mass to body mass (1.039) com- centration among herbivorous, predaceous, and pared with the exponent relating cuticle mass to detritivorous arthropods after accounting for the body mass (1.135) may reflect this dampening. allometric effects of body mass. Trophic level did not appear to generally affect arthropod %N. This Nitrogen Contents in Orders or Suborders contradicts the overall difference in N concentration between herbivorous and predaceous arthro- Exoskeleton composition may have contrib- pods detected by Fagan et al. (2002). Different re- uted to different N concentrations among orders sults may have been due to statistical methodol- of spiders and insects (Fagan et al. 2002). Arthro- ogy. Fagan et al. controlled for body length and pod cuticle is composed primarily of protein and taxonomic group, to account for phylogeny, chitin (Neville 1975), and concentrations of N are whereas I controlled only for body mass. Control- higher in the former. For example, I estimated ling for phylogeny is difficult, because different %N in arthropod cuticular protein from percent- frequencies of herbivores compared with preda- ages of amino acids in pronotal and abdominal cu- tors among taxonomic groups cause trophic level ticles of adult Tenebrio beetles (Andersen et al. and phylogeny to be confounded. Phylogeny and 1973; reported in Table 3.4 in Neville 1975) by as- trophic level cannot be statistically separated. suming the amino acids were bonded into Similar N contents between trophic levels polypeptides. The estimated N concentration of agree with the concept that most insects satisfy cuticular protein (17.4%) exceeded that of chitin nutrient requirements by adjusting food intake (6.89%). Based on the maximum range of chitin (Waldbauer 1968; reviewed in Simpson et al. concentration (10-60% of dry mass) in insect cuti- 1995). An example in riparian arthropods may be cle (Richards 1978; see also Table 1 in Hackman found in the 2 suborders of Hemiptera, insects 1974), and assuming cuticle is entirely chitin and with piercing-sucking mouthparts. Phytophagous protein, N concentrations in insect exoskeleton Heteroptera, such as Lygus leaf bugs (Backus et may vary from 11.1% to 16.4%. al. 2007), typically rupture, dissolve with saliva, Greater concentrations of protein in arthro- and ingest mesophyll from a variety of plant pod cuticle, producing higher N contents, have structures. All Homoptera are herbivorous, and been associated with concentrations of resilin many homopterans feed on phloem which is high (Andersen 1979). Resilin is a flexible, elastic in water and carbohydrates but low in other nu- protein that occurs in cuticle in near-pure con- trients including N. The Opsius stactogalus Fie- centrations or combined with other proteins ber leafhoppers collected here increase food in- and chitin (Richards 1978). I estimated as take, concentrate nutrients within their filter- above that resilin contains 19.0% N from per- chamber digestive tracts (Wiesenborn 2004), and centages of amino acids in resilin from Schisto- void excess water and sugars. Concentrations of cerca grasshoppers (Andersen 1966; reported in N in Homoptera, phytophagous Heteroptera, and Table 3.4 in Neville 1975). Various mechanical predaceous Heteroptera were similar despite dif- structures in arthropods are elastic due to resi- ferent diets and physiologies. lin (Table 2.1 in Neville 1975). Resilin is espe- An exception was Diptera. Herbivorous flies, cially prevalent in the wing tendons and hinges all Tephritidae, contained lower N concentra- of Odonata and Orthoptera (Andersen & Weis- tions than predaceous or detritivorous flies after Fogh 1964), primitive orders with synchronous considering body mass. Fagan et al. (2002) com- flight muscles. Andersen and Weis-Fogh also de- pared phylogenetic categories of herbivorous in- tected resilin in the abdominal sclerites of sects and found lower N concentrations in Schistocerca grasshoppers, presumably allow- Diptera and Lepidoptera, combined as the recent ing the abdomen to stretch. Abundances of resi- lineage Panorpida, after accounting for body lin in riparian Odonata and Orthoptera may length. The database analyzed by Fagan et al. in- have contributed to their high N contents. Al- cluded the herbivorous flies Bibionidae, Chlorop- though resilin has not been found in spiders idae, and Drosophilidae, each in a different su- (Andersen & Weis-Fogh 1964), the high degree perfamily separate from Tephritidae. The diver- of abdominal stretching by spiders (Browning sity of phytophagous Diptera found to contain 1942) suggests their cuticles contain a similar low N concentrations suggests N contents in flies elastic protein. Cuticles of Coleoptera are likely generally vary by trophic level. Fagan et al. less elastic. A dominant feature of beetles is the (2002) suggested several explanations for lower elytra, hardened front-wings that act only to N contents in herbivores than in predators. cover the folded hind-wings and abdomen. The These included the direct effects of diet N, indi- likely absence of resilin and resultant high con- rect effects of trophic niche unrelated to diet, and centrations of chitin, in elytra may have low- selection for low body N in response to low diet N. ered %N in Coleoptera. The A. picturata tephritids that I collected de- Wiesenborn: Nitrogen Contents in Riparian Arthropods 79 velop as larvae in the flower heads of Pluchea The importance of prey N-concentration to in- spp. (Foote et al. 1993), corresponding with the sectivorous birds that feed on more-diverse prey flowering P. sericea at the study site. Infestations is less clear. An example is the southwestern wil- by A. picturata reduce seed production (Alyokhin low flycatcher (Empidonax traillii (Audubon) ssp. et al. 2001), suggesting larvae eat ovaries or extimus Phillips), a migrant that winters in Cen- seeds. The species does not appear to concentrate tral America and breeds in southwestern U.S. ri- N from food, because its N concentration (5.1%) is parian habitats. Adult flycatchers ate mostly het- within the range (1-7% of dry mass) reported for eropterans, flies, and beetles but fed more odo- seeds (Mattson 1980). The structural or biochem- nates and beetles to nestlings (Drost et al. 2003). ical features correlated with low N concentration Diet N may be increased by including odonates, in A. picturata and other plant-feeding flies are especially dragonflies due to their large biomass. unknown. Low exoskeleton mass in tropical, her- Diets of nestling flycatchers in other localities bivorous beetles has been attributed to low diet contained more Diptera than those of adults N, short larval-development time, and high fe- (Durst et al. 2008) or prey compositions similar to cundity (Rees 1986). Equivalent N concentra- adults (Wiesenborn & Heydon 2007). The high-N tions in predaceous or parasitic flies and detritiv- orders of Araneae, Odonata, and Hymenoptera, orous flies suggest their diets contain similar taken together, were eaten with similar frequency amounts of N. by flycatchers at different localities and habitats. These orders comprised 21% of prey in California Arthropod Nitrogen as a Nutrient for Birds (Drost et al. 2003), 31% of prey in Arizona (Durst et al. 2008), and 21% of prey at 3 localities in Ar- Not all N in arthropods is digested by insectiv- izona and Nevada (Wiesenborn & Heydon 2007). orous birds. Bird diets are frequently determined In summary, N concentrations in riparian ar- by identifying undigested fragments of exoskele- thropods are primarily dependent on body mass ton in fecal samples (e.g., Wiesenborn & Heydon and order and less dependent on trophic level. 2007). Digestion of arthropod cuticle by verte- Variation in prey N concentration may affect for- brates likely depends on its sclerotization (Kara- aging by insectivorous birds and the qualities of sov 1990). Sclerotized proteins are bonded to- food they obtain. gether, frequently with chitin, forming an irre- versibly-hardened cuticle that cannot be hydro- ACKNOWLEDGMENTS lyzed into amino acids (Richards 1978). Unsclerotized proteins, like resilin, can be hydro- I am grateful to A. Stephenson, USBR Lower Colo- lyzed (Richards 1978). Relative proportions of rado Regional Laboratory, for measuring ammonia con- sclerotized and unsclerotized proteins vary centrations. I appreciate the help identifying insects provided by S. L. Heydon, L. S. Kimsey, and T. J. Zavort- greatly among species (Richards 1978) producing ink at the Bohart Museum of Entomology, and C. A. cuticles with different digestibilities. Arthropod Tauber and P. S. Ward at the Entomology Department, orders with high amounts of elastic protein, such UC Davis. I am grateful to J. E. O’Hara at Agriculture as Odonata and Orthoptera and probably Ara- and Agrifood Canada for identifying tachinids and to D. neae, may provide insectivorous birds with high Ubick for identifying spiders. I thank J. Allen of the U.S. concentrations of digestible protein. Fish and Wildlife Service for the collection permit. This Riparian arthropods presented insectivorous work was funded by the Lower Colorado River Multi- birds with prey containing a range (5.1-14.0%) of Species Conservation Program. N concentrations. Foraging by insectivorous birds in relation to prey N concentration can be difficult REFERENCES CITED to discern, because birds frequently forage in response to prey availability which is transitory ALYOKHIN, A. V., MESSING, R. H., AND DUAN, J. J. 2001. Utilization of the exotic weed Pluchea odorata and hard to estimate. 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Paraiso et al.: Egg Parasitoids of the Cactus Moth 81

EGG PARASITOIDS ATTACKING CACTOBLASTIS CACTORUM (LEPIDOPTERA: PYRALIDAE) IN NORTH FLORIDA

OULIMATHE PARAISO1, STEPHEN D. HIGHT2, MOSES T. K. KAIRO1 AND STEPHANIE BLOEM3 1Center for Biological Control, College of Engineering Sciences, Technology and Agriculture, Florida Agricultural & Mechanical University, Tallahassee, FL 32307

2USDA, ARS, CMAVE, Tallahassee, FL 32308

3USDA, APHIS, PPQ, CPHST, Plant Epidemiology and Risk Analysis Laboratory, Raleigh, NC 27606

ABSTRACT

Interest in the natural enemies of Cactoblastis cactorum (Berg) has increased since the moth was found in Florida in 1989. Previous surveys for natural enemies in Argentina identified egg parasitoids in the family Trichogrammatidae as potentially important control agents of C. cactorum. A study was conducted in north Florida to identify and to assess occurrence of egg parasitoids attacking this invasive moth in its new homeland. Surveys undertaken at 6 locations in north Florida from Jul 2008 to Dec 2009 revealed that eggsticks of C. cactorum were attacked by egg parasitoids from the Trichogramma genus: T. pretiosum Riley, T. fuentesi Torre, and an additional unidentified Trichogramma species belonging to the T. pretiosum group. In order to assess the importance of these egg parasitoids, the fate of individual C. cactorum eggsticks was determined during weekly visits to each site. This assessment showed that the combined level of parasitism of C. cactorum eggsticks was very low with less than 0.2% of host eggs attacked at any one of the 6 sites. While parasitoids attacked smaller eggsticks, there was no correlation between the numbers of eggs in an eggstick attacked with increasing number of eggs/eggstick. Comparing the mean number of eggs/eggstick, there was no difference between the 3 flight periods of C. cactorum, but there was a difference between the 6 sites. Based on these results, the use of Trichogramma wasps as an inundative biological control agent, complementary to the Sterile Insect Technique application, is discussed.

Key Words: Cactoblastis cactorum, cactus moth, Trichogramma, egg parasitoids, North Florida

RESUMEN

El interés en los enemigos naturales de Cactoblastis cactorum (Berg) ha aumentado desde que esta especie fue encontrada en el estado de Florida en 1989. Busquedas de enemigos naturales de C. cactorum hechas en años pasados en Argentina identificaron a parasitoides de huevos de la familia Trichogrammatidae como enemigos naturales de posible importancia para esta especie. Llevamos a cabo un estudio en seis localidades en el norte del estado de Florida con el objetivo de identificar y evaluar la ocurrencia de parasitoides de huevos ata- cando a esta especie en su nueva área de distribución. La busqueda de parasitoides llevada a cabo entre julio del 2008 y diciembre del 2009 indico que los bastoncitos de huevos de C. cactorum son atacados por parasitoides del genero Trichogramma: T. pretiosum Riley, T. fuentesi Torre, y una especie adicional no identificada perteneciente al grupo taxonómico de Trichogramma pretiosum. Para evaluar la importancia de estos parasitoides en el control de C. cactorum, seguimos el destino de bastoncitos individuales a través de visitas semanales a cada una de las localidades. Estas observaciones demostraron que el grado de parasitismo en esos basoncitos es muy bajo, con menos de 0.2% de los huevos parasitados en cualquiera de las seis áreas. Mientras que observamos que los parasitoides atacaron bastoncitos de huevos de tamaño pequeño, no hubo correlación entre el número de huevos parasitados por bastoncito y el tamaño del mismo. Comparando el numero promedio de huevos por bastoncito, no detectamos diferencia en los bastoncitos ovipositados en las tres generaciones anuales de C. cactorum pero detectamos diferencias dependiendo del área. Basado en estos resultados, discutiremos el uso de parasitoides del genero Trichogramma como agentes inundativos de control biológico, complementando la aplicación de la Técnica del Insecto Estéril contra C. cactorum.

Translation provided by the authors.

The cactus moth, Cactoblastis cactorum (Berg) fect example of a successful weed biological con- (Lepidoptera: Pyralidae), is often cited as the per- trol agent (Moran & Zimmermann 1984). In 1925,

82 Florida Entomologist 94(1) March 2011 the cactus moth was introduced from its native pedalis Cresson) and 1 unidentified egg parasi- Argentina into Australia to control prickly pear toid from the family Trichogrammatidae attack- cactus, Opuntia spp., which had originally been ing C. cactorum in Florida. Logarzo et al. (2008) brought into Australia for commercial purposes found the larval parasitoid Apanteles alexanderi (Dodd 1940; Mann 1970). The cactus had become Brethes (Hymenoptera: Braconidae) and the egg invasive and made large tracts of rangeland unfit parasitoid Trichogramma pretiosum Riley (Hy- for grazing cattle. Within a few years after the in- menoptera: Trichogrammatidae) attacking C. troduction of C. cactorum into Australia, US $6 cactorum in Argentina. million worth of rangeland was restored, equiva- Trichogrammatid egg parasitoids have been lent to more than US $60 million in today’s dol- used successfully for inundative biological control lars (Dodd 1940; Williamson 2009). Based on against major lepidopteran pests such as corn these promising results, C. cactorum was im- borers, sugarcane borers, and cotton bollworm ported from Australia to South Africa, Mauritius, (Li-Ying 1994; van Lenteren 2000). Egg parasi- and Hawaii to manage other non-native and inva- toids are easy to rear in mass quantity in labora- sive Opuntia spp. (Moran & Zimmermann 1984). tory conditions and to release over wide areas. Bi- In 1957, C. cactorum was introduced into several ological control can be used to complement and Caribbean islands (Nevis, Montserrat, and Anti- synergize the application of SIT (Gurr & Kve- gua) to control non-native as well as native Opun- daras 2010). Recent studies showed that the com- tia spp. (Simmonds & Bennett 1966). Unfortu- bination of both techniques was more efficient in nately, the implementing agencies did not fully controlling pest population of the codling moth, consider the potentially injurious environmental Cydia pomonella (L.) (Bloem et al. 1998). Syner- impacts of C. cactorum if this insect were to move gistic interactions between SIT and fruit fly bio- to neighboring countries where some species of logical control with parasitoids increased the sup- Opuntia are important native species and some pression of pest fruit flies, even leading to eradi- are commercially important (Stiling et al. 2004). cation (Sivinski 1996; Rendon et al. 2006). SIT The first record of C. cactorum in the U.S. was and biological control have been successfully com- from Bahia Honda Key, Florida, in Oct 1989 bined to combat several lepidopteran pests, in- (Dickel 1991). It is uncertain how the moth ar- cluding C. pomonella (Bloem et al. 1998) and rived in Florida, but several interceptions of Car- painted apple moth, Orgyia anartoides (Walker) ibbean ornamental Opuntia spp. infested with C. (Suckling et al. 2007). Radiation doses for steril- cactorum were found at ports of entry in south izing C. cactoblastis adults have been determined Florida during the 1980s and 1990s (Pemberton to produce partially sterile but more fit males 1995; Zimmermann et al. 2001; Stiling 2002; Si- which, when mated with wild females, generate monsen et al. 2008). Since its appearance in Flor- sterile offspring (Carpenter et al. 2005). The com- ida, C. cactorum has become a threat to native bination of egg parasitoid releases and SIT has Opuntia spp. in North America. Current manage- the advantage that parasitoids manage high pest ment options include the use of Pherocon 1-C densities, while SIT works best at low pest densi- Wing traps (Trécé Incorporated, Salinas, CA) ties. In addition, release of sterile insects provides baited with a 3-component synthetic sex lure (Su- an egg resource for egg parasitoids increasing the terra, LLC, Bend, OR) to identify the presence of ratio of natural enemies to adult hosts. Egg para- the moth, coupled with removal of infested plants sitoids and sterile insects have the characteristic to reduce C. cactorum populations (Bloem et al. of being self dispersing and consequently are able 2005; Hight & Carpenter 2009). Complementary to cover wide areas (Sivinski 1996). to the detection, monitoring, and removal efforts, We conducted field surveys in order to identify implementation of the Sterile Insect Technique egg parasitoids already established in North Flor- (SIT) is being used to slow the geographic expan- ida that attack C. cactorum. Cactoblastis cac- sion of C. cactorum in the U.S. (Hight et al. 2002; torum adults have 3 annual flight periods in Bloem et al. 2005; Bloem et al. 2007). In Mexico, north Florida (Apr-May, Jul-Aug, and Oct-Nov) localized invasions of C. cactorum on 2 islands (Hight et al. 2005; Hight & Carpenter 2009). We were eradicated in 2008 with a program of phero- report on the distribution, seasonality, and para- mone traps, host removal, and SIT (NAPPO 2006; sitism parameters of the Trichogramma species NAPPO 2008; NAPPO 2009). attacking C. cactorum in northern Florida. The Bennett & Habeck (1995) suggested biological number of eggs/eggstick was compared between control as an additional control option that should different flight periods and sites to assess host be considered for C. cactorum. Pemberton & egg resource for egg parasitoids. The effect of C. Cordo (2001) reported that several larval and pu- cactorum eggstick size on level of parasitism was pal parasitoids attacked the cactus moth in South evaluated by comparing number of eggs from par- America, including species of Hymenoptera (Bra- asitized versus un-parasitized eggsticks. These conidae, Chalcidae, and Ichneumonidae), and 1 data will be beneficial in promoting discussions Diptera (Tachinidae). They also reported on 2 on possible implementation of biological control chalcid species (Brachymeria ovata (Say) and B. for the cactus moth and, in particular, assessing

