Wetterer & O’Hara: of the Dry Tortugas 303

ANTS (: FORMICIDAE) OF THE DRY TORTUGAS, THE OUTERMOST FLORIDA KEYS

JAMES K. WETTERER AND BRANDON C. O’HARA HONORS COLLEGE, FLORIDA ATLANTIC UNIVERSITY, 5353 PARKSIDE DR., JUPITER, FL 33458

ABSTRACT

We examined the distribution of ants on the Dry Tortugas, the outermost of the Florida Keys. These small islands are important nesting grounds for sea turtles and sea birds. We sampled ants on the five vegetated islands: Garden Key, Loggerhead Key, Bush Key, Long Key, and East Key. We found 17 species, seven of which had not been recorded from the Dry Tortu- gas. Paratrechina longicornis was common on four of five islands. Otherwise, the five islands had strikingly different ant faunas. On Garden Key, Solenopsis geminata was dominant, and Pheidole megacephala was absent. On Loggerhead Key and Bush Key, Ph. megacephala was dominant, and S. geminata was absent. On Long Key, vegetated primarily with mangrove, the most common ant was the arboreal elongatus. On sparsely-vegetated East Key, all ants were uncommon. Twenty ant species are now known from the Dry Tortugas: eight New World and 12 Old World species. Only Solenopsis globularia is an undisputed Florida native. Florida specimens previously identified as C. tortuganus are actually Camponotus zona- tus; true C. tortuganus are Bahamian. The two dominant ant species, Ph. megacephala and S. geminata, may pose a threat to native fauna, including sea turtle and sea bird nestlings.

Key Words: Camponotus tortuganus, exotic species, Solenopsis geminata, tramp ants, Pheidole megacephala

RESUMEN

Examinamos la distribución de hormigas en las Dry Tortugas, las islas mas exteriores de las Florida Keys. Estas islas pequeñas son areas importantes para nidos de las tortugas del mar y de los pájaros del mar. Muestreamos hormigas en las cinco islas vegetadas: Garden Key, Loggerhead Key, Bush Key, Long Key, y East Key. Encontramos 17 especies hormigas; siete de estas no fueron registrados de Dry Tortugas. Paratrechina longicornis eran comunes en cuatro de cinco islas. De otras maneras, las cinco islas tenían llamativo diversos faunas de la hormiga. En Garden Key, Solenopsis geminata era dominante, y Pheidole megacephala es- taba ausente. En Loggerhead Key y Bush Key, Ph. megacephala era dominante, y S. gemi- nata estaba ausente. En Long Key, vegetada sobre todo con el mangle, la hormiga más co- mún era la arboreal Pseudomyrmex elongatus. En East Key, una isla sin mucha vegetación, las hormigas en general eran raro. Viente especies de las hormigas estan conocidas de las Dry Tortugas ahora: ocho especies del mundo viejo y 12 del mundo nuevo. Solamente Sole- nopsis globularia esta nativa de la Florida sin disputación. Los especímenes de la Florida identificada previamente como C. tortuganus es realmente Camponotus zonatus; C. tortug- anus verdadero es de las Bahamas. Las dos especies dominantes de la hormiga, Ph. megace- phala y S. geminata, pueden plantear una amenaza a la fauna nativa, incluyendo las juveniles de la tortuga marinas y del pájaro del mar.

The Dry Tortugas (24°33’-24°44’N; 82°46- and magnificent frigate birds (Fregata magnifi- 83°15’W) are the outermost of the Florida Keys, cens) (Robertson 1964; Lenihan 1997). The is- located 100-112 km West of Key West, Florida. lands are far from pristine and have numerous These islands and the surrounding waters were non-indigenous plants and (Stoddart & declared a wildlife refuge in 1908, a national mon- Fosberg 1981), such as wetland nightshade ument in 1935, and a national park in 1992. Most (Solanum tampicense; Fox & Bryson 1998) and biological research in the Dry Tortugas area has the Old World house gecko (Hemidactylus concerned the diverse marine biota in the coral mabouia; Meshaka & Moody 1996). In the reefs surrounding the keys (see Schmidt & Pikula present study, we examined the distribution of 1997 for an annotated bibliography of research). native and exotic ants on the Dry Tortugas. The Dry Tortugas are also important ecologically Hurricanes periodically reshape the Dry Tor- as the feeding and nesting grounds for loggerhead tugas, altering the size and even the number of turtles (Caretta caretta), green sea turtles (Chelo- keys (Stoddart & Fosberg 1981). Currently, there nia mydas), and numerous sea birds, including are seven keys: (from west to east) Loggerhead masked boobies (Sula dactylatra), brown boobies Key, Garden Key, Bush Key, Long Key, Sand Key, (Sula leucogaster), sooty terns (Sterna fuscata), Middle Key, and East Key. Garden Key and Log-

304 Florida Entomologist 85(2) June 2002 gerhead Key are the largest and the only inhab- (Table 1; Emery 1895; Mayor 1922; Wheeler 1932; ited islands. At present, about ten park staff MacArthur & Wilson 1967; Deyrup et al. 1988). members live on these two keys. Garden Key is The questionable record from the Dry Tortugas the site of Fort Jefferson, a 19th century structure is the earliest. Emery (1895) described Campono- that occupies most of the island. Plans are being tus tortuganus based on specimens that he be- made to expand tourist facilities on Garden Key lieved were collected by T. Pergande in the Dry to accommodate the large number of people who Tortugas. Mackay (pers. comm.), however, found visit the Dry Tortugas each year (almost 90,000 in that the collection locality for Emery’s C. tortuga- 1999; Dustin 1999). Loggerhead Key has a Coast nus holotype specimen was labeled in Emery’s Guard Station and lighthouse and is the former handwriting as Egg Island, Bahamas. Also, Mackay site of Tortugas Laboratory, a research station ad- (pers. comm.) has received additional specimens ministered by the Carnegie Institute from 1904 to from the Bahamas that match the C. tortuganus 1939 (Mayer 1902; Schmidt & Pikula 1997). Bush holotype. In contrast, Mackay (pers. comm.) Key is an important breeding site for many sea- found that all “C. tortuganus” specimens from birds. It is currently connected by a sand spit to Florida that he examined are actually Campono- Long Key, which is largely vegetated with man- tus zonatus (formerly Camponotus conspicuus groves. East Key is a small, sparsely vegetated is- zonatus). Mackay (pers. comm.) suggested that land far to the east of the other keys. Finally, the Emery erroneously attributed the Dry Tortugas smallest two islands, Sand Key and Middle Key, as the type locality for C. tortuganus and that true are simple sand bars without vegetation. C. tortuganus is Bahamian. The aim of my study was to establish a base- The next mention of ants from the Dry Tortu- line of information on the distribution of ants in gas come from Cole (1906), who worked at Tortu- the Dry Tortugas, as a first step in evaluating the gas Laboratory on Loggerhead Key, and reported potential impact of ants on native species on the that he “studied the reactions of ants” there. It ap- islands. pears that this work was never published, and we could not find any Cole specimens. MATERIALS AND METHODS Mayor (1922) studied an Old World exotic ant, Monomorium destructor, on Loggerhead Key, de- In addition to compiling published records on scribing it as “a great pest in the wooden build- ants, we surveyed the ants on the five vegetated ings of Tortugas Laboratory, making its nests in islands of the Dry Tortugas, Loggerhead Key, crevices of the woodwork. So voracious are these Garden Key, Bush Key, Long Key, and East Key, that we are obliged to swing our beds from on 17-19 March 2000 during daylight hours, with the rafters and to paint the ropes with a solution the help of undergraduate assistants from the of corrosive sublimate, while all tables must have Honors College of Florida Atlantic University. We tape soaked in corrosive sublimate wrapped did not sample on the unvegetated Sand Key and around their legs if ants are to be excluded from Middle Key because these islands are periodically them. These pests have the habit of biting out submerged for extended periods, and thus have small pieces of skin, and I have seen them kill no permanent ant populations. within 24 hours rats which were confined in To evaluate the overall variety of ant species cages.” Mayor (1922) mentioned no other ant spe- present on each island, we conducted intensive cies on the island. hand-collecting, searching accessible surfaces Wheeler (1932), in his review of ant records and turning over rocks and logs. from Florida, listed 91 species, including only two To evaluate the relative dominance of different species from the Dry Tortugas, both collected by ant species on Garden Key, we surveyed ants at Pergande (again, presumably from Garden Key): 40 bait stations, each with two bait cards. A bait Camponotus tortuganus and card consisted of a 7.6 × 12.7 cm index card with guineense (now Tetramorium bicarinatum). approximately 2 g of tuna bait placed at its center. Wilson’s (1964) survey of the ants of the Flor- We folded each card in half and placed it flush ida Keys did not include any records from the Dry with the ground about one meter from the road, Tortugas. MacArthur & Wilson (1967) stated that trail, transect, or building, using a small rock or 13 ant species “have succeeded in colonizing the branch to weigh it down. We returned to each sta- Dry Tortugas,” but only mentioned two species by tion 1.5-2.0 hours later and placed the cards, name, Paratrechina longicornis, and Pseudo- along with all ants on the bait, in plastic bags. myrmex elongatus. MacArthur & Wilson (1967) reported that on the Dry Tortugas, Paratrechina RESULTS longicornis was “an overwhelmingly abundant ant and has taken over nest sites that are nor- Published Ant Records mally occupied by other species in the rest of southern Florida: tree-boles, usually occupied by In addition to one questionable record, we found species of Camponotus and Crematogaster, which reports of 14 ant species from the Dry Tortugas are absent from the Dry Tortugas; and open soil,

Wetterer & O’Hara: Ants of the Dry Tortugas 305

TABLE 1. ANTS OF THE DRY TORTUGAS.

Key

Loggerhead Garden Bush Long East Status

Brachymyrmex obscurior Forel f2 NW/N? *Camponotus tortuganus Emery a* ? Camponotus zonatus Emery f0 f NW/N? Cardiocondyla ectopia Snelling f f OW/E Cardiocondyla emery Forel df0 OW/E Cardiocondyla venustula Wheeler f1 f OW/E Cardiocondyla wroughtoni (Forel) f0 OW/E Hypoponera opaciceps (Mayr) d NW?/N? Monomorium destructor (Jerdon) b OW/E Monomorium floricola (Jerdon) f df6 f OW/E Paratrechina bourbonica (Forel) f df5 f f OW/E Paratrechina guatemalensis (Forel) e f NW/E Paratrechina longicornis (Latr.) f cdef14 f f OW/E Pheidole megacephala (Fabr.) f de f OW/E Pseudomyrmex cubaensis Forel f NW/N? Pseudomyrmex elongatus (Mayr) df0 cf f f NW/N? Solenopsis geminata (Fabr.) def28 NW/N? Solenopsis globularia (Smith) f f1 f NW/N Tapinoma melanocephalum (Fabr.) def3 OW/E Tetramorium bicarinatum Nylander a OW/E Tetramorium caldarium (Roger) df6 f OW/E

Collectors/Authors: a* = Pergande (in Emery 1895) a = Pergande (in Wheeler 1932); b = Mayor (1922); c = MacArthur & Wilson (1967); d = Winegarner (in Deyrup et al. 1988); e = Carlin (in Deyrup et al. 1988); f = present study (including # of Garden Key bait stations with the species present); * = ap- parently erroneous record. Status (from Deyrup et al. 1988, 2000): NW = New World origin; OW = Old World origin. N = native; E = exotic; ? = status in question. normally occupied by crater nests of Conomyrma recorded from the Dry Tortugas. One species, Bra- [now Dorymyrmex] and Iridomyrmex [now Fore- chymyrmex obscurior, belongs to a genus in which lius], which genera are also absent from the Dry species boundaries for Florida specimens have Tortugas.” MacArthur & Wilson (1967) also re- not been adequately defined (Deyrup 1991). We ported that Pseudomyrmex elongatus was “the did not find four species recorded in previous only member of the arboreal assemblage that has studies: Camponotus tortuganus (see discussion colonized the Dry Tortugas, where it has a red above), Tetramorium bicarinatum, Monomorium mangrove swamp on Bush Key virtually to itself. destructor, and Hypoponera opaciceps. Yet it is still limited primarily to thinner twigs in Paratrechina longicornis was common on four the canopy and, unlike Paratrechina longicornis, of the five islands. Otherwise, the five islands had has not increased perceptibly in abundance.” strikingly different ant faunas. On Garden Key, Deyrup et al. (1988), in their review of the ants Solenopsis geminata and Paratrechina longicor- of the Florida Keys, included records from recent nis were by far the most common ants at baits, collections in the Dry Tortugas by C. Winegarner found, respectively at 28 and 14 of the 40 bait sta- (collected 12 April 1983; no island site locality, tions (Table 1). Solenopsis geminata was the dom- presumably from Garden Key) and by N. F. Carlin inant ant on the ground, while Paratrechina (no island site locality, presumably from Garden longicornis was the most common ant up in trees. Key). These two collections recorded a total of 11 On Loggerhead Key and Bush Key, Pheidole ant species from the Dry Tortugas, bringing the megacephala and Paratrechina longicornis were total number of previously published ant records the most common ants. Solenopsis globularia, to 14 species (Table 1). Although Deyrup et al. which we found only once on Garden Key, was (1988) reported Pheidole dentata specimens col- common on Loggerhead and Bush Key. On Long lected by Winegarner, these were actually mis- Key, vegetated primarily with mangrove, the identified Pheidole megacephala specimens most common ant was the mangrove-inhabiting (M. Deyrup, pers. comm.). Pseudomyrmex elongatus. On sparsely vegetated East Key, all ants were uncommon. New Ant Collection We collected Camponotus specimens on both We collected 17 ant species on the Dry Tortu- Garden Key and on Long Key. On Garden Key, we gas (Table 1). Seven of these were not previously found Camponotus workers foraging in trees and 306 Florida Entomologist 85(2) June 2002 we collected dead Camponotus workers and of St. Thomas a few miles to the east, had exter- males from the windowsills of a park warden’s minated all the other ants which must have pre- house. On Long Key, we found Camponotus work- viously inhabited Culebrita. The absence of ers inside a bamboo pole near the beach. Mackay megacephala on Culebra is perhaps explained by (pers. comm.) examined all my Camponotus spec- the presence of the equally prolific and pugna- imens and found that they did not match Emery’s cious fire-ant” (Wheeler 1908). Mutually exclu- C. tortuganus holotype, but instead were Cam- sive distributions between pairs of dominant ant ponotus zonatus. species has also been noted for many other spe- cies, such as between Ph. megacephala and Line- DISCUSSION pithema humile on Madeira (Schmitz 1896) and Bermuda (Haskins & Haskins 1965). Excluding Camponotus tortuganus, which ap- Paratrechina longicornis coexisted at high den- pears to be an erroneous record, 20 ant species sities with both Solenopsis geminata (on Garden are recorded from the Dry Tortugas (Table 1), 12 Key) and Ph. megacephala (on Loggerhead Key of Old World origin and eight from the New World. and Bush Key). An Old World native, P. longicornis Only one species, Solenopsis globularia, is an un- now has a pantropical distribution and commonly disputed Florida native. Of the other seven New occurs at high densities in disturbed habitats (e.g., World species, Deyrup et al. (2000) considered see Wetterer 1998, Wetterer et al. 1999). Paratrechina guatemalensis as an exotic in Flor- Remarkably, Monomorium destructor, found in ida and listed Brachymyrmex obscurior, Cam- such overwhelming abundance on Loggerhead ponotus tortuganus (= Camponotus zonatus), Key by Mayor (1922), was not found on any island Solenopsis geminata, Pseudomyrmex cubaensis, in the Dry Tortugas by later collectors. It is possi- and Pseudomyrmex elongatus as “dubious na- ble that M. destructor only dominated the Tortu- tives.” In addition, although Deyrup et al. (1988) gas Lab buildings which have subsequently been considered Hypoponera opaciceps as native to the torn down. Deyrup et al. (1988) wrote that M. de- Florida Keys, Dlussky (1994) has proposed that structor “is not common in Florida, and may be on this species is actually of Old World origin. the decline in the Keys, except in Key West where We found a strikingly different ant fauna on it is a dominant urban ant.” Deyrup (1991) wrote each of the five vegetated keys of the Dry Tortu- that M. destructor “is spectacularly common on gas. On Garden Key, Solenopsis geminata was the Key West.” It is possible that Ph. megacephala dominant ant species, occurring at 70% of the bait and/or S. geminata competitively excluded M. de- stations, usually with several hundred workers structor from the Dry Tortugas. on each bait card. Their presence on the island The two dominant ant species of the Dry Tortu- was noted by visitors who were often stung on the gas, S. geminata and Ph. megacephala, may pose a feet by these fire ants. Although Pheidole mega- threat to the native fauna of the Dry Tortugas. cephala may have occurred at one time on Garden Pheidole megacephala, an African native, is now a Key, we found none. In contrast, we found no pantropical pest. In lowland Hawaii, it has been S. geminata on any other key in the Dry Tortugas. implicated in the extermination of much of the en- Instead, on Loggerhead Key and Bush Key, demic fauna (Zimmerman 1948). Zimmerman Ph. megacephala was the dominant ant. (1948) reported that the “voracious immigrant Wheeler (1908) similarly observed mutually ant,” Ph. megacephala, eliminates most endemic exclusive distributions of Ph. megacephala and insects throughout its range. Solenopsis geminata S. geminata on two Caribbean islands: “I devoted is an important exotic pest on Pacific islands (Wet- ten days to a careful study of the ant-fauna of the terer 1997). On the Dry Tortugas, this species may little island of Culebra off the eastern coast of prey on hatchling sea turtles and birds (e.g., see Porto Rico without seeing a single specimen of Kroll et al. 1973; Wetterer & Wood 2001). Ph. megacephala. This island is, however, com- Two highly-destructive exotic ant species pletely overrun with a dark variety of the vicious known from the Florida Keys that also pose im- fire-ant (Solenopsis geminata). One day, on visit- portant threats to native species (e.g., see Vinson ing the island of Culebrita, which is separated by 1994; Jourdan 1997; Wetterer & Wood 2001) are a shallow channel hardly a mile in width from the the red imported fire ant, Solenopsis invicta, and eastern coast of Culebra, I was astonished to find the little fire ant, Wasmannia auropunctata (Dey- it completely overrun with Ph. megacephala. This rup et al. 1988, Horvitz 1997). These species are ant was nesting under every stone and log, from not yet known from the Dry Tortugas. Construc- the shifting sand of the sea-beach to the walls of tion of new tourist facilities on the Dry Tortugas the light-house on the highest point of the island. may result in the introduction of these and other The most careful search failed to reveal the pres- exotic ant species, arriving with building material ence of any other species, though the flora and or heavy machinery during the building stage, or physical conditions are the same as those of Cul- accompanying visitors in their camping supplies ebra. It is highly probable that Ph. megacephala, and picnic baskets. On the Dry Tortugas, a rela- perhaps accidentally introduced from the island tively small effort in preventing the further inva- Wetterer & O’Hara: Ants of the Dry Tortugas 307

sion of exotic ants could have a great effect in KROLL, J. C., K. A. ARNOLD, AND R. F. GOTIE. 1973. An protecting the native inhabitants of these ecolog- observation of predation by native fire ants on nest- ically important islands. ling Barn Swallows. Wilson Bull. 85: 478-479. LENIHAN, D. J. 1997. The Tortuga Triangle. Natural Hist. 106: 36-41. ACKNOWLEDGMENTS MACARTHUR, R. H., AND E. O. WILSON. 1967. The The- ory of Island Biogeography. Princeton Univ. Press. We thank S. Paquette, N. McClain, L. Eurich, J. Chang, MAYER, A. G. 1902. The Tortugas, Florida as a station for and L. Arencibia (students from Florida Atlantic Univer- research in biology. Science 17: 190-192. sity’s Honors College) for field assistance; S. Cover (Har- MAYOR, A. G. 1922. The tracking instinct in a Tortugas vard University) for ant identification; W. Mackay (Texas ant. Papers Tortugas Lab. 18: 101-107. Tech) for unpublished information concerning Campo- MESHAKA, W. E. JR., AND B. A. MOODY. 1996. The Old notus; A. Wetterer, M. Wetterer, and M. Deyrup (Archbold World tropical house gecko (Hemidactylus mabouia) Field Station) for comments on this manuscript; S. Bass, on the Dry Tortugas. Florida Scientist 59: 115-117. P. Taylor, and W. Meshaka (U.S. National Park Service) ROBERTSON, W. B., JR. 1964. The terns of the Dry Tor- for logistic support; Florida Atlantic University and the tugas. Bull. Fla. State Mus. 8(1): 1-93. National Save the Sea Turtle Foundation for financial SCHMIDT, T. W., AND L. PIKULA. 1997. Scientific studies support. on Dry Tortugas National Park: an annotated bibli- ography. Atoll Res. Bull. 449: 2-9. REFERENCES CITED SCHMITZ, E. 1896. As formigas da Madeira. Ann. Sci. Nat. 3: 55-58. COLE, L. J. 1906. Ant Studies. Carnegie Inst. Washing- STODDART, D. R., AND F. R. FOSBERG. 1981. Topographic ton, Year Book 5: 110. and floristic change, Dry Tortugas, Florida, 1904- DEYRUP, M. A. 1991. Exotic ants of the Florida Keys. 1977. Atoll Res. Bull. 253: 1-55. Proc. 4th Symp. Nat. Hist. Bahamas. pp. 15-22. VINSON, S. B. 1994. Impact of the invasion of Solenopsis DEYRUP, M., N. CARLIN, J. TRAGER, AND G. UMPHREY. invicta (Buren) on native food webs, pp. 240-258. In 1988. A review of the ants of the Florida Keys. Flor- D. F. Williams [ed.], Exotic ants. Biology, impact, and ida Entomol. 71: 163-176. control of introduced species. Westview Press, Boul- Deyrup, M., L. Davis, and S. Cover. 2000. Exotic ants in der, CO. 322 pp. Florida Trans. Amer. Entomol. Soc. 126: 293-326. WETTERER, J. K. 1997. Alien ants of the Pacific islands. DLUSSKY, G. M. 1994. Zoogeography of southwestern Aliens 6: 3-4. Oceania, pp. 48-93. In Y. G. Puzatchenko, S. I. Golo- WETTERER, J. K. 1998. Ants on Cecropia trees in urban vatch, G. M. Dlussky, K. N. Diakonov, A. A. Zakharov, San José, Costa Rica. Florida Entomol. 81: 118-121. and G. A. Korganova [eds.], population of the WETTERER, J. K., S. E. MILLER, D. E. WHEELER, C. A. islands of Southwestern Oceania. Nauka Publishers, OLSON, D. A. POLHEMUS, M. PITTS, I. W. ASHTON, Moscow, Russia. A. G. HIMLER, M. YOSPIN, K. R. HELMS, E. L. DUSTIN, D. 1999. The curious history of Dry Tortugas HARKEN, J. GALLAHER, C. E. DUNNING, M. NELSON, National Park. Parks Recreat. 34: 126-133. J. LITSINGER, A. SOUTHERN, AND T. L. BURGESS. EMERY, C. 1895. Beitäge zur Kenntniss der nordameri- 1999. Ecological dominance by Paratrechina longicor- kanischen Ameisenfauna. Zoolog. Jahr. Abtheil. Sys- nis (Hymenoptera: Formicidae), an invasive tramp tem. Geogr. Biol. Tiere 8: 257-360. ant, in Biosphere 2. Florida Entomol. 82: 381-388. FOX, A. M., AND C. T. BRYSON. 1998. Wetland night- WETTERER, J. K., AND L. D. WOOD. 2001. Distribution shade (Solanum tampicense): a threat to wetlands in and impact of ants on a sea turtle nesting beach in the United States. Weed Technol. 12: 410-413. Palm Beach County, Florida. Proc. 21st Ann. Symp. HASKINS, C. P., AND E. F. HASKINS. 1965. Pheidole mega- Sea Turtle Biol. Conserv. NOAA Tech. Mem. NMFS- cephala and Iridomyrmex humilis in Bermuda—equi- SEFSC, in press. librium or slow replacement? Ecology 46: 736-740. WHEELER, W. M. 1908. The ants of Porto Rico and the Vir- HORVITZ, C. C. 1997. The impact of natural distur- gin Islands. Bull. Am. Mus. Nat. Hist. 24: 117-158. bances, pp. 63-74. In D. Simberloff, D. C. Schmitz, WHEELER, W. M. 1932. A list of the ants of Florida with and T. C. Brown [eds.], Strangers in Paradise. Impact descriptions of new forms. J. New York Entomol. Soc. and management of nonindigenous species in Flor- 40: 1-17. ida. Island Press, Washington, D.C. 467 pp. WILSON, E. O. 1964. Ants of the Florida Keys. Breviora JOURDAN, H. 1997. Threats on Pacific islands: the spread 210: 1-14. of the tramp ant Wasmannia auropunctata (Hymenop- ZIMMERMAN, E. C. 1948. Insects of Hawaii. Volume 1. tera: Formicidae). Pacific Conserv. Biol. 3: 61-64. Introduction. Univ. Hawaii Press, Honolulu, HI.

308 Florida Entomologist 85(2) June 2002

INSECT POLLINATORS OF THREE RARE PLANTS IN A FLORIDA LONGLEAF PINE FOREST

THERESA L. PITTS-SINGER1, JAMES L. HANULA1 AND JOAN L. WALKER2 1U.S.D.A. Forest Service, Forestry Sciences Laboratory, 320 Green St., Athens, GA 30602

2U.S.D.A. Forest Service, Department of Forest Resources, Clemson University, Clemson, SC 29634

ABSTRACT

As a result of human activity, longleaf pine (Pinus palustris Miller) forests in the southern United States have been lost or drastically altered. Many of the plant species that histori- cally occupied those forests now persist only as remnants and are classified as threatened or endangered. In order to safeguard such species, a better understanding of their pollination ecology is needed. We identified visitors and potential pollinators of Harperocallis flava (McDaniel) (Amaryllidaceae), Macbridea alba Chapman (Lamiaceae) and Scutellaria floridana Chapman (Lamiaceae) that occur in longleaf pine habitat on the Apalachicola Na- tional Forest in Florida. We observed that potential pollinators of H. flava were Halictidae, of M. alba were bumble bees (Apidae: Bombus), and of S. floridana were Megachilidae and Halictidae. However, the rates at which these insects visited the flowers were very low. Our results raise important concerns about how forest management practices affect the survival of rare plants, as well as their pollinators.

Key Words: Harperocallis flava (McDaniel), Macbridea alba Chapman, Scutellaria floridana Chapman, Pinus palustris Miller, threatened species, endangered species

RESUMEN

Como resultado de la actividad humana, los bosques de pino de hoja larga (Pinus palustris Miller) del sureste de los Estados Unidos han desaparecido o han sido drasticamente altera- dos. Muchas de las especies de plantas que historicamente ocupaban estos bosques persisten en la actualidad como restos y estan clasificadas como amenazadas o en peligro de extincíon. Para salvaguardar estas especies es necesitario entender la ecologia de su polinizacíon. Para ello, identificamos los insectos visitadores y polinizadores potenciales de Harperocallis flava (McDaniel) (Amaryllidaceae), Macbridea alba Chapman (Lamiaceae) y Scutellaria flori- diana Chapman (Lamiaceae) que existen en el habitat del pino de hoja larga del Bosque Na- cional de Apalachicola en Florida. Observamos que los polinizadores potenciales de H. flava eran Halictidae, de M. alba eran abejorros (Apidae: Bombus), y de S. floridiana eran Mega- chilidae y Halictidae. Sin embargo, los niveles de visitacion de estos insectos a las flores era muy bajo. Nuestros resultados crean dudas importantes acerca de como practicas de mante- nimiento de los bosques afectan a la supervivencia de plantas amenazadas, asi como a sus polinizadores.

The longleaf pine ecosystem is a conservation ical and ecological requirements of these plants in priority area within the U.S. Department of Agri- order to develop appropriate recovery plans. culture Conservation Reserve Program (Food Se- Studies of potential management effects on rare curity Act of 1985, Title XII). Longleaf pine (Pinus plants in some fire-disturbed habitats have consid- palustris Miller) forests once occupied >24 million ered the response of plants to management prac- ha in the southern United States. Today, <1.3 mil- tices e.g., Hessl & Spackman 1995; Brewer 1999; lion ha remain as small isolated parcels (Outcalt Lesica 1999), but not the effects on their pollinator & Sheffield 1996). The diversity of groundcover systems. One reason for this lack of information is plants per unit area (e.g., 140 vascular plant spe- simply that the effective pollinators are unknown. cies/1000 m2 in mesic longleaf woodlands) illus- Pollinators are critical to the long-term survival of trates the remarkable species richness of longleaf many flowering plants because they provide a pine ecosystems (Peet & Allard 1993). At least 30 mechanism for ensuring seed set and develop- endangered or threatened plant species now re- ment, and often facilitate gene flow between plants side in the few remnant understory communities and plant populations. In return for pollination of longleaf pine forest, and populations of at least services, the flowers provide vital floral resources 191 taxa of vascular plants have been reduced to for the foraging insects (Proctor et al. 1996). low levels (Hardin & White 1989; Walker 1993). A worldwide pollination crisis is at hand, and Thus, it is important to understand the physiolog- environmental degradation from habitat destruc-

Pitts-Singer et al.:Pollinators of Rare Plants 309 tion, modification, or fragmentation can disrupt during the growing season and usually occurs plant-pollinator interactions and jeopardize their from May to July (U.S. Fish & Wildlife Service existence (Rathcke & Jules 1993; Kearns et al. 1992, Madsen 1999). Some information available 1998). Tepedino et al. (1997) express a need for concerning this flower species relies on insect pol- “extended care” in conservation. When rare linators. Madsen (1999) found that when insects plants are imperiled, their extended families of were excluded from M. alba flowers, almost no pollinators also must be considered. It is impor- seeds were produced. tant to “maintain the integrity of ecosystems by Scutellaria floridana was also placed on the preserving interactions between plants and their federal threatened species list in 1992 (U.S. Fish pollinators” (Tepedino et al. 1997). & Wildlife Service 1992). It is found in Franklin, Our study investigates the pollinator-plant Liberty, and Gulf Counties, Florida. The stem is relationships of Harper’s beauty, Harperocallis simple or sparingly branched, and its solitary flava (McDaniel) (Amaryllidaceae), white birds- purple flowers are well separated in the axils of in-a-nest, Macbridea alba Chapman (Lamiaceae), short, leafy bracts. The corolla (2.5 cm in length) and Florida skullcap, Scutellaria floridana Chap- has 2 lips, the lower one being white in the mid- man (Lamiaceae). All of these plants are federally dle. The preferred habitat of S. floridana is simi- listed species and are endemic to the longleaf pine lar to that of M. alba, although it is more ecosystem in the Apalachicola lowlands of the restricted (Kral 1983; U.S. Fish & Wildlife Service Florida panhandle (Kral 1983; Walker 1993). They 1992). This perennial herb is reported to flower in are fire-dependent species (Kral 1983; Walker May or June (Kral 1983). 1993), and the local USDA Forest Service uses We monitored visitors to flowers of prescribed fires to help maintain their habitats. these plants, identified them, and evaluated their For H. flava pollination, insects may not be as importance as pollinators. The results of our important as for the other species in this study, if study are an important first step towards under- they are important at all. Allozyme studies have standing the interactions between these rare indicated that H. flava individuals and popula- plants and their pollinators in this Florida tions are genetically uniform (Godt et al. 1997). longleaf pine ecosystem. Further, a preliminary study by Wagner & Spira (1996) indicated that H. flava flowers are self- MATERIALS AND METHOD compatible and capable of selfing. Mature fruits were produced from flowers that were open-polli- All study sites were located on the Apalachi- nated, cross-pollinated, and self-pollinated. cola National Forest near Sumatra and Wilma, Harperocallis flava was first listed as an en- Florida. Field observations were made to deter- dangered species in 1979 (U.S. Fish & Wildlife mine which insects frequently visited flowers and Service 1992). It occurs in Franklin and Liberty how they behaved. In the summers of 1999 and Counties, Florida, in open pineland bogs and 2000, we selected and tagged 10-25 flowers (or in- along moist roadside ditches. This perennial spe- florescences) at each site, and then monitored the cies has a single yellow flower with 6 tepals (each flowers for 3- or 5-min periods throughout the day being 9-15 mm in length) produced atop a stalk for 2-5 days. Arthropod activity and the time they that emerges from stiff, grasslike leaves. Flower- were present on the flowers were recorded during ing occurs from mid-April through May and fruits the observation period. The visits of to mature in July. Kral (1983) reported that the den- flowers were counted only when they occurred sity of H. flava has declined since its discovery in during the observation period for that flower, al- 1965 primarily because of lack of fire in the area. though visits to other tagged flowers in the vicin- Presently, the USDA Forest Service manages a ity could be seen during this same time. Franklin County location and carries out periodic Observations were not made at the same time controlled burns to help maintain the open habi- each day. Because the 1999 sites did not produce tat required by this species (U.S. Fish & Wildlife enough flowers for study in 2000, sites chosen in Service 1992). 2000 were not the same as those used in 1999. In Macbridea alba was first listed as a federally summer 2000, we also used a video camcorder to threatened species in 1992 (U.S. Fish & Wildlife record insect activity on focal flowers. Service 1992). It occurs in open savannahs as well The video equipment consisted of a tripod- as in drainage areas in pine stands in Bay, Gulf, mounted Sony CCD-VX3 Video Recorder that was Franklin, and Liberty Counties, Florida (Kral auto-controlled via a Dell Latitude Xpi Laptop 1983; U.S. Fish & Wildlife Service 1992). This pe- computer with custom-developed “VideoSpy” soft- rennial herb usually has only one stem, which ware (Mark Evans, John Deere Commercial Prod- may be branched. Brilliant white flowers are clus- ucts, Inc., unpublished). The custom software tered in terminal compressed thyrses on erect allowed us to set a filming schedule for each day stems. Each flower has a green calyx (1 cm in using “record” times (3 or 5 min) and “wait” times length) and a white, 2-lipped corolla (3 cm in (10-15 min) so that up to 2 h of video could be re- length). Flowering of M. alba is stimulated by fire corded throughout the day. The system was pow-

310 Florida Entomologist 85(2) June 2002

ered by a 12-volt marine deep cycle battery. Video- tention on one or two anthers for pollen collection, imaging allows pollinator visits to be reviewed re- occasionally crawling across the centrally located peatedly or in slow-motion. Furthermore, this stigma. Duration of visits by the halictids ranged technique eliminates any effect of human pres- from 30 s to 7 min on a single flower. ence (e.g., scent and motion) on the behavior of In summer 2000 at a different site located ap- the visitors. From both researcher and video mon- proximately 5.1 km from the 1999 site, an ob- itoring, we recorded the identity of a visitor, the server monitored 12 H. flava flowers on May 16, duration of its visit, and its behavior on a flower. and 13 flowers on May 17, for a total of 6 h 15 min From field observations, we identified insects (range 0830-1656 h EDT). Observations were that were potentially pollinators, herbivores, nec- made for 5-min periods (instead of 3-min). The tar robbers, or incidental visitors. We also deter- video camcorder recorded 3 h 56 min of videotape mined the time of day potential pollinators were over the same 2 days noted above. Four visits (2 present. Visitation rates were calculated from data by halictids, 1 by a mordellid, and 1 by a syrphid) recorded by the field observer while watching focal were recorded on one flower using the video cam- flowers and also from data recorded on individual corder. Arthropods that were recorded visiting flowers by the camcorder. Rate of visitation (visits H. flava by both the human observer and by video per flower per min) was calculated as number of camcorder are shown in Table 1. Again, halictid arthropod visits to flowers per number of flowers bees were the only insects that gathered pollen observed per total observation time. When flowers from the flowers (Fig. 1), and their visits lasted were observed over more than one day, a visitation from 1 s to 10 min (i.e., some visits exceeded the 5- rate was calculated for each day, and then an aver- min period). Halictid visits occurred throughout age rate for those days was determined. the day (0945-1345 h EDT), but not in the early On two separate occasions in 1999, a halictid morning or late afternoon (Fig. 1). Another small bee was collected after it had visited an H. flava yellow flower, Xyris baldwiniana Roemer & flower. Each bee was washed in a vial containing Schultes (Xyridaceae), was also present at this deionized water to remove pollen from its body, H. flava site and was visited by halictid bees. and then it was released unharmed. The pollen Xyris baldwiniana flowers were only open during was cleaned and slide-mounted according to a po- the morning, and their presence could have af- tassium hydroxide-acetolysis procedure provided fected the rate at which bees visited H. flava. No by Jean Porter at the University of Georgia Paleo- halictids were collected in 2000 for identification ecology Laboratory, Athens, Georgia (see also Erdt- to the generic level. man 1969). Jean Porter also prepared a reference In both 1999 and 2000, average insect visita- slide of H. flava pollen from pollen extracted from tion rates to H. flava were very low (Table 2). a specimen in the University of Georgia Herbar- Based on human observations, the rate was ium. We used the reference slide for determining higher in 2000 than in 1999. Rates from the video if H. flava pollen was present on the slides we camcorder used in 2000 on a focal flower were made from field collections. higher than that recorded by the human observer.

RESULTS Macbridea alba

Harperocallis flava Twenty inflorescences of M. alba at each of two sites (distance between sites = approx. 4.4 km) In 1999, 25 H. flava flowers at one roadside were observed for 34 h over 5 days (June 24-25, study site and 20 flowers at a nearby site (1 km June 29-July 1; range 0724-1805 h EDT). Nine in- north of other site, approximately 20-40 m off the sect and spider species made 70 visits (Table 1), road) were tagged and then monitored at 3-min but only bumblebees (Bombus) were large enough periods for a total of 20 h of observation over 3 to contact the reproductive structures of the days (May 18-20; range 0830-1555 h EDT). Al- flower. No bumblebees were collected in order to though many bees, wasps, and beetles were seen determine their specific scientific identity during on nearby Ilex glabra L. (Aquifoliaceae), Hyperi- this study. Bumblebee visits to each flower were cum sp. (Clusiaceae), and Iris sp. (Iridaceae), only very short (1-9 s), but they usually visited more 5 different insect species visited H. flava (Table 1), than one flower per inflorescence (Fig. 2) and vis- resulting in 8 floral visits. Of all the insect visitors, ited throughout the day (0800-1645 h EDT). The halictid bees (n = 3) were the only ones that spent bees landed on the lower lip (petal) of the flower time on the flowers gathering pollen (Fig. 1). One and immediately pushed their heads down into of the halictids was collected, and later identified the corolla, presumably to obtain nectar. With its by T. L. Pitts-Singer as Dialictus sp. (using key in head in the flower corolla, the bee’s thorax con- Mitchell 1960). The specimen was retained for ref- tacted anthers and stigma. Occasionally, just be- erence at the U.S. Department of Agriculture For- fore leaving the flower, the bee appeared to probe est Service, Forestry Sciences Laboratory, Athens, the anthers with its proboscis for ~1 s. The purpose Georgia. While on a flower, each bee focused its at- of the latter behavior is not clear. Overall, visita-

Pitts-Singer et al.:Pollinators of Rare Plants 311

TABLE 1. LIST OF VISITORS TO MARKED FLOWERS OF H. FLAVA, M. ALBA AND S. FLORIDANA IN SUMMER 1999 AND 2000 AT THE APALACHICOLA NATIONAL FOREST, FLORIDA, AND TOTAL OBSERVATION TIME AND NUMBER OF VISITS.

Plant species; observation time and no. visits Visitor scientific name Common name

H. flava 1999 1999 1999 obs. time = 20 h Halictidae: Dialictus* sweat bee visits = 8 Apidae: Bombus bumble bee Mordellidae tumbling flower beetle Tettigoniidae katydid nymph Diptera fly 2000 2000 2000 human obs. = 8 h 25 min Halictidae: sp. #1* sweat bee video = 3 h 56 min Halictidae: sp. #2* sweat bee visits = 8 Mordellidae tumbling flower beetle Syrphidae bee fly M. alba 1999 1999 1999 obs. time = 34 h Apidae: Bombus* bumble bee visits = 70 Formicidae: Paratrechina ant Curculionidae weevil Melandryidae false darkling beetle Lycaenidae gossamer-winged butterfly Tettigoniidae katydid Oxyopidae lynx spider Thomisidae crab spider S. floridana 1999 1999 1999 obs. time = 6 h Apidae: Bombus* bumblebee visits = 9 Megachilidae* leafcutter bee Halictidae: Dialictus sweat bee Halictidae?: sp. #2 bee Syrphidae? syrphid fly Acrididae grasshopper Thomisidae crab spider S. floridana 2000 2000 2000 human obs. = 6 h Megachilidae* leafcutter bee video = 2 h 43 min Halictidae* sweat bee visits = 10 Tettigoniidae small katydid Thomisidae crab spider

*Indicates insects that were seen to collect pollen and are probably important pollinators. ?Researcher is not positive of family-level identification.

tion rates were higher for M. alba than for H. flava Scutellaria floridana (Table 2). Bumble bees also visited Rhexia alifa- nus Walter (Melastomaceae) and Hibiscus aculea- In 1999, 10 inflorescences of S. floridana at one tus Walter (Malvaceae) that occurred at the same site were monitored over 2 days (September 14- sites. The bees spent more time collecting pollen 15; range 0825-1632 h EDT) for 6 h of observa- on these common flowers than they did on M. alba tion, during which 7 insect and spider species collecting nectar. made 9 visits (Table 1). Visitations occurred be- In 2000, very few M. alba flowers were pro- tween 1000 and 1530 h (Fig. 3). Of the visitors duced, presumably because of drought and other recorded during observation periods, megachilid unfavorable environmental conditions. Thus, we bees and possibly halictid bees displayed behav- were unable to make further observations of visi- ior that may have resulted in pollination of S. tors to this flower species. floridana flowers (Fig. 3). Megachilids landed on 312 Florida Entomologist 85(2) June 2002

Fig. 1. Visitation by halictids on H. flava flowers recorded by human observer and by video camcorder in May 1999 and 2000 at the Apalachicola National Forest, Florida. Bars represent time frame in which 3- or 5-min obser- vation periods occurred. Symbols represent pollinator visits. the flower, clung to the flower hood in an upside- they appeared to collect only nectar and did not down position, and then kicked into the hood contact the anthers. Compared to the other 2 through the seam so that pollen fell onto the plants in this study, the overall visitation rate scopa on the venter of the gaster. Durations of was high (Table 2). megachilid visitations were 21-23 s. Although In 2000 at a site located approximately 16.25 halictids (Dialictus sp.) also visited these flowers km from the 1999 site, 10 flowers of S. floridana (Fig. 3), they did not appear to come into contact were monitored by a human observer for 6 h on with the stigma and, therefore, may have been April 20-21 (range 0850-1754 h EDT) in 2000. The pollen “robbers.” The small halictids entered into video camcorder captured 2 h 43 min of tape, re- the hood of the flower, out of the observer’s view. vealing visits by 4 megachilids and 1 halictid. Al- The halictids presumably collected pollen before together, 4 insect species made 10 visits to flowers exiting the flower. Their visits lasted from 2 s to 1 (Table 1). Megachilids (visit duration = 10-15 s) min. In addition to the megachilid and halictid and halictids (visit duration = 5-31 s) were the bees, the observer noted that one bumblebee vis- most frequent visitors to S. floridana (Fig. 3). Be- ited a flower before the study began (Fig. 3). This havioral observations of megachilids and halic- bee spent a few seconds on a flower and used flo- tids, which were larger halictids than those seen ral sonication to dislodge pollen from the anthers. in 1999, indicated that these bees probably are ef- A few (not on marked flowers) and a fective pollinators of S. floridana. These bees fly also were observed on the flowers, although clung to the outside of the flower hood, sometimes Pitts-Singer et al.:Pollinators of Rare Plants 313

TABLE 2. AVERAGE VISITATION RATES (ARTHROPOD VISITS PER FLOWER PER MIN) AND NUMBER OF VISITS (N) OVER 1- 5 DAYS OF OBSERVATIONS OF MARKED FLOWERS AT THE APALACHICOLA NATIONAL FOREST, FLORIDA IN SUM- MERS OF 1999 AND 2000.

Visitation rate (× 10-2)

Flower species: visitors—method 1999 2000

H. flava: halictids—observer 0.01 (n = 3) 0.06 (n = 3) H. flava: all visitors—observer 0.03 (n = 8) 0.06 (n = 3) H. flava: halictids—video n.a. 0.85 (n = 2) H. flava: all visitors—video n.a. 2.1 (n = 5) M. alba: bumblebees—observer 0.07 (n = 21) n.a. M. alba: all visitors—observer 0.18 (n = 70) n.a. S. floridana: bees*—observer 0.14 (n = 5) 0.06 (n = 3) S. floridana: all visitors—observer 0.25 (n = 9) 0.15 (n = 5) S. floridana: bees*—video n.a. 3.72 (n = 5) S. floridana: all visitors—video n.a. 3.72 (n = 5)

*“Bees” includes megachilids and halictids.

touching the stigma, as they forced their heads U.S.D.A. Forest Service, Bristol, FL, pers. comm.), and thoraces into the hood. Pollinator visitations we observed no honeybees on these flowers. occurred from morning until late afternoon (Fig. Arthropod visits to the rare flowers were quite 3). Visitation rates were lower in 2000 than they infrequent (Table 2) (Figs. 1-3). Unfortunately, we were in 1999. Carpenter bees (Xylocopa sp.) were found no other pollinator visitation data in the lit- seen to rob nectar by piercing the base of the co- erature for flowers in the Apalachicola National rolla. Other flowers such as Aletris lutea Small Forest or vicinity for making comparisons. How- (Liliaceae), Balduina uniflora Nuttall (Aster- ever, pollinator studies performed on flowers from aceae), and Pityopsis graminifollia (Michaux) other habitats may offer an idea of potential bee (Asteraceae) were also at this site, attracting visitation rates. For example, a study on Hibiscus bumblebees and butterflies. moscheutos in Maryland revealed average visita- tions by bumblebees and anthophorid bees of 27- Pollen from Halictid Bees 13 visits per flower per min × 10-2 (Spira et al. 1992). Another study in Michigan on catnip, We found pollen on only 1 of the 2 slides pre- Nepeta cataria L., found average visitations by pared from washes of the 2 halictid bees captured bumblebees to be 8.6 visits per flower per min × 10- on H. flava flowers. There were 7 pollen grains of 2 and by Halictidae to be 7.4 visits per flower per H. flava, 1 grain of 3 other types, and 3 grains of min × 10-2 (Sih & Baltus 1987). Compared to these a fourth type. The absence of pollen on the second studies, bee visitation rates to the flowers we stud- slide does not preclude that pollen was not ied are indeed quite low (range = 0.01 × 10-2 to 3.72 present on the bee. The pollen may have been lost × 10-2 per min) (Table 2). However, we saw these in the insect net during collection, the water wash same insects visiting other flowers in the area. may have not removed pollen from the bee, or Regardless of whether or not these rare flow- maybe the pollen was lost in the process of ex- ers are imperiled because of low pollinator fre- tracting pollen out of water in the vial. quency, our studies showed that certain insects collect pollen and nectar from them and may be DISCUSSION important in their pollination ecology. Bumble- bees were the only insects observed whose size We found that native bees were the likely pol- and behavior was efficient for pollinating M. alba. linators of all 3 of the threatened or endangered Observations of M. alba flowers in the South flowers we studied in the Apalachicola longleaf Carolina Botanical Garden suggest that bumble- pine forest. Bumblebees are probably important bees are very frequent visitors and efficient polli- pollinators of M. alba and possibly of S. floridana, nators (J. L. Walker, pers. obs.). Continued studies but megachilids and halictids may be the primary of pollinators of this flower species should deter- pollinators of S. floridana. Halictids may play a mine how many individual bumble bees are visit- significant role in the pollination system of ing the rare flowers or other flowers in the area H. flava. Although honeybee (Apis mellifera L.) and if these bees are important for out-crossing in hives are present near the forest (Louise Kirn, this species. 314 Florida Entomologist 85(2) June 2002

Fig. 2. Visitation by bumblebees on M. alba inflorescences recorded by human observer in June-July 1999 at the Apalachicola National Forest, Florida. Bars represent time frame in which 3-min observation periods occurred. Symbols represent pollinator visits.