Paraiso et al.: Egg Parasitoids of the Cactus Moth 83 the potential of an inundative biological control parasitoids. The number of eggs per eggstick was program against C. cactorum in North America. determined either by a direct count or by a correlated estimate of eggstick length to egg number MATERIALS AND METHODS (2.62 ± 0.013 eggs/mm). The ratio of eggstick length to egg number was calculated in this study Field surveys were carried out at 6 locations by counting the number of eggs in a segment of (Fig. 1) in north Florida from Jul 2008 to Dec eggstick, replicated on 20 eggsticks. Eggstick 2009. The selection of study sites was based on ex- length was estimated in situ by placing a plastic isting records of infestations from the literature, string next to the eggstick and cutting a piece of personal observations from preliminary surveys, equivalent length. The length of the piece of and information provided by experts. Female C. string was then measured to the nearest 0.01 mm cactorum place their eggs end to end to form a with a metric micrometer. Measurements of egg- chain that looks like a short “stick”, and the egg sticks were obtained so that the number of eggs mass is referred to as an eggstick. Although no exper eggstick could be estimated if the eggstick tensive field surveys were conducted from May to was lost before it could be collected and directly Jul 2008 at St. Marks and St. George Island, egg- counted. The fate of each eggstick was deter- sticks with eggs that appeared parasitized were mined by making weekly visits to each site to collected and held in laboratory conditions until evaluate the status of previously tagged egg- parasitoids emerged. At survey locations, 20 to 30 sticks. The fate of each eggstick was categorized healthy Opuntia spp. plants were chosen with no as follows: eggstick lost; predated (visible chew- to minor feeding damage by cactus moth larvae ing damage) eggs in the eggstick versus non pre- and an average of at least 50 pads per plant. Dur- dated eggs; or parasitized eggs in the eggstick ing weekly visits throughout all 3 flight periods, (black eggs formed before C. cactorum larvae suc- any new eggstick was identified by plant, pad, cessfully developed). Eggsticks were collected if and its general location on the plant so the egg- they were damaged during evaluation or mea- stick could be found during subsequent checks. A surement, eggs of the eggstick had hatched, or mark was made on the plant at the base of the eggs appeared predated or parasitized. Eggsticks eggstick with a felt tip pen and a red tape “flag” with viable eggs were collected and held in small affixed to an insect pin placed near the eggstick to plastic cups (30 mL) under laboratory conditions aid in finding the eggstick. The flag was labeled (25 ± 1 C°, 16:8 L:D and 40-60% RH) to record with a unique number to identify each eggstick. hatch rate. Eggsticks with parasitized eggs were The oviposition preferences of C. cactorum fe- collected and monitored in the laboratory to de- males on host plants were recorded by classifying termine the emergence rate, number of eggs/egg- the attachment of the eggstick to either a glochid stick attacked by parasitoids, number of parasi- at an areole, to a spine, or on the fruit. Observa- toids emerging per parasitized egg, and to ascer- tions on plant habitat and host eggstick distribu- tain the identity of the parasitoids. Parasitoid tion within the surveyed site and within the se- specimens were submitted to R. Stouthamer, De- lected plant were collected to provide additional partment of Entomology, University of California, information on the host finding behavior of egg Riverside, for molecular identification. The se- quencing of ribosomal DNA Internal Transcribed Spacer 2 (ITS 2) was used to identify the different species of egg parasitoids.

Data Analysis

The numbers of eggs/eggstick at different ﬂight periods for each surveyed location and the average number of eggs/eggstick at each site were log transformed before analyses to satisfy the as- sumptions of the analysis of variance. One way analysis of variance (PROC GLM) was applied to the log transformed data and the separation of means was made with the least signiﬁcant difference (LSD) test. Comparison of number of eggs/ eggstick that was parasitized versus number of eggs/eggstick not parasitized was also evaluated with a one-way analysis of variance (PROC GLM). Since only a few eggsticks with parasitized eggs were collected in this study (see text below), Fig. 1. Locations and their coordinates surveyed for egg comparisons between eggsticks with parasitized parasitoids of Cactoblastis cactorum in North Florida. eggs were made against the same number of ran-

84 Florida Entomologist 94(1) March 2011 domly selected eggsticks with un-parasitized Marks. In addition, heavy rainfall may have sep- eggs. Variation between the number of eggs for arated the eggsticks from plants, but we do not parasitized eggsticks and the number of eggs for have any data on the severity of the rain storms the randomly selected un-parasitized eggsticks at different study sites. Cactoblastis cactorum life was analyzed by a folded F test (Davis 2007). Be- table studies in Argentina (Logarzo et al. 2009) cause the variances in numbers of eggs for egg- and South Africa (Robertson & Hoffmann 1989) sticks with parasitism and number of eggs in egg- identified rain and wind as major factors contrib- sticks without parasitism were not significantly uting to mortality of eggs. different, means of these 2 groups were compared Surveyed sites and oviposition periods were with a two-sample t-test. A Pearson’s Correlation analyzed to evaluate their influence on number of Coefficient (r) was calculated to determine eggs/eggstick. Eggsticks were collected for multi- whether the numbers of eggs parasitized were de- ple oviposition periods at 3 sites (St. George Is- pendent on the number of eggs/eggstick. The SAS land, St. Marks, and Okaloosa Island) (Table 2). Statistical Software Version 9.2 (SAS Institute, The numbers of eggs/eggsticks for the different Cary, North Carolina) was used to perform the oviposition periods were not significantly differ- statistical analyses. Estimates of central tenden- ent for St. George Island (F = 1.84, df = 1, P = cies were reported as mean ± standard error of 0.18), St. Marks (F = 93.86, df = 3, P = 0.07), or mean. Okaloosa Island (F = 0.22, df = 3, P = 0.88). Be- cause the numbers of eggs/eggstick for multiple RESULTS AND DISCUSSION oviposition periods were not different, eggsticks from all flight periods were pooled to calculate the Although host plant species of Opuntia stricta means for those sites (St. George Island (62 ± 2.8), (Haworth) Haworth, O. humifusa (Rafinesque) St. Marks (53 ± 2.8), and Okaloosa Island (59 ± Rafinesque, and O. ficus-indica (L.) P. Miller var- 1.8)). The pooled eggsticks were used to compare ied among the different geographic regions sur- the number of eggs/eggstick between all 6 sites veyed, the oviposition preferences of C. cactorum and significant differences were found (F = 11.44, females was similar on the various species (Table df = 5, P < 0.0001) (Table 2). 1). In this study, parasitized eggsticks of C. cac- Female C. cactorum laid similar numbers of torum appeared mostly on the areole/glochid eggs/eggstick for each of the 3 oviposition periods structure of the pads (Table 1). but not at all 6 survey sites along the Florida pan- Altogether, 1,527 eggsticks with 91,013 C. cac- handle. The longest eggsticks were observed at torum eggs, not including 344 eggsticks missing St. George Island, Pensacola Beach, and Oka- from the field or lost during collection, were loosa Island (Table 2). Significantly smaller egg- tagged on plants of Opuntia spp. (Table 2). Of all sticks were recorded at St. Marks and Mexico the eggsticks checked, 62% were collected on Oka- Beach (Table 2). Panacea had significantly loosa Island and had a mean of 59 (+/- 1.83) eggs/ smaller number of eggs/eggstick than all other eggstick. The proportion of eggsticks examined in sites (Table 2). The cause of differences between the laboratory as percentage of all eggsticks sur- eggsticks at the various sites was unclear. Studies veyed at the 6 field sites ranged from 53 to 100%, in South Africa identified differences in total fe- except for summer 2008 at St. George Island and cundity of C. cactorum due to host plant species, St. Marks National Wildlife Refuge (NWR) in the flight period when eggs were laid, and the which only 30% and 24%, respectively, of the mon- temperature during oviposition (Robertson 1989). itored eggsticks were examined (Table 2). The We did not distinguish eggsticks collected from majority of the eggsticks from these 2 locations different host plants (Table 1). While South Afri- for this flight period were recorded as lost (Table can female C. cactorum had significantly higher 2). The cause for this high number of lost egg- fecundity during the summer flight (Robertson sticks is not clear. Several biotic and abiotic fac- 1989), our study did not show any difference in tors could have contributed to the high number of number eggs/eggstick between flight periods in lost eggsticks. During summer 2008, 23% of egg- north Florida. Cactoblastis cactorum has a ten- sticks examined from St. Marks had eggs that dency to oviposit on plants with high nitrogen were preyed upon compared with less than 3% in (Myers et al. 1981; Robertson 1987), but we have other locations. Although not directly observed at no direct measurements of plant quality at our St. George Island or St. Marks, substantial preda- sites. tion of C. cactorum eggs by ants has been recorded Comparing the number of eggs/eggstick for in South Africa (Robertson 1984). Because the eggsticks that were parasitized (38 ± 13.7) (Table plants surveyed at St. Marks were located within 3) against un-parasitized eggsticks (61 ± 13.1) re- 100 m of the waters of the Gulf of Mexico, strong vealed a significant difference (pooled t test = winds characteristic of coastal regions could have 3.14, df = 12, P = 0.0085). Although the number of knocked eggsticks off the plants. All other study eggs/eggstick was highly variable, the variation of sites were along the Gulf Coast; in none of them the number of eggs/eggstick for parasitized ver- were the plants as close to the water as at St. sus the randomly selected un-parasitized group Paraiso et al.: Egg Parasitoids of the Cactus Moth 85 2 INFORMA-

1 ADDITIONAL

Location AND . SP

Percent Eggsticks at Attachment Attachment at Eggsticks Percent PUNTIA Areole/ Glochid Spine Fruit Missing O

9 78 0 22 0 45 80 13 7 0 28 89 1147 0 88 0 2 0 10 2965 n/a n/a n/a n/a n/a n/a n/a n/a 280 77 19 2 2 151 n/a n/a n/a n/a 120105 50 34 63 16 30 0 7 0 151 n/a n/a186 n/a n/a 81 18.5 0.5 0 308 79 15 1 4 Eggsticks Eggsticks Evaluated EGGSTICKS

Total Number Total CACTORUM 18 25 20 23 30 30 27 35 35 10 18 n/a n/a

Plant Examined Number Host ACTOBLASTIS C

Plant OF

Species of Opuntia Host O. humifusa O. O. stricta O. humifusa O. ficus-indica O. stricta O. stricta O. O. ficus-indica O. stricta O. ficus-indica O. ficus-indica O. PARASITOIDS

3 3 18 20 10 10 20 18 13 21 14 EGG Surveys

Total Number Total RICHOGRAMMA . T

FOR

Surveyed Dates Eggsticks Dates Eggsticks (Apr 08-Jul 08, 09) 08, (Apr 08-Jul Summer 2009 (Jul 01-Sep 25, 09) 01-Sep 25, (Jul 2009 Fall (Sep 18, 09-Jan 12, 10) 12, 09-Jan (Sep 18, Fall 2008 Fall Fall 2008 Fall 09) 25, 08-Feb (Sep 25, Summer 2008 09) 25, 08-Feb (Oct 01, Spring 2009 09) 15, (Apr 17-Jul 2009 Fall (Oct 07, 09-Jan 12, 10) 12, 09-Jan (Oct 07, 2009 Fall 2008 Fall Spring 2009 LORIDA PREFERENCE

F NORTH

GPS OVIPOSITION

Coordinate W84.18242 08) 15,-Sep 12, (Jul W87.48928 08) 10-Sep 10, (Jul W84.9120 08) (Jul17-Sep19, N30.07772 W85.40636N30.03127 09) (Oct 21-Nov12, W84.39353N30.08674 W84.39353 W86.37807 09) 27, 08-Feb (Oct 08, MOTH

SURVEYED

ITES TION 1. S Attachment locations of eggsticks that were not determined is indicated by “n/a”. failed to be recorded. attachment Information about eggstick 1 2 ABLE Site St. George IslandSt. N29.65068 Summer 2008 Marks (NWR) St. Pensacola BeachPensacola N30.33525 Summer 2008 Mexico BeachPanacea N29.94133 2009 Fall Okaloosa Island T 86 Florida Entomologist 94(1) March 2011 1 EGGSTICK

PER

53 ± 2.8 b 62 ± 2.8 a 62 ± 1.5 a 52 ± 3.0 b 59 ± 1.3 a 45 ± 1.9 c Eggstick Eggstick EGGS ± SE at Each Site ± SE at Each

Overall Mean Eggs/ OF

NUMBER

± SE 60+/-1.1 68+/-2.9 57+/-3.3 54+/-3.1 57+/-1.2 64+/-2.0 58+/-3.6 61+/-1.2 62+/-1.5 46+/-1.4 52+/-3.0 60+/-1.4 45+/-1.9 MEAN

Eggs/Eggstick Eggs/Eggstick Mean Number AND , 513 2,561 8,638 6,685 1,614 3,088 6,917 7,402 1,522 2,892 LABORATORY

17,126 20,527 11,118 Examined Moth Eggs Total Number Total THE

EXAMINED 55 54 92 99 30 53 24 42 99 71 , . 100 100 100 Percent Eggsticks Eggsticks Examined FIELD

PERIODS

THE

LOST , OVIPOSITION 4 (44) 1 (0.6) 0 (0) 0 (0) 3 (1) 0 (0)

23 (46) 21 (8) 84 (70) 13 (46) 35 (77) 69 (58) 61 (29) (Percent) (Percent) Total Number Total Eggsticks Lost Eggsticks COLLECTED

DIFFERENT

FOR

9 P < 0.05). 47 45 28 29 65 151 151 280 105 120 308 186 EGGSTICKS

LORIDA Total Number Total F Eggsticks Tagged Eggsticks NORTH CACTORUM

SITES

Fall 2009 Fall Fall 2008 Fall Spring 2009 2009 Fall Fall 2008 Fall Summer 2009 Fall 2009 Fall Spring 2009 ACTOBLASTIS C

DIFFERENT

UMBER SE ± 2. N Means with different letter are significantly ( 1 ABLE St. Marks (NWR)St. Summer 2008 St. George IslandSt. Summer 2008 Mexico Beach 2009 Fall Pensacola BeachPensacola Summer 2008 Panacea Site Flight Period Okaloosa Island 2008 Fall T Paraiso et al.: Egg Parasitoids of the Cactus Moth 87 , 1 0.2 0.1 0.1 EGGSTICK n/a CACTORUM

Level of C.

PER

Parasitism (%) Parasitism EGGS

40 70 73 71 62 56 89 77 ATTACKING

Percent Females NUMBER , PARASITOIDS

SPECIES 7 9 5

10 11 16 63 18 EGG

Emerged OF

Number Parasitoids Number Parasitoids PARASITOID LEVEL

6 (33) 2 (10) 6 (11) 3 (6) 8 (18) PARASITISM IDENTITY 10 (24) 13 (52) 19 (22)

, AND , Eggs/Eggstick # (%) Eggs/Eggstick Number Parasitized Number Parasitized RATIO

COLLECTED

WAS

18 42 20 56 52 25 44 7888 17 (22) 34 85 73 5 (7) 8 75 n/a FEMALE Eggstick , Number Eggs/ EGGSTICK

sp. EMERGED

CACTORUM

T. pretiosum T. pretiosum T. T. fuentesi T. fuentesi T. unknown fuentesi T. T. pretiosum T. T. pretiosum T. T. pretiosum T. PARASITOIDS

Trichogramma OF

ACTOBLASTIS C

NUMBER Flight Period , EGGS

PARASITIZED

. Date DATE

08/13/08 10/16/08 10/16/08 11/03/08 10/23/09 09 Fall 08/06/08 Summer 08 05/15/08 Collection PARASITIZED LORIDA