Although some evidence has indicated that noted that the newly opened flowers were more H. flava flowers are self-compatible and undergo readily approached and subsequently visited selfing, we observed several halictid bees collect- than 2-day-old flowers. In 2000, the flowers ing pollen from H. flava flowers, especially in 2000 bloomed at the beginning of the summer (May) (Tables 1 and 2) (Fig. 1). The pollen-collecting be- and not at the end of the summer (September) as havior of halictids on H. flava flowers could have in 1999. These different flowering dates may have facilitated pollination. For a hermaphroditic spe- had an effect on the diversity and abundance of cies like H. flava, visitation by bees may improve insects that visited the flowers. Further investi- out-crossing; or the activity of bees on anthers gation of pollinators of this species should include may dislodge pollen for better wind dispersal insect exclusion experiments on S. floridana and (Cane et al. 1992). Nonetheless, if out-crossing comparative studies with another common skull- were evolutionarily important for this species in cap, S. integrifolia L., which occurs in the area. the past, it may now be ineffectual as a result of Another useful outcome from this study is in the low genetic variability in H. flava. Though pol- the methodology. Using the video camcorder to linator services may not benefit H. flava, the collect visitation data may have had an advan- availability of this pollen may be a useful resource tage over human observation (Table 2). By focus- for the solitary bees that harvest it. ing on only 1 flower throughout the day, up to 2 Our study provides the first documentation of hours of observable data could be gathered with- the pollination ecology of S. floridana. The behav- out human intervention. A higher visitation rate iors of megachilids and halictid bees on the flow- was recorded for both H. flava and S. floridana ers were appropriate for pollination. We also when this method was used. Pitts-Singer et al.:Pollinators of Rare Plants 315

Fig. 3. Visitation by bumblebees, megachilids, and halictids on S. floridana flowers recorded by human observer and video camcorder in September 1999 and April 2000 at the Apalachicola National Forest, Florida. Bars repre- sent time frame in which 3-min observation periods occurred. Symbols represent pollinator visits.

Early in the study, we planned to collect in- available. Thus, we must also understand the life sects that visited the rare flowers in order to cre- cycle of important pollinators, and, further, ate a reference collection and to identify pollen whether fire has an effect on the availability of samples from the insects. However, because visi- food and nesting materials. tation rates were low, we decided that removal of At least three requirements must be met for ef- potential pollinators might have been detrimen- fective conservation of pollinators in any ecosys- tal to the reproductive success of the plants, and tem and of bee pollinators in the Apalachicola we abandoned this effort. ecosystem. They require proper nesting sites, Each of the plants we studied responds to fire nest-building material, and a sufficient amount of by increased growth and flowering (Madsen 1999; food at appropriate times both for adults (who Louise Kirn, USDA Forest Service, Apalachicola need nectar) and for larvae (who need pollen) Ranger Station, Bristol, FL, pers. comm.). This (Westrich 1996). Thus, it is imperative to identify response suggests that fire timing is important to the specialized needs of each pollinator. Such in- ensure that flowering occurs when pollinators are formation will help in determining which, if any, 316 Florida Entomologist 85(2) June 2002

forest management practices affect availability of KEARNS, C. A., D. W. INOUYE, AND N. M. WASER. 1998. these resources; or if improvements can be made Endangered mutualisms: the conservation of plant- in certain areas to maintain suitable habitat. pollinator interactions. Ann. Rev. Ecol. Syst. 29: 83- The pollination ecology of some communities 112. has been neglected, and more information concern- KEVAN, P. G. 1975. Pollination and environmental con- servation. Environ. Conserv. 2: 293-298. ing the plant-pollinator interactions of rare and KEVAN, P. G., E. A. TIKHMENEV, AND M. USUI. 1993. In- endangered plants is needed (Kevan 1975; Kevan sects and plants in the pollination ecology of the bo- et al. 1993; Buchmann & Nabhan 1996; Kearns et real zone. Ecol. Res. 8: 247-267. al. 1998). Researchers must first have basic knowl- KRAL, R. 1983. A report on some rare, threatened, or en- edge of a system before making hypotheses at an dangered forest-related vasular plants of the south. ecosystem level and designing experiments to test U.S.D.A. Forest Service Tech. Pub. R8-TP, Vols. 1&2. them. Researchers must perform the necessary KWAK, M. M., O. VELTEROP, AND E. J. M. BOERRIGTER. studies to determine the importance of insect (and 1996. Insect diversity and the pollination of rare plant other animal) visitors in pollination (Kwak et al. species, pp. 116-124. In A. Matheson, S. L. Buch- mann, C. O’Toole, P. Westrich, and I. Williams [eds.], 1996). Conservation plans need to be backed by The Conservation of Bees. Academic Press, London. solid, scientific evidence. Our results are a starting LESICA, P. 1999. Effects of fire on the demography of the point for understanding insect pollinators of three endangered, geophytic herb Silene spaldingii rare plants in a longleaf pine ecosystem and pro- (Caryophyllaceae). Amer. J. Bot. 86(7): 996-1002. vide a basis for future examination and conserva- MADSEN, D. L. 1999. Seed production and germination tion of these plants and their pollinators. studies of Macbridea alba. Master’s Thesis, Clemson University. MITCHELL, T. B. 1960. Bees of the Eastern United ACKNOWLEDGMENTS States, Vol. I. North Carolina Agricultural Experi- We thank personnel of the Apalachicola National ment Station, Tech. Bul. No. 141. Forest Ranger Station for their support of this project, PEET, R. K., AND D. J. ALLARD. 1993. Longleaf pine- especially Louise Kirn (USDA Forest Service) who kept dominated vegetation of the southern Atlantic and us informed of when flowers were in bloom, guided us to eastern Gulf Coast region, USA. Proc. Tall Timbers field sites, and helped us identify various flowers at field Fire Ecol. Conf. #18, pp. 45-81. sites. David Jenkins (University of Georgia) was a valu- PROCTOR, M., YEO, P. AND A. LACK. 1996. The Natural able field and laboratory assistant in summer 1999, and History of Pollination: Timber press, Portland, OR. his observations and suggestions are much appreciated. OUTCALT, K. W. AND R. M. SHEFFIELD. 1996. The We thank Robert W. Matthews, Christopher J. Fettig, longleaf pine forest: trends and current conditions. and James P. Pitts (Department of Entomology, Univer- U.S. Forest Service Resource Bulletin SRS-RB-9. sity of Georgia, Athens, Georgia), Kerry O. Britton RATHCKE, B. J. AND E. S. JULES. 1993. Habitat fragmen- (USDA Forest Service, Athens, Georgia), and Paul tation and plant-pollinator interactions. Current Smith (USDA Forest Service, Asheville, North Caro- Science 65: 273-277. lina) and 3 anonymous reviewers for their critical as- SIH, A. AND M.-S. BALTUS. 1987. Patch size, pollinator sessments of this manuscript. behavior, and pollinator limitation in catnip. Ecol- ogy 68: 1679-1690. SPIRA, T. P., A. A. SNOW, D. F. WHIGHAM, AND J. LEAK. REFERENCES CITED 1992. Flower visitation, pollen deposition, and pol- BREWER, J. S. 1999. Effects of competition, litter, and len-tube competition in Hibiscus moscheutos (Mal- disturbance on an annual carnivorous plant (Utricu- vaceae). Am. J. Bot. 79: 428-433. laria juncea). Plant Ecol. 140: 159-165. TEPEDINO, V. T., S. D. SIPES, J. L. BARNES, AND L. L. BUCHMANN, S. L., AND G. P. NABHAN. 1996. The Forgot- HICKERSON. 1997. The need for “extended care” in ten Pollinators. Island Press/Shearwater Books: conservation: examples from studies of rare plants Washington, D.C. in the Western United States, pp. 245-248. In K.W. CANE, J. H., S. L. BUCHMANN, AND W. E. LABERGE. Richards [ed.], Seventh International Symposium on 1992. The solitary bee Melissodes thelypodii thelypo- Pollination, Acta Hort 437, ISHS. dii Cockerell (Hymenoptera: Anthophoridae) collects U.S. FISH AND WILDLIFE SERVICE. 1992. Endangered pollen from wind-pollinated Amaranthus palmeri and Threatened Species of the Southeastern United Watson. 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Effects of fire on European bees and the problems of partial habitats, threatened and endangered plants: an annotated bib- pp. 1-16. In A. Matheson, S. L. Buchmann, C. O’Toole, liography. Information and Technology Report 2. U.S. P. Westrich, and I. Williams [eds.], The Conserva- Department of the Interior, Washington, DC. 55 pp. tion of Bees. Academic Press, London.

Fedorka & Mousseau: Nuptial Gifts in a Cricket 317

TIBIAL SPUR FEEDING IN GROUND CRICKETS: LARGER MALES CONTRIBUTE LARGER GIFTS (: GRYLLIDAE)

KENNETH M. FEDORKA AND TIMOTHY A. MOUSSEAU Department of Biological Sciences, University of South Carolina, 700 Sumter Street, Columbia, SC 29208

ABSTRACT

Many male insects provide somatic nuptial gifts that may strongly influence reproductive fitness by insuring an effective copulation or by increasing paternal investment. In the striped ground cricket, Allonemobius socius (Scudder), females receive a nuptial gift by chewing on a specialized spur on the male’s hind tibia during copulation. Using a series of no-choice trials, we attempted to quantify gift magnitude and to determine the relationships between male size, gift contribution, and male mating success. Tibial spur chewing duration

was a significant predictor of gift contribution (F1,17 = 17.02, P < 0.001) and the magnitude of the gift ranged between 0.2% and 8% of the male’s body mass, implying that females receive mostly hemolymph. Large males produced bigger gifts than small males (2.52 ± 0.59 mg vs. ± 1.33 0.28 mg, t17 = 1.88, P < 0.05, respectively) and females were more likely to mate with

larger males (F1,39 = 4.76, P < 0.05). If gift size is shown to influence female reproductive fit- ness, then nuptial gifts may play a large role in the evolution of male body size.

Key Words: nuptial gift, tibial spur, body size, Allonemobius socius

RESUMEN

El macho en muchos insectos provee un regalo nupcial somático que puede influenciar fuer- temente la adaptabilidad óptima reproductiva (fitness) al asegurar una cópula eficaz o al in- crementar la inversión paternal. En el grillo Allonemobius socius, las hembras reciben un regalo nupcial al morder un espolón especializado en la tibia posterior del macho durante la cópula. Utilizando una serie de pruebas de no alternativas, tratamos de cuantificar la mag- nitud del regalo y determinar la relación entre el tamaño del macho, la contribución del re- galo, y el éxito de la cópula del macho. La duración de las mordidas del espolón de la tibia fué

un predictor significativo de la contribución del regalo (F1,17 - 17.02, P < 0.001) y la magnitud del regalo abarcó entre el 0.2% y el 8% del peso del cuerpo del macho, indicando que las hem- bras reciben principalmente hemolinfa. Los machos grandes produjeron regalos más grandes ± ± que los machos pequeños (2.52 0.59 mg vs 1.33 0.28 mg,t17 =1.88, P < 0.05) y era más pro- bable que las hembras copularan con los machos más grandes. Si el tamaño del regalo in- fluencia la adaptabilidad óptima reproductiva de las hembras, entonces los regalos nupciales pueden actuar un papel importante en la evolución del tamaño del cuerpo de los machos.

In many birds and insects, males offer nuptial rate (Green and Krebs 1995). Regardless of the gifts to females prior to and/or during copulation precise functional significance, these associations (Wiggins and Morris 1986; Gwynne and Brown suggest that variation in nuptial gift mass may 1994; Simmons 1995; Neuman et al. 1998). These have strong fitness implications for both sexes. offerings include captured prey, somatic tissue, Nuptial gifts are generally transferred to fe- synthesized secretions and suicidal food transfers males via external, intermediate packages such (see Andersson 1994 and references therein). The as prey items or self-contained somatic secretions function of these gifts may vary, serving as a form (Andersson, 1994), permitting an easy quantifica- of male mating effort by increasing fertilization tion of their mass. However, in some animal gen- success (Alexander & Borgia 1979; Sakaluk 1984) era the gift is internally transferred making the and/or as paternal investment by increasing repro- magnitude and the fitness implications of such ductive fitness (e.g., increasing egg size, offspring gifts difficult to assess. For instance, in the cricket number or offspring viability; Gwynne 1984; Rein- genus Allonemobius (Orthoptera: Gryllidae), hold 1999). For instance, in the hangingfly, Hylo- males internally deliver a nuptial gift through a bittacus apicalis (Byers), nuptial prey functions to specialized spur on each hind tibia that is chewed increase mating effort by increasing copulation du- by the female during copulation (Fig. 1). Although ration, which is in turn associated with the volume chewing damages the spur, males can mate mul- of sperm transferred (Thornhill 1976). In contrast, tiple times and females do not discriminate nuptial feeding in the osprey, Pandion haliaetus among males based on the spur’s condition (un- (Linnaeus), acts as paternal investment since it published data). This somatic gift has previously was positively associated with offspring growth been described as a limited glandular contribu-

318 Florida Entomologist 85(2) June 2002

Fig. 1. The tibial spur. This specialized spur delivers a somatic nuptial gift directly to the female through court- ship feeding. The spur on the right has been chewed by a female whereas the left is still intact indicating a male’s previous mating success. tion contained within the spur that is exuded once 1989). All crickets used in this experiment were the tip is removed (Fulton 1931; Mays 1971; For- second-generation lab-reared individuals origi- rest 1991). However, a preliminary study sug- nating from a single population near Asheville, gested that, when the spur tip is artificially North Carolina. Crickets were maintained in 10 × amputated and a capillary tube attached, the vol- 10 × 8 cm plastic cages containing ground cat ume of the extract exceeds the volume of the spur food, a carrot slice, water vial and strips of brown (unpublished data). Thus, the spur may provide paper towel for cover. Every three days the food, females with direct access to the male’s hemo- carrot and paper towel were replaced. Cages were lymph, making male mass a potential limiting kept in a constant environment at 28°C and a factor in gift contribution. This is important since 12:12 [L:D] photoperiod provided by a Percival in- the evolutionary trajectory of male body size and cubator. The age of all experimental crickets was sexual size dimorphism may be modified if posi- held constant at 12 ± 2 days post eclosion. tive selection on the gift exists, coupled with a ge- A. socius males perform a calling song used to netic correlation between body and gift size. attract distant females. Once a potential mate is Unfortunately, no mass measure of this or any encountered, males switch to a courtship song other internally transferred nuptial gift exists. and dance that culminates with the male orient- Using the striped ground cricket, Allonemobius ing his abdomen toward the stationary female. If socius (Scudder), we attempted to quantify the the female is receptive, she will briefly mount the magnitude of an internally transferred gift. We male in a “mock copulation” lasting only a few sec- predicted that male A. socius donate more than a onds. Once an effective mock copulation is limited glandular contribution, and may offer fe- achieved (this may take several attempts) the males a continuous supply of hemolymph. Fur- male will cease courting and begin to form a sper- thermore, we predicted that male contribution is matophore (approximately 20 min). When com- constrained by male mass, with the largest males plete, he will renew his courtship behavior, again capable of offering the largest gift. Therefore, fe- enticing the female to mount. At this time, the males should prefer large males if gift mass is pos- couple will adjoin abdomens as the male adheres itively related to male mass and to female fitness. the spermatophore to the female’s genitalia. The male will then bring his hind tibia forward allow- MATERIALS AND METHODS ing the female to chew on his spur until the couple separates (upwards of 30 min). Once apart, the fe- A. socius is a small chirping ground cricket male will remove and consume the spermato- found throughout the southeastern United phore. If a spermatophore is formed but not States, with closely related sister taxa ranging attached, no spur chewing will take place and the throughout North America (Alexander & Thomas spermatophore will be removed and consumed by 1959; Howard & Furth 1986; Mousseau & Roff the male (Alexander & Thomas 1959; Mays 1971).

Fedorka & Mousseau: Nuptial Gifts in a Cricket 319

Virgin males and females were randomly paired mating trial, t. A second independent estimate of

(n = 41 pairs), placed into a mating arena (6 cm di- the nuptial gift, G[f], was based on female mass ameter petri dish) and allowed to mate once. Al- gain and obtained by substituting the change in though crickets mate multiply in the wild (Walker female mass, (∆f), for (∆m) into equations one and 1980; Sakaluk and Cade 1983), only one mating two and dropping s. All data analyses were per- bout is needed to detect the mass changes used in formed using SAS 8.1. estimating the size of the nuptial gift. Two groups were created depending on the outcome of the mat- RESULTS ing trial. Males who attached their spermatophore to the female were grouped as successful. Males Since the ejaculate contained within the sper- who formed a spermatophore, but failed to re-at- matophore that is transferred directly to the fe- tract a female for copulation were grouped as un- male genitalia may confound our estimate of gift successful. In both cases, spermatophores were mass, we compared the mass of the successful collected after removal to avoid consumption. male’s depleted spermatophore (mean ± se: 1.03 ± Male, female and petri dish mass were recorded be- 0.01 mg; n = 19) with the unsuccessful male’s in- fore and after each mating trial. The mass gain in tact spermatophore (0.99 ± 0.07 mg; n = 22) to es- the petri dish gave us a measure of mass loss due to timate ejaculate contribution. No difference defecation. In addition, spermatophore mass, trial existed between these two groups (one tailed t- duration, spermatophore attachment duration, tib- test: t39 = -0.93, NS), suggesting that the ejaculate ial spur chewing duration, and post-chewing sper- mass was negligible. There was no effect of male matophore attachment duration (i.e., the amount body size on spermatophore mass (F = 2.07, NS) of time it takes the female to remove the spermato- 1,39 or estimated respiratory mass loss (F1,17 = 0.17, phore after chewing the tibial spur) were recorded. NS). Average male mass was 77.55 ± 1.24 mg and Since by nature of the mating ritual, a positive as- ± the average G[m] was 1.89 0.33 mg. Thus, the av- sociation between chewing duration and spermato- erage nuptial gift was approximately 2.44% of the phore attachment seems inherent, post-chewing male’s initial body mass. When coupled with aver- spermatophore attachment duration was exam- age spermatophore size (1.03 ± 0.01 mg) males in- ined to assess whether larger gifts satiate females vested approximately 3.77% of their total mass to where they will postpone removal and consump- into a single copulation. This is a large invest- tion of the spermatophore. Mass was measured us- ment considering that males are promiscuous ing a Sartorius scale (Bohemia, NY) accurate to and that a substantial proportion of their mass is 0.01 mg. Trial duration was defined as the time attributable to exoskeleton. elapsed from the onset of male courtship to the re- When we replaced ∆m with ∆f in equations one moval of the spermatophore by the female (suc- ± and two, average G[f] was estimated to be 1.01 cessful) or the male (unsuccessful). 0.45 mg, providing a second, independent mea- The mass of the gift contribution was esti- sure of gift size. Although this estimate is lower mated for each successful trial by taking the total than the male estimate, they are not significantly change in male mass (∆m) and subtracting out different (two tailed t-test: t36 = 1.53, NS). Six G[f] the mass lost to defecation (∆p), spermatophore estimates were negative (Fig. 2), and were most mass (s), and respiration (r), such that, likely the result of the error inherent in the meth- odology used to estimate the mass lost to respira- tion, r. All six negative G estimates were well G = ∆m – ()∆p ⁄ 2 – s – r (eq. 1) [f] []m within one standard deviation in r away from a positive estimate. As a consequence, our estimate of the average G is conservative. The term ∆p was halved since we assumed that [f] the rate of defecation was equal between the Since no difference existed between the suc- sexes. To estimate r, we first estimated the aver- cessful and unsuccessful groups with regard to spermatophore mass (s), mass lost to defecation age mass lost per minute, R, from the unsuccess- ± ± ful male data while controlling for defecation and (0.58 0.02 mg and 0.67 0.02 mg, respectively; ∆p two tailed t-test: t = 0.52, NS), and the dura- spermatophore production, 39 tion of the trial (two tailed t-test: t39 = -0.72, NS), n we calculated two additional estimates of gift mass, G and G , using the least squares means ()∆ ()∆ ⁄ ⁄ [lm] [lf] ∑ m – p 2 – s t ∆ ∆ (eq. 2) difference in m and f between the two groups, R = ------1 - respectively. These estimates were calculated be- n cause they are free of the error attributed to esti- mating mass lost to respiration and defecation. where t is the duration of the trial. We assumed Since initial male mass was significantly associ- ∆ that R was equal for successful and unsuccessful ated with m (F1,39 = 4.41, P < 0.05), we used an males. Thus, for each successful trial, r was esti- analysis of covariance with initial male mass as

mated by multiplying R by the duration of the the covariate (ANCOVA: F2,38 = 14.75, P < 0.001;

320 Florida Entomologist 85(2) June 2002

Chewing duration covaried with both the male

estimate (Fig. 2a; G[m]: F1,17 = 17.02, P < 0.001) and

the female estimate (Fig. 2b; G[f]: F1,17 = 6.67, P < 0.05) of the nuptial gift implying that females who chewed longer received a larger gift contribu- tion. The size of the gift ranged from 0.2% to 8% of initial male mass. Although, the rate of male gift contribution and female gift gain (Fig. 2) were not

significantly different (F1,16 = 2.12, NS), G[f] ex-

ceeded G[m] in four trials. In three of these trials the differences were small and, as with the nega-

tive estimates of G[f], fell well within 1 standard deviation of our estimate of r. The remaining trial discrepancy showed female mass gain to be 3.94 mg greater than the male contribution, and can- not be reconciled with the previous argument suggesting that it may be the result of measure- ment error. However, when this observation was removed, the relationship between chewing dura-

tion and G[m] and G[f] remained unchanged (F1,17 2 =15.68, P < 0.001, R = 0.50 and F1,17 = 10.21, P < 0.01, R2 = 0.40, respectively). In addition, male mass was significantly asso-

ciated with spur chewing duration (Fig. 3: F1,17 = 4.24, P = 0.05) implying that larger males provide a larger gift. In turn, spur chewing duration was significantly associated with spermatophore at-

tachment duration (F1,17 = 4.51, P < 0.05), which is expected since the spermatophore is attached at the onset of spur chewing. However, chewing du- ration was not associated with post-chewing sper-

matophore attachment duration (F1,17 = 0.06, NS), implying that females do not delay removal of the spermatophore if a larger gift is received.

Fig. 2. Nuptial gift contribution as a function of spur chewing duration. A) As females chewed on the tibial spur, male gift contribution increased (y = 0.1496x + 0.2444, R2 = 0.50). B) The relationship between female mass gain and courtship feeding also provided a rate of male contribution (y = 0.1547x - 0.6922, R2 = 0.28). Both male (G[m]) and female (G[f]) estimates of the nuptial gift were controlled for mass lost to defecation, respiration and spermatophore production. The arrows indicate a potential outlier in the female G[f] estimate and the cor- responding G[m] estimate.

no interaction between covariate and group). Us- ing this method G[lm] was estimated to be 1.75 mg.

Likewise, G[lf] was estimated to be 1.28 mg. Con- sidering that the tibial spur weighs less than 0.001 mg, these estimates suggest that the gift far exceeds the capacity of the spur. Moreover, these Fig. 3. Spur chewing duration as a function of male gift estimates exceed the weight of the entire male mass. Large males were chewed longer than small tibia (0.75 ± 0.05 mg), implying that the gift is males (y = 564.18x - 35.419, R2 = 0.20) suggesting that comprised mostly or entirely of hemolymph. male size may ultimately constrain gift size. Fedorka & Mousseau: Nuptial Gifts in a Cricket 321

Successful males were significantly larger luk 1998; Reinhold 1999). However, an associa-

than unsuccessful males (F1,39 = 4.76, P < 0.05). tion between male mass and gift mass is not This may be the result of female choice or a me- (Wedell 1997), though an association between chanical constraint of small male size. In crickets, male mass and a spermatophylax gift/sperm am- copulation may be impeded if the size difference pulla complex has also been shown in some spe- of a mating pair is too great, making proper geni- cies (Gwynne 1982; Sakaluk 1985). To elucidate talic alignment and spermatophore transfer diffi- the implications of the nuptial gift on male size cult (Sakaluk, pers. comm.). However, there was evolution in this system, we are presently exam- no distinction between successful and unsuccess- ining the impact of gift mass on female fitness ful males with regard to male and female size dif- along with the selective pressures and underlying

ferences (F1,39 = 1.28, NS), suggesting that large genetic architecture surrounding these traits. male success was not due to small male inability Currently, it is unknown whether gift mass is in passing a spermatophore, but perhaps to fe- shaped through male mating effort (e.g., by in- male preference for larger mates. creasing sperm transfer) or paternal investment Although chewing duration was related to (i.e., by increasing the number and fitness of the both male mass and estimated gift mass, male gift givers offspring). Our data suggest that mat- mass and gift mass were not directly related. To ing effort is a plausible hypothesis since gift size further investigate the relationship between body (i.e., chewing duration) was highly associated and gift mass, we separated the successful males with spermatophore attachment duration. These into two groups, large and small, based on their results run contrary to a previous study by Bi- mass relative to the mean (82.30 ± 1.27 mg). The dochka and Snedden (1985) that examined ten males who fell below the mean were placed mating behavior in a closely related sister spe- into the ‘small male’ group, and the nine males cies, Allonemobius. fasciatus. In this study female who fell above the mean were placed into the access to the tibial spur was manipulated through ‘large male’ group. On average, larger males were three treatments (spur covered, surrounding spur chewed upon twice as long and provided nearly area covered—spur uncovered, and spur uncov- twice as much gift as did smaller males (chewing ered) and subsequent copulation duration (an ap- ± ± duration: 15.46 2.38 min vs. 7.01 1.20 min, t17 proximation of chewing duration and hence = - 3.27, P < 0.01; gift donation: 2.52 ± 0.59 mg vs. nuptial gift size) and spermatophore attachment ± 1.33 0.28 mg, t17 = 1.88, P < 0.05, respectively. T- duration recorded. They found that females who tests were one tailed). Using this analysis, male were denied access to the spur had a significantly mass was significantly associated with gift mass, shorter copulation duration than the other two suggesting that the magnitude of the nuptial gift treatments. In addition, they concluded no associ- may be constrained by male size. ation between treatment and spermatophore at- tachment duration. However, if we compare the DISCUSSION published summary statistics for the covered (16.72 ± 3.59 min, n = 30) and uncovered treat- In ground crickets (Neonemobius sp.), females ments (29.26 ± 5.35 min, n = 19) only, they are sig-

are attracted to larger males and this preference nificantly different (two tailed t-test: t47 = -2.023, was previously speculated to be based on the P < 0.05), suggesting that spermatophore attach- male’s ability to provide a larger nuptial gift (For- ment duration may be related to copulation dura- rest et al. 1991). In this study, we have shown that tion in A. fasciatus, supporting our results. larger males are more successful at attracting In the wild, male and female ground crickets and copulating with females. More importantly, are promiscuous and mate with numerous indi- we have shown that male mass was positively as- viduals throughout the breeding season, exceed- sociated with the magnitude of gift contribution, ing the mating rate necessary to continually providing a mechanism for the maintenance of fe- produce offspring prior to senescence. Promiscu- male preference. This body size and gift size rela- ous behavior often carries associated costs includ- tionship coupled with sexual selection on gift size ing increased time and energy expenditure via female choice may have profound evolution- (Thornhill & Alcock 1983), increased predation ary implications. risk (Arnqvist 1989), increased disease suscepti- In insects, the common pattern of female- bility (Hurst et al. 1995), and/or caustic seminal biased size dimorphism is usually attributed to the fluids that potentially reduce fitness (Fowler & reproductive advantages of being large (Shine Partridge 1989; Rice 1996). The large size of the 1988). However, if gift mass is related to female nuptial gift in A. socius described here provides fitness, then the strength of size selection on males the opportunity for these costs to be offset by in- may surpass females, eventually increasing male creasing female fitness through increasing repro- size and modifying degree of dimorphism (e.g., ductive rate, reproductive longevity or fecundity. Leimar et al. 1994). An association between gift Associations between gift size and female repro- mass and female fitness is common in Orthoptera ductive fitness components are common in insects (Gwynne 1983, 1984; Brown 1997; Calos & Saka- (Gwynne 1984; Andersson 1994). 322 Florida Entomologist 85(2) June 2002

Currently, the contents of the gift in A. socius GREEN, D. J., AND E. A. KREBS. 1995. Courtship feeding are unclear though our data suggest it is mostly in ospreys Pandion hallaetus—a criterion for mate hemolymph and not simply a limited glandular assessment. Ibis. 137: 35-43. secretion. Other systems have shown that nuptial GWYNNE, D. T. 1982. Mate selection by female katydids gifts may contain oviposition inducing hormones (Orthoptera: Tettigoniidae, Conocephalus nigropleu- rum). Animal Behav. 30: 734-738. (Friedel & Gilliot, 1977), or act as indicators of GWYNNE, D. T. 1983. Male nutritional investment and important male chemical resources that may af- the evolution of sexual differences in Tettigoniidae fect offspring fitness (Eisner et al. 1996). Consid- and other Orthoptera, pp. 337-366. In D. T. Gwynne ering the relationship between gift size, male size and G. K. Morris (eds.), Orthopteran mating sys- and large male mating success in our data, gift tems: Sexual competition in a diverse group of in- quantity may be more important than gift quality. sects. Boulder, Colorado: Westview Press. GWYNNE, D. T. 1984. Courtship feeding increases female reproductive success in bushcrickets. Nature 307: ACKNOWLEDGMENTS 361-363. GWYNNE, D. T., AND W. D. BROWN. 1994. Mate feeding, We thank Jacqueline Litzgus, Scott Sakaluk and offspring investment, and sexual differences in katy- Wade Winterhalter for their insightful comments on this dids (Orthoptera, Tettigoniidae). Behavioral Ecology manuscript. This research was supported in part by a 5: 267-272. grant from the National Science Foundation (NSF HOWARD, D. J., AND D. G. FURTH. 1986. Review of the 0090177) to T.A.M. and by a GAANN fellowship to K.M.F. Allonemobius faciatus (Orthoptera: Gryllidae) com- plex with the description of two new species sepa- rated by electrophoresis, songs, and morphometrics. 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K. 1998. Paternity of off- veluchianus bushcrickets: beneficial effects of nup- spring in multiply mated female crickets: the effect tial feeding on offspring viability. Behav. Ecol. Socio- of nuptial food gifts and the advantage of mating biol. 45: 293-299. first. Proc. Royal Soc. London, 265: 2191-2195. SAKALUK, S. K., AND W. CADE. 1983. The adaptive sig- EISNER, T., S. R. SMEDLEY, D. K. YOUNG, M. EISNER, B. nificance of female multiple matings in house and ROACH, AND J. MEINWALD. 1996. Chemical basis of field crickets, pp. 319-336. In D. T. Gwynne and G. K. courtship in a beetle (Neopyrochroa flabellata): Morris (eds.), Orthopteran mating systems: Sexual cantharidin as precopulatory ‘enticing’ agent. Proc. competition in a diverse group of insects Boulder, Nat. Acad. Sci., USA, 93: 6499-6503. Colorado: Westview Press. FORREST, T. G., J. L. SYLVESTER, S. TESTA, S. W. SMITH, SAKALUK, S. K. 1984. Male crickets feed females to en- A. DINEP, T. L. CUPIT, J. M. HUGGINS, K. L. ATKINS, sure complete sperm transfer. Science 223: 609-610. AND M. EUBANKS. 1991. Mate choice in ground crick- SAKALUK, S. K. 1985. Spermatophore size and its role in ets (Gryllidae: Nemobiinae). Florida Entomol. 71: the reproductive behaviour of the cricket, Gryllodes 74-80. supplicans (Orthoptera: Gryllidae). Canadian J. Zool. FOWLER, K., AND L. PARTRIDGE. 1989. A cost of mating 63: 1652-1656. in female fruit flies. Nature 338: 760-761. SHINE, R. 1988. The evolution of large body size in fe- FRIEDEL, T., AND C. GILLOTT. 1977. Contribution of males: a critique of Darwin’s ‘fecundity advantage’ male-produced proteins to vitellogenesis in Melano- model. American Naturalist 131: 124-131. plus sanguinipes. J. Insect Physiol. 23: 145-151. SIMMONS, L. W. 1995. Correlates of male quality in the FULTON, B. B. 1931. A study of the genus Nemobius. field cricket, Gryllus campestris L: Age, size and (Orthoptera: Gryllidae). Ann. Entomol. Soc. Amer. 24: symmetry determine pairing success in field popula- 205-237. tions. Behav. Ecol. 6: 376-381. Fedorka & Mousseau: Nuptial Gifts in a Cricket 323

THORNHILL, R. 1976. Sexual selection and paternal in- WALKER, T. J. 1980. Reproductive behavior and mating vestment in insects. American Naturalist 110: 153- success of male short-tailed crickets: differences 163. within and between demes. Evol. Biol. 13: 219-260. THORNHILL, R., AND J. ALCOCK. 1983. The evolution of WEDELL, N. 1997. Ejaculate size in bushcrickets: the insect mating systems. Harvard University Press, importance of being large. J. Evol. Biol. 10: 315-325. Cambridge. WIGGINS, D. A. AND R. D. MORRIS. 1988. Courtship feed- VAHED, K. 1998. The function of nuptial feeding in insects: ing and copulatory behavior in the common tern a review of empirical studies. Biol. Rev. 73: 43-78. Sterna hirundo. Ornis Scandinavica 19: 163-165.

324 Florida Entomologist 85(2) June 2002

ATTRACTION OF MATED FEMALE CODLING MOTHS (LEPIDOPTERA: TORTRICIDAE) TO APPLES AND APPLE ODOR IN A FLIGHT TUNNEL

H. C. REED1 AND P. J. LANDOLT2 1Dept. Biological Sciences, Oral Roberts University, Tulsa, OK 74171

2USDA, ARS, Yakima Agriculture Research Laboratory, 5230 Konnowac Pass Rd., Wapato, WA 98951

ABSTRACT

In a flight tunnel, mated female codling moths, Cydia pomonella L., were attracted (upwind flight with zigzagging flight patterns) to cold-stored thinning apples. Greater numbers of cod- ling moths were attracted to apples infested with codling moth larvae than to uninfested apples. However, codling moth response to piped odor from cold-stored thinning apples infested with larvae was not significantly greater than that of moths to piped odor from uninfested apples. In a flight tunnel, significant numbers of mated female codling moths were captured in traps baited with fresh-picked immature apples or in traps through which odor from such apples was piped. Also, more codling moths were captured in traps baited with infested versus un- infested apples, and more were captured in traps with odor from infested apples compared to odor from un-infested apples. These studies demonstrate upwind attraction by flying fe- male codling moths to apple fruit and odors from apple fruit and show increased response by moths to odors of fruit that are infested with codling moth larvae. It is suggested that this heightened response to infested apples may be due to increased apparency of infested fruit that may release greater amounts of volatile odorants.

Key Words: codling moth, host-finding, attraction, apple, kairomone

RESUMEN

En un túnel de vuelo, hembras apareadas de Cydia pomonella L. fueron atraídas (vuelo con- tra el viento en un patrón zigzag) a manzanas inmaduras y refrigeradas. Un mayor número de palomillas fueron atraídas a manzanas infestadas con larvas que a manzanas no infesta- das. Sin embargo, la respuesta de las palomillas al olor de manzanas inmaduras refrigeradas e infestadas con larvas introducido por un tubo no fué significativamente mayor que al de manzanas no infestadas. En el túnel de vuelo, un número significativo de hembras apareadas fueron capturadas en trampas cebadas con manzanas inmaduras recién cosechadas o en trampas con el olor introducido de tales manzanas. También se capturaron más palomillas en trampas cebadas con manzanas infestadas que no infestadas, y se capturaron más palomillas en trampas con el olor de manzanas infestadas que con el olor de manzanas no infestadas. Estos estudios demuestran la atracción de las palomillas hembra de Cydia pomonella en vuelo contra el viento hacia manzanas y hacia el olor de manzanas y demuestra una res- puesta mayor de las palomillas hacia los olores de frutos infestados con larvas de C. pomone- lla. Esta respuesta mayor a manzanas infestadas puede ser debido a una mayor apariencia de las frutas infestadas por que tal vez emiten mayores cantidades de olores volátiles.

Adult codling moths are attracted to host fruit wind in tubes with apple odor compared to tubes and codling moth attraction responses to apple without apple odor. Using a Y-tube olfactometer odorants are thought to be important host-finding design, Hern and Dorn (1999) demonstrated an behavior (Wearing et al. 1973, Wearing and ambulatory orientation response by codling moth Hutchins 1973, Sutherland et al. 1974, Hern and females to the apple odorant (E,E)-α-farnesene. Dorn 1999, Yan et al. 1999). Although Wearing et Attraction of flying codling moths to apple odor al. (1973) could not show upwind movement by has yet to be shown, although Light et al. (2001) moths in an olfactometer in response to apple trapped male and female codling moth with the odor, they and others (Wearing and Hutchins pear chemical ethyl (E,Z)-2,4-decadienoate. Un- 1973, Sutherland et al. 1974) speculated that ap- derstanding such behavior is critical to the devel- ple odor, as well as the apple odorant α-farnesene, opment of appropriate assays to explore codling may be an attractant for the adult female codling moth host-finding and to isolate and identify host moth. An ambulatory upwind response to apple plant kairomones that codling moths use to locate odor by adult codling moths was recently demon- and select oviposition sites. strated by Yan et al. (1999). In that study, both Adult moth host-finding behavior can include virgin and mated female moths moved farther up- chemoanemotactic flight in response to host odor

Reed & Landolt: Attraction of Codling Moth to Apple 325

(Phelan and Baker 1987, Landolt 1989, Tingle et commercial apple orchard in June of 1999 and al. 1989, Rojas and Wyatt 1999). Such responses were placed in cold storage at 2°C until used in appear to be similar to those documented for male experiments from December 1999 through Febru- moth responses to female pheromone, a combina- ary 2000. Apples were infested by placing 2 neo- tion of upwind chemoanemotaxis with self- nate codling moth larvae with individual apples steered countering (Baker 1989). We investigated in paper cups with lids for 5 to 12 days. Infesta- whether codling moths, like other moths, respond tion was evident by physical damage to the fruit to host (apple) odor with zigzagging upwind and the presence of frass. Apples were 30 to 45 flights that lead them to the odor source. mm in diameter. Attraction or orientation responses of phyto- Fresh-picked Granny Smith apples were ob- phagous insects to host plant odor may be en- tained from commercial apple orchards in June hanced or increased with injury to the plant. This and July of 2000 and in July of 2001. Apples were has been noted in flight tunnel studies of cabbage picked at 800 to 900 h the morning of the day they looper moth, Trichoplusia ni Hübner, attraction were used in flight tunnel experiments. Apples to cotton plants, but not to cabbage or potato were infested by confining 3rd instar codling plants (Landolt 1993, 2001), and in both wind moth larvae with sleeved fruit on trees for 5 days tunnel and olfactometer ambulatory assays of prior to picking of fruit and their use in assays. Colorado potato beetle, Leptinotarsa decemlin- Again, successful infestation of fruit was con- eata Say, responses to potato plants (Bolter et al. firmed by the presence of physical damage and 1997, Landolt et al. 1999), and larval codling frass. Fresh-picked apples were 30 to 50 mm in di- moth attraction to apple fruit (Landolt et al. 1998, ameter, depending on the date. 2000). Particularly because of the increased re- sponse of codling moth larvae to apple fruit in- Observational Experiments fested with other codling moth larvae (Landolt et al. 2000), adult codling moths may also respond A set of experiments was conducted as obser- more strongly to the odors of infested apple fruit, vational assays of moth responses in a flight tun- compared to un-infested fruit. Additional experi- nel. The flight tunnel was a 1 m wide × 1 m tall × ments were conducted to test this hypothesis, us- 2 m long plexiglas box with a blower motor push- ing a flight tunnel assay. ing air into the upwind end and a second blower motor pulling air through the downwind end. Air- MATERIALS AND METHODS flow was balanced to produce a slight positive pressure in the tunnel, with an in-tunnel air speed General of 22 cm/sec. Charcoal-coated filters were installed at both ends of the flight tunnel. Moths were Moths used in assays were obtained as pupae tested one at a time, released from a horizontal from a laboratory colony maintained on an artifi- polystyrene vial near the center of the downwind cial diet (Toba and Howell 1991) at the Yakima end of the tunnel, during the 2nd and 3rd h of the Agricultural Research Laboratory, Wapato, Wash- scotophase. Moths were observed for 3 min and ington. Pupae were sorted by sex and were placed were scored for upwind oriented flight and con- in plastic-screened cages in a controlled environ- tact with the odor source (apples or airflow vent) ment room. The room was at 23°C, 50-70% RH, at the center of the upwind end of the tunnel. Up- with both a red incandescent light on 24 h per day wind oriented flights were zigzagging upwind and white fluorescent lamps on a reversed 14 h flights within the likely odor plume downwind of light:10 h dark photocycle. As adults emerged, pu- the fruit or pipe vent. These flights are here re- pae were moved to new cages daily to provide ferred to as attraction, or upwind oriented flights. males and females of known ages. Cages of moths The first experiment compared moth re- were provided water on cotton. sponses to 3 cold-stored thinning apples that were To obtain mated female codling moths for bio- not infested with a codling moth larva, 3 cold- assays, 15-20 females (2-3 day old) were placed in stored thinning apples infested with a codling one cage with 20-25 males (2-5 day old) for one moth larva, and a control (no apples). Apples were scotophase (dark period of the light cycle). Fe- placed on a petri plate on a ring stand at the cen- males were removed during the following photo- ter of the upwind end of the flight tunnel. On each phase (light period) and were then used in flight of 11 days, five moths were tested per treatment, tunnel tests during the next 2 scotophases. Fe- with the 3 treatments rotated in the treatment males used in bioassays were dissected to confirm sequence each day. Totals of 55 moths were tested the presence of spermatophores in the bursa cop- per treatment in this experiment. Treatment ulatrix. Data sets for groups of females that were means were separated by Tukey’s test following a <90% mated were rejected (one group). significant ANOVA F value (DataMost 1995). For flight tunnel experiments involving obser- The second experiment compared mated fe- vation of moth responses to cold-stored thinning male codling moth responses to 1) airflow that apples, Red Delicious apples were obtained from a was passed over 6 un-infested cold-stored thin-

326 Florida Entomologist 85(2) June 2002 ning apples in a glass jar, 2) airflow passed over 6 an empty jar and into a second trap that served as such apples each infested by a codling moth larva, an experimental control. Twenty to 25 female cod- or 3) airflow through a “system blank” consisting ling moths were released at the downwind end of of the jar and plumbing but with no apples. On the tunnel at the end of the second hour of the sco- each of 10 days, 5 females were tested per treat- tophase and traps were checked 4 h later to count ment, with the 3 treatments rotated in the treat- moths captured on the sticky insides of the traps. ment sequence each day. Totals of 50 moths were The fourth trapping experiment compared moth tested per treatment. Treatment means were sep- response to airflow with and without uninfested arated by Tukey’s test following a significant apples in the jar. This test was replicated five ANOVA F value (DataMost 1995). times over five days. The fifth trapping experi- ment compared moth response to airflow with Flight Tunnel Trapping Assays and without apples infested with codling moth in the jar. This test was also replicated five times Two sticky traps containing a bait or odor over five days. The sixth trapping experiment source were placed near the center of the upwind compared moths trapped in response to infested end of the flight tunnel at the beginning of the apples compared to un-infested apples. This test scotophase. Traps were triangular cardboard tent was replicated 10 times over 10 days. Apples were traps (10 cm wide, 15 cm tall and 10 cm deep) infested in the field by placing 3rd instar codling painted yellow and coated on all 3 inside panels moth larvae on fruit in sleeves on the tree, five with adhesive. Two traps were set up for each as- days prior to assays. say, with one serving as a control and the other Data for moths captured in traps were ana- containing an apple treatment. Groups of 20-25 lyzed as percentages of released moths recap- female codling moths were released from five tured in traps. Differences between treatments or 20-ml clear plastic vials hung horizontally at the between treatments and controls were deter- center of the downwind end of the flight tunnel at mined using a paired t-test (DataMost 1995) at p the end of the second hour of the scotophase. Vials ≤ 0.05. were open on the upwind end and screened on the downwind end to permit moth escape upwind into RESULTS the tunnel. Traps were 20 cm apart and 30 cm from the tunnel walls. Four hours after the re- Flight Tunnel Observational Assays lease of codling moths into the flight tunnel, traps were checked to count codling moth captured in Mated female codling moths were attracted to the traps Three experiments were conducted to and contacted cold-stored thinning apples pre- evaluate codling moth responses to two or three sented in the flight tunnel (Table 1). Percentages fresh-picked, immature apples, using this experi- of moths exhibiting upwind oriented flights to- mental design. Data were analyzed as mean per- wards either un-infested or infested apples were centages of released moths recaptured in traps. significantly greater than percentages of moths Treatment and control means were compared by responding to the control. Also, percentages of a paired t-test (DataMost 1995) to determine if moths contacting un-infested or infested apples they were significantly different. were significantly greater than percentages of The first trapping experiment evaluated moth moths contacting the control. Percentages of response to two un-infested apples in a trap com- moths attracted to infested apples were signifi- pared to a control trap (no apples). The assay was cantly greater than percentages of moths at- conducted six times, with releases of moths in the tracted to un-infested apples (Table 1), but source flight tunnel made on each of six days. The second contact was similar on infested and un-infested trapping experiment evaluated moth response to apples in this test. two infested apples in a trap compared to a con- Mated female codling moths were also at- trol trap (no apples). This assay was conducted tracted to odors of cold-stored thinning apples. five times, with releases of moths in the flight Percentages of moths exhibiting upwind oriented tunnel made on each of five days. The third trap- flights towards the pipe that vented odor of un- ping experiment compared moth response to two infested or infested apples were significantly un-infested apples and to two infested apples, in a greater than the percentages of moths responding trap. This assay was conducted six times, with re- to the control airflow (Table 1). Also, percentages leases of moths in the flight tunnel made on each of moths contacting the odor source (pipe vent) of six days. were greater in response to odor from over un- Three additional trapping tests evaluated infested or infested apples compared to the system moth response to airflow passed over fresh picked control (Table 1). Percentages of moths respond- immature apples. For each test replicate, three ing (attraction or contact) to odor of infested apples were placed in a glass jar and air was apples were not statistically different than per- pumped through the jar and into a trap in the centages of moths responding to odor of un- flight tunnel. Airflow was also pumped through infested apples.

Reed & Landolt: Attraction of Codling Moth to Apple 327

TABLE 1. MEAN (±SEM) PERCENTAGES OF MATED FEMALE CODLING MOTHS RESPONDING TO COLD-STORED THINNING APPLES IN A FLIGHT TUNNEL.

Treatment n % Upwind oriented flight % Source contact

Experiment 1 Blank 55 0.0 + 0.0 a 0.0 + 0.0 a Un-infested apples 55 18.2 + 3.3 b 12.7 + 3.0 b Infested apples 55 38.2 + 8.3 c 18.2 + 6.3 b Experiment 2 Blank airflow 50 0.0 + 0.0 a 0.0 + 0.0 a Un-infested apple airflow 50 10.0 + 3.3 b 8.0 + 0.0 b Infested apple airflow 50 8.0 + 4.4 b 6.0 + 4.3 b

Within a column and within an experiment, means followed by a the same letter are not significantly different by Tukey’s test, at p ≤ 0.05.

Flight Tunnel Trapping Assays DISCUSSION

Significantly more moths were captured in These results demonstrate attraction re- flight tunnel traps baited with immature apples, sponses of flying mated female codling moths to compared to moths captured in un-baited traps, apple fruit and to apple fruit odor. Previous stud- when those apples were infested with a codling ies had shown attraction by walking codling moth moth larva (Table 2). In a direct comparison of un- adult to apple fruit, apple odor, and the apple infested and infested apples, significantly more odorant E,E-α-farnesene (Hern and Dorn 1999, moths were captured in traps baited with infested Yan et al 1999), but not flight responses. Flight re- apples (Table 2). sponses by moths to host plants have been shown Female codling moths were captured in traps for the noctuids T. ni (Landolt 1989), Heliothis into which odor (as airflow) from over fresh- subflexa Tingle et al. 1989), and Mamestra bras- picked apples was introduced, indicating attrac- sica L. (Rojas and Wyatt 1999), and the plutellid tion to the apples (Table 2). This response was sig- Plutella xylostella (L.) (Palaniswamy et al. 1986, nificant in comparison to traps without odor from and Pivnick et al. 1990). Additionally, the codling apples, both using apples infested with codling moth has been captured in traps baited with the moth and using apples that were not infested. In pear volatile ethyl-(Z, E)-2,4-decadienoate (Light a direct comparison of odor from infested apples et al. 2001). Such flight responses indicate a po- and odor from un-infested apples, significantly tential for long distance host-finding by the co- more moths were captured in traps baited with dling moth, with a significant role of host odor in odor of infested apples (Table 2). host finding.