AND

F OF

NORTH

OCATION IN NUMBER 3. L Indicates that the level of parasitism was not determined. 1 ABLE Okaloosa Island 10/16/08 08 Fall Pensacola BeachPensacola 04/22/08 Spring 08 unknown St. Marks St. 05/15/08 Spring 08 Site T 88 Florida Entomologist 94(1) March 2011 was similar (folded F test = 1.10, df = 6, P = 0.91), cea) (Hübner), corn earworm (Helicoverpa zea) suggesting that the difference found between the (Boddie), tomato pinworm (Keiferia lycopersi- two groups was not driven by unequal or extreme cella) (Walshingham), sugarcane borers (Di- variation. However, there was not a significant atraea spp.), and cabbage looper (Trichoplusia correlation between the number of eggs/eggstick ni) (Hübner) (Pinto et al. 1986; Hassan 1993; Li- and number of eggs parasitized by Trichogramma Ying 1994; Monje et al. 1999). Trichogramma fu- spp. (n = 7, r = -0.16, P = 0.74). Therefore, while fe- entesi have been recorded in countries in South male Trichogramma spp. parasitized eggsticks America (Argentina, Columbia, Mexico, Peru, with fewer eggs, they did not parasitize more eggs and Venezuela) and in the U.S. (Alabama, Cali- as the number of eggs in an eggstick increased. fornia, Florida, Louisiana, New Jersey, South The average number of eggs parasitized in an Carolina and Texas) (Fry 1989, Pinto 1999). Its eggstick was 9 (±5.8). primary hosts are species from the Noctuidae Ten eggsticks were found parasitized at 3 of family such as H. zea and Heliothis virescens (F.) the 6 sites surveyed (Pensacola Beach, St. Marks, and from the Pyralidae family such as Diatrea and Okaloosa Island). Five of the parasitized egg- saccharalis (F.), Ephestia kuehniella Zeller, and sticks were found at Okaloosa Island. Parasitized Ostrinia nubilalis (Hübner) (Fry 1989; Wilson & eggsticks were found during all 3 oviposition pe- Durant 1991; Pintureau et al. 1999; Querino & riods of C. cactorum females: the spring flight (St. Zucchi 2003). Trichogramma parasitoids also are Marks and Pensacola Beach), summer flight widely used for pest control in orchards (Pensacola Beach), and fall flight (Okaloosa Is- (Olkowski & Zang 1990). The observed low inci- land). Of the 496 eggs in the 10 parasitized egg- dence of the wasps in natural areas might be ex- sticks, a total of 89 eggs (or 18%) were parasit- plained by unfavorable environmental factors or ized, resulting in the emergence of 181 adult par- natural plant chemicals (Smith 1996; Romeis et asitoids with a sex ratio of 70% (±14) females (Ta- al. 1997, 1999). However, contrary to other natu- ble 3). The level of parasitism by Trichogramma ral enemies, Trichogramma can be easily and spp., relative to the total number of eggs exam- cheaply mass-reared for the implementation of ined during the different flight periods for each an inundative biological control program. site, was less than 0.2% of total C. cactorum eggs The potential for inundative releases of Tri- collected (Table 3). We did not observe any para- chogramma spp. as a strategy against C. cac- sitized eggsticks at St. George Island, Mexico torum is currently being investigated with sus- Beach, or Panacea. tainable laboratory colonies of T. fuentesi origi- Two species of Trichogramma were reared nating from field collected insects reared from from C. cactorum eggsticks in north Florida (Ta- parasitized C. cactorum eggsticks. Biological ble 3) and identified by differences in IST2 se- characteristics (sex ratio, egg load, and longevity) quences. Trichogramma pretiosum was collected and different behavioral mechanisms (influence at St. Marks, Pensacola Beach, and Okaloosa Is- of parasitoid age, density, and host age on parasit- land, while T. fuentesi Torre was recovered only ism) involved in host finding of T. fuentesi reared from Okaloosa Island. It was not possible to iden- on C. cactorum eggs are being evaluated. The in- tify 2 collections of Trichogramma spp. from Oka- undative releases of Trichogramma wasps could loosa Island; one because a good molecular se- be integrated in the current pest management quence could not be obtained and for the other the system based on SIT applications during the 3 sequence was not in the database and possibly flight periods by building Trichogramma popula- represents a new species in the T. pretiosum tions. This field survey was useful in identifying a group (R. Stouthamer, UC—Riverside, personnel potential inundative biological control agent that communication). could be integrated within a pest management More than 15 million ha of agriculture and strategy against C. cactorum. forestry worldwide are treated annually with Trichogramma egg parasitoids (van Lenteren ACKNOWLEDGMENTS 2000). Trichogrammatid wasps have been used successfully in inundative release programs We thank Shalom Benton (FAMU) for field collection against lepidopteran pests in greenhouses and and laboratory assistance and Chris Albanese, Michael crop production worldwide (Smith 1996). Inun- Getman, and John Mass (USDA-ARS-CMAVE, Talla- dative releases of Trichogramma spp. have been hassee) for field assistance. We thank Stuart Reitz implemented in Florida to control major lepi- (USDA-ARS-CMAVE, Tallahassee, FL) and Jim Nation dopteran pests of collards, cabbages, soybeans, (University of Florida) for comments on earlier drafts of bell peppers, tomatoes, corn, and tobacco produc- this manuscript. This work is funded under the FAMU- tion (Martin et al. 1976). Trichogramma pretio- USDA APHIS Cooperative Agreement, 07-10-8100- 0755-CA. Mention of trade names or commercial prod- sum is commonly found in the Western hemi- ucts in this publication is solely for the purpose of pro- sphere. This Trichogramma species has been re- viding specific information and does not imply leased commercially against major lepidopteran recommendation or endorsement by the U.S. Depart- pests such as cotton leafworm (Alabama argilla- ment of Agriculture. Paraiso et al.: Egg Parasitoids of the Cactus Moth 89

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Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata 91

HOST STATUS OF PURPLE PASSIONFRUIT FOR THE MEDITERRANEAN FRUIT FLY (DIPTERA: TEPHRITIDAE)

JOSE A. RENGIFO1, JAVIER G. GARCIA1, JOHN F. RODRIGUEZ1 AND KRIS A. G. WYCKHUYS2 1Colombian Institute for Agriculture and Livestock, ICA, Bogota, Colombia

2International Center for Tropical Agriculture CIAT, Cali, Colombia

ABSTRACT

The Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is a key pest of a wide range of fruit crops and the focus of rigid quarantine restrictions and eradication measures in several countries. In Colombia, the susceptibility of purple passionfruit (Pasiflora edulis f. edulis Sims; Violales: Passifloraceae) to C. capitata is uncertain. Field collections of fruit were made to evaluate natural infestation. Forced infestation studies were conducted in the laboratory with punctured and intact fruit to determine the acceptability of fruit at different stages of maturity and physiological suitability of fruit to development. No C. capitata larvae were found and no adults emerged from a total of 976 hand-picked fruit and 623 fallen fruit. In the meantime, trap data indicated that C. capitata is not present in the principal passionfruit production regions. For intact fruit, C. capitata females oviposited exclusively in fruit of maturity level zero, with 41.67% of fruit accepted for oviposition and an average of 183.1 ± 33.8 eggs per fruit. No oviposition was recorded in fruit of maturity levels 2 and 4. For punctured fruit, C. capitata oviposited a total of 84,410 and 84,250 eggs into fruit of maturity levels 0 and 2, respectively, but no C. capitata adults emerged from fruit at either maturity level. Laboratory tests suggest that purple passionfruit is a non-host for C. capitata.

Key Words: quarantine pest, Ceratitis capitata, host status, risk analysis, fruit ﬂy

RESUMEN

La mosca del Mediterráneo Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) es una plaga clave de una amplia gama de frutales y es el foco de estrictas restricciones cuarente- narias y medidas de erradicación en varios países. En Colombia, la susceptibilidad del ma- racuyá morado (Pasiflora edulis f. edulis Sims; Violales: Passifloraceae) a C. capitata es incierta. Se hicieron colectas de frutos en campo para evaluar el nivel de infestación. En el laboratorio se desarrollaron estudios de infestación forzada con frutos perforados e intactos para determinar la aceptabilidad del fruto en los diferentes estados de maduración e idonei- dad fisiológica del desarrollo de los frutos. No se encontraron larvas de C. capitata ni adultos emergidos en un total de 976 frutos recogidos manualmente y 623 frutos caídos. Mientras tanto, los datos de captura indicaron que C. capitata no está presente en las principales re- giones de producción del maracuyá. Para frutos intactos, las hembras de C. capitata ovipo- sitaron exclusivamente frutos de nivel de maduración cero, con 41.67% de aceptación de frutos para oviposición y en un rango de 183.1 ± 33.8 huevos por fruto. No se registró ovipo- sición en frutos con niveles de maduración 2 y 4. Para frutos perforados, C. capitata ovipositó un total de 84,410 y 84,250 huevos dentro de frutos con nivel de maduración 0 y 2 respectivamente, pero no emergieron adultos de C. capitata de los frutos en ningún nivel de madu- ración. Las pruebas de laboratorio sugieren que el maracuyá morado no es hospedero para C. capitata.

Translation provided by the authors.

Tephritid fruit flies are key pests of a wide va- (Wiedeman), a destructive pest of multiple fruit riety of fruit species, affecting crop yield, quality crops worldwide (Liquido et al. 1991). In assess- of harvested produce, and (international) market ing risk of C. capitata arrival in the U.S. and de- access (e.g., Robinson & Hooper 1989; Aluja & veloping associated quarantine protocols, su- Mangan 2008). Given the polyphagous nature of preme precaution is taken to avoid entry of poten- many fruit fly species, quarantine restrictions are tial host fruits of this pest. Listings of the status in place to avoid their introduction in certain of particular fruits as hosts of C. capitata are the countries or geographical regions. A key quaran- cornerstone of quarantine restrictions (Liquido et tine pest for the continental United States is the al. 1991). However, current restrictions include Mediterranean fruit fly, Ceratitis capitata fruit species for which there is poor information

92 Florida Entomologist 94(1) March 2011 regarding C. capitata host status. Hence, re- pal purple passionfruit production regions, lo- search is needed to revise and update C. capitata cated in the departments of Boyacá, Cundi- host information and thereby improve quarantine namarca, Tolima, and Huila (Colombia). During decision making (Aluja et al. 2004; Peña et al. each sampling event, 9-16 different purple pas- 2006; Jenkins & Goenaga 2008; Staub et al. 2008; sionfruit orchards were visited and fruit was col- De Graaf 2009; Follett et al. 2009). lected from each orchard. Fruit samples con- Purple passionfruit (Passiflora edulis f. edulis sisted of hand-picked fruit of different maturity Sims) is one of several tropical fruits that is well- levels (i.e., fruit harvested from vines) and fallen positioned in local markets and gradually becom- fruit, collected from the ground. Fruit was sam- ing popular internationally (Ocampo 2007; Wyck- pled in a random fashion, and the number of huys et al. in press). In Colombia, purple passionfruit collected from each orchard depended upon fruit is mainly grown by small-scale, resource- phenological stage of the crop. We collected a to- poor farmers on a total area of 100-400 ha. It is a tal of 405, 285, 183, and 113 hand-picked fruit profitable crop and fresh fruit is increasingly be- from Boyaca, Cundinamarca, Huila, and Tolima, ing exported to northern Europe and Canada respectively. Respective numbers of fallen fruit (Wyckhuys, unpublished data). Entry of fresh collected from each department were 345, 124, fruit into the continental U.S. is not permitted 96, and 58. currently, based upon its presumed suitability as Fruit samples were counted, weighed and a host for Anastrepha spp. and C. capitata. taken to the Horticulture Research Center CIAA Liquido et al. (1991) list C. capitata as a poten- (Chia, Colombia) in ventilated plastic containers tial pest of P. edulis, but provide no evidence of (70 × 50 × 50 cm) for further laboratory process- adult fly emergence from field-collected fruit. ing. In the laboratory, fruit samples were kept at Other reports indicate C. capitata is an occasional 22.0 ± 2.0°C, 65% RH and 12:12 L:D. Within 1 pest of Passiflora sp., without specifying the exact week following the collection, containers were crop species, botanical form or variety (Thomas et screened for presence of fruit fly puparia, and al. 2001). Yellow passionfruit (P. edulis f. fla- fruit were dissected to assess presence of te- vicarpa Degener) is reported as a possible host of phritid larvae. Larvae were subsequently trans- C. capitata in Hawaii (Akamine et al. 1954), while ferred to ventilated plastic Petri dishes with many tephritids attack certain Passiflora species moistened vermiculite. Petri dishes were checked in Brazil (Aguiar-Menezes et al. 2002). In Colom- daily for adult emergence. We recorded the num- bia, national pest survey records for C. capitata ber of tephritid larvae and C. capitata adults for maintained since 1986 have not detected this pest each sampling event and production region. in the principal production regions of purple pas- Simultaneous with field collections, McPhail sionfruit (ICA, 2009). As a note of caution, it is im- traps (baited with protein hydrolysate; Cebofrut, portant to indicate that climate change could AgroBiologicos SAFER, Medellin, Colombia) were cause altitudinal range shifts of pest tephritids deployed in orchards in each production region and may eventually bring C. capitata into those and visited bi-weekly to record the number of C. production regions in the future (Hill et al. 2011). capitata adults. A total of 6 traps were deployed Considering a lack of scientific information re- per orchard, of which 5 were placed within the or- garding purple passionfruit host status for C. cap- chard itself and a sixth trap was placed outside itata and the importance of its production as the orchard in the dominant surrounding habitat source of income for rural smallholders, we at- type. To check trap attractiveness, we recorded tempted to determine the host suitability of Co- captures of other tephritids. lombia-grown purple passionfruit for C. capitata using standard methods (Cowley et al. 1992). This Laboratory Experiments information can be used to re-evaluate the quarantine status of this fruit for market access to the Insect material was collected from coffee fruit United States. (Coffea arabica L.) in commercial orchards in Fre- donia (Antioquia, Colombia), at 1,400 m altitude, MATERIALS AND METHODS and Medellín (Antioquia), at an altitude of 1,493 m. Upon field collection, fruits were transferred to All methodologies for host status screening the ICA Entomology Laboratory in Bello (Antio- were adopted from Cowley et al. (1992), taking quia). Each fruit was dissected and any tephritid into account parameters set by RSPM No. 30 larvae were allowed to pupariate in vermiculite. (NAPPO 2008) and APPPC RSPM No. 4 (FAO Puparia of C. capitata were subsequently taken to 2005; Follett & Hennessey 2007). the Quarantine Treatment Laboratory of the Co- lombian Institute for Agriculture and Lifestock Field Collections ICA in Mosquera (Cundinamarca) for further ex- perimenting. Adults from field collected puparia Between Sep 2008 and May 2010, sampling were exposed to mango (Mangifera indica L.), a was done during 4 distinct events in the princi- preferred host of C. capitata (NAPPO 2008).

Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata 93

Adult flies were maintained within mesh cages replicates of each fruit type and maturity level, (25x25x25 cm), allowed ample access to water and simultaneous trials were conducted. In total, and fed ad libitum with torula yeast and sugar. 990 purple passionfruit and 495 mango fruits All insect developmental stages were maintained were subjected to an infestation pressure of 10.9 within climate-controlled rearing chambers at 25 C. capitata females per fruit. ± 1ºC, 65 ± 5% RH and 12:12 L:D. Second genera- Over the course of 15 d, fruits within each cage tion C. capitata adults were then used for host were replaced on a daily basis, and subsequently status trials. Laboratory experiments were car- kept within ventilated plastic containers. In a ried out between Oct 2008 and May 2009. Voucher random fashion, a subsample of 45 fruits of either specimens of study insects were kept at the ICA species or maturity degree was dissected upon re- laboratory. moval from experimental cages to assess the All fruits used in the experiment were selected number of C. capitata eggs. Remaining fruits and harvested in several purple passionfruit or- were kept at 25 ± 1ºC, 65 ± 5% RH and 12:12 L:D chards in Venecia (Cundinamarca) or mango or- and were checked daily for larval emergence, pu- chards in La Mesa (Cundinamarca). Fruit of dif- paria formation, or adult eclosion. After 15 d, all ferent maturity levels were selected based on fruits were dissected and C. capitata larvae (per commonly-used color tables for either mango or fruit) were counted and placed within vermiculite purple passionfruit (ICONTEC 1999; Pinzón et to allow pupariation. al. 2007). Prior to use in experiments, fruit was disinfected by immersion in a 0.05% sodium hy- RESULTS pochlorite solution for 10 min. Subsequently, each fruit was dried and stored in plastic containers to Field Collections use in host status trials. Fruit was used for exper- imenting within 72 h of harvest. From 2008 up to 2010, a total of 976 purple passionfruit were hand-picked and 623 fallen Oviposition Preference Assay fruit were collected. No C. capitata adults emerged from any fruit. Diptera larvae were A total of 120 C. capitata pairs, aged 14 d, were found within (immature) fruit; all of which suc- placed within a mesh cage (70 × 50 × 50 cm) (Vidal cessfully developed into lonchaeid adults. No C. et al. 2005) and allowed access to water and ad li- capitata adults were caught in McPhail traps de- bitum torula yeast and sugar. Within each cage, ployed in or near orchards in any of the produc- we placed 8 purple passionfruit of each of 3 matu- tion regions. Trap effectiveness was confirmed rity levels (i.e., maturity 0, 2, and 4; see Pinzon et through capture of Lonchaeidae (Diptera: Tephri- al. 2007). Purple passionfruit are approximately 5 toidea) at all locations. cm in diameter. After 24 h, fruit was removed from the cages and dissected to determine the to- Laboratory Experiments: Oviposition Preference Assay tal number of C. capitata eggs. Over the course of 3 d, fruit were placed within each cage and sub- The number of C. capitata eggs significantly ject to the same ovipositing C. capitata females. differed between fruit of distinct maturity de- The experiment was carried out with 3 replicates, grees (F = 18.84, df = 2, P < 0.0001). The highest thus screening 72 fruit per maturity level. The number of eggs per fruit (183.7 ± 33.8; mean ± SE) number of eggs within fruit of differing maturity was oviposited in purple passionfruit of maturity level was compared by one-way analysis of vari- level 0, while no eggs were laid in maturity levels ance (ANOVA). For all analyses, the statistical 2 and 4. package SAS was used. Host Status Trials Host Status Trials Ceratitis capitata successfully completed its Based upon results of the previous assay, fur- development on the preferred host mango, but no ther trials were conducted to determine purple adults emerged from punctured fruit of maturity passionfruit host status to C. capitata. To stimu- levels 0 and 2 (Table 1). Few C. capitata larvae late fly oviposition, fruit was punctured with developed in passionfruit, with larval weights standard dissection pins (10 pinholes 1-2 mm into ranging from 2.5 to 3.2 mg. In mango, the weight the fruit) before placing them within experiment of third instars ranged from 9.7 to 10.3 mg. Of the cages (FAO 2005; NAPPO 2008). Purple passion- 194 C. capitata that were obtained from passionfruit of maturity levels 0 and 2 were included in fruit (maturity 0), <10% successfully pupariated. trials, while mango fruit (maturity degree 2 or 3) Puparial weights of individuals developing on was used as a positive control. We placed 11 fruit passionfruit ranged from 2.3 to 3.1 mg, compared per cage (70 × 50 × 50 cm) with 120 C. capitata to C. capitata puparia from mango that weighed pairs, aged 14-19 d and provided with water and between 9.3 and 10.2 mg. Also, most C. capitata ad libitum torula yeast and sugar. There were 3 puparia that developed from passionfruit were 94 Florida Entomologist 94(1) March 2011

TABLE 1. OVIPOSITION AND SUBSEQUENT DEVELOPMENT OF C. CAPITATA ON MANGO AND PURPLE PASSIONFRUIT (PPF) OF 2 MATURITY LEVELS UNDER LABORATORY CONDITIONS. DATA REPRESENT CONSOLIDATED NUMBER OF INDI- VIDUALS WITHIN EACH C. CAPITATA DEVELOPMENT STAGE ON A TOTAL OF 990 PPF FRUIT OR 495 MANGO FRUIT.

C. capitata development stages

Tested commodity Eggs Larvae Puparia Adults

Mango 139,410* 64,990 53,854 46,920 PPF - maturity degree 0 84,410 194 18 0 PPF - maturity degree 2 84,250 0 0 0

*The total number of eggs was determined by counting the number of C. capitata eggs on 10% of (dissected) fruits, and extrap- olating this for all tested fruits. malformed. No adults eclosed from purple pas- cropping regions and equally shift current passionfruit puparia, whereas 46,920 adults sionfruit production zones to higher altitudes emerged from infested mangos. (Hill et al. 2011). At present however, natural field infestation data remain inconclusive with re- DISCUSSION spect to passionfruit host status. Forced infestation trials under laboratory con- Fruit fly host status determination lies at the ditions proved critical in delineating purple pas- basis of trade and can help connect small-scale sionfruit host status to C. capitata. Even though fruit producers in the developing world to lucra- C. capitata females oviposited in intact fruit (ma- tive export markets. To aid developing nations in turity degree 0) as in punctured fruit of different the process of assessing whether a given fruit is a maturity degrees, larval development was very host to a particular fruit fly species, well-defined poor and no adults emerged. No adult emergence protocols and experimental guidelines have been from fruit under laboratory conditions is either defined (FAO 2005; Hennessey 2007; Aluja & indicative of its character as non-host under ex- Mangan 2008; NAPPO 2008). Natural field infes- perimental conditions (NAPPO 2008) or as non- tation trials and a set of screen-house or labora- host overall (FAO 2005). Nevertheless, we need to tory experiments all help determine whether a indicate that adult development from purple pas- given fruit crop is natural host, non-host or condi- sionfruit could have been affected by dissecting tional host (e.g., Jenkins & Goenaga 2008; De infested fruit 15 d after oviposition. On less suit- Graaf 2009). These protocols have been adopted able hosts, C. capitata likely develop slow and for a wide range of fruit crops, such as mamey sa- take longer to complete larval development. How- pote (Pouteria sapota (Jacq.)), litchi (Litchi chin- ever, fruit was dissected according to its deterio- ensis Sonn.), rambutan (Nephelium lappaceum ration status (see FAO 2005; NAPPO 2008), while L.), avocado ‘Hass’ (Persea americana (Mill.) taking into account an upper C. capitata egg-lar- ‘Hass’), highbush blueberry (Vaccinium corymbo- val development time of 15 d (EPPO 2010). In sum L.), green mango (Mangifera indica L. conclusion, even though early dissection of purple ‘Tommy Atkins’ and ‘Keitt’), and others. passionfruit may have affected pupation and Although data from natural field infestation adult eclosion, the poor larval development and trials provide the most accurate assessment of lack of emergence of adults from 18 ºC capitata host status of a given fruit (NAPPO 2008), a key puparia clearly indicate the poor suitability of limitation of these trials is that one cannot con- this fruit. trol variability in fruit fly abundance. In our ex- For intact fruit, maturity level 0 was preferred, periments, no C. capitata adults were reared from while fruit of more advanced maturity were not field-collected passionfruit in the principal pro- accepted for oviposition by C. capitata. Fruit ma- duction regions of Colombia. However, McPhail turity state can greatly affect its acceptability as trapping in orchards and surrounding habitats an oviposition substrate by certain fly species also did not encounter any wild C. capitata popu- (Armstrong 2001; Willink & Villagran 2007). Cer- lations in any of these zones. Purple passionfruit tain physical stimuli determined by fruit matu- crops are located at 2016.1 ± 250.9 m (mean ± SD) rity level (e.g., color) influence C. capitata accep- above sea level (Wyckhuys et al. in press), while tance or rejection of fruit of particular maturity C. capitata has not been reported above 1,600 m levels (Prokopy et al. 1984; Suarez et al. 2007). (ICA 2009). Thus, under the current altitudinal Also, fruit maturity level can affect physical resis- and geographic distribution of C. capitata in Co- tance to oviposition and interfere with successful lombia it is very unlikely that this species affects C. capitata oviposition (Gould & Hallman 2001). purple passionfruit orchards. Climate change To circumvent such, C. capitata tend to oviposit in could eventually bring C. capitata into actual existing oviposition holes, bird pecks or crevices Rengifo et al.: Host Status of Purple Passionfruit for Ceratitis capitata 95

(Aluja & Mangan 2008). This could further ex- 390 In J. E. Peña, J. L. Sharp, and M. Wyoski [eds.], plain high degrees of oviposition in punctured Tropical Fruit Pests and Pollinators: Biology, Eco- fruits and low acceptability of intact fruit, more so nomic Importance, Natural Enemies and Control. at advanced maturity degrees at which purple CABI Publishing, Wallingford, UK. passionfruit has an exceptionally firm epicarp. AKAMINE, E. K., HAMILTON, R. A., NISHIDA, T., SHER- MAN, G. D., AND STOREY, W. B. 1954. Passion Fruit Fruit fly oviposition in hosts that are inade- Culture. University of Hawaii, Extension Circular quate for larval development is commonly ob- 345. 23 p. served (Joachim-Bravo et al. 2001). Especially for ALUJA, M., AND MANGAN, R. L. 2008. Fruit fly (Diptera: highly polyphagous species such as C. capitata, Tephritidae) host status determination: critical con- behavioral adaptations cause oviposition in a ceptual, methodological and regulatory consider- wide range of fruit crops (Aluja & Mangan 2008). ations. Annu. Rev. Entomol. 53: 473-502. Additionally, under highly artificial conditions, ALUJA, M., DIAZ-FLEISCHER, F., AND ARREDONDO, J. time-limited gravid females may accept a broad 2004. Nonhost status of commercial Persea america- range of substrates for oviposition (see Robacker na ‘Hass’ to Anastrepha ludens, Anastrepha obliqua, Anastrepha serpentina and Anastrepha striata & Fraser 2002). A high level of acceptance for ovi- (Diptera: Tephritidae) in Mexico. J. Econ. Entomol. position of intact and punctured fruit does not 97: 293-309. necessarily imply suitability of the infested fruit ARMSTRONG, J. W. 2001. Quarantine security of bananas for further larval development or adult emer- at harvest maturity against Mediterranean and Ori- gence. Increased mortality, poor larval develop- ental fruit flies (Diptera: Tephritidae) in Hawaii. J. ment and reduced puparia size or weight all are Econ. Entomol. 94: 302-314. indicative of antibiosis and biochemical defenses COWLEY, J. M., BAKER, R. T., AND HARTE, D. S. 1992. (Greany et al. 1983) that cannot be detected by Definition and determination of host status for mul- ovipositing females. Passiflora species are cyano- tivoltine fruit fly (Diptera: Tephritidae) species. J. Econ. Entomol. 85: 312-317. genic and liberate hydrogen cyanide in fruits or DE GRAAF, J. 2009. Host status of avocado (‘hass’) to leaves when under (insect) attack (Spencer & Sei- Ceratitis capitata, Ceratitis rosa and Ceratitis cosyra gler 1983). Possibly, these compounds disrupt lar- (Diptera; Tephritidae) in South Africa. J. Econ. Ento- val development in passionfruit. mol. 102: 1148-1459. As presence of low numbers of larvae in fruit is EPPO. 2010. Ceratitis capitata. Datasheet on Quaran- not indicative that it is an acceptable host (Gould tine Pests. Accessed online from www.eppo.org on & Hallman 2001; Jenkins & Goenaga 2007; Will- November 22, 2010. ink & Villagran 2007), we can conclude that Co- FAO. 2005. The Asia and Pacific Plant Protection Com- lombia-grown purple passionfruit is a non-host mission (APPPC RSPM No 4). Regional Standards for Phytosanitary Measures; Guidelines for the Con- under the experimental conditions used in these firmation of non-host Status of Fruit and Vegetables tests and may be a non-host in the field. Since C. to Tephritid Fruit Flies; Rap Publication 2005/27. capitata is currently not established in the princi- FOLLETT, P. A., ARMSTRONG, J. W., AND ZEE, F. T. 2009. pal growing areas in Colombia, it is very unlikely Host status of blueberry to invasive tephritid fruit that this pest will infest purple passionfruit un- flies in Hawaii. J. Econ. Entomol. 120: 1859-1863. der natural conditions. There may therefore be FOLLETT, P. A., AND HENNESSEY, M. K. 2007. Confi- significant potential for the establishment of pest dence limits and sample size for determining non- free areas to allow exports to the United States or host status of fruits and vegetables to tephritid fruit a systems approach based upon low C. capitata flies as a quarantine measure. J. Econ. Entomol. 100: 251-257. prevalence and poor host status. GOULD, W. P., AND HALLMAN, G. 2001. Host status of mamey sapote to Caribbean fruit fly (Diptera: Thep- hritidae). Florida Entomol. 84: 370-375. ACKNOWLEDGMENTS GREANY, P. D., STYER, S. C., DAVIS, P. L., SHAW, P. E., AND CHAMBERS, D. L. 1983. Biochemical resistance We are grateful to Catherine Varela, Maribel Hur- of citrus to fruit flies. Demonstration and elucidation tado, Jaime Abelló, Juan Camilo Rodríguez, Paola Urón, of resistance to the Caribbean fruit fly, Anastrepha Liliana Cárdenas, Nubia Esmeralda Guerrero, Rubén suspensa. Ent. Exp. Appl. 34: 40-50. Molina, José Rey, Gloria Palma, Oscar Menjura, Jorge HENNESSEY, M. K. 2007. Guidelines for the Determina- Enrique Arias, and Yaneth Bernal at the ICA Quaran- tion and Designation of Host Status of a Commodity tine Treatment Laboratory for help in fruit dissections for Fruit Flies (Tephritidae). USDA-CPHST, Orlan- and fruit fly colony maintenance. We are grateful to Glo- do, Florida. ria Marlene Vidal (USDA-APHIS) for help with data in- HILL, J. K., GRIFFITHS, H. M., AND THOMAS, C. D. 2011. terpretation and research conceptualization. This Climate Change and Evolutionary Adaptations at research was financed by the Colombian Ministry of Ag- Species’ Range Margins. Annu. Rev. Entomol. 56: riculture and Rural Development, with project grant 143-159. MADR 2008L6772-3445 to KW. ICONTEC, 1999. Frutas Frescas. Mango. Especifica- ciones. NTC 5210. Bogota - Colombia. REFERENCES CITED ICA (INSTITUTO COLOMBIANO AGROPECUARIO). 2009. Mosca del Mediterráneo (Ceratitis capitata) en Co- AGUIAR-MENEZES, E. L., MENEZES, E. B., CASSINO, P. lombia Año 2008-2009. Plan Nacional Mosca de las C., AND SOARES, M. A. 2002. Passion Fruit, pp. 361, Frutas PNMF. Boletin Epidemiologico. 96 Florida Entomologist 94(1) March 2011

JENKINS, D. A., AND GOENAGA, R. 2007. Host status of ROBACKER, D. C., AND FRASER, I. 2002. Attraction of mamey sapote, Pouteria sapota (Sapotaceae), to the Mexican fruit flies (Diptera: Tephritidae) to grape- West Indian fruit fly, Anastrepha obliqua (Diptera: fruit: enhancement by mechanical wounding of and Tephritidae) in Puerto Rico. Florida Entomol. 90: experience with grapefruit. J. Ins. Behavior 15: 399- 384-388. 413. JENKINS, D. A., AND GOENAGA, R. 2008. Host status of ROBINSON, A. S., AND HOOPER, G. 1989. Fruit Flies: litchi and rambutan to the West Indian fruit fly Their Biology, Natural Enemies and Control. World (Diptera: Tephritidae) Florida Entomol. 91: 228-231. Crop Pests, Vol. 3A. Elsevier Amsterdam. JOACHIM-BRAVO, I. S., FERNANDES, O. A., DE BORTOLI, SPENCER, K. C., AND SEIGLER, D. S. 1983. Cyanogenis S. A., AND ZUCOLOTO, F. S. 2001. Oviposition behav- of Passiflora edulis. J. Agric. Food Chem. 31: 794- ior of Ceratitis capitata Wiedemann (Diptera: Te- 796. phritidae): association between oviposition prefer- STAUB, C. G., DE LIMA, F., AND MAJER, J. D. 2008. De- ence and larval performance in individual females. termination of host status of citrus fruit against the Neotropical Entomol. 30: 559-564. Mediterranean fruit fly, Ceratitis capitata (Wiede- LIQUIDO, N. J., SHINODA, L. A., AND CUNNINGHAM, R. T. mann) (Diptera: Tephritidae). Australian J. Ento- 1991. Host Plants of the Mediterranean Fruit Fly mol. 47: 184-187. (Diptera: Tephritidae): An Annotated World Review. SUAREZ, L., MOLINA, A., MURUA, F., ACOSTA, J. C., MOY- Misc. Publ. No. 77, Entomol. Soc. America. ANO, B., AND ESCOBAR, J. 2007. Evaluación de colores NAPPO. 2008. Guidelines for the Determination and para la oviposición de Ceratitis capitata (Diptera, Te- Designation of Host Status of Fruits and Vegetables phritidae) en Argentina. Revista Peruana de Bi- for Fruit Flies (Diptera: Tephritidae). RSPM No. 30. ología 14: 291-293. OCAMPO, J. A. 2007. Study of the Diversity of the Genus THOMAS, M. C., HEPPNER, J. B., WOODRUFF, R. E., Passiflora L. (Passifloraceae) and its Distribution in WEEMS, H. V., STECK, G. J., AND FASULO, T. R. 2001. Colombia. Ph.D Dissertation, Centre International Mediterranean fruit fly—Ceratitis capitata. http://ent- d’Études Supérieures en Sciences Agronomiques - nemdept.ufl.edu/creatures/fruit/mediterranean_fruit_ Montpellier Sup. Agro. 268 pp. fly.htm, revised on July 24, 2010. PEÑA, J. E., GOULD, W. P., HENNESSEY, M. K., HALL- VIDAL, C. G., ABELLO, S. J., AND AREVALO, P. E, 2005. MAN, G. J., AND CRANE, J. H. 2006. Laboratory and Multiplicación de la mosca del Mediterráneo Cerati- field infestation studies on immature green ‘Tommy tis capitata W. En condiciones controladas de labora- Atkins’ and ‘Keitt’ mangoes to determine host status torio. Revista ICA 32: 25-30. to the Caribbean fruit fly (Diptera: Tephritidae). WILLINK, E., AND VILLAGRAN, M. E. 2007. Evaluación Proc. Florida State Hort. Soc. 119: 16-20. del riesgo cuarentenario de introducción de Ceratitis PINZON, I. M., FISCHER, G., AND CORREDOR, G. 2007. De- capitata en la palta Hass de Argentina. Actas VI terminación de los estados de madurez del fruto de Congreso Mundial del Aguacate. Viña Del Mar, gulupa (Passiflora edulis Sims). Agronomía Colombi- Chile. 12-16 Nov. 2007. ana 25: 83-95. WYCKHUYS, K. A. G., LOPEZ ACOSTA, F., ROJAS, M., AND PROKOPY, R. J., MCDONALD, P. T., AND WONG, T. T. Y. OCAMPO, J. D. 2011. Do farm surroundings and local 1984. Inter-population variation among Ceratitis infestation pressure relate to pest management in capitata flies in host acceptance pattern. Ent. Exp. cultivated Passiflora species in Colombia? Interna- Appl. 35: 65-69. tional J. Pest Management, in press.

Salas et al.: Population Dynamics of Greenidea on Guava and Ficus 97

POPULATION DYNAMICS OF TWO SPECIES OF GREENIDEA (HEMIPTERA: APHIDIDAE) AND THEIR NATURAL ENEMIES ON PSIDIUM GUAJAVA (MYRTACEAE) AND FICUS BENJAMINA (MORACEAE) IN CENTRAL MEXICO

MANUEL DARÍO SALAS-ARAIZA1, ROBERT W. JONES2, ALEJANDRO PEÑA-VELASCO1, OSCAR ALEJANDRO MARTÍNEZ-JAIME1 AND EDUARDO SALAZAR-SOLÍS1 Departamento de Agronomía, División Ciencias de la Vida, Universidad de Guanajuato, Mexico [email protected]

Facultad de Ciencias Naturales, Universidad Autónoma de Querétaro, Juriquilla, Queretaro, Mexico

ABSTRACT

Greenidea psiidi van der Goot and Greenidea ficicola Takahashi (Hemiptera: Aphid- idae), are Asiatic species that feed on guava, Psidium guajava and Ficus spp.; both of these aphids were reported as exotic pests in Florida in 2002 and in Mexico in 2003. The present study characterized the population dynamics of both aphid species and their natural enemies on guava and ornamental figs in the Bajio region of Central Mexico. This report represents the first record of G. psiidi on Ficus sp. in Mexico and the first report of the presence of both species in the state of Guanajuato. Greenidea psiidi and G. ficicola were detected on guava in Mar 2007 and on fig trees during the same year in Apr near Irapauto, Guanajuato. Populations of both alate and apterous forms of G. psiidi in Apr were greater on guava than on fig trees (W = 119.0; P = 0.0122), which coincided with new vegetative growth after leaf loss in winter on guava. In Apr populations of apterous forms of both species were significantly greater than winged forms on both guava and figs. No correlation was found between temperature changes and population densities of aphids. The indigenous predators, Chrysoperla comanche Banks, Chrysoperla exotera (Navás) and Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), fed readily on the aphids and were found on both guava and fig trees, although densities of all 3 species were in greater numbers on Ficus. The combined population densities of the 3 predators had a positive correlation with that of G. ficicola (r = 0.74), with a best fit found with a quadratic model of simple regression: y (densities of Chrysoperla spp.) = 1.2479x2 - 4.3073x + 9.6493, and R2 = 0.703. Nine species of coccinelid beetles (Coleoptera: Coccinel- lidae) were identified, the most common being of the genus Scymnus. Results suggest that non-deciduous ornamental fig trees may serve as reservoirs of beneficial insects for deciduous guava trees. Results from the present study provide basic biological data to aid in management of these 2 exotic species of Greenidea on guava in central Mexico.

Key Words: guava, aphids, Greenidea, Psidium guajava, Ficus benjamina

RESUMEN

Greenidea psiidi van der Goot y Greenidea ficicola Takahashi (Hemiptera: Aphididae), son de origen asiático, en 2002 fueron reportados como plaga exótica en Florida y en México en 2003. En el presente estudio, se describe la dinámica poblacional de estas dos especies de áfi- dos y sus enemigos naturales en guayaba y Ficus en la región de El Bajío en el centro de México. Es el primer reporte de G. psiidi en Ficus spp. en México y el primer reporte de ambas especies en el estado de Guanajuato. Greenidea psiidi y G. ficicola fueron observadas en guayaba desde marzo y en Ficus desde mediados de abril, en el área de estudio. Las poblaciones de áfidos alados y ápteros de G. psiidi fueron más altas en abril en guayaba que en Ficus (W = 119.0; P = 0.0122), esto coincide con los nuevos brotes después de que el árbol de guayaba pierde las hojas. Las poblaciones de las formas ápteras de ambas especies fueron significativamente mayores que las formas aladas tanto en guayaba como en Ficus. No se en- contró correlación entre la temperatura y las poblaciones de áfidos. Los enemigos naturales Chrysoperla comanche Banks, Chrysoperla exotera (Návas) y Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), se alimentan de estos áfidos y fueron encontrados tanto en guayaba como en Ficus, aunque la densidad de las tres especies fueron más altas en Ficus. Las poblaciones combinadas de las tres especies de depredadores presentaron una correlación positiva con la especie G. ficicola (r = 0.74), lo cual se explica con la ecuación cuadrática de regresión simple: y (densidades de Chrysoperla spp.) = 1.2479x2 - 4.3073x + 9.6493, donde el coeficiente R2 = 0.703. Se identificaron nueve especies de Coccinellidae, siendo el más común el género Scymnus. Estos resultados sugieren que los árboles de Ficus cuyo follaje es pe- renne, puede servir como refugio de insectos benéficos para los árboles de guayaba, cuyo há-

98 Florida Entomologist 94(1) March 2011

bito es caducifolio. Los resultados de esta investigación aportarán conocimientos sobre la biología de estas especies exóticas de áfidos lo que ayudará en el manejo de la plaga en huer- tas de guayaba en la región central de México.