TABLE 2. MEAN (±SEM) PERCENTAGES OF MATED FEMALE CODLING MOTHS CAPTURED IN FLIGHT TUNNEL TRAPS BAITED WITH FRESH PICKED IMMATURE GRANNY SMITH APPLES.

% Upwind oriented flight

Treatment N Apples in trap N Apple airflow into trap

Experiment 1 Blank 6 1.3 ± 0.8 a 5 2.0 ± 1.2 a Un-infested apples 3.3 ± 1.0 a 7.0 ± 1.2 b Experiment 2 Blank 5 1.6 ± 1.6 a 5 1.4 ± 1.4 a Infested apples 18.0 ± 2.7 b 9.0 ± 2.1 b Experiment 3 Un-infested apples 5 0.9 ± 0.9 a 10 2.0 ± 0.8 a Infested apples 7.5 ± 1.6 b 5.5 ± 1.6 b

Means within an experimental comparison that are followed by the same letter are not different by a paired t-test at p ≤ 0.05.

328 Florida Entomologist 85(2) June 2002

These results also indicate that host-attraction host attractants for the codling moth. Qualitita- responses of adult codling moth, like that of neo- tive and quantitative comparisons of the odor of nate larvae, are enhanced by prior infestation of un-infested and infested apple fruit may provide fruit by the codling moth. Both anemotactic and clues as to which volatile chemicals are important klinotactic responses by neonate larvae of the cod- to codling moth host finding and host selection. ling moth are enhanced when larvae are in the air- stream with odor from apple fruit infested with ACKNOWLEDGMENTS other codling moth larvae, compared to un-in- fested fruit (Landolt et al. 2000). Similar enhanced Technical assistance was provided by J. F. Alfaro, orientation responses by herbivores to damaged or L. L. Biddick, J. A. Brumley, J. R. Dedlow, and D. Lovelace. induced plant odors have been shown for the cab- Helpful comments to improve the manuscript were made bage looper moth and for the Colorado potato bee- by W. L. Yee and R. S. Zack. This work was supported by tle (Landolt 1993, Landolt et al. 1999). Although it funding from the Washington State Tree Fruit Research might appear disadvantageous for a phytopha- Commission, by a USDA, NRI Research Career Enhance- gous insect to respond to an infested plant as an ment grant to Oral Roberts University, and by a sabbati- cal leave from Oral Roberts University to H. C. Reed. oviposition site, because the moth’s offspring would likely face immediate competition for food, there are other explanations for the observed be- REFERENCES CITED havior. If host plants occur in patches, locating an infested plant or fruit may provide proximity to BAKER, T. C. 1989. Sex pheromone communication in the Lepidoptera: New research progress. Experientia 45: other, un-infested, plants or fruits. In flight tunnel 248-262. studies of cabbage looper attraction to cotton BOEVE, J. L., U. LENGWILER, L. TOLLSTEN, S. DORN, AND plants, moths were likely to fly to plants damaged T. C. J. TURLINGS. 1996. Volatiles emitted by apple by conspecific larvae but were also likely to then fruitlets infested by larvae of the European apple oviposit on undamaged adjacent plants, when sawfly. Phytochemistry 42: 373-381. given that choice (Landolt 1993). Plants, including BOLTER, C. J., M. DICKE, J. J. A. VAN LOOP, J. H. VIS- apple and pear, that are injured or induced, often SER, AND M. A. POSTHUMUS. 1997. Attraction of Col- produce, or produce increased amounts of, particu- orado potato beetle to herbivore-damaged plants lar odorants (Boeve et al. 1996, Landolt et al. 2000, during herbivory and after its termination. J. Chem. Ecol. 23: 1003-1023. Scutareanu et al. 1997) and may be more chemi- DATAMOST. 1995. StatMost statistical analysis and cally apparent to a plant-seeking insect compared graphics. DataMost Corporation. Salt Lake City, UT. to an un-injured plant. Perhaps the codling moth HERN, A., AND S. DORN. 1999. Sexual dimorphism in the cannot easily detect apple fruit from a distance un- olfactory orientation of adult Cydia pomonella in less it is injured or infested. response to alpha farnesene. Entomol. Exp. Appl. 92: The attraction response rates of codling moths 63-72. in these flight tunnel assays were not high (up to LANDOLT, P. J. 1989. Attraction of the cabbage looper to 35%), compared to male moth responses to phero- host plants and host plant odor in the laboratory. En- mones, but are not out of line with studies of other tomol. Exp. et Appl. 53: 117-124. LANDOLT, P. J. 1993. Effects of host plant leaf damage moths and their responses to host plants. Rates of on cabbage looper moth attraction and oviposition. attraction to cabbage plants by cabbage looper Entomol. 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Attraction of Colorado potato beetle (Coleop- codling moth and to understand the role of vola- tera: Chrysomelidae) to damaged and chemically in- tile chemicals in host finding and host selection duced potato plants. Environ. Entomol. 28: 973-978. behavior of this insect. If moths use flight re- LANDOLT, P. J., J. A. BRUMLEY, C. L. SMITHHISLER, L. L. sponses to apple odor as a means of locating ovi- BIDDICK, AND R. W. HOFSTETTER. 2000. Apple fruit position sites, then flight attraction assays might infested with codling moth are more attractive to ne- be useful in studying codling moth host finding onate codling moth larvae and possess increased behavior and also in isolating those chemicals amounts of (E,E)-alpha farnesene. J. Chem. Ecol. 26: 1685-1699. emitted from apple fruit that attract codling LIGHT, D. M., A. L. KNIGHT, C. A. HENRICK, D. RAJA- moth. Additionally, if infested fruit are more PASTA, B. LINGREN, J. C. DICKENS, K. M. REYNOLDS, strongly attractive to codling moth adults and lar- R. G. BUTTERY, G. MERRILL, J. ROITMAN, AND B. C. vae, then the odors of these fruits might best be CAMPBELL. 2001. A pear-derived kairomone with used in studies to isolate and identify improved pheromonal potency that attracts male and female Reed & Landolt: Attraction of Codling Moth to Apple 329

codling moth, Cydia pomonella (L.). Naturwissen- attraction of anthocorid predators. J. Chem. Ecol. 23: schaften 88: 333-338. 2241-2260. PALANISWAMY, P., C. GILLOT, AND G. P. SLATER. 1986. SUTHERLAND, O. R. W., R. F. N. HUTCHINS, AND C. H. Attraction of diamondback moth, Plutela xylostella WEARING. 1974. The role of the hydrocarbon α-farne- (L.) (Lepidoptera: Plutellidae), by volatile com- sene in the behavior of codling moth larvae and pounds of canola, white mustard, and faba bean. adults, pp. 249-263. In Browne, L. B. (ed.), Experi- Can. Entomol. 118: 1279-1285. mental Analysis of Insect Behaviour. Springer-Ver- PHELAN, P. L., AND T. C. BAKER. 1987. An attracticide for lag, New York. control of Amyelois transitella (Lepidoptera: Pyral- TINGLE, F. C., R. R. HEATH, AND E. R. MITCHELL. 1989. idae) in almonds. J. Econ. Entomol. 80: 779-783. Flight responses of Heliothis subflexa (Gn.) (Lepidop- PIVNICK, K. A., B. J. JARVIS, G. P. SLATER, C. GILLOT, tera: Noctuidae) to an attractant from ground cherry, AND E. W. UNDERHILL. 1990. Attraction of the dia- Physalis angulata L. J. Chem. Ecol. 218: 168-170. mondback moth (Lepidoptera: Plutellidae) to vola- TOBA, H. H., AND J. F. HOWELL. 1991. An improved sys- tiles of oriental mustard: the influence of age, sex tem for mass-rearing codling moths. J. Entomol. Soc. and prior exposure to mates and host plants. Envi- B. C. 3: 625-631. ron. Entomol. 19: 704-709. WEARING, C. H., AND R. F. N. HUTCHINS. 1973. α-farne- ROJAS, J. C. 1999. Influence of age, sex and mating sta- sene, a naturally occurring oviposition stimulant for tus, egg load, prior exposure to mates, and time of the codling moth, Laspeyresia pomonella. J. Insect day on host-finding behavior of Mamestra brassicae Physiol. 19: 1251-1256. (Lepidoptera: Noctuidae). Environ. Entomol. 28: WEARING, C. H., P. J. CONNOR, AND K. D. AMBLER. 1973. 155-162. Olfactory stimulation of oviposition and flight activ- ROJAS, J. C., AND T. D. WYATT. 1999. Role of visual cues ity of the codling moth Laspeyresia pomonella, using and interaction with host odour during the host-find- apples in an automated olfactometer. N. Z. J. Science ing behavior of the cabbage moth. Entomol. Exp. et 16: 697-710. Appl. 91: 59-65. YAN, F., M. BENGTSSON, AND P. WITZGALL. 1999. Behav- SCUTAREANU, P., B. DRUKKER, J. BRUIN, M. A. POSTHU- ioral response of female codling moths, Cydia MUS, AND M. W. SABELIS. 1997. Volatiles from Psylla pomonella, to apple volatiles. J. Chem. Ecol. 25: infested pear trees and their possible involvement in 1343-1351.

330 Florida Entomologist 85(2) June 2002

SUSCEPTIBILITY OF DIAPREPES ABBREVIATUS (COLEOPTERA: CURCULIONIDAE) TO A COMMERCIAL PREPARATION OF BACILLUS THURINGIENSIS SUBSP. TENEBRIONIS

A. A. WEATHERSBEE III,1 Y. Q. TANG,2,3 H. DOOSTDAR3 AND R. T. MAYER1 1USDA, ARS, U.S. Horticultural Research Laboratory, 2001 South Rock Road, Fort Pierce, FL 34945

2Visiting scientist, Biological Control Research Institute, Fujian Agricultural University, Fuzhou, Fugian 35002 China

3Visiting scientist, USDA, ARS, U.S. Horticultural Research Laboratory, 2001 South Rock Rd., Fort Pierce, FL 34945

ABSTRACT

A commercial preparation of the microbial entomopathogen, Bacillus thuringiensis subsp. tenebrionis (Btt) was evaluated for biological activity against the Diaprepes root weevil, Dia- prepes abbreviatus (L.). A reduction in survival was observed for neonatal larvae exposed to insect diet incorporated with Btt and in potted citrus treated with a Btt soil application. A treatment-induced, weight gain reduction for neonates was indicated only in the diet assay. Larvae exposed at 5 weeks old to diet treated with Btt demonstrated a dose-dependent mor- tality response. The mean ages for larval death ranged from 111 to 128 days among treat-

ments. The LC50 for larvae in this age group was 6.2 ppm [AI] and the slope of the probit line was 2.29. The mortality response of larvae exposed at 12 weeks old also was dose dependent

and the mean ages for larval death ranged from 130 to 141 days among treatments. The LC50 for larvae in this age group was 25.4 ppm [AI] and the slope of the probit line was 2.75. The delayed patterns of mortality that we observed among larvae treated at 5 and 12 weeks old indicates that disease is slow to develop in older larvae but that death occurs before matura- tion is completed.

Key Words: Diaprepes abbreviatus, Bacillus thuringiensis, entomopathogen, citrus, Diaprepes root weevil

RESUMEN

Se evaluó una formulación comercial del entomopatógeno microbial, Bacillus thuringiensis subsp. tenebrionis (Btt) por su actividad biológica en contra del gorgójo Diaprepes abbrevia- tus (L.). Se observó una reducción en la sobrevivencia de larvas neonatas expuestas a una dieta artificial que contenia Btt, y de larvas en potes con arboles tratados con Btt. Una re- ducción del peso ganado por las larvas neonatas fue inducida por el tratamiento solamente en el caso del ensayo con dieta artificial. Larvas expuestas a dieta tratada a las 5 semanas de edad manifestaron una mortalidad proporcional a la dosis. El rango del promedio del nú-

mero de dias hasta la muerte de las larvas fue de 111 a 128 dias entre tratamientos. La DL50 para las larvas de este grupo fue de 6.2 ppm (IA) y el pendiente de la linea probit fue de 2.29. La mortalidad de larvas expuestas a las 12 semanas de edad fue dependiente de la dosis y

la edad promedia de la muerte varió entre 130 y 141 dias entre tratamientos. La DL50 para las larvas de este grupo fue de 25.4 ppm (IA) y el pendiente de la linea probit fue de 2.75. El patron de mortalidad que observamos entre larvas tratadas a las 5 y a las 12 semanas de edad indica que la enfermedad se desarrolló lentamente en las larvas mayores pero la muerte ocurrió antes de la maduración.

The most economically important insect pest ranean larvae extensively damage host plant root of citrus in Florida is the Diaprepes root weevil, systems on which they feed. Larval feeding in cit- Diaprepes abbreviatus (L.). Losses to citrus grow- rus predisposes root tissues to infections by ers are estimated in excess of $75 million yearly pathogens such as Phytophthora spp. (Rogers et (Anonymous 1997). This root weevil also attacks al. 1996), induces a decline in tree health and sugar cane, vegetable crops, and ornamental fruit production, and may eventually kill the tree plantings in areas that are infested. The subter- (Schroeder & Sutton 1977). In severe cases of un- controlled weevil populations, entire citrus groves are destroyed (pers. obs.).

Mention of a trademark or proprietary product does not con- Entomopathogens have received considerable stitute a guarantee or warranty of the product by the U.S. De- partment of Agriculture and does not imply its approval to the attention as potential biocontrol agents of root exclusion of other products that may also be suitable. weevil larvae. Beavers et al. (1983) and Román

Weathersbee et al.: Susceptibility of D. abbreviatus to B. thuringiensis 331 and Beavers (1983) initially demonstrated the in a heated water bath before incorporating treat- pathogenicity and seasonal prevalence of several ments. A commercial preparation of B. thuring- naturally occurring fungi and nematodes attack- iensis subsp. tenebrionis (Novodor 3% [AI], [30 mg ing D. abbreviatus. Since then, many others have spores and δ-endotoxin crystals per ml product]) demonstrated the potential for biological control of was incorporated into the diet at rates of 0, 0.3, root weevil larvae by entomopathogens (Duncan et 3.0, 30, and 300 ppm (µg AI/ml diet). Treatments al. 1996; Figueroa & Román 1990; Quintella & Mc- were incorporated into the agar-laden diet with Coy 1997, 1998; Schroeder 1987, 1994). Nematode the aid of a heated, stirrer plate. The resulting applications were recommended for control as re- mixtures were pipetted into diet cups (15 ml diet cently as 1999, but the insecticidal compound, per cup), allowed to solidify and cool to room tem- bifenthrin, is currently the only material listed in perature, and then were closed with a lid. All the Florida Citrus Pest Management Guide for steps after heating of the diet were performed in a control of larval stages (Futch et al. 2001). laminar flow, clean bench to avoid contamination. Bacterial entomopathogens have not been pre- viously investigated as potential biocontrol Evaluation of Btt Activity agents for Diaprepes root weevil. The microbial pathogen, Bacillus thuringiensis subsp. tenebrio- The biological activity of Novodor was evalu- nis (Btt), was shown to have activity against rep- ated initially against neonatal larvae exposed to resentatives of the order Coleoptera including the treated insect diet. Neonates were briefly surface families Chrysomelidae, Bostrichidae, and Cur- sterilized in a 5% bleach solution and rinsed with culionidae (Herrnstadt et al. 1986; Krieg et al. sterile, deionized water prior to being placed in 1983, 1987; Beegle 1996; Saade et al. 1996). The diet cups. The treatments included the 5 rates of bacterium produces a crystalline δ-endotoxin dur- treatment-incorporated diet described above and ing the process of sporulation (Hofte & Whiteley there were 3 replications of each treatment. A 1989). Upon ingestion by a susceptible insect treatment comprised 18 diet cups, each infested host, the endotoxin is proteolytically activated in with 5 neonatal larva. The numbers and weights the midgut and causes feeding inhibition, septi- of surviving larvae in each diet cup were assessed caemia, and eventual death (Knowles 1994). We after 6 weeks of exposure to treated diet. investigated a commercially available Btt prod- The activity of Novodor also was evaluated uct, Novodor® (Novo Nordisk, North Chicago, IL), against 5-week old weevil larvae. Larvae used in to determine if it had biological activity against the experiment weighed ~110 mg before exposure D. abbreviatus, envisioning that a demonstration to treatments. Treatments were prepared as of activity may provide the impetus to develop above and replicated 6 times. Each treatment this and possibly other B. thuringiensis subsp. for comprised 18 diet cups, each containing 1 larva. field application. Mortality, age at death, and weights of eclosed adults were recorded twice weekly. The experi- MATERIALS AND METHODS ment was terminated after 5 months when most larvae had either died or matured to adults. Insect Rearing Novodor activity also was evaluated against 12-week old larvae that weighed ~510 mg before Neonatal, 5-week-old, and 12-week-old larvae exposure to treatments. The treatments were in- of D. abbreviatus were obtained from a laboratory corporated in diet at the rates described above. colony maintained by the U.S. Horticultural Re- There were 3 replications of treatments, each search Laboratory, Fort Pierce, FL. Larvae were comprising 30 larvae confined individually to diet reared on a commercially-prepared insect diet cups. Mortality, age at death, and weights of (Product No. F1675, Bio-Serv, Frenchtown, NJ) eclosed adults were recorded twice weekly. The placed within sealed diet cups (PC100, 30 ml cups experiment was terminated after 3 months when and lids, Jet Plastica, Harrisburg, PA). Prepara- most larvae had either died or matured to adults. tion of insect diet and rearing of larvae were sim- Another experiment was conducted to deter- ilar to the methods described by Beavers (1982) mine the effect of Novodor suspensions applied as with temperature and moisture content of diet op- soil treatments against neonatal larvae feeding timized for larval development (Lapointe 2000, on potted citrus roots. The citrus plants used in Lapointe & Shapiro 1999). Weevil larvae used in the study were 1 year old Cleopatra Mandarin the bioassays were maintained in diet cups stored (Citrus reshni Hort., ex Tan.) rootstock potted in in plastic trays in a growth chamber at 26°C and 473 cm3 containers of potting soil (Metromix 500, a 24-h dark cycle. Scotts, Marysville, OH). The treatments included 4 rates of Novodore (0, 3.0, 30, and 300 ppm [(g AI/ Diet Incorporation of Btt ml DI water]) applied as 50 ml suspensions to the soil of each container. Two drench applications Prepared insect diet was heated to 90°C for 15 were made 14 days apart. There were 6 replica- min., covered with foil, and allowed to cool to 56°C tions of each treatment. A treatment comprised 1

332 Florida Entomologist 85(2) June 2002 potted citrus plant infested with 20 neonates im- in mortality of larvae exposed at the age of 5 mediately after the first drench application. All weeks. Mortality of larvae exposed to the 3 ppm treatments were maintained in a growth chamber treatment (44%) was significantly (P ≤ 0.05) at 26°C with a photoperiod of 14:10 (L:D) h. The greater than that of the controls (15%). Larval numbers and weights of surviving larvae were as- mortality in the 30 ppm treatment exceeded 50%

sessed after 6 weeks. (Table 2). The calculated LC50 for larvae in this age group was 6.2 (95% FL = 2.7-13.2) ppm [AI]. Data Analyses and Statistics The slope of the probit line was 2.29 (SE = 0.31) (χ2 = 43.07; df = 1; P < 0.0001). Treatments also Percentage data were adjusted for control mor- caused a significant reduction (F = 6.25; df = 4, 20; tality using the Abbott (1925) formula and trans- P = 0.0020) in the mean death ages of larvae. The formed (arcsine) before analyses. Data were death ages for larvae maintained on treated diets analyzed by the General Linear Models Proce- ranged from 111 to 128 d while that of control lar- dure, and differences among treatment means vae was 154 d. Larvae in the 30 and 300 ppm were determined by Tukey’s studentized range treatments died significantly (P ≤ 0.05) earlier test (SAS Institute 1990). Differences among than the control larvae (Table 2). The mortality means were considered significant at a probabil- ≤ response we observed occurred later than is typi- ity level of 5 percent (P 0.05). Probit analyses cal for insects susceptible to B. thuringiensis; nev- were conducted using the Probit Procedure (SAS ertheless, the response was consistent, dose- Institute 1990) to generate LC values and slopes 50 dependent, and occurred earlier than that ob- of probit lines for larval mortality due to treat- served for the controls. We observed that disease ments. Untransformed means were presented in and death occurred in treated insects during the the data tables. later stages of larval development and early phases of pupation (Fig. 1). The presence of Btt RESULTS was confirmed by isolating the bacteria from in- ternal tissues of dead larvae. Bacterial isolates Neonatal Larvae were cultured in 40 ml LB broth in a 125 ml baf- fled flask maintained in an incubator shaker at The survival of root weevil larvae exposed as ° neonates to diet treated with Novodor was signif- 28 C and 200 rpm until sporulation phase. Cul- icantly reduced (F = 9.36; df = 4, 262; P < 0.0001) tures were examined using Hoffman contrast microscopy at 1000× magnification to identify Btt after 6 weeks. While 2 of 5 larvae survived after 6 δ weeks in the control group, only 1 larvae survived endospores and -endotoxin crystals. The mean on average in the 300 ppm treatment (Table 1). fresh weights of adults that eclosed on treated di- The low survival rate observed for control larvae ets also were significantly reduced (F = 3.39; df = is addressed in the discussion section. The fresh 4, 20; P = 0.0286). Adult weevils that survived the weights of surviving larvae were significantly re- 300 ppm treatment weighed 15% less than con- duced (F = 8.46; df = 4, 262; P < 0.0001) by treat- trol adults, indicating that inhibition of feeding or ments indicating that larval feeding or nutrient disruption of nutrient assimilation may have oc- assimilation may have been inhibited. Surviving curred among survivors of treatments (Table 2). larvae in the 300 ppm treatment weighed 53% less than those in the control group (Table 1). 12-Week Old Larvae

5-Week Old Larvae A significant increase (F = 8.82; df = 4, 8; P < 0.0050) in mortality was observed for 12-week old Insect diet treated with Novodor caused a sig- larvae fed Novodor treated diet. Mortality in the nificant increase (F = 76.57; df = 4, 20; P < 0.0001) 30 ppm treatment (53%) was significantly (P ≤

TABLE 1. SURVIVAL AND AVERAGE FRESH WEIGHT OF D. ABBREVIATUS LARVAE EXPOSED AS NEONATES (5 PER DIET CUP) FOR 6 WEEKS TO INSECT DIET INCORPORATED WITH NOVODOR.

Treatment Number of surviving Weight (mg) of surviving (ppm AI)a larvae ± SE (n = 3)b larvae ± SE (n = 3)b

0.0 2.0 ± 0.1 a 306.3 ± 22.4 a 0.3 1.9 ± 0.1 a 251.8 ± 16.7 a 3.0 1.8 ± 0.1 a 236.1 ± 21.5 a 30.0 1.6 ± 0.1 a 231.1 ± 20.5 a 300.0 1.0 ± 0.1 b 144.0 ± 20.9 b

aAI refers to the concentration (µg/ml) of active ingredient, comprising spores and δ-endotoxin of B. thuringiensis var. tenebrionis, in prepared diet. bMeans within a column sharing the same letter were not significantly different (P > 0.05, Tukey’s studentized range test [SAS Institute 1990]). Weathersbee et al.: Susceptibility of D. abbreviatus to B. thuringiensis 333

TABLE 2. MORTALITY AND AGE AT DEATH OF LARVAE, AND FRESH WEIGHT OF ECLOSED ADULTS, FOR D. ABBREVIATUS EXPOSED AS 5-WEEK-OLD LARVAE TO INSECT DIET INCORPORATED WITH NOVODOR.

Treatment % Mortality ± SE Death age (d) ± SE Adult wt. (mg) ± SE (ppm AI)a (n = 6)b,c (n = 6)b,c (n = 6)b,c

0.0 15.3 ± 4.4 a 154.3 ± 14.5 a 315.0 ± 12.0 a 0.3 30.3 ± 4.3 a 127.9 ± 10.0 ab 329.2 ± 23.8 ab 3.0 43.7 ± 5.8 b 126.9 ± 8.6 ab 323.2 ± 15.2 ab 30.0 58.1 ± 4.9 c 114.4 ± 8.1 b 294.3 ± 9.0 ab 300.0 71.0 ± 4.4 d 110.6 ± 2.9 b 266.9 ± 19.5 b

aAI refers to the concentration ((g/ml) of active ingredient, comprising spores and δ-endotoxin of B. thuringiensis var. tenebrionis, in prepared diet. bMortality, age at death, and fresh weights of eclosed adults were assessed twice weekly during the experiment. cMeans within a column sharing the same letter were not significantly different (P > 0.05, Tukey’s studentized range test [SAS Institute 1990]).