Translation provided by the authors.

Mexico contributes 25% of the world produc- tivated hosts is crucial in the development of tion of guava (Psidium guajava L.; Myrtaceae), of management strategies. The objective of the which the states of Michoacan, Aguascalientes, present study was to determine the population and Zacatecas of the central Altiplano region are dynamics of Greenidea psidii, G. ficicola and their the principal producers (González-Gaona et al. natural enemies on Psidium guajava and Ficus 2002). In 2008, the exotic aphid pests, Greenidea benjamina in the state of Guanajuato, Mexico. psidii van der Goot and Greenidea ficicola Taka- The data generated from this study will help eval- hashi were observed on guava and ornamental fig uate the importance of these aphids within the trees, Ficus benjamina (L.), in the central Altipl- pest complex attacking guava in the Bajío Region, ano state of Guanajuato (Salas-Araiza, unpub- as well as aid in the development of integrated lished data). There is concern among producers pest management strategies. and agricultural researchers of central Mexico that these new pests may pose a threat to guava MATERIALS AND METHODS production in Mexico. The genus Greenidea Schouteden belongs to The study site was conducted at the experi- the subfamily Greenideinae within the Aphididae mental field station of the Division of Life Sci- and includes approximately 45 species (Pérez ences of the University of Guanajuato at the Ex- Hidalgo et al. 2009). The natural distribution of Hacienda El Copal (101°01’01”N, 20°49’49”O) at the genus is Asiatic and species are found to favor 1,750 masl in the municipality of Irapuato, Gua- young foliage of plants of the families, Fagaceae, najuato, Mexico. The region has a mean annual Betulaceae, Juglandaceae, Myrtaceae, Rosaceae, precipitation of 750 mm, a mean temperature of and Rubiaceae (Blackman & Eastop 1994). The 19°C and mean relative humidity of 56% (INEGI presence of Greenidea in the New World may be 2009). Two orchards, 1 guava (P. guajava), and the result of importation of infested ornamental the other ornamental figs, (F. benjamina) were fig trees which are widely commercialized chosen for the study sites. throughout the world, and they have been impli- All trees were 10 years old and the 2 orchards cated as vehicles for the introduction of exotic were separated by approximately 1 km. Weekly pests (O’Donnell & Parrella 2005). samples were made of aphids (alate and apterous Greenidea psidii was first discovered in the forms) and predators (adults of Coccinellidae and New World in1916 on guava and its relative, Psid- larvae of Chrysopidae) from 10 trees of each host ium cattleianum Sabine in Brazil (Noemberg- species. For each tree, samples consisted of 20 Lazzari et al. 2006). Halbert (2004) reported that beats of a 1-m wooden rod on the branches of the G. ficicola was collected in Florida in 2002, and to- trees, from which insects fell onto a 1-m2 beating gether with G. psidii were found to feed on guava sheet. All insects on the sheet were collected and and ornamental fig trees (Ficus benjamina (L.)). placed in a labeled vial with 70% alcohol. The sam- Pérez Hidalgo et al. 2009 reported the presence of ple period was from 23 Mar to 22 Jun 2007 which G. psidii in Costa Rica in 2009. The first reports of corresponded to the early spring growth period of Greenidea in Mexico come from Peña-Martínez et leaves of guava and to the reproductive activity of al. (2003), who recorded G. psidii feeding on aphids. Samples were not taken on subsequent guava in the states of Hidalgo, Morelos, Guerrero, dates because aphids had ceased reproductive and the Federal District, and G. ficicola feeding growth and individuals were virtually undetect- on ornamental fig trees in the state of Guana- able. All material from each tree was preserved in juato, Mexico. Trejo-Loyo et al. (2004) reported G. individual vials containing 70% alcohol, with a cor- ficicola occurs on Myrtaceae in Cuernavaca, Mo- responding label and brought to the laboratory. relos. Although further data is lacking, it is prob- The collected specimens were identified in the able that both G. psidii and G. ficicola are widely Entomology Laboratory of University of Guana- distributed in Mexico. juato, with a compound and stereo microscope. The biology of species of Greenidea on guava Aphids were mounted for species determination and other host plants is poorly known (Halbert following techniques given by Peña-Martínez 2004; Sousa-Silva et al. 2005; Noemberg-Lazzari (1995). The keys of Blackman & Eastop (1994, et al. 2006). Northfield et al. (2008) noted that 2000) were used for species identification of many insect pests feed on alternative host plants aphids. For the neuropteran and coccinellid pred- and that the understanding of the relationships of ators, the keys of López-Arroyo et al. (2008) and pest populations on wild hosts with those on cul- Gordon (1985) were used, respectively. Meteoro-

Salas et al.: Population Dynamics of Greenidea on Guava and Ficus 99 logical data was obtained from a weather station the abundance of G. psiidi was notably greater maintained at the study site located 200 m from during Mar and early Apr. This appearance and the orchards. All identified specimens were depos- population growth coincided with the emergence ited in the Entomological Collection “Leopoldo Ti- of new shoots and leaves on guava, the timing of noco Corona” of the University of Guanajuato. which corresponds to that described by Damián- Data were analyzed with the statistical soft- Nava et al. (2004). Colonizing alate aphids are of- ware program SAS (SAS 1995). Abundance mea- ten attracted to specific volatiles of host plants sures were calculated for G. psiidi and G. ficicola (Chapman et al. 1981; Nottingham et al. 1991; and the various predator species on guava and Powell & Hardie 2001). Because G. psiidi and G. figs. Due to non-linearity of data determined by ficicola are found initially on young shoots and the Shapiro-Wilks test, mean comparisons of leaves, it is probable that initial alate colonizers aphid and predator numbers between hosts and are attracted to volatiles associated with new among samples dates were made with Mann- growth of guava plants. Whitney non-parametric procedures. In addition, It important to note that the 2 tree species correlation analyses were conducted between studied have marked differences in leafing pat- population numbers of aphids with those of the terns and the appearance of new growth. Guava various predators and also with climatic vari- is deciduous, with complete leaf loss occurring ables (temperature and precipitation). generally in Nov with new growth beginning in Mar in the study area. This is in contrast to F. RESULTS AND DISCUSSION benjamina which is a non-deciduous tree that produces new growth apparently in response to Species of Greenidea in Guava and Figs environmental factors. However, both G. psiidi and G. ficicola appeared first on guava, and then Both G. psidii and G. ficicola were present on on F. benjamina (Fig. 1, A, B, C), although foliage guava and the ornamental fig, F. benjamina (Figs. was available on the latter host throughout win- 1, A, B, C). This is the first report of G. psiidi on ter months. These data suggest that G. psiidi and the widely planted F. benjamina from Mexico, and G. ficicola first colonize and establish on guava for this species for the state of Guanajuato. Al- and then later move to F. benjamina. This behav- though the habitual host for G. ficicola is Ficus ior was expected for G. psiidi for which guava is spp. (Noemberg-Lazzari et al. 2006), Halbert considered a habitual host, but not for G. ficicola (2004) reports that this aphid species also occurs which Ficus is considered the habitual host. Fur- on guava, as confirmed in the present study. Al- ther study is needed to establish the initial colo- though the apparent habitual hosts of G. psidii is nization behavior of these 2 species and whether guava, and that of G. ficicola is Ficus spp., it is un- the behaviors are the result of greater attractive- clear how populations on the habitual and other ness of guava in comparison to F. benjamina and/ infrequent host plants interact and which of the or the leaf quality of F. benjamina is inadequate infrequent hosts can maintain viable, reproduc- until late Apr. tive populations in the absence of the habitual The peak abundance of both species (alate and host plants. apterous forms) occurred in mid and late Apr (7.5 Both species of Greenidea were aggregated on and 6.0 aphids/sample, for G. psiidi and G. fici- new shoots and leaves of their hosts, a feeding pref- cola, respectively) (Fig. 1, A, B). The densities of erence previously reported by Pérez Hidalgo et al. G. psiidi were greater on guava than on F. ben- (2009). On guava, G. psiidi and G. ficicola were jamina during the first 5 sample periods, and sig- present on new leaf buds and on either side of young nificantly so at peak densities during the third leaves, whereas on F. benjamina they were found week of Apr (w = 119.0; P = 0.0122; Fig. 1, A). principally on the underside of young leaves. The By mid May, densities of G. psiidi and G. fici- preference by aphids for new plant growth is a com- cola were barely detectable, and remained at very mon behavior in aphids. Gould et al. (2007) state low densities through late Jun. These low densi- that certain stages of aphids have preferences for ties are presumably the result of the maturation specific tissues of host plants. For example, Chaito- of leaves and the deterioration of the physical and phorus populicola Thomas, preferably feeds on new nutritional requirements for these species (Fig. 2, growth with diverse and high levels of amino acids. A). These authors also note that high sugar levels and The appearance and abundance of alate forms low amino acid concentration in leaves increases in relation to apterous forms of G. psiidi and G. the production of winged individuals. ficicola followed patterns expected from observations reported by Noemberg-Lazzari et al. (2006). Population Dynamics of Greenidea psidii and Greenidea Alates were found in very low numbers initially ficicola on both guava and F. benjamina and differences between the 2 forms were not significantly differ- Both G. psiidi and G. ficicola were first re- ent until the third week of Apr (Fig. 2, B, C). At corded on guava trees in late Mar 2007, although that time, the abundance of apterous forms in- 100 Florida Entomologist 94(1) March 2011 Mean number aphids/sample

Fig. 1. Population dynamics of apterous and alate forms of Greenidea on 14 samples dates in Irapuato, Guana- juato, México. Means with the same letter not signiﬁcantly different based on Mann-Whitney non parametric test (A: 04/20/07, w = 119.0, P =0.0122). Salas et al.: Population Dynamics of Greenidea on Guava and Ficus 101 Mean number aphids/sample

Fig. 2. Population dynamics of apterous and alate forms of Greenidea on 14 samples dates in Irapuato, Guana- juato, Mexico. 2007. Means with same letter not significantly different based on Mann-Whitney non parametric test (B: 04/13/07, w = 138.5, P = 0.0179; 04/20/07, w = 100.5, P = 0.002; 04/27/07, w = 35.0, P = < 0.001; 05/04/07, w = 88.0, P = < 0.0003; 05/11/07, w = 118.0, P = 0.0037; C: 04/20/07, w =127.5, P = 0.016; 04/27/07, w = 17, P = < 0.0001). 102 Florida Entomologist 94(1) March 2011 creased considerably (16-18 fold) for both species. primarily in Jun, and thus well after populations Significant differences between the abundance of had already declined in late May. It could be ar- alate and apterous forms of G. psiidi on guava gued that the life cycle of G. psiidi and G. ficicola and F. benjamina were found from mid Apr is adapted to avoid the unfavorable effects of the through mid May (Fig. 2, B). Alate forms were seasonal rains by completing their life cycle dur- also most abundant during this period, and con- ing the dry season. However, Noemberg-Lazzari tinued to be found into Jun. It is assumed that et al. (2006) reported that in Brazil both species many of these alate individuals dispersed from are present year round (G. psiidi on guava and G. guava and F. benjamina when leaf and shoot ficicola on Ficus sp.), suggesting that the life cycle quality declined after Apr. No correlation was of the 2 insects is facultative and probably depen- found between the mean ambient temperature dant more on host quality than on specific sea- nor mean precipitation with the population sonal variations in climate. changes of G. psiidi and G. ficicola on guava and F. benjamina (r = 0.15 for temperature; r = -0.21 Natural Enemies for precipitation for G. psidii; r =0.16 for temperature; and r = -0.18 for precipitation for G. fici- Chrysopidae. Three species of Chrysopidae cola), This was expected given that there was lit- (Neuroptera) were identified on guava and F. ben- tle change in either temperature or precipitation jamina: (1) Chrysoperla comanche (Banks), (2) during the sample period (Fig. 3). Even with Chrysoperla exotera (Návas), and (3) Chrysoperla greater temperature variation, however, little carnea (Stephens). The most abundant species correlation would be expected given that temper- was C. comanche and the present work represents ature is often not a principal factor in aphid pop- the first report of this species for the state of Gua- ulation growth. For example, Desneux et al. najuato. This chrysopid is cited by Ramírez-Del- (2006) reported that temperature was not posi- gado et al. (2006) as the predominant species in tively correlated with population densities of pecans and grapes in the states of Coahuila and Aphis glycines Matsumura on soybeans, and Durango, suggesting that it is one of the most im- some species, such as Schizaphis graminum portant aphid predators of the central highlands (Rondan) on wheat produce more nymphs at of northern Mexico. lower temperatures (Pendleton et al. 2009). Dif- The combined populations of the 3 species of ferences in precipitation also were probably not Chrysoperla were present from 20 Apr 2007 to 15 an important factor in the changes in populations Jun 2007 in guava and ficus. Chrysoperla was that were observed because the only noteworthy first recorded on F. benjamina on 20 Apr of 2007 change in precipitation occurred between the last (Fig. 4, A). In both guava and F. benjamina, an in- two samples. Although precipitation has been ob- crease of individuals of Chrysoperla spp. coin- served to physically dislodge aphids from plants cided with an apparent increase in the popula- and increase infection by pathogens (Nielson & tions of the apterous forms for both aphid species Barnes 1961), in our study precipitation occurred on 20 Apr 2007 (Fig. 1, A, B, C). Populations of the

Fig. 3. Mean temperature (°C) and cumulative precipitation (mm) at El Copal, Irapuato, Guanajuato, México, 2007. Salas et al.: Population Dynamics of Greenidea on Guava and Ficus 103 Mean number predators/sample

Fig. 4. Population dynamics of the combined totals of 3 species of Chrysoperla and Scymnus spp on 14 sample dates in Irapuato, Guanajuato, Mexico. 2007. Means with the same letter are not signiﬁcantly different based on the Mann-Whitney non parametric test (B: 05/11/07, w =77.5, P = 0.016; 05/25/07, w = 92.0, P = < 0.001; 06/01/07, w = 95.0, P = < 0.001; 06/15/07, w = 83.0, P = < 0.006; 06/22/07, w = 83.0, P = < 0.006).

combined densities of Chrysoperla spp. peaked sities of Chrysoperla spp.) = 1.2479x2 - 4.3073x + first on F. benjamina on 27 Apr 2007 (0.32 individ- 9.6493, grade 2 polynomial with R2 = 0.703 uals/sample) and later on guava on 11 May 2007 (Fig. 5). This suggests that there was a numerical (0.15 individuals/sample). Densities of these response in the populations of Chrysoperla spe- predators gradually decreased from mid-May to cies in conjunction with density changes in G. fici- mid Jun. cola. There was no apparent correlation found be- Combined populations of the 3 species of tween populations of G. psiidi with Chrysoperla Chrysoperla were positively correlated with pop- on either guava or F. benjamina. ulations of G. ficicola (r = 0.74) on guava and F. Coccinellidae. Species of Coccinellidae (Co- benjamina and the relationship defined by y (den- leoptera) on guava and F. benjamina were: Hippo- 104 Florida Entomologist 94(1) March 2011

ural enemies of the pest aphids in the region, than as an alternative host plant from which Greenidea colonizes guava. The movement of predators from one host to another is well documented, in particular for aphidophagous generalists such as chrysopids and coccinellids (Sloggett et al. 2008). The following study lays the groundwork for the management of Greenidea psidii and G. fisi- cola in guava in the region. Indigenous predators were found to readily feed on G. psiidi and G. ficicola and are apparently an important component in reducing densities of the pests. Further work is needed to study in detail the colonization behavior of G. psiidi and G. ficicola in relation to guava and other hosts, and to investigate methods to op- timize the action of native predators on these aphids. Fig. 5. Correlation and simple regression model between number of individuals of Greenidea ficicola and number of individuals of 3 species of Chrysoperla on 14 REFERENCED CITED sample dates in Irapuato, Guanajuato, México, 2007. BLACKMAN, R. L., AND EASTOP, V. F.1994. Aphids on the World’s Trees. An Identification and Information Guide. CAB International, Oxon. 8+988 pp and 16 damia convergens Guérin-Méneville, Harmonia plates axyridis Pallas, Zoglobra spp., Coccinella spp., BLACKMAN, R. L., AND EASTOP, V. F. 2000. Aphids on the Olla v-nigrum (Mulsant), Cycloneda sanguinea World’s Crops: An Identification Guide. John Wiley L., Stethorus spp., Hyperaspis quadrioculata & Sons. New York. 466 pp. (Motschulsky), and Scymnus spp. On F. benjam- CHAPMAN, R. F., BERNAYS, E. A., AND SIMPSON, S. J. ina, Scymnus spp. were the most abundant with 1981. Attraction and repulsion of the aphids Cavar- iella aegopodii, by plant odors. J. Chem. Ecol. 7: 881- 167 total individuals found compared with 126 889. captured in guava. Individuals of Scymnus spp. DAMIÁN-NAVA, A., GONZÁLEZ-HERNÁNDEZ, V. A., first appeared in F. benjamina on 20 Apr 2007 and SÁNCHEZ-GARCÍA, P., PEÑA-VALDIVIA, C. B., LIVERA- began increasing by 11 May 2007, reaching a MUÑOZ, M., AND BRITO-GUADARRAMA, T. 2004. Crec- maximum abundance on 1 Jun 2007 of approxi- imiento y fenología del guayabo (Psidium guajava mately 6 individuals/sample and maintained den- L.) cv. “Media China” en Iguala, Guerrero. Rev. Fito- sities of 2.5-4 individuals/sample until the end of tec. Mexicana 27(4): 349-358. the sample period. The overall population densi- DESNEUX. N., O’NEIL. R. J., AND YOO, H. J. S. 2006. Sup- ties of Scymnus spp. was higher on F. benjamina pression of population growth of the soybean aphid, Aphis glycines Matsumura, by predators: The identi- than on guava (11.9 vs 9 individuals/sample, re- fication of key predators and the effects of prey dis- spectively), and on 5 of the 10 sample dates, and persion, predators abundance, and temperature. En- these predators had significantly higher densities viron. Entomol. 35(5): 1342-1349. on F. benjamina than on guava (Fig. 4, B). Results GORDON, R. D. 1985. The Coccinellidae (Coleoptera) of suggest that other prey present on F. benjamina America North of Mexico. J. New York Entomol. Soc. were more important in maintaining Scymnus 93(1): 1-912 spp. than the Greenidea species because these GOULD, G. G., JONES, C. G., RIFLEMAN, P., PÉREZ, A., predators appeared after major populations AND COLEMAN, J. S. 2007. Variation in eastern cot- peaks of Greenidea, and increased after mid-May tonwood (Populus deltoides Bartr.) phloem sap content caused by leaf development may affect feeding in the virtual absence of these aphids. site selection behavior of the aphid, Chaitophorus Aphid mummies were not observed in the populicola Thomas (Homoptera: Aphididae). Envi- present study and there are no reports of parasit- ron. Entomol. 36(5): 1212-1225. ism of Greenidea suggesting that predators are HALBERT, S. E. 2004. The genus Greenidea (Rhynchota: the principal insect biological control agents of Aphididae) in the United States. Florida Entomol. Greenideae in the region. Although F. benjamina 87(2): 159-163. was a host plant of Greenidea, both aphid species INEGI. 2009. Instituto Nacional de Estadística were found first on guava, and then reached Geografía e Informática. Información Geográfica. higher densities on guava than on F. benjamina. Mapa de Climas. (3 January 2009) (http://mapaserv- er.inegi.org.mx/geografía/español/ estados/gto/ However, following aphid colonization, chrysopid clim.cfm?c=444&e=07) and coccinelid predators developed greater densi- LÓPEZ-ARROYO, J. I., DE LEÓN-HERNÁNDEZ, T., ties on F. benjamina. These data suggest that F. RAMÍREZ-DELGADO, M., AND LOERA-GALLARDO, J. benjamina is more important as a refuge for nat- 2008. Especies de Chrysoperla (Neuroptera: Salas et al.: Population Dynamics of Greenidea on Guava and Ficus 105