0.05) greater than that for the control group (6%) Soil Treatments

(Table 3). The calculated LC50 for larvae in this age group was 25.4 (95% FL = 12.5-60.0) ppm Survival of neonates on potted citrus was sig- [AI]. The slope of the probit line was 2.75 (SE = nificantly reduced (F = 4.87; df = 3, 15; P < 0.0146) 0.44) (χ2 = 47.37; df = 1; P < 0.0001). The mean age by soil treatments with Novodor. Larval survival at which mortality occurred also was significantly was significantly (P ≤ 0.05) less in all treatments reduced by treatments (F = 4.54; df = 4, 8; P = as compared to the controls after 6 weeks expo- 0.0330). The mean death age for larvae in the 300 sure to treated soils and citrus roots (Table 4). ppm treatment (130 d) was significantly (P ≤ 0.05) The low rate of survival for control larvae is ad- less than that for control larvae (153 d) (Table 3). dressed in the discussion section. In contrast to Treatments did not significantly affect the fresh the results of the diet bioassay against neonates, weights of eclosed adults in this age group (F = the fresh weights of surviving larvae were not sig- 1.34; df = 4, 8; P = 0.3341), likely because larvae nificantly reduced (F = 1.02; df = 3, 15; P = 0.4098) used in the experiment had attained near maxi- in the potted citrus bioassay (Table 4). Some lar- mum weights before exposure to treatments. vae may have been able to avoid exposure to Btt where soil drenches resulted in unequal disper- sion of treatments.

DISCUSSION

The results of our experiments indicate that neonatal, 5-week-old, and 12-week-old larvae of D. abbreviatus were susceptible to the biological effects of Novodor. Activity for B. thuringiensis against older larval stages of insects is not nor- mally expected; nevertheless, the dose-dependent mortalities we observed for larvae exposed at 5 and 12 weeks old were reproducible. The low survival rates we observed for neo- nates in our controls are typical in experiments such as this, where multiple larvae are collec- tively grouped to challenge plants or generate data for bioassays of diet-incorporated materials. Low survival rates for neonatal D. abbreviatus have been reported by others (Schroeder & Sie- burth 1997, Quintella & McCoy 1997), and are due to natural mortality factors including aggres- sive interactions among larvae confined together (Lapointe & Shapiro 1999). The response we observed for older larvae to Novodor may be attributed to the finding that midgut trypsin activity increases to a maximum Fig. 1. Healthy (left) and diseased (right) larvae and in D. abbreviatus larvae at the age of approxi- pupae of D. abbreviatus due to feeding on diet incorpo- mately 7 weeks (Yan et al. 1999). Yan et al. also rated with Novodor. inferred that the midgut of D. abbreviatus larvae 334 Florida Entomologist 85(2) June 2002

TABLE 3. MORTALITY AND AGE AT DEATH OF LARVAE, AND FRESH WEIGHT OF ECLOSED ADULTS, FOR D. ABBREVIATUS EXPOSED AS 12-WEEK-OLD LARVAE TO INSECT DIET INCORPORATED WITH NOVODOR.

Treatment % Mortality ± SE Death Age (d) ± SE Adult Wt. (mg) ± SE (ppm AI)a (n = 3)b,c (n = 3)b,c (n = 3)b,c

0.0 5.6 ± 5.6 a 153.1 ± 7.4 a 353.0 ± 6.5 a 0.3 21.1 ± 13.1 ab 132.0 ± 14.9 ab 331.2 ± 22.2 a 3.0 31.1 ± 21.5 abc 141.3 ± 7.8 ab 317.6 ± 32.3 a 30.0 53.3 ± 15.4 bc 134.8 ± 10.8 ab 338.5 ± 1.3a 300.0 67.8 ± 12.5 c 130.1 ± 9.4 b 374.6 ± 13.5 a

aAI refers to the concentration (µg/ml) of active ingredient, comprising spores and δ-endotoxin of B. thuringiensis var. tenebrionis, in prepared diet. bMortality, age at death, and fresh weights of eclosed adults were assessed twice weekly during the experiment. cMeans within a column sharing the same letter were not significantly different (P > 0.05, Tukey’s studentized range test [SAS Institute 1990]). is probably alkaline, as shown for many Lepi- oped slowly, the mean ages at death were consis- doptera and Diptera, since the enzyme activity tently earlier among treated as compared to observed was most active at a pH of 10.4. Alkaline control larvae. Also, the fresh weights of adults solubilization and proteolytic cleavage by serine that survived treatments were reduced compared proteases, such as trypsin, are required for acti- to those of the control, indicating that midgut vation of many B. thuringiensis δ-endotoxins (Pie- damage or feeding inhibition may have occurred trantonio et al. 1993) including those produced by among larvae that survived treatments. Btt. The Cry3a toxin of Btt may undergo substan- A dose-dependent mortality response also was tial cleavage at the N-terminus without loss of observed for larvae treated at the age of 12 weeks. biological activity (Carroll et al. 1989). It has been Larvae treated at this stage of development often suggested that these additional protease cleavage died as pupae, further supporting the contention sites may facilitate insertion of a portion of the that disease was slow to develop. The fresh protein into the target membrane of the midgut weights of the adults that developed from larvae epithelium (Knowles 1994) and subsequent pore treated at 12 weeks old were not affected by treat- formation. The mortality responses we observed ments. Lapointe (2000) reported that larval for larvae treated at 5 and 12 weeks old may have weights of D. abbreviatus were maximum at 12 been a product of appropriate levels of pH and weeks old, and then declined during further de- serine proteases in midgut tissues during those velopment, thus explaining the lack of a weight stages of development. The delayed period to lar- response for larvae in this age group. val death may have been a function of the ability There is much yet to be learned about the acti- of D. abbreviatus to survive for long periods with- vation and binding patterns of B. thuringiensis out food or water (personal observation) and the endotoxins, and the subsequent onset of disease slow development of disease. Any future evalua- in susceptible insects. We observed that disease tion of Btt on larval D. abbreviatus, and possibly was slow to develop in D. abbreviatus. The results other insects, should consider the importance of of our experiment indicate that there may be age- suitable midgut environments for endotoxin acti- related factors that influence larval D. abbrevia- vation and the general hardiness of the insect. tus response to Btt endotoxins. There may also be We observed dose-dependent responses for other endotoxins awaiting discovery that are bet- mortality, age at death, and fresh weights of adult ter adapted to activation and binding by larval survivors from larvae treated at the age of 5 D. abbreviatus. This report provides the impetus weeks. Although the mortality response devel- to further explore these processes in the Dia-

TABLE 4. SURVIVAL AND MEAN FRESH WEIGHT OF D. ABBREVIATUS LARVAE EXPOSED AS NEONATES (20 PER PLANT) FOR 6 WEEKS TO POTTED CITRUS TREATED WITH NOVODOR.

Treatment Number of surviving Weight (mg) of surviving (ppm AI)a larvae ± SE (n = 6)b larvae ± SE (n = 6)b

0.0 4.2 ± 1.1 a 52.0 ± 22.5 a 3.0 1.7 ± 0.3 b 40.7 ± 9.9 a 30.0 1.5 ± 0.4 b 19.8 ± 6.6 a 300.0 1.3 ± 0.3 b 41.2 ± 12.1 a

aAI refers to the concentration (µg/ml) of active ingredient, comprising spores and δ-endotoxin of B. thuringiensis var. tenebrionis, in prepared diet. bMeans within a column sharing the same letter were not significantly different (P > 0.05, Tukey’s studentized range test [SAS Institute 1990]). Weathersbee et al.: Susceptibility of D. abbreviatus to B. thuringiensis 335

prepes root weevil and other insects. Our demon- KRIEG, V. A., A. M. HUGER, G. A. LANGENBRUNCH, AND stration of Novodore activity against different W. SCHNETTER. 1983. Bacillus thuringiensis var. larval stages of D. abbreviatus is encouraging tenebrionis, a new pathotype effective against larvae enough to warrant additional testing. of Coleoptera. Z. Angew. Entomol. 96: 500-508. LAPOINTE, S. L. 2000. Thermal requirements for the de- velopment of Diaprepes abbreviatus (Coleoptera: ACKNOWLEDGMENTS Curculionidae). Environ. Entomol. 29: 150-156. LAPOINTE, S. L., AND J. P. SHAPIRO. 1999. Effect of soil The authors thank Ginny Schmit and Karin Crosby moisture on development of Diaprepes abbreviatus (USDA, ARS, U.S. Horticultural Research Laboratory, (Coleoptera: Curculionidae). Florida Entomol. 82: Fort Pierce, FL) for technical support and assistance 291-299. with insect rearing and preparation of insect diets. PIETRANTONIO, P. V., B. A. FEDERICI, AND S. S. GILL. 1993. Interaction of Bacillus thuringiensis endotox- REFERENCES CITED ins with the insect midgut epithelium, pp.55-80. In N. E. Beckage, S. N. Thompson, and B. A. Federici ABBOTT, W. S. 1925. A method for computing the effective- (eds.), Parasites and Pathogens of Insects, Vol 2. Ac- ness of an insecticide. J. Econ. Entomol. 18: 265-267. ademic Press, New York. ANONYMOUS. 1997. Diaprepes Task Force Minutes, July QUINTELLA, E. D., AND C. W. MCCOY. 1997. Pathogenic- 17, 1997. University of Florida. Lake Alfred, FL. ity enhancement of Metarhizium anisopliae and Beau- BEAVERS, J. B. 1982. Biology of Diaprepes abbreviatus veria bassiana to first instars of Diaprepes abbreviatus (Coleoptera: Curculionidae) reared on artificial diet. (Coleoptera: Curculionidae) with sublethal doses of Florida Entomol. 65: 263-269. imadacloprid. Environ. Entomol. 26: 1173-1182. BEAVERS, J. B., C. W. MCCOY, AND D. T. KAPLAN. 1983. QUINTELLA, E. D., AND C. W. MCCOY. 1998. Synergistic Natural enemies of subterranean Diaprepes abbre- effect of imadacloprid and two entomopathogenic viatus (Coleoptera: Curculionidae) larvae in Florida. fungi on the behavior and survival of larvae of Dia- Environ. Entomol. 12: 840-843. prepes abbreviatus (Coleoptera: Curculionidae) in BEEGLE, C. C. 1996. Efficacy of Bacillus thuringiensis soil. J. Econ. Entomol. 91: 110-122. against Lesser Grain Borer, Rhyzopertha dominica ROGERS, S., J. H. GRAHAM, AND C. W. MCCOY. 1996. In- (Coleoptera: Bostrichidae). Biocontrol. Sci. Tech. 6: sect-plant pathogen interactions: Preliminary stud- 15-21. ies of Diaprepes root weevil injuries and CARROLL, J., J. LI, AND D. J. ELLAR. 1989. Proteolytic Phytophthora infections. Proc. Fla. State Hort. Soc. processing of a coleopteran-specific δ-endotoxin pro- 109: 57-62. duced by Bacillus thuringiensis var. tenebrionis. Bio- ROMÁN, J., AND J. B. BEAVERS. 1983. A survey of Puerto chem. J. 261: 99-105. Rican soils for entomogenous nematodes which at- DUNCAN, L. W., C. W. MCCOY, AND A. C. TERRANOVA. tack Diaprepes abbreviatus (L.) (Coleoptera: Curcu- 1996. Estimating sample size and persistence of en- lionidae). J. Agric. Univ. P.R. 67: 311-316. tomogenous nematodes in sandy soils and their effi- SAADE, F. E., G. B. DUNPHY, AND R. L. BERNIER. 1996. cacy against the larvae of Diaprepes abbreviatus in Response of the Carrot weevil, Listronotus oregonen- Florida. J. Nematol. 28: 56-67. sis (Coleoptera: Curculionidae), to strains of Bacillus FIGUEROA, W., AND J. ROMÁN. 1990. Biocontrol of the thuringiensis. Biological Control. 7: 293-298. sugarcane rootstalk borer, Diaprepes abbreviatus SAS INSTITUTE. 1990. SAS/STAT user’s guide for per- (L.) (Coleoptera: Curculionidae), with entomophilic sonal computers, vol. 6. SAS Institute, Cary, NC. nematodes. J. Agric. Univ. P.R. 74: 395-404. SCHROEDER, W. J. 1987. Laboratory bioassays and field FUTCH, S. H., J. L. KNAPP, AND C. W. MCCOY. 2001. Fact trials of entomogenous nematodes for control of Di- Sheet ENY-611, 2001 Florida Citrus Pest Manage- aprepes abbreviatus (Coleoptera: Curculionidae) in ment Guide, FCES, IFAS, University of Florida. Re- citrus. Environ. Entomol. 16: 987-989. vised: October 2000. SCHROEDER, W. J. 1994. Comparison of two steinerne- HERRNSTADT, C., G. G. SOARES, E. R. WILCOX, AND D. L. matid species for control of the root weevil Diaprepes EDWARDS. 1986. A new strain of Bacillus thuringien- abbreviatus. J. Nematol. 26: 360-362. sis with activity against coleopteran insects. Bio- SCHROEDER, W. J., AND P. J. SIEBURTH. 1997. Impact of technology. 4: 305-308. surfactants on control of the root weevil Diaprepes HÖFTE, H., AND H. R. WHITELEY. 1989. Insecticidal crys- abbreviatus larvae with Steinernema riobravis. J. tal proteins of Bacillus thuringiensis. Microbiol. Rev. Nematol. 29: 216-219. 53: 242-255. SCHROEDER, W. J., AND R. A. SUTTON. 1977. Citrus root KNOWLES, B. H. 1994. Mechanism of action of Bacillus damage and the spatial distribution of eggs of Di- thuringiensis insecticidal δ-endotoxins. Adv. Insect. aprepes abbreviatus. Florida Entomol. 60: 114. Physiol. 24: 275-308. YAN, X. H., H. L. DE BONDT, C. C. POWELL, R. C. BUL- KRIEG, V. A., A. M. HUGER, AND W. SCHNETTER. 1987. LOCK, AND D. BOROVSKY. 1999. Sequencing and char- “Bacillus thuringiensis var. San Diego” strain M-7 is acterization of the citrus weevil, Diaprepes identical to the formerly in Germany isolated B. thu- abbreviatus, trypsin cDNA: Effect of Aedes trypsin ringiensis var. tenebrionis strain BI 256-82. J. Appl. modulating oostatic factor on trypsin biosynthesis. Entomol. 104: 417-424. Eur. J. Biochem. 262: 627-636.

336 Florida Entomologist 85(2) June 2002

HOST SELECTION BY (ORTHOPTERA: ACRIDIDAE) INHABITING SEMI-AQUATIC ENVIRONMENTS

JASON M. SQUITIER AND JOHN L. CAPINERA Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611-0620

ABSTRACT

Through laboratory choice tests involving 19 plant species, we assessed the host selection behavior of six grasshopper species: Stenacris vitreipennis (Marschall) (glassywinged tooth- pick grasshopper), Leptysma marginicollis (Serville) (cattail toothpick grasshopper), Gymnoscirtetes pusillus Scudder (little wingless grasshopper), clavuliger (Serville) (olivegreen swamp grasshopper), Scudder (Atlantic grasshopper), and Romalea microptera (Beauvois) (eastern lubber grasshopper). This grasshopper assemblage is commonly associated with semi-aquatic habitats in the southeastern United States. These poorly studied species display both graminivorous (S. vetreipennis and L. marginicollis) and mixed graminivorous-forbivorous feeding habits (the remaining species), the nature of which are fairly predictable based on examination of mouthpart morphology, but not entirely consistent with the tendency of cyrtacanthacridine species to feed on forbs.

Key Words: host preference, plant acceptance, mouthpart morphology

RESUMEN

Por medio de pruebas de selección de laboratorio utilizando 19 especies de plantas, evalua- mos el comportamiento de selección de hospederos de seis especies de saltamontes: Stenacris vitreipennis (Marschall), Leptysma marginicollis (Serville), Gymnoscirtetes pusillus Scudder, Paroxya clavuliger (Serville), Paroxya atlantica Scudder, y Romalea microptera (Beauvois). Este grupo de saltamontes, esta comúnmente asociado con habitats semiacuáticos, en el su- reste de los Estados Unidos. Estas especies poco estudiadas demuestran hábitos de alimen- tación graminívora (S. vetreipennis y L. marginicollis) y de alimentación mixta graminívora y de hierbas (las especies restantes), la naturaleza de los cuales son suficientemente prede- cibles basándose sobre la examinación de la morfología del aparato bucal, pero no totalmente consistentes con la tendencia de las especies cirtacanthacridines para alimentarse de plan- tas herbaceas.

Although often viewed as polyphagous herbi- ing. Such tests commonly are used to construct vores, most grasshoppers are selective to some de- preference hierarchies (Lewis and van Emden gree, exhibiting definite plant preferences 1986), but are constrained by experimental de- (Mulkern 1967). In previous studies, it has been sign. The investigator must have the wisdom to shown that grasshoppers are conveniently classi- present the correct array of plants, which should fied as grass-feeders (graminivorous), forb-feed- be based on the habitat in which the insect is ers (forbivorous), or a mix of the two (ambivorous found. Nevertheless, even good designs are sub- or mixed feeders) (Isely 1944). Phylogenetic dif- ject to faulty interpretation, as lack of a “pre- ferences exist among grasshoppers in relation to ferred” host among the array of choices may force host plant preferences (Dadd 1963, Joern 1979). a hungry individual to feed on non-preferred For example, members of the acridid subfamily plants which, in nature, might be accepted only if Gomphocerinae tend to have a preference for faced with starvation. grasses, Cyrtacanthacridinae (Melanoplinae in A very general method to determine the diet of part) prefer forbs, and Oedopodinae eat both a grasshopper is by the morphology of the grass- grasses and forbs (Dadd 1963, Joern & Lawlor hopper’s mandibles (Mulkern 1967, Patterson 1980, Otte 1981). Joern (1986) points out that 1984). The morphological characters of the man- most monophagous and polyphagous species are dibles, incisor and molar surfaces are useful in la- forb-feeders while oligophagous species are grass beling grasshoppers as grass- or forb-feeders feeders. (Chapman 1964, Bernays & Barbehenn 1987, Information on dietary habits of Florida’s Kang et al. 1999) though most species with forb- grasshoppers is growing, though still far from feeding mandibles feed on a mixture of grasses complete. Preference tests (Capinera 1993, and forbs. Isley (1944) suggested that the study of Scherer 1997) have been conducted for some of mandibular morphology would aid in under- Florida’s upland plants and crops, though infor- standing grasshopper ecology and their role in mation on most of Florida’s grasshoppers is lack- terrestrial communities.

Squitier & Capinera: Grasshopper Host Selection 337

We evaluated host selection behavior by grass- when 81-100% consumed. The feeding trials were hoppers among the plant species abundantly conducted under the same environmental condi- found inhabiting semi-aquatic habitats. Most re- tions mentioned earlier. The temperature and hu- search on grasshopper feeding behavior has been midity maintained in the laboratory during this conducted in arid environments, where grasshop- experiment was approximately the optimal feed- pers most often attain high and damaging levels ing range for most grasshoppers (Chapman 1957, of abundance. Very little is known concerning spe- Mulkern 1967). cies that inhabit moist or wet environments be- The number of grasshoppers per cage was ad- cause such species usually do not become pests. justed to reflect the individual appetites of the The host selection behavior displayed by grass- grasshoppers, thereby allowing measurable con- hoppers in choice tests was compared with host sumption without exhausting any of the plant preference predictions based on mouthpart (man- material. Thus, there were 5 Romalea microptera, dible) morphology. 6 Paroxya clavuliger, 6 Leptysma marginicollis, 6 Stenacris vitreipennis, 10 Paroxya atlantica, and MATERIALS AND METHODS 12 Gymnoscirtetes pusillus per cage. These grass- hopper populations resulted in relatively the Grasshoppers were collected from several wet same amount of plant material eaten between habitats: freshwater marshes, lakesides, flat- species over the 24-h test period, an average of woods and ditchbanks. The grasshopper species about 30%. included in this study were Stenacris vitreipennis We took steps to assure that the grasshoppers (Marschall) (glassywinged toothpick grasshopper), had ample opportunity to explore the cages and Leptysma marginicollis (Serville) (cattail tooth- host plants before registering acceptance of hosts pick grasshopper), Gymnoscirtetes pusillus Scud- by prolonged feeding. We maintained relatively der (little wingless grasshopper) Paroxya clavuliger low densities, and in no case was consumption (Serville) (olivegreen swamp grasshopper), Par- high enough on one plant species to influence con- oxya atlantica Scudder (Atlantic grasshopper), sumption of another plant. The cages were small, and Romalea microptera (Beauvois) (eastern lub- allowing the grasshoppers to encounter most or ber grasshopper). The grasshoppers were field- all plant species with relatively little movement. collected as large nymphs or adults and main- Therefore, we believe that the grasshoppers were tained in the laboratory during the experimental fully capable of assessing the host options, and period. The insects and choice tests were held in registered “preference” by their host consumption an insect growth room at 30-32°C with a photope- behavior. Observation of the insects confirmed riod of 14:10 (L:D) and a relative humidity of 40 ± that grasshoppers moved freely and often sam- 10%. Between study repetitions, the grasshop- pled plants without continuing to feed. pers fed Romaine lettuce, a plant that seems to The 19 plants that were evaluated in this have nearly universal acceptance by acridids. study were collected from the same habitats as The plant species were presented to the grass- the grasshoppers, and represented the most hoppers in a series of four-choice tests. The plants abundant floral elements in the semi-aquatic were clipped into equal sized portions, the stems habitats sampled. The study plants included wrapped in cotton, and the cuttings were placed Typha spp. (cattail) (Typhaceae); Eichhornia into vials filled with water in order to maintain crassipes (floating water hyacinth), and Ponte- plant turgidity. The 19 plant species were ran- deria cordata (pickerel weed) (Pontederiaceae); domly assembled into 6 clusters of 4 plants each, Urochloa mutica (para grass), Leptochloa spp. with each cluster containing cattail as a common (sprangletop), Panicum repens (torpedograss), plant, and the components not varying among Sacciolepis striata (American cupscale) and grasshoppers species or replicate tests. Each clus- Chasmanthium sessiliflorum (long leaf spike- ter was replicated 7 times, with replicates occur- grass) (Gramineae); Polygonum punctatum (dot- ring on different days. Each of the four plants was ted smartweed) and Polygonum hirsutum (hairy randomly placed at equal distances from the oth- smartweed) (Polygonaceae); Hydrocotyle spp. ers and from the sides of screen cages measuring (pennywort) and Cicuta mexicana (water hem- 0.3 m on each side. lock) (Umbelliferae); Ludwigia octovalvis (long The grasshoppers were exposed to the host fruited primrose willow) and Ludwigia suffruti- plants for approximately 24 h followed by estima- cosa (headed seedbox) (Onagraceae); Sagittaria tion of the amount consumed. The scale for deter- latifolia (common arrowhead) (Alismataceae); mining grasshopper consumption was taken from Cyperus compressus (poorland flatsedge) and Capinera (1993): the consumption was deter- Cyperus surinamensis (tropical flatsedge) (Cyper- mined by a visual estimate of the remaining plant aceae); Juncus efusus (softrush) (Juncaceae); and material and assigning it a value of 1 to 5. A value Sesbania macrocarpa (hemp sesbania) (Legumi- of 1 would be assigned when 0-20% of the plant nosae). For the purposes of this study the monocot was eaten, 2 when 21-40% consumed, 3 when 41- families of Typhaceae, Pontederiaceae, Gramineae, 60% consumed, 4 when 61-80% consumed and 5 Alismataceae, Cyperaceae and Juncaceae were

338 Florida Entomologist 85(2) June 2002 considered grasses and the dicot families Poly- gonaceae, Umbelliferae, Leguminosae and Ona- graceae were considered forbs. The mean consumption values for each plant from the 6 plant clusters were calculated for each grasshopper species using Graph Pad Software (Instat 1993) one-way analysis of variance (ANOVA). The individual means were then com- pared in each grasshopper species trial using the Tukey-Kramer multiple comparison test. Simul- taneously, mandibles of the grasshoppers were ex- amined visually from preserved specimens and classified as forb- or grass-feeding types according to the descriptions and drawings of Isley (1944).