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106 Florida Entomologist 94(1) March 2011

TRANSMISSION OF THE MYCOPATHOGEN, HIRSUTELLA SPP., TO NYMPHS AND ADULTS OF THE GLASSY-WINGED SHARPSHOOTER, HOMALODISCA VITRIPENNIS (=COAGULATA), IN THE GREENHOUSE

VERENA-ULRIKE LIETZE1, RUSSELL F. MIZELL III2 AND DRION G. BOUCIAS1 1Entomology and Nematology Department, University of Florida, Gainesville, FL 32611

2North Florida Research and Education Center, University of Florida, Quincy, FL 32351

The mycopathogen Hirsutella spp. produces on water agar to record mycosis. After 3 weeks, regular epizootics in Florida populations of the each plant was examined for mycosed cadavers, glassy-winged sharpshooter (GWSS), Homalo- and surviving GWSS were subjected to disca vitripennis, a polyphagous, xylem-feeding hemolymph examination. Statistical analyses cicadellid native to the southeastern U.S. and were conducted using the Proc Genmod procedure northeastern Mexico (Boucias et al. 2007). The and Ls means statement of the Statistical Analy- GWSS transmits a lethal phytopathogenic bacte- sis System (SAS) for Windows to compare mortal- rium, Xylella fastidiosa, and thereby threatens ity or infection responses by logistic regression the production of economically important plants (Neter et al. 1990; SAS 2004). (Redak et al. 2004). Recent introduction of GWSS For topical application of Hirsutella spores, into southern California, French Polynesia, Tahiti healthy adults were treated in three replicate as- and Hawaii has stimulated interest in identifying says by touching their ventral surface to either potential biocontrol agents against invasive pop- sporulating in vitro colonies of strain 3A (main- ulations (Boucias et al. 2007; Hunnicutt et al. tained at the UF Entomology/Insect Pathology 2008). During a field survey in Quincy, FL, a new lab, Gainesville, FL) (total n = 46 adults), to species of Hirsutella, H. homalodiscae nom. prov., GWSS cadavers displaying spores of Hirsutella (n was detected in ~50% of mycosed sharpshooters = 33), or to a nutrient agar plate (control, n = 35). (Boucias et al. 2007). Virtually nothing is known Mortality in control, in vitro, and cadaver treat- about the transmission of Hirsutella within ments was similar with 56 ± 11%, 74 ± 12%, and GWSS populations. Our field observations have 60 ± 8%, respectively (χ2= 2.48, P = 0.1155). Con- shown that Hirsutella-infected sharpshooters at- tact with agar, in vitro colonies or cadavers pro- tach via rhizoids to the plant bark in the summer duced 0%, 13 ± 6% and 42 ± 5% infection, respec- and often remain attached throughout the winter. tively, and only the latter treatment produced my- Placed in a moist environment, overwintered ca- cosis (28 ± 6%). Infection transmitted from cadav- davers produce conidiospores suggesting they ers was significantly higher than that serve as microhabitats protecting Hirsutella and transmitted from in vitro colonies (χ2 = 8.02, P = facilitating transmission to the new generation of 0.0046). GWSS. This study examined potential transmis- To examine whether co-existence of healthy sion routes of Hirsutella under greenhouse condi- and mycosed GWSS on a plant would result in tions. Experimental treatments included topical disease transmission, either overwintered cadav- spore application and choice-exposure to sporu- ers collected in Jan (stored at -80°C) or new ca- lating cadavers on host plants. davers collected in Jul/Aug were pinned to plants For insect rearing, field-collected, healthy (10 per plant) and groups of healthy nymphs or adults without hyphal bodies (HBs) in their adults were maintained on each plant for up to 3 hemolymph were identified by non-destructive weeks. Controls were conducted on plants with- antennal bleeding (Breaux 2005) and maintained out cadavers. The majority of pinned cadavers on caged, potted plants (soybean, Glycine max, displayed sporulating Hirsutella mycelium cotton, Gossypium hirsutum, and cowpea, Vigna within one week. New cadavers developed an un- unguiculata) in a greenhouse (temperature 26- usually thick, white mycelium overgrowing the 30°C, 14:10 h light:dark photoperiod). Leaves entire insect (Fig. 1A). Disease transmission was with egg masses were removed and hatching neo- observed to varying degrees (Table 1). Dead ex- nates were transferred to caged lemon basil (Oci- posed nymphs and adults (Fig. 1B and C), at- mum basilicum). Soil was watered to saturation tached to the plant and displaying Hirsutella-in- once daily. In each bioassay replicate, individual duced mycosis, were seen as early as 7 and 12 d cotton or basil plants housing groups of 10-20 after exposure, respectively. Hemolymph-borne adults or nymphs, respectively, were covered with HBs were detected as early as 8 d after exposure. clear, gauze-covered acryl cylinders (15 cm diam- New cadavers were more efficient in disease eter × 45 cm high). Mortality and infection were transmission when compared with overwintered recorded daily by removing dead individuals from cadavers (Table 1). When nymphs were exposed the soil surface and examining their hemolymph to new cadavers, no survivors were found and all for HB propagation. Cadavers were maintained dead insects were overgrown with thick mycelium

Scientiﬁc Notes 107

Fig. 1. Cadaver exposure experiments with Homalodisca vitripennis. (A) Part of a lemon basil plant spiked with new cadavers 3 weeks after introduction of nymphs. Note the white, thick mycelium overgrowing the pinned cadaver (pc) in the center and the introduced, mycosed nymphs (ic). (B and C) Mycosed nymph and adult, respectively, displaying thin Hirsutella mycelium (m) 3 weeks after release on a plant spiked with overwintered cadavers.

(Fig. 1A). Exposure of nymphs to overwintered ca- Potentially, Hirsutella is one of the major mor- davers induced 13 ± 8% mortality and 8 ± 5% in- tality factors in endemic GWSS populations (Bou- fection producing a light and flat mycelium (Fig. cias et al. 2007). Our results clearly demonstrated 1B). Adult mortality after exposure to new and that disease transmission from mycosed cadavers overwintered cadavers was 97 ± 2% and 76 ± 8%, to healthy conspecifics efficiently occurs in undis- respectively, and significantly higher than in the turbed, small host populations. Identifying at- controls (47 ± 14%) (Table 1). Infection of adults tractive cues that would cause foraging sharp- induced by new cadavers (60 ± 9%) was signifi- shooters to contact infectious spores will be the cantly higher than that induced by overwintered next step for developing this pathogen as micro- cadavers (13 ± 9%). bial control agent against H. vitripennis.

TABLE 1. MEAN (±SE) PERCENT MORTALITY AND INFECTION OF HOMALODISCA VITRIPENNIS 3 WEEKS AFTER INTRO- DUCTION TO CAGED PLANTS HARBORING HIRSUTELLA-MYCOSED CADAVERS.

Life stage exposed Cadavers Mortalitya,b Infectiona Induced mycosisc

Nymphs None (control) 0 ± 0 a (139) 0 ± 0 a No Overwintered 13 ± 8 b (68) 8 ± 5 b Yes (2) New 100 ± 0 f (53) 100 ± 0 d Yes (53)

Adults None (control) 47 ± 14 c (103) 0 ± 0 a No Overwintered 78 ± 6 d (61) 13 ± 5 b Yes (1) New 97 ± 2 e (61) 60 ± 9 c Yes (30)

aNumbers followed by different letters indicate significant differences (SAS proc genmod and lsmeans statement, P < 0.05). bNumbers in parentheses indicate the total number of GWSS exposed in ≥4 replicate assays. cNumbers in parentheses indicate the number of GWSS displaying mycosis within the 3-week observation period.

108 Florida Entomologist 94(1) March 2011

SUMMARY native range. University of Florida (Master’s thesis), Gainesville, 108 pp. For the first time, transmission of Hirsutella HUNNICUTT, L. E., MOZORUK, J., HUNTER, W. B., from mycosed, sporulating H. vitripennis to healthy CROSSLIN, J. M., CAVE, R. D., AND POWELL, C. A. conspecifics was demonstrated. In addition, meth- 2008. Prevalence and natural host range of Homalo- ods were established to amplify infectious material disca coagulata virus-1 (HoCV-1). Arch. Virol. 153: in vivo for potential inoculative release. 61-67. NETER, J. WASSERMAN, W., AND KUTNER, M. H. 1990. Applied linear statistical models: regression, analy- REFERENCES CITED sis of variance, and experimental designs. Richard Irwin Inc., Illinois. BOUCIAS, D. G., SCHARF, F. W., BREAUX, S. E., PURCELL, REDAK, R. A., PURCELL, A. H., LOPES, J. R. S., BLUA, M. D. H., AND MIZELL, R. E. 2007. Studies on the fungi as- J., MIZELL, R. F., III, AND ANDERSEN, P. C. 2004. The sociated with the glassy-winged sharpshooter Homalo- biology of xylem fluid-feeding insect vectors of Xytel- disca coagulata with emphasis on a new species Hirsu- la fastidiosa and their relation to disease epidemiol- tella homalodiscae nom. prov. Biocontrol 52: 231-258. ogy. Annu. Rev. Entomol. 49: 243-270. BREAUX, S. E. 2005. Fungi associated with the glassy SAS. 2004. User’s guide, version 9.1. SAS Institute, winged sharpshooter, Homalodisca coagulata in its Cary, North Carolina.

Scientiﬁc Notes 109

EXAMINATION OF METHODS FOR FORMOSAN SUBTERRANEAN TERMITE (ISOPTERA: RHINOTERMITIDAE) FECES RECOVERY

TIMOTHY J. ARQUETTE1 AND JOSE M. RODRIGUEZ2 1Department of Entomology, Mississippi State University, Mississippi State, MS 39762

2State Chemical Laboratory, Mississippi State, MS 39762

Chemical assay of undigested food from termite fecal material has been reported for Formo- san subterranean termites (Coptotermes formosanus Shiraki) and other species (Itakura et al. 1995; Hyodo et al. 1999; Mishra & Sen-Sarma 1979; Katsumata et al. 2007). Unlike drywood termites, which produce solid fecal pellets with a conspicuous appearance (Lewis et al. 2010), subterranean termites produce liquid feces (Gouge et al. 2001) that is potentially difficult to recognize and obtain. The current study describes the out- come of attempts at recovery of C. formosanus feces from laboratory arenas in which no food had been placed, and from arenas containing filter paper or wood. Termites used for this study were collected at Audubon Park, New Orleans, LA. Arenas without food were maintained and processed 2 different ways. For the first set of arenas, 18 groups of approximately 50 workers that had been fed southern yellow pine (Pinus sp.) for at least 72 h were transferred to pre-weighed bowls shaped from aluminum foil. Bowls containing termites were placed in polystyrene boxes (32 cm × 25 cm × 10 cm) (Tri-State Plas- tics, Latonia, KY) lined with saturated paper towels. The boxes were covered and maintained at 23°C. At 3, 6, 20, 24, 40, and 48 h, workers were removed from three randomly selected bowls by allowing them to crawl up a filter paper disc. After removal of dead insects and body parts, bowls were dried 15 min at 70°C, and folded closed immediately upon removal from heat. Fecal dry weight was determined to 0.01 mg from the difference between the initial and final weight of the bowl. For the second set of Fig. 1. Mean (+SEM) dry weight of C. formosanus fe- arenas, 3 groups of approximately 500 workers ces recovered from laboratory arenas without food. fed southern yellow pine for at least 72 h were Graph A: mean feces recoveries from 3 randomly se- transferred to polystyrene boxes lined with alu- lected foil bowls holding 50 workers. Graph B: repeated minum foil over saturated paper towels. Con- feces collection from containers that each held 500 workers and soldiers. tainers were covered and maintained at 23°C. After 4, 8, 12, 28, and 38 h, termites were transferred to clean polystyrene boxes. Following each transfer, dead termites were removed, and als were loosely capped and maintained side- air-dried feces transferred to pre-weighed alu- ways at 28°C in the dark. After 1 week insects minum foil bowls with a watercolor brush. Fe- were removed and collection of fecal material ces were further dried 15 min at 70°C before attempted by scraping vials and paper with a weighing. For arenas containing filter paper, 3 razor blade and spatula. For laboratory arenas groups of 50 workers were placed in plastic containing wood blocks, 5 groups of approxi- screw-top vials (40 mm × 15 mm diameter) mately 200 to 500 workers and soldiers were along with a piece of moistened Whatman 1 fil- placed in clear polystyrene containers lined ter paper (Whatman Inc., Piscataway, NJ). Vi- with a piece of aluminum foil over saturated pa-

110 Florida Entomologist 94(1) March 2011

Fig. 2. Cemented structures constructed by C. formosanus termites in a laboratory arena containing southern yellow pine blocks. per towels. Southern yellow pine blocks (either minute quantity of feces recovered from arenas 50 mm × 20 mm × 5 mm, or 20 mm3) that had without food (Fig. 1), little fecal material ap- been soaked in distilled water for 1 h were pears to have been available for use in construc- added to arenas. Containers were covered and tion of the structures. Feces were not visually stored at 28°C. Accumulated debris on foil and discernible from any arena containing wood wood was periodically collected from 4 arenas blocks. with a spatula and razor blade, while a fifth Observations from the current study under- arena was left undisturbed for 6 weeks. score the need for caution in collection of C. formo- The weight of fecal material recovered from sanus fecal material for assay because it can easily arenas that had not contained food averaged be contaminated by foreign particles or unrecover- approximately 0.03 mg to 0.2 mg feces (dry able if food is present. Furthermore, collection of weight) from foil bowls, and about 1 mg total fe- feces for assay from arenas without food may be ces (dry weight) after repeated collections from impractical because of low yield (Fig. 1). For in- the same groups of insects (Fig. 1). Feces in stance, this species digests wood carbohydrate these arenas were apparent either as liquid very efficiently (Yoshimura 1995), so large num- droplets that dried to dark solid specks, or col- bers of termites would be needed to produce fecal lections of small, sticky black piles mixed with material with enough undigested sugars for detec- dead termites, body parts, and trapped live ter- tion by conventional chromatographic methods. mites. No feces were recovered from arenas con- We appreciate the helpful assistance and ad- taining filter paper, although liquid fecal mate- vice from Ms. S. Parikh and Dr. J. Sun (Missis- rial was apparent from dark spots on the paper. sippi State University), and thank Dr. B. Layton For arenas with wood, solid debris was appar- and Dr. G. Baker (Mississippi State University) ent on wood blocks and arena surfaces within for reviewing an early draft of this manuscript. 72 h. Debris was formed into branched structures in the single undisturbed arena (Fig. 2). SUMMARY The structures were soft and friable immediately upon removal from arenas, and when pul- The current study attempted different meth- verized dried to a fine powder. Considering the ods for collection of Formosan subterranean ter-

Scientiﬁc Notes 111

mite feces. When food was present, the liquid fe- ITAKURA, S., UESHIMA, K., TANAKA, H., AND ENOKI, A. cal material was not recoverable either due to 1995. Degradation of wood components by subterra- absorption into food, or occlusion by food parti- nean termite, Coptotermes formosanus Shiraki. cles that were used instead for construction Mokuzai Gakkaishi 41: 580-586. purposes. Uncontaminated fecal material was KATSUMATA, K. S., JUN, Z., HORI, K., AND IIYAMA, K. 2007. Structural changes in lignin of tropical woods recoverable from arenas without food only in during digestion by the termite, Cryptotermes brevis. minute quantity. J. Wood Sci. 53: 419-426. LEWIS, V. R., NELSON, L. J., HAVERTY, M. I., AND BALD- REFERENCES CITED WIN, J. A. 2010. Quantitative changes in hydrocar- bons over time in fecal pellets of Incisitermes minor GOUGE, D. H., SMITH, K. A., OLSON, C., AND BAKER, P. may predict whether colonies are alive or dead. J. 2001. Drywood termites. Cooperative Extension, Col- Chem. Ecol. (in press). lege of Agriculture & Life Sciences, The University of MISHRA, S. C., AND SEN-SARMA, P. K. 1979. Studies on de- Arizona. Available from: http://ag.arizona.edu/pubs/ terioration of wood by insects III. Chemical composi- insects/az1232/ tion of faecal matter, nest material and fungus comb of HYODO, F., AZUMA, J.-I., AND ABE, T. 1999. Estimation some Indian termites. Mater. Organismen 14: 1-14. of effect of passage through the gut of a lower ter- YOSHIMURA, T. 1995. Contribution of the protozoan fau- mite, Coptotermes formosanus Shiraki, on lignin by na to nutritional physiology of the lower termite, Solid-State CP/MAS 13C NMR. Holzforschung 53: Coptotermes formosanus Shiraki (Isoptera: Rhino- 244-246. termitidae). Wood Res. 82: 68-129.