RESULTS AND DISCUSSION Of the 6 species collected from the wetland plant habitats, 5 were in the subfamily Cyrtacan- thacridinae whereas Romalea microptera, though closely related to Cyrtacanthacridinae, is placed Fig. 1. Diagrams of the left mandible of the grass- in the subfamily Romaleinae or in the family hopper species: (a) Romalea microptera Beauvois, (b) Romaleidae. Based on subfamily taxonomy, such Paroxya clavuliger (Serville), (c) Paroxya atlantica cyrtacanthacridines might be expected to be forb- Scudder, (d) Leptysma marginicollis (Serville), (e) Sten- feeders. acris vitreipennis (marschall), (f) Gymnoscirtetes pusil- lus Scudder. Examination of the mandibles revealed that not all species were morphologically equipped to feed on forbs (Fig. 1). Three of the cyrtacanthridines, Paroxya clavuliger, Paroxya atlantica, and Gym- species tested. Other plant species were often noscirtetes pusillus possessed toothed mandibles, consumed about as readily, or more readily, than suggesting a tendency to feed on forbs. However, cattail. Gymnoscirtetes pusillus had low con- Leptysma marginicollis and Stenacris vitreipenni sumption values but showed preferences for cat- were found to possess blunt-toothed mandibles, tail, long fruited primrose willow, headed seedbox characteristics of grass-feeding species. Romalea and sprangletop. Paroxya atlantica showed pref- microptera displayed mandibles suitable for forb erence for cattail, pennywort, common arrow- feeding (Isley 1944, Patterson 1984). head, poorland flatsedge and water hemlock. The grouping of grasshoppers by mandible Paroxya clavuliger showed preference for cattail, type to predict host preference was, for the most water hyacinth, dotted smartweed, pennywort, part, confirmed with the food choice experiments water hemlock and sprangletop. Stenacris vit- (Table 1). The cyrtacanthridine species with forb- reipennis showed preference for cattail and poor- feeding mandibles proved to be mixed feeders, ac- land flatsedge. Leptysma marginicollis showed cepting both grasses and forbs. Gymnoscirtetes preference for cattail, pickerel weed, long fruited pusillus displayed a mixed preference by readily primrose willow and softrush. Romalea mi- consuming 2 grasses and 2 forbs. Paroxya atlan- croptera showed preference for cattail, common tica also displayed a mixed preference by selecting arrowhead, poorland flatsedge, headed seedbox, 3 grasses and 2 forbs, and Paroxya clavulger chose sprangletop and hairy smartweed. 2 grasses and 2 forbs. Romalea microptera also This study provides the first documentation of displayed a mixed preference, preferring 2 forbs the host selection behavior of the grasshopper as- and 4 grasses. The two species with grass-feeding semblage commonly associated with semi-aquatic mandibles, Stenacris vitreipennis and Leptysma habitats in the southeastern United States. These marginicollis, displayed strong preference for poorly studied species display both graminivorous grasses. Although in a small number of cases and forbivorous feeding habits, the nature of forbs were consumed, in no case was a forb con- which is fairly predictable based on examination sumed by these latter species more than a grass. of mouthpart morphology, but not entirely consis- Typha spp. (cattail) was used as a standard in tent with the tendency of cyrtacanthridine species each trial because it is a very common aquatic to feed on forbs. The graminivorous feeding behav- plant and relative consumption of this species ior of Stenacris vitreipennis and Leptysma mar- would allow comparison to the other plants. Con- ginicollis is also reflected in modified body form. sumption values for cattail ranged from 1.3 to 5.0 Both species display unusually long, thin bodies in the various tests. Overall, cattail is one of the that allow them to blend with narrow emergent most readily accepted hosts for the grasshopper grass vegetation. This crypsis undoubtedly makes

Squitier & Capinera: Grasshopper Host Selection 339 rt, FPdf 0.8 b 10.3 0.0002 3,24 0.4 b 16.3 0.0001 3,24 1.8 ab 4.5 0.0126 3,24 0.0 c 15.8 0.0001 3,24 1.0 ab 16.7 0.0001 3,24 0.9 ab 6.3 0.0027 3,24 0.8 b 10.0 0.0003 3,24 0.5 b 201.7 0.0001 3,24 1.9 a 2.9 0.0589 3,24 0.4 b 7.0 0.0015 3,24 1.1 a 2.3 0.1071 3,24 1.5 b 16.1 0.0001 3,24 0.0 a 1.6 0.2476 3,12 0.5 a 1.0 0.4262 3,12 0.0 b 14.3 0.0003 3,12 0.0 a 3.7 0.0439 3,12 0.5 a 2.1 0.1522 3,12 0.0 a 3.9 0.0369 3,12 0.4 b 8.1 0.0007 3,24 0.5 a 1.7 0.1846 3,24 0.0 b 25.5 0.001 3,24 0.0 b 4.6 0.0113 3,24 0.4 b 10.2 0.0002 3,24 0.0 b 23.5 0.0001 3,24 0.0 b 3.8 0.0242 3,24 0.0 b 13.4 0.0001 3,24 1.1 b 9.7 0.0002 3,24 0.4 b 3.0 0.0491 3,24 1.0 ab 4.1 0.0170 3,24 0.4 b 19.5 0.0001 3,24 0.4 b 5.9 0.0036 3,24 0.0 b 5.6 0.0047 3,24 0.4 a 1.0 0.4098 3,24 0.4 a 2.7 0.0760 3,24 0.5 a 4.3 0.0145 3,24 0.0 b 15.0 0.0001 3,24 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± . TESTS a

SD ± ater) hemlock, h(hemp) sesbania, torpedograss, h(headed) seedbox, t(tropical) flatsedge, CHOICE

0.0 b spikegrass 1.3 1.1 b flatsedge t. 1.1 1.1 a sesbania 2.9 0.9 bc cupscale a. 1.0 0.4 c willow p. 4.0 0.0 b smartweed d. 2.9 1.6 a spikegrass 1.5 0.0 a flatsedge t. 1.3 0.8 a sesbania 3.3 1.5 a cupscale a. 1.1 1.9 a willow p. 4.6 0.0 b smartweed d. 1.6 0.0 a spikegrass 1.0 0.5 a flatsedge t. 1.8 0.0 b sesbania 1.0 0.6 a cupscale a. 1.0 0.5 a willow p. 1.3 0.5 a smartweed d. 1.0 0.0 b spikegrass 1.1 1.0 a flatsedge t. 1.3 0.0 b sesbania 1.0 0.8 ab cupscale a. 1.0 0.4 b willow p. 1.1 0.5 b smartweed d. 1.0 0.4 ab spikegrass 1.0 1.1 b flatsedge t. 1.0 1.0 a sesbania 2.7 0.5 ab cupscale a. 1.1 0.5 b willow p. 2.1 0.0 b smartweed d. 1.1 0.0 b spikegrass 1.1 0.9 a flatsedge t. 1.0 0.4 a sesbania 1.1 0.5 a cupscale a. 1.1 0.0 b willow p. 1.7 0.0 b smartweed d. 1.0 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± LABORATORY

IN

GRASSHOPPERS

1.3 a smartweed h. 1.0 0.0 b seedbox h. 2.1 1.1 b hemlock 4.4 0.8 b flatsedge p. 1.9 0.4 a weed p. 1.1 1.7 ab para grass 1.0 1.5 a smartweed h. 4.3 0.5 b seedbox h. 5.0 1.6 a hemlock 4.5 1.2 a flatsedge p. 3.4 1.9 a weed p. 2.3 1.6 b para grass 1.0 0.5 a smartweed h. 1.0 0.5 a seedbox h. 1.3 0.0 a hemlock 1.0 0.0 a flatsedge p. 1.5 0.0 a weed p. 1.3 0.0 a para grass 1.3 0.4 b smartweed h. 1.0 0.5 a seedbox h. 1.7 0.4 b hemlock 1.0 0.0 b flatsedge p. 1.7 0.0 b weed p. 1.1 0.4 b para grass 1.7 1.3 ab smartweed h. 1.1 0.4 b seedbox h. 1.6 0.5 b hemlock 4.3 0.5 ab flatsedge p. 1.3 1.3 a weed p. 1.3 0.5 b para grass 1.0 1.0 a smartweed h. 1.0 0.0 b seedbox h. 1.8 0.0 a hemlock 1.1 0.0 a flatsedge p. 1.6 0.4 ab weed p. 1.0 0.0 b para grass 1.0 = 0.05) by Tukey-Kramer Multiple Comparison Test. Cattail, floating w(water) hyacinth, paragrass, d(dotted) smartweed, pennywo ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± P BY

PLANTS

AQUATIC - SEMI

OF

2.0 a sprangletop 4.0 1.4 a torpedograss 1.0 1.8 ab softrush 1.6 1.0 a arrowhead 2.3 1.7 b pennywort 4.9 1.7 a hyacinth w. 2.0 0.0 a sprangletop 3.8 0.0 a torpedograss 1.3 0.8 a softrush 2.7 1.6 a arrowhead 3.9 1.7 a pennywort 3.6 0.0 a hyacinth w. 2.3 0.6 a sprangletop 1.3 0.5 a torpedograss 1.3 0.5 b softrush 2.0 0.8 a arrowhead 1.0 0.5 a pennywort 1.0 0.8 a hyacinth w. 1.0 1.3 a sprangletop 1.4 1.8 a torpedograss 1.3 1.1 a softrush 1.1 1.5 a arrowhead 1.0 1.0 a pennywort 1.0 1.1 a hyacinth w. 1.1 1.5 a sprangletop 2.0 1.4 a torpedograss 1.1 1.4 b softrush 1.4 1.1 a arrowhead 1.6 1.5 ab pennywort 3.4 1.4 a hyacinth w. 1.4 1.0 ab sprangletop 2.4 0.5 ab torpedograss 1.0 0.8 a softrush 1.0 0.5 a arrowhead 1.0 0.5 ab pennywort 1.1 0.5 a hyacinth w. 1.0 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± CONSUMPTION

cattail 3.4 cattail 4.0 cattail 2.4 cattail 3.7 cattail 3.0 cattail 3.9 cattail 5.0 cattail 5.0 cattail 4.7 cattail 3.7 cattail 3.9 cattail 5.0 cattail 1.5 cattail 1.3 cattail 1.3 cattail 2.0 cattail 1.8 cattail 2.0 cattail 2.6 cattail 2.4 cattail 3.3 cattail 2.4 cattail 2.4 cattail 3.6 cattail 2.6 cattail 3.7 cattail 2.0 cattail 2.1 cattail 2.4 cattail 3.7 cattail 1.7 cattail 1.7 cattail 1.4 cattail 1.4 cattail 1.3 cattail 1.7 OMPARATIVE SD within rows followed by the same letter are not significantly different ( ± 1. C Means a ABLE clavuliger Paroxya microptera Romalea marginicollis Leptysma vitreipennis Stenacris atlantica Paroxya pusillus p(pickerel) weed, long fruited p(primrose) willow, common arrowhead, p(poorland) flatsedge, a(American) cupscale, softrush, w(w sprangletop, h(hairy) smartweed and long leaf spikegrass. Gymnoscirtetes Grasshopper species species and associated mean consumption values Plant T 340 Florida Entomologist 85(2) June 2002

them difficult to detect and presumably safer from ISLEY, F. B. 1944. Correlation between mandibular mor- predation. Interestingly, these grass-feeding cyrt- phology and food specificity in grasshoppers. Ann. acanthacridine species have very slanted faces, Entomol. Soc. Am. 37: 47-67. thereby physically resembling grass-feeding gom- JOERN, A. 1979. Resource utilization and community phocerines more than their close relatives, the structure in assemblages of arid grassland grasshop- pers (Orthoptera: Acrididae). Trans. Am. Entomol. forb-feeding cyrtacanthacridines. Soc. 105: 253-300. JOERN, A. 1986. Resource partitioning by grasshopper ACKNOWLEDGMENT species from grassland communities. Proc. Triennial Mtg. Pan Am. Acrid. Soc. 4: 75-100. This research was supported by Florida Agricultural JOERN, A., AND L. R. LAWLOR. 1980. Food and microhab- Experiment Station, and published as Journal Series itat utilization by grasshoppers from arid grass- number R-08174. lands: comparisons with neutral models. Ecology 61: 591-599. KANG, L., Y. GAN, AND S. L. LI. 1999. The structural ad- REFERENCES CITED aptation of mandibles and food specificity in grass- hoppers on Inner Mongolian grasslands. J. Orth. BERNAYS, E. A., AND R. BARBEHENN. 1987. Nutritional Res. 8: 257-269. ecology of grass foliage-chewing insects, pp. 147-175. LEWIS, A. C., AND H. F. VAN EMDEN. 1986. Assays for in- In F. Slansky and J. G. Rodriguez (eds.), Nutritional sect feeding, pp. 95-119. In J. R. Miller and T. A. Ecology of Insects, Mites, Spiders, and Related In- Miller (eds.), Insect-Plant Interactions. Springer- vertebrates. John Wiley & Sons, New York. Verlag, New York. CAPINERA, J. L. 1993. Host-plant selection by Schisto- MULKERN, G. B. 1967. Food selection by grasshoppers. cerca americana (Orthoptera: Acrididae). Environ. Annu. Rev. Entomol. 12: 59-78. Entomol. 22: 127-133. OTTE, D. 1981. The North American Grasshoppers. Vol- CHAPMAN, R. F. 1957. Observations on the feeding of ume I: Acrididae. Gomphocerinae and Acridinae. adults of the red locust (Nomadacris septemfasciata Harvard Univ. Press, Cambridge. 275 pp. (Serville)). Brit. J. Anim. Behav. 5: 60-75. PATTERSON, B. D. 1984. Correlation between mandibu- CHAPMAN, R. F. 1964. The structure and wear of the lar morphology and specific diet of some desert mandibles in some African grasshoppers. Proc. Zool. grassland Acrididae (Orthoptera). Am. Midl. Nat. Soc. London 142: 107-121. 11: 296-303. DADD, R.H. 1963. Feeding behavior and nutrition in grass- SCHERER, C. W. 1997. Response of grasshoppers (Orthop- hoppers and locusts. Adv. Insect Physiol. 1: 47-109. tera: Acrididae) to different forest restoration tech- INSTAT. 1993. Graph Pad Instat Mac Instat Statistics. niques in a Florida sandhill community. Unpublished Graph Pad Software, San Diego, CA. 110 pp. M.S. Thesis. Univ. of Florida.

Sourakov & Mitchell: Laboratory Biology of C. scutellaris 341

LABORATORY BIOLOGY OF CHETOGENA SCUTELLARIS (DIPTERA: TACHINIDAE), A PARASITOID OF NOCTUIDAE, REARED ON FALL ARMYWORM AND CABBAGE LOOPER

A. SOURAKOV1 AND E. R. MITCHELL2 1Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611

2Center for Medical, Agricultural and Veterinary Entomology, U.S. Department of Agriculture Agricultural Research Service, P.O. Box 14565, Gainesville, FL 32604, USA

ABSTRACT

The tachinid parasitoid Chetogena scutellaris (Wulp) was reared from southern armyworm, Spodoptera eridania (Cramer), a new host record. When reared in cabbage looper, Tricho- plusia ni (Hübner), (a new host) and in fall armyworm, Spodoptera frugiperda (J. E. Smith), in the laboratory, C. scutellaris developed successfully ca. 30% of the time. Success of para- sitism depended on the numbers of eggs laid per host, host age, and host species. Parasitoid development was not synchronized with host development. C. scutellaris developed mostly as a solitary parasitoid. Female flies preferred fifth instar hosts for oviposition. Cabbage looper was a better host than fall armyworm for mass rearing of this parasitoid.

Key Words: biological control, parasitoid development, southern armyworm, Spodoptera fru- giperda, Spodoptera eridania, Tachinidae, Trichoplusia ni

RESUMEN

El parasitoide taquínido Chetogena scutellaris (Wulp) fué criado de Spodoptera eridania (Cramer), un nuevo registro de hospedero. Cuando fué criado en el gusano medidor de repo- llo, Trichoplusia ni (Hübner), (un nuevo registro de hospedero) y en el gusano cogollero, Spo- doptera frugiperda (J. E. Smith), en el laboratorio, C. scutellaris se desarrolló exitosamente ca. 30% del tiempo. El éxito de parasitismo dependió del número de los huevos puestos en cada hospedero, la edad del hospedero, y la especie del hospedero. El desarrollo del parasi- toide no fué sincronizado con el desarrollo del hospedero. C. scutellaris se desarrolló princi- palmente como un parasitoide solitario. Las moscas hembras prefirieron ovipositar en el quinto estadío larval de los hospederos. El gusano medidor de repollo fué un mejor hospedero para criar el parasitoide en masa que el gusano cogollero.

The tachinid fly Chetogena scutellaris (Wulp) nia (Cramer), from lambsquarters, Chenopodium is largely known by its synonym Euphorocera album (L.), in a corn field near Bunnell, Florida. floridensis Townsend (Aldrich & Webber 1924). Approximately 25% of late instar larvae had one or Two families of Coleoptera (Cerambycidae and more tachinid eggs on their surface, and laboratory Coccinellidae) and 11 families of Lepidoptera colony was established from the resulting C. scutel- (Arctiidae, Citheroniidae, Ctenuchidae, Geometri- laris adults. Two major noctuid pests were used as dae, Hesperiidae, Noctuidae, Notodontidae, Sphin- hosts: cabbage looper, Trichoplusia ni (Hübner), gidae, Zygaenidae, Pierdae, and Saturniidae) are and fall armyworm. This study was conducted to listed as hosts for this species (Arnaud 1978). assess the biology of C. scutellaris in the laboratory C. scutellaris was recorded from seven noctuid and evaluate its potential for mass rearing. species that are crop pests: velvetbean caterpillar, Anticarsia gemmatalis Hübner; in Georgia, cot- MATERIALS AND METHODS ton leafworm, Alabama argillacea (Hübner), true armyworm, Pseudoletia unipuncta (Hawitson.), Host third-fifth instar larvae were maintained and corn earworm, Helicoverpa zea (Boddie); in on a pinto bean diet (Guy et al. 1985) and offered North Carolina, green cloverworm, Plathypena in groups of 20-30 larvae to C. scutellaris main- scabra (F.); in S. Carolina, Florida and Maryland, tained inside 20-cm3 cages with netting sides. fall armyworm, Spodoptera frugiperda (J. E. C. scutellaris tend to parasitize larger larvae, thus Smith); in Mississippi, soybean looper, Pseudo- to obtain parasitism in younger larvae they had to plusia includens (Walker). It is also known from be offered separately from the old ones. The time Costa Rica and Peru (Arnaud 1978). that larvae were exposed to flies varied, though In June, 2000, we collected ca. 100 late-instar usually flies attack larvae right away, so within an larvae of southern armyworm, Spodoptera erida- hour larvae were removed and checked for eggs.

342 Florida Entomologist 85(2) June 2002

Initially, flies were maintained in groups on escaped parasitism and developed into healthy honey and water, and then some were randomly adult moths. chosen for the experiments (freshly emerged cop- In 10 trials of 12, flies parasitized older larvae ulating pairs were carefully removed from the at a higher rate (fifth instars, 6.6 ± 0.4%; fourth communal cage into solitary ones, insuring that instars, 5.0 ± 0.5%, P < 0.05), which suggests that the female is young, mated and that it has a male older larvae are preferred for oviposition. The to- partner with it for further matings). Parasitized tal number of eggs laid also was higher in older larvae were identified by observing the eggs that larvae (fifth instars, 11.5 ± 1.1 eggs; fourth in- were laid on them. Host larvae with one or more stars, 6.5 ± 0.7 eggs, P < 0.05). The probability of tachinid eggs were transferred into individual successful parasitism increased from 29 ± 8.2% to 200-ml waxed paper cups with plastic lids, sup- 53.1 ± 4.1% (P < 0.05), when number of eggs ovi- plied with a diet cube, and checked daily for par- posited was more than one per host (Fig. 1). A sin- asitoid emergence. Thus, the development rates gle maggot emerged from 81% (N = 228) of the from egg to pupa, and from pupa to adult were hosts, though two-thirds of these hosts had multi- recorded. The study was repeated three times, ple parasitoid eggs laid on them. In 16% of the using flies and hosts of three consecutive genera- hosts, two parasitoids per host emerged and, only tions. Overall, 705 parasitized larvae were reared in 3% of the hosts did three or more (maximum of individually. six) parasitoids emerge. The latter parasitoids To estimate host age preference, 20 fall army- were smaller (e.g., the dry weight of an average worm larvae (10 fourth and 10 fifth instar) were fly was 10.7 mg, exceeding eight times the weight exposed for a period of five minutes to several 10- of a fly that developed gregariously in a group of d-old female flies in a similar cage. Larvae were six (1.3 mg)). When five or six parasitoids came then checked for parasitoid eggs, easily noticeable from a single host, half of the flies never emerged on the larval surface, to compare egg numbers from their puparium. Our data concerning influ- laid on hosts of different stages. This experiment ence of host age and number of eggs laid on suc- was repeated 12 times and was analyzed using cess of parasitoid development correspond with ANOVA and t-tests (“JMP”, SAS Institute 1995). similar studies on other species of Tachinidae Standard error values are provided for all means. (Konotie & Paolo 1992). The host stage from which the parasitoid RESULTS AND DISCUSSION emerged depended on the stage at which it was parasitized. When fourth-instar host larvae were Substantial variation was observed among in- attacked, 85% of maggots emerged from larvae dividual C. scutellaris females in their parasitism and 15% emerged from pupae (N = 65). In con- (emergence of maggot(s)) in hosts of different trast, when hosts were attacked as fifth instars, stages and species (Table 1). Higher percentage of 13% of maggots emerged from larva while 87% successful parasitism was observed when fifth- emerged from pupae (N = 139). Thus, this parasi- instar versus fourth-instar cabbage loopers were toid’s development does not appear to be synchro- attacked (46.5 ± 5% vs. 22.7 ± 4%, P < 0.05 (Means nized with development of its host. C. scutellaris’ ± SD)). In fall armyworm, differences in parasit- development time from egg to pupa at 21°C was ism were not statistically significant among lar- 12.3 ± 0.2 days (N = 145) with no significant dif- val stages. The decline in success of parasitism ference (P > 0.05) found between different hosts. towards the end of the fourth instar probably emerged from pupae in 13.3 ± 0.6 days. should be attributed to moulting prior to hatching C. scutellaris reproduce easily in the labora- of parasitoid eggs. Parasitized third-instar hosts tory on hosts fed artificial diet. Cabbage looper, a produced no parasitoids in either host species. species that has not been previously recorded as a Thus, majority (close to 70%) of attacked larvae host of C. scutellaris, would probably be attacked

TABLE 1. SUCCESSFUL PARASITISM OF CABBAGE LOOPER AND FALL ARMYWORM LARVAE BY CHETOGENA SCUTELLARIS.

Successful parasitism (% ± SE)

Stage Attacked Cabbage looper N Fall armyworm N

Early 4th 24.0 ± 5.0 23 49.0 ± 8.4 92 Mid-4th 30.5 ± 4.5 38 — 0 Late 4th 13.5 ± 8.5 30 2.9 ± 1.5 68 Early 5th 48.3 ± 16.3 56 50.0 ± 0.0 16 Mid-5th 47.3 ± 3.9 89 15.0 ± 8.0 93 Late 5th 44.0 ± 12.5 164 31.0 ± 11.6 36 Mean 34.5 ± 8.5 400 29.5 ± 5.9 305

Sourakov & Mitchell: Laboratory Biology of C. scutellaris 343

pate in soil. When these hosts were used, collect- ing pupae of C. scutellaris was labor intensive because they had to be extracted from the diet.