112 Florida Entomologist 94(1) March 2011

TRACHYAPHTHONA NIGRITA AND TRACHYAPHTHONA SORDIDA (COLEOPTERA: CHRYSOMELIDAE) REJECTED AS POTENTIAL BIOLOGICAL CONTROL AGENTS OF PAEDERIA FOETIDA L. (RUBIACEAE), AN INVASIVE WEED IN HAWAII AND FLORIDA

ROBERT W. PEMBERTON1 AND GLORIA L. WITKUS United States Department of Agriculture, Agricultural Research Service, Invasive Plant Research Laboratory, 3225 College Ave., Ft. Lauderdale, FL 33314

[email protected]

Skunk vine, Paederia foetida L. (Rubiaceae), is range testing for T. nigrita and T. sordida initially an invasive weed native to Asia but a problem in concentrated on 3 subfamilies within the Rubi- Florida and other areas of the southern USA, and aceae. Test plants were native and ornamental in Hawaii (Pemberton & Pratt 2002). In the members of this family selected to represent dif- southern states of the USA it is mostly a problem ferent taxonomic tribes. Host range testing on in natural areas, whereas in Hawaii the plant is a plants outside of the Rubiaceae consisted of spe- problem in both natural areas and agriculture. cies selected to represent related orders including Skunk vine is both a Category I weed (Florida Ex- the Cornales, Gentianales, Lamiales, and the So- otic Pest Plant Committee 2009) and a Florida lonales. In total, 34 species in 7 families and 5 or- Noxious Weed (USDA, Natural Resource Conser- ders were tested. vation Service). Host range tests on both beetles were con- Biological control research began with field ducted with a no-choice test design, and each in- surveys in Asia in 1997 (Pemberton & Pratt dividual test plant was exposed to 3 beetles for 1 2002). A lace bug, Dulinius conchatus Distant month. Control plants of P. foetida and P. crudda- (Tingidae), and a leaf beetle, Sphenoraia rutilans siana were exposed to 3 beetles for 1 month. Be- Hope (Chrysomelidae) have undergone host spec- cause research on T. nigrita spanned 2 seasons, ificity testing but neither demonstrated the nar- each experiment was replicated 5 times. Research rowness of host range needed in a skunk vine bio- on T. sordida was conducted for 1 season so each control agent (Pemberton et al. 2005; Pemberton test was replicated 3 times. Host range testing for & Witkus unpublished data). Two Trachyaph- T. nigrita and T. sordida was conducted on thona spp. flea beetles were identified as promis- branches of whole potted plants in the quarantine ing biological control candidates during surveys greenhouse. Clear acrylic tubes, 15.2 cm long × in Japan (Pemberton & Pratt 2002). Both species 3.8 cm diameter (6” × 1.5” diameter), each with 2 have larvae that feed on roots and adults that mesh ventilation holes, were placed on the end of consume leaves. The biology and preliminary host a small branch or stem of each test plant. A slit ranges of Trachyaphthona sordida (Baly) and T. sponge bung was placed around the base of the nigrita Ohno were studied in Japan (Okamato et stem and up into the bottom of the tube. The stem al. 2008). and bung juncture was wrapped tightly with Adult host range testing on T. nigrita was con- parafilm. Another bung was used to seal the top of ducted in our Fort Lauderdale quarantine labora- the tube. Due to the saltatory nature of the flea tory during 2007 and 2008, with field-collected beetles, an aspirator was used to gently transfer 3 beetles shipped by Japanese cooperators during tiny beetles through the bung slit and into the each Jun of those years. Adult host range testing tubes and onto each test branch. The 3 beetles on field-collected T. sordida beetles was done in consisted of 1 mating pair and another beetle of 2009 after Japanese cooperators shipped the bee- undetermined gender due to the difficulty in de- tles during Jun of that year. Adults of both species termining gender in this flea beetle. Beetles were were set up in wooden screened sleeve cages, 50 monitored for 1 month, and at the end of each cm × 50 cm × 53 cm, and fed cut vines of the con- week, all leaves with feeding damage were trol plant, Paederia foetida, and its invasive rela- clipped, pressed and then scanned into jpeg files tive Paederia cruddasiana Pain. Paederia crud- to enable feeding to be measured with SigmaS- dasiana, native to Nepal, Burma, China, and can® Pro. Thailand (Puff 1991), is a naturalized weed in Mi- The most important finding of the research ami-Dade County, Florida. The beetles mated was the nearly equal or greater feeding by both readily and laid eggs throughout the sleeved rear- Trachyaphthona species on the native Florida ing cages. test plant Diodia virginiana L. (Table 1). Tra- The most likely potential non-target plants of chyaphthona nigrita adults ate a mean of 3.75 ± these beetles are the many native and economic 1.1 mm2 leaf material of D. virginiana, compared members of the Rubiaceae in Florida. The host to 4.76 ± 1.53 mm2 of leaf material on P. foetida,

Scientiﬁc Notes 113 SEA- HREE 2 . T OVER

FEEDING

****** CONDUCTED )SDSE

2 RECEIVED

* * * THAT

REPLICATES Trachyaphthona sordida Trachyaphthona ) mean (mm 2 5 PLANTS

DUE

ARE SHOWING

FEEDING BEETLES

. FLEA

* * 0.00 0.00 0.00 0.00 ** * ** 0.11* 0.00 * 0.04 * 0.00 0.04 0.04 0.00 0.01 0.02 0.01 0.00 0.01 0.01 BOLD

ADULT IN AMOUNTS

) SD SE total (mm 2 ARE

TOTAL

1 SORDIDA IN

NATIVES

Trachyaphthona nigrita Trachyaphthona ) mean (mm 2 LORIDA IFFERENCES 0.010.030.11 * * 3.33 * 0.02 0.671.150.012.79 0.37 * 0.230.00 * 0.56 0.22 0.18 0.00 0.16 0.10 0.45 0.00 0.09 0.00 0.67 0.15 0.12 0.06 0.21 0.22 0.12 0.28 0.16 F 16.5323.82 3.3118.76 4.76 0.57 3.75 1.53 0.33 1.10 0.88 0.64 4.15 4.84 9.70 1.38 1.61 0.63 3.23 0.82 0.36 0.50 0.47 0.29 . . D total (mm RACHYAPHTHONA T

SORDIDA SPECIES AND

EACH

WITH

NIGRITA

FOR

Species MONTH 1 REPLICATES 3 Acuba japonica lanceolata Pentas Morinda citrifolia cruddasiana Paederia foetida Paederia Serissa foetida Diodea virginiana Ernodia littoralis Mitchella repens Spermacoce assurgens Spermacoce tetraquetra Spermacoce verticillata FOR

RACHYAPHTHONA T

VERSUS OF , TESTED

WERE

NIGRITA TESTING

BEETLES

RANGE WITH

OST ADULT SONS 1. H Test feeding was documented in only 1 replicate of each species. 1 ABLE Cornales – Garryaceae Rubiales - Rubiaceae Hedyotideae Order—Family Tribe Order—Family Spermacoceae Spermacoceae Morindeae Paederieae Spermacoceae Spermacoceae Spermacoceae Paederieae Paederieae Spermacoceae T

114 Florida Entomologist 94(1) March 2011 and 3.31 ± 0.57 mm2 on P. cruddasiana. Tra- longs (Paederieae). These beetles, therefore, lack chyaphthona sordida adults consumed a mean of the appropriate level of host specificity, and this 3.23 ± 0.5 mm2 leaf material on D. virginiana, eliminates them from further consideration as po- compared to 1.61 ± 0.82 mm2 on P. foetida, and tential biological controls of Paederia foetida, 1.38 ± 0.63 mm2 on P. cruddasiana. Moreover, D. skunk vine. virginiana belongs to a taxonomic group within the Rubiaceae different than that of skunk vine, ACKNOWLEDGMENTS i.e., the tribe Spermacoceae rather than the tribe Paederieae for skunk vine. In addition, T. nigrita We thank Junichi Yukawa, Fukuoka University (re- caused what is considered to be more than test tired), and his students for collecting the Trachyaph- feeding on Spermacoce tetraquetra A. Rich., an- thona beetles in Japan and shipping them to Florida. other member of the Spermacoceae. Trachyaph- Rachel Taylor provided technical assistance and Luke thona nigrita adults consumed a mean of 0.56 ± Kasarjian grew the test plants. The South West Florida 2 Water Management District and the Florida Depart- 0.16 mm of S. tetraquetra, compared to 1.61 ± ment of Environmental Protection funded this research. 0.82 mm2 on P. foetida, and 1.38 ± 0.63 mm2 on P. cruddasiana. To prevent harm to native and im- REFERENCES CITED portant economic members of the Rubiaceae, the feeding of these flea beetles would need to be lim- FLORIDA EXOTIC PEST PLANT COMMITTEE. 2009. Florida ited to plants in the skunk vine tribe, the Paed- Exotic Pest Plant Council’s 2009 list of invasive spe- erieae. cies. Wildland Weeds 12: 13-16. It may be possible that the larvae of these bee- OKAMOTO, C., TUDA, K., YAMAGUCHI, D., SATO, S., PEM- tles could have greater specificity than the adults. BERTON, R. W., AND YUKAWA, J. 2008. Life history Two factors influenced our decision to cease re- traits and host specificity of Japanese Trachyaphtho- search on these Trachyaphthona species and to na species (Coleoptera: Chrysomelidae), candidates shift to other candidate insects of skunk vine. as biological control agents against skunk vine, Paed- eria foetida (Rubiaceae), in Southeastern United Most importantly, we have not successfully States and Hawaii. Entomol. Sci. (Japan) 11: 143-152. reared these univoltine beetles, which are, like PEMBERTON, R. W., AND PRATT, P. D. 2002. Skunk vine many root-feeding flea beetles, difficult to culture (Paederia foetida), pp. 343-351 In R. Van Driesche, B. in captivity. Secondly, our previous testing with Blossey, M. Hoddle, S. Lyon, and R. Reardon [eds.], the leaf beetle Sphenoraia rutilans found that it Biological Control of Invasive Plants in the Eastern could complete its development both on Diodia United States, U.S. Forest Service, Forest Health virginiana and Spermacoce tetraquetra, indicat- Technology Enterprise Team-2002-04, Morgantown, ing that these plants have an attractiveness and West Virginia. vulnerability to some Paederia feeding insects. PEMBERTON, R. W., MURAI, K., PRATT, P. D., AND TERA- MOTO, K. 2005. Dulinius conchatus (Hemiptera: Tingidae), considered and rejected as a potential bi- SUMMARY ological control agent of Paederia foetida, an invasive weed in Hawaii and Florida. Proc. Hawaiian En- Trachyaphthona nigrita and Trachyaphthona tomol. Soc. 37: 81-83. sordida, 2 potential biological control agents of PUFF, C. 1991. Revision of the genus Paederia in Asia, Paederia foetida L. (Rubiaceae), were collected pp. 207-289 In C. Puff [ed.], The Genus Paederia L. from Japan and brought into quarantine for adult (Rubiaceae–Paederieae): A Multidisciplinary Study, Opera Botanica Belgica 3. National Botanic Garden host specificity testing. Testing indicated that of Belgium. Meise, Belgium. they fed significantly on Florida native plants in USDA NATURAL RESOURCE CONSERVATION SERVICE. the tribe Spermacoceae, a different tribe from 2010. http://plants.usda.gov/java/noxious?rptType= that to which the target weed Paederia foetida be- State&statefips=12. Last accessed on 29 Oct 2010.

Scientiﬁc Notes 115

RICH MICROBIAL COMMUNITY ASSOCIATED WITH THE NEST MATERIAL OF RETICULITERMES FLAVIPES (ISOPTERA: RHINOTERMITIDAE)

THOMAS CHOUVENC1*, MONICA L. ELLIOTT2 AND NAN-YAO SU1 1Department of Entomology and Nematology, Ft. Lauderdale Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, 3205 College Ave, Ft. Lauderdale, FL 33314, USA 2Department of Plant Pathology, Ft. Lauderdale Research and Education Center, University of Florida, Institute of Food and Agricultural Sciences, 3205 College Ave, Ft. Lauderdale, FL 33314, USA *Corresponding author; E-mail: tomchouv@ufl.edu Subterranean termites live in a soil environment that can contain a rich community of microorganisms and the nest material can be a site of enhanced microbial activity (Holt & Lepage 2000). By using feces and saliva as building materials for nest construction and gallery systems, termites can alter the qualitative and quantitative composition of the resident soil microorganism community from adjacent soils (Jouquet et al. 2005). Keya et al. (1982) suggested that a significant part of the altered microorganism community was mostly composed of cellulose decompos- ers, but function and origin of the microbial community associated with termite nests remains unclear. In this report we evaluate whether subterranean termites have the ability to import various microorganisms from distant soils or Fig. 1. A group of termites in a two-dimensional from their own gut microbial community into a arena for 90 d. Arrows show sampling sites for micro- microorganism-free soil environment. bioal isolation. Note the dark shade in the gallery wall In Chouvenc et al. (2008), we investigated the showing the presence of deposited fecal material. Bar = survivorship of groups of 960 Reticulitermes flavi- 1 cm. pes (Kollar) from large two-dimensional arenas filled with sterile sand (washed with 70% ethanol, rinsed with sterile deionized water and oven and no termites to confirm the absence of mi- dried at 60°C for 24 h). At the end of the 90-d ex- crobes in the sterile sand. Growth of microbes for periment, 4 arenas from 2 different termite colo- each dilution and each selective medium was nies were dismantled to obtain 2 types of nest ma- quantified to estimate the overall microbial struc- terial: (1) gallery material including sand mixed ture of the 2 nest materials (Table 1). Individual with termite fecal material from termites galler- microbe colonies were selected and transferred on ies, and (2) non-gallery material including undis- 1/5 strength potato dextrose agar. Based on col- turbed sand (not tunneled by the termite) at least ony morphology of more than 2,000 isolates, we 3 cm away from any termite gallery (Fig. 1). A to- found 432 morphologically distinct isolates in- tal of 12 soil samples per arena (10 g per sample) cluding 399 aerobic bacterial isolates, 11 actino- including 6 samples for each type of nest material bacterial isolates, and 22 fungal isolates. The mo- were collected. In order to estimate the microbial lecular identification of these isolates and their communities in the 2 types of nest material, 1 g potential function will be presented elsewhere. sub-sample from each original sample was pro- Samples from the control arena with no termites cessed following the general procedure described showed no microbial growth. in Elliott & Des Jardin (1998). Serial dilutions of Our results showed that when groups of 960 the sub-samples were made (10-1 to 10-7) with ster- termites are allowed to forage and build a com- ile water and the diluted sub-samples were plated plex gallery system in sterile sand, a large micro- on 5 different selective media to isolate various bial community establishes in the sand wall gal- groups of microbes including (1) overall aerobic leries mixed with fecal material. Because the bacteria, (2) fluorescent pseudomonads, (3) acti- sampling method cannot take into account the nobacteria, (4) overall fungi and, (5) ento- non-culturable microbes, our results suggest that mopathogenic fungi. All selective media were pre- the actual microbial diversity within the termite’s viously described in Elliott & Des Jardin (1998) gallery system is probably greater than what we except the medium for entomopathogenic fungi could observe. After 90 d in the presence of ter- (Veen & Ferron 1966). Soil samples collected from mites in the galleries, this microbial community a control arena were established with only sand moved away from the tunnels into the undis-

116 Florida Entomologist 94(1) March 2011

TABLE 1. COLONY FORMING UNITS (CFUS) OF VARIOUS MICROBE GROUPS GROWN ON DIFFERENT SELECTIVE MEDIA, PER GRAM OF SAMPLE OF TERMITE-GALLERY MATERIAL AND NON-GALLERY MATERIAL (MEAN ± SE).

Microbe Group na Gallery material Non-gallery material t-testb

Overall aerobic bacteria 24 3.5 × 107 ± 7.9 x 106 3.9 × 105 ± 5.2 x 104 P < 0.001 Fluorescent Pseudomonads 24 7.2 × 102 ± 3.0 x 102 8 ± 6 P < 0.001 Actinobacteria 24 1.4 × 105 ± 2.8 x 104 1.1 × 102 ± 5.2 x 101 P < 0.001 Overall fungi 24 6.1 × 102 ± 1.1 x 102 4.3 × 101 ± 1.6 x 101 P < 0.001 Entomopathogenic fungi 12 0 ± 0 0 ± 0 N/A

aTotal number of samples plated on a given selective media per concentration from serial dilutions. bFor a given microbe group based on selective media, comparison of the colony forming units between gallery material and non- gallery material. turbed sand, although the overall amount of bac- 90 d, the tunnel walls made of sand mixed with fe- teria remains 100-fold less than the fecal-sand cal material, and sand from areas not disturbed mix. Previous studies have shown that actinobac- by termites were compared for the presence of mi- teria can be isolated from the gut of various ter- crobes. We show that a rich microbial community mites (König et al. 2006). Here we demonstrate associated with the termites can colonize the ter- that actinobacteria are also part of the microbe mite nest environment and is primarily associ- community that can colonize the termite nest ated with the tunnels. structure. Fungi represented a very small frac- tion of the overall microbes enumerated, support- REFERENCES CITED ing the observation by Chouvenc et al. (2008, 2009) that termite gut contents, i.e., fecal mate- CHOUVENC, T., AND SU, N.-Y. 2010. Apparent synergy rial, used as building material for the gallery sys- among defense mechanisms in subterranean ter- tem possess highly fungistatic properties, and do mites (Rhinotermitidae) against epizootic events - not allow fungi to colonize the termite nest envi- The limits and potential for biological control. J. ronment. The apparent absence of entomopatho- Econ. Entomol. 103: 1327-1337. CHOUVENC, T., SU, N.-Y., AND ELLIOTT, M. L. 2008. In- genic fungi support that termites have mecha- teraction between the subterranean termite Reticu- nisms to prevent the spread of harmful fungi litermes flavipes (Isoptera: Rhinotermitidae) and the within the termite nest (Chouvenc et al. 2010). entomopathogenic fungus Metarhizium anisopliae Since all the arenas were constructed of sterile in foraging arenas. J. Econ. Entomol. 101: 885-893. sand, it is assumed all the microorganisms origi- CHOUVENC, T., SU, N.-Y., AND ROBERT, A. 2009. Inhibi- nated from the termite guts or from the surface of tion of Metarhizium anisopliae in the alimentary the cuticle of the termites. This indicates that a tract of the eastern subterranean termite Reticuli- complex microbial community is associated with termes flavipes. J. Invertebr. Pathol. 101: 130-136. the termites themselves before they were intro- ELLIOTT, M. L., AND DES JARDIN, E. A. 1998. Compari- son of media and diluents for enumeration of aerobic duced into the arena. We assume that most of the bacteria from bermudagrass golf course putting cuticular microbes were acquired from the field greens. J. Microbiol. Methods 34: 193-202. nest where they were originally collected, al- HOLT, J. A., AND LEPAGE, M. 2000. Termites and soil though some could be strict termite-associated mi- properties, pp. 389-407 In T. Abe, D. E. Bignell, and crobes and therefore ectosymbionts transmitted M. Higashi [eds.], Termite: Evolution, Sociality, vertically. In addition, the gut can be a reservoir Symbioses, Ecology. Kluwer Academic Publishers, that temporarily hosts various microbes, which Dordrecht. are later inoculated into various part of the gal- JOUQUET, P., RANJARD, L., LEPAGE, M., AND LATA, J. C. lery system. Because fungi were sampled in very 2005. Incidence of fungus-growing termites (Isoptera, Macrotermitinae) on the structure of soil small quantities, we hypothesize that the termite microbial communities. Soil Biol. Biochem. 37: 1852- fecal material within the tunnel wall acts as an in- 1859. hibitor against specific types of organisms. A KEYA, S. O., MURERIA, N. K., AND ARSHAD, M. A. 1982. broader sampling of termite colonies from diverse Population dynamics of soil microorganisms in rela- origin and molecular identification of the microbes tion to proximity of termite mounds in Kenya. J. Ar- will provide a better understanding of the origin id Environ. 5: 353-359. and the function of some of these microbes. KÖNIG, H., FRÖHLICH, J., AND HERTEL, H. 2006. Diver- sity and lignocellulolytic activities of cultured microorganisms, pp. 271-301 In H. König and A. Varma SUMMARY [eds.], Intestinal Microorganisms of Termites and Other Invertebrates. Springer, New York. Field-collected subterranean termites were VEEN, K. H., AND FERRON, P. 1966. A selective medium held in groups of 960 individuals for 90 d in two- for the isolation of Beauveria tenella and Metarrhi- dimensional arenas filled with sterile sand. After zium anisopliae. J. Inverteb. Pathol. 8: 268-269.

Scientiﬁc Notes 117

GUT CONTENT ANALYSIS OF SOUTHERN AND TAWNY MOLE CRICKETS (ORTHOPTERA: GRYLLOTALPIDAE: SCAPERTISCUS)

D. E. SILCOX AND R. L. BRANDENBURG Department of Entomology, North Carolina State University, Campus Box 7613, Raleigh, NC 27695

Two introduced species of mole crickets, Scap- only, or presence of plant and insect material. Con- teriscus borellii Giglio-Tos, the southern mole tent was determined to be plant material if it was cricket and S. vicinus Scudder, the tawny mole fibrous, green, or light brown in color and if it had cricket cause economic damage to turfgrass in a blade-like appearance (Castner & Fowler 1984). North Carolina and throughout the southeastern Content was determined to be animal material if it U.S. Previous studies indicate that Scapteriscus was dark brown or black, obviously sclerotized, or borellii primarily feed on insect material and Scap- if they were a recognizable structure such as, tarsi, teriscus vicinus feed on plant material (Ulagaraj antennae, legs, etc (Castner & Fowler 1984). The 1975; Taylor 1979; Matheny 1981; Fowler et al. gut content was thoroughly examined and ana- 1985). There have been no additional research ef- lyzed until all pieces were identified and catego- forts to examine the gut content of these 2 pests as rized. The contents were then removed from the their range has expanded over the past 25 years to petri dish with a bulb-pipette and placed in the their northern-most habitat (North Carolina) with vial with the cricket from which it was extracted. changes in ecology, host range, or diet. This re- Data were analyzed by Chi-square analysis search was conducted to determine the current through use of Statistical Analysis System version feeding preferences of North and South Carolina 9.1 program (SAS Institute 2003). populations of these 2 mole cricket species. Of the 25 S. borellii nymph alimentary canals Nymph and adult S. borellii and S. vicinus examined 28% ± 0.46% contained only plant mate- were collected during the spring and summer of rial, 4% ± 0.20% contained only insect material, 2009 with soapy water flushing (Short & Koehler and 68% ± 0.48% contained plant material and in- 1979) and individual crickets were immediately sect material (Fig. 1). Of the 25 S. borellii adult al- preserved in 70% ethyl alcohol. Scapteriscus imentary canals examined 56% ± 0.51% contained borellii nymphs were collected from Belvedere only plant material, 20% ± 0.41% contained only Country Club (Pender Co., NC, 34.3675, insect material, and 24% ± 0.44% contained plant 77.710833) on 15 Sep and adults were collected material and insect material (Fig. 1). There is sig- from Olde Fort Golf Course (Brunswick Co., NC, nificant difference in the overall gut contents be- 34.0857, 78.0536) on 5 May. Scapteriscus vicinus tween S. borellii nymphs and S. borellii adults nymphs were collected from Scotch Meadows (Chi-square = 10.2609, P = 0.0059). Of the 25 S. Country Club (Scotland Co., NC, 34.4553, vicinus nymph alimentary canals examined 60% ± 79.2813) on 8 Oct and adults were collected from 0.5% contained only plant material, 4% ± 0.2% con- High Tech Turf (Horry Co., SC, 33.5097, 79.322) tained only insect material and 36% ± 0.49% con- on 6 May and 12 May. All collected individuals tained plant material and insect material (Fig. 1). were examined for species characteristics to en- Of the 25 S. vicinus adult alimentary canals exam- sure proper identification (Potter 1998). ined, 96% ± 0.2% contained only plant material, The alimentary canals (crop, proventriculous, 0% contained only insect material, and 4% ± 0.2% and hindgut) of 25 late instars (large nymphs) and contained plant material and insect material (Fig. 25 adults for each species were removed and exam- 1). There is significant difference in the overall gut ined. Each cricket was placed in a petri dish (8.89 content between adult S. vicinus and nymph S. cm diameter, Fisher Scientific, Pittsburgh, PA) vicinus (Chi-square = 9.4769, P = 0.0088). There is ventral side up, and an X-Acto knife (x3201, Elmer’s Products Inc., Columbus, OH) was used to remove the sterna to expose the alimentary canal. A pair of forceps was used to remove the alimentary canal. The cricket body cavity was placed into a plastic vial filled with 70% ethyl alcohol and labeled to identify each specimen. The alimentary canal remaining in the petri dish was covered with 70% ethyl alcohol to prevent desiccation. Forceps were used to tease open the crop, proventriculous, and hindgut. All gut content was noted for each cricket. The content was examined under a binocu- lar microscope used in the 7X-30X power range and the contents were categorized as presence of Fig. 1. The frequency of various gut contents for S. plant material only, presence of insect material borellii and S. vicinus nymphs and adults.

118 Florida Entomologist 94(1) March 2011

TABLE 1. THE PERCENT HERBIVORY, CARNIVORY, AND OMNIVORY FOR S. BORELLII AND S. VICINUS POPULATIONS IN BRAZIL AND FLORIDA (OTHER STUDIES) AND NORTH AND SOUTH CAROLINA (THIS STUDY).

Species Location % Herbivory % Carnivory % Omnivory Reference

S. borellii Florida 4% 44% 8% Ulagaraj (1975) S. vicinus Florida 53% 15% 21% Ulagaraj (1975) S. borellii Florida 14% 90% — Taylor (1979) S. vicinus Florida 50% 61% — Taylor (1979) S. borellii Florida 6% 70% 43% Matheny (1981) S. vicinus Florida 72% 1% 10% Matheny (1981) S. borellii Brazil 18% 32% 12% Fowler et al. (1985) S. vicinus Brazil 44% 6% 10% Fowler et al. (1985) S. borellii North Carolina 42% 12% 46% NC 2009 S. vicinus North and South Carolina 78% 2% 20% NC/SC 2009

also significant difference (P < 0.05) in the gut con- Carolina mole cricket populations as compared to tent between S. borellii (nymphs and adults) and previous studies in other locations S. borellii S. vicinus (nymphs and adults) (Chi-square = nymphs were found to primarily consume plant 14.8027, P = 0.0006). and insect materials, while S. borellii adults were We determined that the gut content of North found to primarily consume only plant materials. and South Carolina populations of S. borelli and Scapteriscus vicinus nymphs and adults were S. vicinus is similar to previous findings for South found to primarily consume only plant material. America and Florida populations (Table 1). Mod- The results of this study were similar to those of est changes in feeding preferences were apparent previous studies conducted more than 25 years as the southern mole cricket in North Carolina ago in Brazil and Florida. appeared to be more herbivorous than previous studies in other locations indicated in the past. The differences in length between S. borelli and S. REFERENCES CITED vicinus alimentary canals could be related to dif- CASTNER, J. L., AND FOWLER, H. G. 1984. Gut content ferences in diet. The alimentary canal of S. vici- analysis of Puerto Rican mole crickets (Orthoptera: nus is significantly longer than in S. borellii (Na- Gryllotalpidae: Scapteriscus). Florida Entomol. tion 1983). The longer alimentary length seen in 67(3): 479-484. S. vicinus may reflect the greater difficulty of di- FORREST, T. G. 1987. Insect size tactics and developmen- gesting plant cells; more area is needed for the tal strategies. Oecologia 73: 178-184. chemical digestion to occur (Nation 1983). FOWLER, H. G., VIEIRA DE CAMARGO, M. T., AND The differences in gut content seen in both spe- CRESTANA, L. 1985. Feeding habits of Brazilian mole cies between nymphs and adults could be attrib- crickets (Orthoptera: Gryllotalpidae: Scapteriscus spp. and Neocurtilla sp.). J. Econ. Entomol. 78: 1076-1078. uted to the different developmental needs of the MATHENY, JR., E. L. 1981. Contrasting feeding habits of life stages (Forrest 1987). We found a higher per- pest mole cricket species. J. Econ. Entomol. 74: 444- centage of herbivory in both species as adults as 445. compared to nymphs. We also observed greater NATION, J. L. 1983. Specialization in the alimentary canal herbivory in the southern mole cricket as com- of some mole crickets (Orthoptera: Gryllotalpidae). In- pared to previous reports. tl. J. Insect Morphol. & Embryol. 12(4): 201-210. Nymphs of the European mole cricket Gryllo- POTTER, D. A. 1998. Destructive turfgrass insects: biolo- talpa gryllotalpa Linnaeus (Ulagaraj 1975) that gy, diagnosis, and control. Ann Arbor Press, Chelsea, were fed on insect food completed development in Michigan. SAS INSTITUTE. 2003. PROC User’s Manual, version 9.1 2 years, while nymphs fed on vegetable matter SAS Institute, Cary, North Carolina. completed development in over 4 years (Ulagaraj SHORT, D. E., AND KOEHLER P. G. 1979. A sampling 1975). The protein found in animal/insect compo- technique for mole crickets and other pests in turf- nents of diet may be important in development. grass and pasture. Florida Entomol. 62: 282-283. TAYLOR, T. R. 1979. Crop contents of two species of mole crickets, Scapteriscus acletus and S. vicinus (Ortho- SUMMARY ptera: Gryllotalpidae). Florida Entomol. 62(3): 278- 279. The alimentary canals of 25 nymph and 25 ULAGARAJ, S. M. 1975. Food habits of mole crickets (Or- adult S. borellii and S. vicinus were dissected to thoptera: Gryllotalpidae: Scapteriscus). J. Georgia determine the gut contents in North and South Entomol. Soc. 10(3): 229-231.

Book Reviews 119

GOLDSMITH, M. R., AND MAREC, F. (EDS.). 2010. Molecular Biology and Genetics of the Lepidoptera (Contemporary Topics in Entomology Series). CRC Press, Boca Raton. xv + 362 pp. ISBN 978-1-4200- 6014, hardback, $129.95.

With more than 150,000 described species, coverpa (Chapter 12). There are also chapters butterflies and moths constitute one of the most on sex chromosome systems and genetics un- diverse orders of insects. The number of actual derlying sexual dimorphism (Chapters 3, 4), de- species of Lepidoptera may be much greater, velopment of butterfly wing patterns and the possibly as high as 500,000. Lepidoptera are genetics underlying them (Chapters 5, 6), mo- thought to have radiated contemporaneously lecular innovations of butterfly eyes (Chapter with flowering plants, and they are prominent 7), and pheromone production and perception in the literature on co-evolution. Caterpillars, (Chapter 10). Many of these chapters contain both as external and internal feeders, are im- fascinating facts, and information that is highly portant components of terrestrial ecosystems specialized. Of interest to me were the chapters worldwide, and many are pests to agriculture on lepidopteran circadian clocks (Chapter 8) while also serving as pollinators. As perhaps and the genetics that influence host ranges in one of the most conspicuous taxa of terrestrial Lepidoptera (Chapter 11). The former chapter invertebrates, Lepidoptera are also of central includes numerous diagrams illustrating the utility in ecosystem assessment and in educat- mechanisms of circadian rhythms. This chapter ing the public in environmental conservation. also includes a section on how the circadian Lepidoptera include numerous model organ- clock plays a role in the migration of monarch isms that have played central roles in biology, butterflies, which I found to be especially com- for studies on physiology, development, evolu- pelling. The latter chapter on host range (Chap- tion, and genetics. ter 11) reviews model taxa (Euphydryas, Heli- Molecular Biology and Genetics of the Lepi- coverpa, Heliothis, Papilio) in which the genet- doptera, edited by Goldsmith & Marec, is a book ics underlying host specificity has been exam- of eighteen chapters that highlights recent devel- ined. As someone interested in host-use opments on the genetics of Lepidoptera model evolution in Lepidoptera, I found this chapter systems. The book is in many ways a follow up to to be an especially informative source. Goldsmith & Wilkins’ (1995). With the advance- Latter chapters focus on another dominant ment of molecular tools and methods, this timely theme, insect control. For instance, there are work serves as an updated compilation, high- chapters on the genetics underlying insecticide lighting lepidopteran model systems that have resistance (Chapter 13), innate insect immune re- become prominent in recent years. sponse (Chapter 14), interhemocoelic toxins for The book begins with an overview of lepi- pest management (Chapter 16), and polydnavi- dopteran relationships, written by thirteen au- ruses in lepidopteran parasitoids (Chapter 17). thors (Chapter 1). The chapter is divided into These chapters are written in great detail to ex- several sections, each representing a separate plain various aspects of insect/pest control, which superfamily, outlining the role of model organ- also contain quite a bit of specialized terminology, isms in these superfamilies (Bombycoidea, Hes- and acronyms such as “PBLEs”, “pro-PAP-3”, and peroidea + Papilionoidea, Noctuoidea, Pyral- “V421M mutation” might overwhelm the average oidea, and Tortricoidea). Each of these “super- Lepidoptera enthusiast. family sections” includes phylogenies compiled In conclusion, Goldsmith & Marec do an ex- from various studies, and the trees are useful cellent job compiling a book written by a broad for identifying the phylogenetic position of mod- range of leading researchers studying various els in each superfamily. The chapter also in- aspects of lepidopteran genetics. The compila- cludes a “backbone” phylogeny of the Lepi- tion is not a coffee-table book, and the average doptera based on Kristensen & Skalski’s (1998) reader may find it a bit hard to follow without a tree from the Handbook of Zoology. Readers background in molecular biology. However, the should be aware that two key studies on the mo- book is an excellent summary of the recent lecular phylogeny of Lepidoptera have ap- progress in the field, and will be an invaluable peared very recently in the literature, namely resource for researchers and driven enthusiasts Regier et al. (2009) and Mutanen et al. (2010) that are interested in learning more about the that are updates to Kristensen’s tree. Research- genetics of Lepidoptera. ers interested in understanding the phylogenetic position of model organisms should refer to these publications for an updated “backbone” REFERENCES CITED phylogeny of the Lepidoptera. Subsequent chapters focus on particular GOLDSMITH, M. R., AND WILKINS, A. S. 1995. Molecular model species or genera, such as Bombyx Model Systems in the Lepidotera. Cambridge Uni- (Chapter 2), Heliconius (Chapter 6), and Heli- versity Press, Cambridge, England.

120 Florida Entomologist 94(1) March 2011

KRISTENSEN, N. R., AND SKALSKI, A. W. 1998. Phyloge- Current address ny and palaeontology, pp. 7-25 In N. P. Kristensen (ed.), Lepidoptera, Moths and Butterflies. Volume 1: Akito Y. Kawahara Evolution, Systematics and Biogeography. Walter de Department of Plant and Gruyter, New York. Environmental Protection Sciences MUTANEN, M., WAHLBERG, N., AND KAILA, L. 2010. University of Hawai’i, Manoa Comprehensive gene and taxon coverage elucidates 3050 Maile Way radiation patterns in moths and butterflies. Proc. Honolulu, HI 96822 USA Royal Soc. of London, Series B 277: 2839-2848. [email protected] REGIER, J. C., ZWICK, A., CUMMINGS, M. P., KAWAHARA, A. Y., CHO, S., WELLER, S., ROE, A., BAIXERAS, J., Future address BROWN, J. W., PARR, C., DAVIS, D. R., EPSTEIN, M., HALLWACHS, W., HAUSMANN, A., JANZEN, D. H., Akito Y. Kawahara KITCHING, I. J., SOLIS, M. A., YEN, S. H., BAZINET, A. McGuire Center for Lepidoptera L., AND MITTER, C. 2009. Toward reconstructing the and Biodiversity evolution of advanced moths and butterflies (Lepi- Florida Museum of Natural History doptera: Ditrysia): an initial molecular study. BMC University of Florida Evolutionary Biology 9: 280. Gainesville, FL 32611

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