ACKNOWLEDGMENTS

We would like to thank Dr. Susan Webb and Dr. Rob Meagher for critical reviews of this manuscript. Dr. James O’Hara and Dr. Gary J. Steck identified the par- asitoids. This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation for its use by USDA

REFERENCES CITED

ALDRICH, J. M., AND R. T.WEBBER. 1924. The North American species of parasitic two-winged flies be- Fig. 1. Success of development of one or more Cheto- longing to the genus Phorocera and allied genera. gena scutellaris flies in relation to the number of eggs Proc. U.S. Natl. Museum No. 2486, Vol. 63, Art. 17, oviposited on a host larva. Increase was significant (P < pp. 1-90. 0.05) when one (29 ± 8.2%) and more than one (53.1 ± ARNAUD, P. H., JR. 1978. A host-parasite catalog of 4.1%) egg per host was laid. North American Tachinidae (Diptera), pp. 1-860. USDA, Misc. Pub. No.1319, Washington, DC. GUY, R. H., N. C. LEPPLA, J. R. RYE, C. W. GREEN, S. L. in the field should both the host and the parasi- BARRETTE, AND K. A. HOLLIEN. 1985. Trichoplusia toid occur synchronously (cabbage looper is ni, pp. 487-494 In Pritam Singh and R. F. Moore mostly a winter pest in the southeastern United [eds.], Handbook of Insect Rearing, Vol. 2, Elsevier States). Field trials are needed to evaluate poten- Science Publishers B. V., Amsterdam. tial of this species as a biological control agent of KONOTIE, C. A., AND F. PAOLO. 1992. Eucelatoria bryani noctuid pests. Cabbage looper also proved to be a Sabr. (Diptera Tachinidae) rearing on the factitious more convenient host for mass rearing C. scutel- host Galleria mellonella L. (Lepidoptera Galleri- idae): Effect of host age at exposure to the parasitoid laris. Cabbage looper larvae make cocoons on top females. Bolletino dell’Istituto di Entomologia of the cage, while maggots pupate on the bottom “Guido Grandi” della Univesita degli Studi di Bolo- where they can be easily collected. In contrast, gna. 46(0): 229-238. southern and fall armyworms pupate inside the SAS INSTITUTE. 1995. JMP, Version 3, SAS Institute diet cakes in the laboratory, as in nature they pu- Inc., Cary, NC.

344 Florida Entomologist 85(2) June 2002

MORPHOLOGY AND LIFE HISTORY CHARACTERISTICS OF MUCRONATUS (HETEROPTERA: )

SHERYL L. COSTELLO1, PAUL D. PRATT1, MIN B. RAYACHHETRY2 AND TED D. CENTER1 1USDA-ARS, Invasive Plant Research Laboratory, 3205 College Ave., Ft. Lauderdale, FL 33314

2Fort Lauderdale Research and Education Center, University of Florida 3205 College Ave., Ft. Lauderdale, FL 33314

ABSTRACT

Podisus mucronatus Uhler is a generalist predator found in Florida and the islands of the Caribbean. Adult P. mucronatus were observed preying on larvae of the Australian weevil Oxyops vitiosa (Pascoe), a biological control agent of Melaleuca quinquenervia (Cav.) S.T. Blake. To facilitate field-based identification of this predator, we present descriptions of eggs, nymphal stages, and adults. Life history traits of P. mucronatus when held with no food or either of two prey species (O. vitiosa and Tenebrio molitor (L.) larvae) are also reported. The potential use of this species as a biological control agent of arthropods and its interference with weed biological control are discussed.

Key Words: Podisus mucronatus, Pentatomidae, developmental rates, predatory stinkbug, Oxyops vitiosa, Melaleuca quinquenervia, Tenebrio molitor

RESUMEN Podisus mucronatus Uhler es un depredador generalista que se encuentra en Florida y en el Caribe. Se observaron adultos de P. mucronatus alimentándose de larvas de Oxypos vitiosa (Pascoe), un gorgojo australiano y un agente de control biológico de Melaleuca quinquenervia (Cav.) S. T. Blake. Se presentan descripciones morfológicas de los huevos, estadías ninfales 1-5, y los adultos, para facilitar la identificación de este depredador en el campo. Se reportan también características de la historia natural de P. mucronatus mantenidas sin alimento o sin las larvas de cualquier de las dos especies presa (O. vitiosa y Tenebrio mollitor (L.)). Se discute el uso potencial de esta especie como un agente de control biológico de artrópodos y su interferencia con el control biológico de malezas.

Species in the genus Podisus (Pentatomidae) are al. (1991) described the attractant pheromone pro- generalist predators, that attack primarily lepi- duced by male Podisus species, including that of dopteran and coleopteran larvae (Aldrich et al. P. mucronatus. Unfortunately, little else is known 1991). Because prey species include important pests concerning this species. in agroecosystems, some Podisus species have re- We recently observed adult P. mucronatus prey- ceived attention as potential biological control ing on larvae of the weed biological control agent agents for agricultural pests (Drummond et al. Oxyops vitiosa (Pascoe) (Coleoptera: Curculion- 1984, Stamopoulos & Chloridis 1994, De Clercq idae) that feed on Melaleuca quinquenervia (Cav.) 2000). Podisus maculiventris (Say), for instance, has S. T. Blake in Lee Co., FL. We wished to quantify been the focus of various life history and compara- the population dynamics of both O. vitiosa and its tive development studies as well as morphological newly acquired generalist predator, P. mucrona- descriptions of eggs, nymphs and adults (Aldrich tus. To date, no morphological descriptions of im- 1986, Decoursey & Esselbaugh 1962, Legaspi & mature stages or life history traits exist for this O’Neil 1993, Legaspi & O’Neil 1994). This attention predator, making accurate identification of eggs is due, in part, to the use of P. maculiventris as a bi- and nymphs difficult. To facilitate proper identifi- ological control agent of the Colorado potato beetle, cation and future understanding of its ecological Leptinotarsa decemlineata (Say) (Drummond et al. relationship with O. vitiosa, we describe herein 1984, Stamopoulos & Chloridis 1994). the basic morphological characteristics and life Only about 10% of the 300 known asopine spe- history traits of P. mucronatus. cies have been studied in detail (De Clercq 2000). For instance, the native Floridian and Caribbean MATERIALS AND METHODS species Podisus mucronatus Uhler has rarely been reported in the literature (J. Eger, pers. comm.). Rearing and Maintenance of P. mucronatus Colonies Occurrences of P. mucronatus attacking agricul- tural and horticultural pests were reported by Ge- Podius mucronatus adults, found in associa- nung (1959) and Genung et al. (1964). Aldrich et tion with O. vitiosa, were collected from coppicing

Costello et al.: Description and Life History of Podisus mucronatus 345

M. quinquenervia stumps at a site near Estero, brush to individual arenas. Twenty randomly se- Florida (N26°25.530’ W81°48.620’). The biological lected first-instar nymphs were assigned to each control agent was originally released at this site of 4 diets: 1) moistened filter paper only, 2) diet 1 in 1998 (Center et al. 2000). Adult P. mucronatus plus the addition of a M. quinquenervia tip, 3) diet were maintained in 30 × 25 × 9 cm diameter plas- 2 plus a single O. vitiosa larva (second-third in- tic containers provisioned with paper towels lin- star) or 4) diet 2 plus a single T. molitor larva (sec- ing the interior of the container. The paper towels ond-fourth instar). Melaleuca quinquenervia tips were used to provide seclusion and oviposition consisted of the terminal 4 cm of a newly devel- sites. Podisus mucronatus colonies were sus- oped, succulent shoot. All petri dishes were tained by feeding nymphs and adults mixed lar- stacked with an additional dish at the top, which val stages of Tenebrio molitor (L.) and O. vitiosa. contained only a moistened filter paper. Stacks of Paper towels and dead prey were removed petri dishes were placed in an environmental weekly. Adult colonies were examined every 12 h chamber at 25 (±1) °C, a photoperiod of 16:8 (L:D), at which time newly deposited eggs were removed and 65 (±10)% relative humidity. Developmental and placed into 10 × 1.5 cm diameter petri dishes. stages and survivorship were assessed every 24 h. Aged cohorts were held separately. Eggs and The presence of exuvia was used to assess molting first-instar nymphs were reared in 10 × 1.5 cm di- between stages. Melaleuca quinquenervia tips ameter petri dishes on filter paper with a small were replaced every 48 h. Oxyops vitiosa survivor- M. quinquenervia terminal vegetative bud (tip) ship was assessed daily by probing each larva for moisture. After the eggs hatched, decapitated with a camel’s hair brush, those not responding T. molitor third and fourth-instar larvae were were considered dead and were replaced. Tenebrio placed in the petri dish. Tenebrio molitor were de- molitor larvae were replaced every 48 h. capitated to facilitate predation by P. mucronatus nymphs. Second and third-instar nymphs were RESULTS then transferred into a 15 × 4 cm diameter plastic container with filter paper, several M. quinquen- Nymph and Adult measurements are presented in Table ervia tips, and live larvae of T. molitor. Fourth 1. and fifth-instar nymphs were reared in 18 × 8 cm diameter plastic containers and maintained in Eggs (Fig. 1A) the same manner. Colonies were held under labo- Egg shape elliptical, height 0.91 mm (±0.08; ± ° ratory conditions at 25 ( 5) C, a photoperiod of mean (SD), and width 0.80 mm (±0.05), laid in ± 16:8 (L:D), and 70 ( 10)% relative humidity. All clusters. Color opaque initially then silver or ma- stages were monitored every 12 h and exuvia roon with a shiny surface before eclosion. An av- were removed to accurately track life stages. erage of 8.25 (±0.90) micropylar processes occurring circularly around the operculum, gen- Morphological Descriptions and Measurements erally curving outward. Micropylar processes white terminating with a spherical structure, Each nymphal and adult stage was examined ranging from white to black. using a Nikon® dissecting microscope (10-50×). Descriptions were based on live individuals First-Instar Nymph whereas measurements were based on specimens preserved in ethyl alcohol. The number of individ- Shape oval, convex, widest at second or third uals measured ranged from 11-35, depending on abdominal segment. Head convex dorsally, widest the number available from each respective cohort. basally, narrowing to tip of tylus. Head brown and Size and number of micropylar processes were eyes red with a metallic appearance. Antennae 4- measured on 44 eggs. Length of each nymph and segmented, segments brown with white annuli. adult was measured from the tip of the tylus to the Antennae covered with increasingly more setae tip of the abdomen. The width of the head was towards apex. Rostrum 4-segmented and brown, measured from the outer margins of the com- apex extending slightly beyond metacoxae. pound eyes. The pronotal width of adults was mea- Thorax narrowest anteriorly, width increasing sured between the humeral spines. Lengths of the posteriorly. Length of pro-, meso-, and metatho- metathoracic leg and femur were also measured. rax medially equal. Dorsal sutures well defined. Dorsal color brown. Thorasic median line visibly Life History Parameters lighter in color. Coxa, femur, and tibia also brown with tarsi and tarsal claws gray. As stadium pro- Life history parameters were assessed in ex- ceeds, first two tarsal segments turn brown and perimental arenas that consisted of 10 × 1.5 cm tarsal claws elongate. Leg segments setose. Brown plastic petri dishes, provisioned with a slightly scent glands present on pro-epimeron and meta- moistened filter paper. Newly hatched (<10 h old) epimeron, becoming obscured after sclerotization. P. mucronatus first-instar nymphs were collected Abdominal sutures between segments distinct. from colonies and transferred using a camel’s hair Lateral margins of abdominal segments I and II

346 Florida Entomologist 85(2) June 2002

TABLE 1. MORPHOLOGICAL MEASUREMENTS OF PODISUS MUCRONATUS NYMPHAL AND ADULT STAGES.

Nymphal stage Morphological characteristic First Second Third Fourth Fifth Adult male Adult female

Body lengtha 1.24 (0.23)d 2.00 (0.22) 4.01 (0.92) 4.77 (0.35) 8.46 (0.80) 10.13 (0.88) 11.46 (0.94) Pronotal widthb 0.91 (0.16) 1.22 (0.17) 2.34 (0.34) 2.73 (0.26) 4.91 (0.35) 6.51 (0.47) 7.05 (0.47) Head widthc 0.56 (0.13) 0.73 (0.11) 1.33 (0.23) 1.34 (0.14) 2.16 (0.13) 2.22 (0.12) 2.28 (0.14) Antennal segment 1 0.07 (0.01) 0.08 (0.03) 0.15 (0.03) 0.12 (0.04) 0.21 (0.05) 0.27 (0.11) 0.20 (0.00) 2 0.20 (0.08) 0.38 (0.08) 0.83 (0.18) 1.02 (0.12) 1.69 (0.24) 1.20 (0.19) 1.27 (0.20) 3 0.18 (0.06) 0.30 (0.07) 0.62 (0.12) 0.71 (0.08) 1.14 (0.14) 0.97 (0.25) 0.99 (0.16) 4 0.28 (0.06) 0.38 (0.05) 0.64 (0.08) 0.67 (0.06) 0.95 (0.12) 1.14 (0.29) 1.19 (0.17) 5 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.00 (0.00) 0.95 (0.21) 1.00 (0.12) Total length 0.74 (0.19) 1.13 (0.18) 2.25 (0.37) 2.48 (0.19) 3.94 (0.40) 4.51 (0.80) 4.46 (0.43) Rostrum segment 1 0.19 (0.03) 0.30 (0.03) 0.44 (0.09) 0.51 (0.05) 1.07 (0.12) 1.03 (0.16) 1.19 (0.14) 2 0.18 (0.06) 0.35 (0.07) 0.64 (0.09) 0.66 (0.11) 1.13 (0.17) 1.22 (0.16) 1.28 (0.16) 3 0.16 (0.06) 0.22 (0.04) 0.35 (0.05) 0.43 (0.10) 0.89 (0.16) 0.96 (0.11) 1.06 (0.14) 4 0.19 (0.04) 0.27 (0.06) 0.49 (0.10) 0.51 (0.04) 0.76 (0.11) 0.93 (0.17) 0.94 (0.11) Total length 0.72 (0.16) 1.15 (0.14) 1.92 (0.27) 2.11 (0.13) 3.86 (0.29) 4.14 (0.30) 4.46 (0.31) Femur length 0.42 (0.08) 0.63 (0.10) 1.19 (0.28) 1.33 (0.24) 2.73 (0.25) 3.22 (0.81) 3.37 (0.36) Total leg length 1.09 (0.20) 1.58 (0.22) 2.92 (0.42) 3.81 (0.40) 7.23 (0.48) 7.80 (0.89) 8.22 (0.88)

aLength of each nymph and adult individual was measured from the tip of tylus to tip of abdomen. bThe pronotal width of adults was measured between the humeral spines. cThe width of the head was measured from the outer margins of the compound eyes. dmm (±SD). curve anteriorly, segments III-X curve posteriorly. next nymphal stage. Degree of expression of hour- Lateral margin of segments V and VI longest, lat- glass shaped plate decreases with each molt. In eral length of segments narrowing anteriorly and fully sclerotized nymphs, orange and brown colors posteriorly. Segments IX and X not distinguish- change to red and black. able on every individual. Abdomen orange with brown medial and lateral plates. Dorsal lateral Second-Instar Nymph (Fig. 1B) plates semi-circular in shape (see Fig. 1C). Ven- tral lateral plates similar except one spiracle Oval, convex, widest at third abdominal seg- present on segments II-XIII, centrally located be- ment. Head black, eyes dark red, similar in ap- tween midline and anterior margin, resulting in a pearance to first-instar nymph. Antennal seg- small emargination in each lateral plate. One tri- ments I,IV black, II,III gray, annuli white. Head chobothria present on segments III-VII, caudad shape and setae same as first-instar nymph. to spiracle, resulting in an additional emargin- Shape and coloration of thorax similar to first ation in lateral plates on venter. Intersegmental instar except as follows: Legs from coxa to tibia sulcus, originating from center of each lateral gray or brown with white annuli. Coxae black plate on venter, extends transversely. Lateral basally. Tarsal claws white, pulvilli gray to black. plates decreasing in size posteriorly, those of seg- Length of pro- and mesothoracic segments equal ment I minute. Dorsal medial pattern varied. medially, metathorax medially shorter than pro- Most common pattern: brown transverse central and mesothoracic segments. band on suture between segments I and II, hour- Abdominal shape and dorsal pattern same as glass shaped plate centered on segments II and first-instar nymph. Second ventral trichobothria III, rectangular plate centered on segments III-V, present on segments III-VII lateral to first tricho- semi-circle plate with arc towards apex centered bothria. Abdominal sutures less distinct than in on segment VI extending slightly onto segment V, first instar and intersegmental sulci not visible. a central oval macule on segments VII and VIII, Medioventral plates vary by size, darkness, and segments IX and X completely brown (see Fig. number of black oval plates. Predominant medio- 1B). Mediodorsal plates with three pairs of scent ventral plates dark red as follows: single large gland orifices and a visible connecting suture oval on segments I-IV, smaller oval on segments V curving posteriorly (see Fig. 1B). Mediodorsal and VI, third smaller oval on segments VII and pattern size increases with each exuviation to VIII. Segments IX and X dark red. Costello et al.: Description and Life History of Podisus mucronatus 347

Fig. 1. Podisus mucronatus adults and instars. A, egg clutch; B, second instar dorsal view; C, third instar dorsal view; D, fourth instar dorsal view; E, fifth instar dorsal view; F, fifth instar ventral view; G, adult dorsal view; H, adult ventral view. dp, dorsal abdominal pattern; edp, expanded dorsal abdominal pattern; ep, eye pattern; sc, semi- circle plate; vp, ventral abdominal pattern. 348 Florida Entomologist 85(2) June 2002

Third-Instar Nymph (Fig. 1C) ing more dense towards thorax, except for small white lateral area anterior to thorax. Head ven- Oval, convex, widest at third abdominal seg- trally white to green with small punctures. Eyes ment. Head shape, setae, and color same as sec- red to dark red and similar to fifth-instar. Anten- ond-instar nymph. Thorax color similar to second- nae 5-segmented, segments I-IV light brown, seg- instar nymph, except tarsi vary in color from ment V brown, annuli tan. Rostrum 4-segmented, brown to gray. each segment, base to apex, increasingly darker Abdominal shape and patterns slightly different from white and green to medium brown. from second-instar. Longest lateral margin varies Thorax dorsally covered with dark brown from segment V-VII. Dorsal abdominal sutures no punctation. Humeral spines dark brown and pro- longer visible, otherwise, dorsal and ventral pat- jecting forward (Thomas 1992). Anterolateral tern and coloration similar to second-instar. margins of pronotum serrate and green to yellow. Pronotum with 2 yellow cicatrices. Scutellum tri- Fourth-Instar Nymph (Fig. 1D) angular, apex and basal angles white to yellow. Oval, convex, widest at second abdominal seg- Corium and clavus red with same dark punctures ment. Head shape and color similar to third- as thoracic dorsum, claval suture white. Hemely- instar nymph. Last segment of rostrum dark tral membrane light brown. Hindwing iridescent brown, all others black. Ocelli dark red. Thorax with brown venation. Thorax gray to brown punc- similar to third-instar nymph except wing pads tured ventrally. Coxa, trochanter, femur, and tibia present on mesothoracic segment and median green with terminus of tibia brown. Three tarsal length of metathoracic segment shortened. segments brown, tarsal claws dark brown, and Abdominal morphology similar to third-instar pulvilli brown to dark brown. Protibia with brown nymph with few exceptions. A ventral interseg- spur two-thirds distance from apex to base. Legs mental sulcus reappears, similar to first-instar setose, setae denser toward apex. Scent gland nymph. Segment VIII with black ventral lateral ostiole located on intercoxal region of meta- pattern. Coloration of some nymphs vary from or- thoracic epimera. Ostiolar ruga white, yellow or ange and black to red and black. green. A white, elliptical callus also occurs on sternum of mesothorax, anterior to mesocoxae. Fifth-Instar Nymph (Fig 1E, 1F) Abdomen eight-segmented, length of segments medially equal except first, which is much shorter. Oval, convex, widest at third abdominal seg- Lateral margin of segment IV usually longest, ment. Head shape consistent with fourth-instar lateral length of segments narrowing anteriorly nymph. Head color varying from red to black, and posteriorly. Ventrally, single spiracle on each base of head usually black. Eyes and ocelli red. side of segments III-VII. Sternum white to green, Antennal segment I red to black depending on concolorously punctate laterally, punctures lack- maturity, segment II gray to black, segments III ing medially. Tergum brown to red with small and IV black, annuli white. Posterior portion of punctures. Connexiva green with larger punctures eye white in color and possibly lacking ommatidia than tergum. Dorsal and ventral sutures light in (see eye pattern, Fig. 1E). Last rostral segment color. Basal spine occurring ventrally on second brown, remaining segments black. abdominal segment and extending to middle of Thorax color variable black, red or orange. metasternum. Ventral posterior margin of male Legs typically with coxa, trochanter, and femur pygophore setose, setae most dense mesially. red, tibia, tarsi and tarsal claws black, annuli white. Legs all black if entire thorax black. Ven- Female Adult (Fig. 1G and 1H) tral pattern as in fourth-instar nymph. Dorsal pattern differing from previous stages. Scutellum Female similar in shape but generally larger and wing pads evident. Wing pads extending than male. Apex of eighth and ninth paratergites approximately to end of abdominal segment III. moderately setose, apex of first gonocoxae Lateral margins of thorax serrate, black and flat- sparsely setose. Other characteristics similar to tened dorso-ventrally. adult male. Abdominal shape and color same as fourth- instar nymph. All sutures and patterns more Life History Parameters defined. Ventral patterns and dorsal patterns Eggs collected from laboratory colonies were similar to fourth-instar nymph, but central dorsal laid in parallel rows, creating circular or elliptical pattern further expanded (see expanded dorsal masses of 28.18 (±10.25) eggs per group. Eye- pattern, Fig. 1E). spots were apparent under the pseudoperculum Adult Male 1-2 days before eclosion. Rates of nymphal devel- opment differed significantly among diets (Table Shape elliptical, wider anteriorly, dorsally flat- 2). When held with only moistened filter paper tened, ventrally convex, widest between humeral (Diet 1), three individuals (15%) survived to the spines. Head dorsally punctate. Punctures becom- second nymphal stage but did not develop further. Costello et al.: Description and Life History of Podisus mucronatus 349

When held with moist filter paper and a freshly Florida. To date the immature stages have never excised melaleuca tip (Diet 2), development of been described. The descriptions provided herein P. mucronatus was extended: 90% of those tested should facilitate future research on predator-prey developed to the second nymphal stage and 15% interactions with P. mucronatus. of these successfully molted to the third nymphal Morphological characteristics described herein stage (Table 2). Developmental times were simi- are useful for distinguishing various life stages of lar for most within stage comparisons when held P. mucronatus from other commonly occurring with prey, except second-instar nymphs held with Podisus species, specifically P. maculiventris and O. vitiosa, which required approximately one ad- P. sagitta (Fab.). For instance, P. mucronatus eggs ditional day to molt into third-instar nymphs. possess 7-11 (x– = 8.25, ±0.90) micropylar pro- When compared over the entire developmental cesses compared to 13-16 on eggs of P. maculiven- period, individuals feeding on T. molitor devel- tris and P. sagitta (De Clercq & Degheele 1990). oped faster than those on O. vitiosa (Table 2). Sur- Nymphal stages 1-4 of both P. sagitta and P. mac- vivorship was similar among both prey diets, with uliventris exhibit distinctive orange and white 5% mortality occurring among fifth-instar patterns on the abdominal tergum, which are not nymphs regardless of prey. Differences in sex ra- present in P. mucronatus (De Clercq & Degheele tios were also observed between diets (P = 0.04, 1990, DeCoursey & Esselbaugh 1962). Unlike F = 4.16, df = 1,37). Nymphs reared with T. moli- P. sagitta or P. maculiventris, adult P. mucronatus tor larvae were male biased (74:26; :; χ2 = 4.26, possess 3 distinctive white callused spots on the P = 0.039) whereas those reared with O. vitiosa scutellum and forward projecting humeral spines were not (53:47; χ2 = 0.053, P = 0.819). The inter- (Thomas 1992). val between last nymphal molt and first mating Consistent with other predatory pentatomids, for females was 2.67 (±0.71) days. Under labora- P. mucronatus successfully completed the first tory conditions, females began ovipositing 4.75 nymphal stage in the absence of prey, although (±0.39) days after mating. individuals were observed imbibing free water from the filter paper or from condensation in the DISCUSSION petri dish. Without prey, survivorship from sec- ond to third nymphal-instars was 0% when reared Generalist predators are integral components with only filter paper and 15% when reared with of most natural ecosystems due to their abilities a fresh M. quinquenervia bud. These findings sug- to regulate population densities of lower trophic gest that the presence of M. quinquenervia foliage levels in the absence of specialist predators increased nymphal survivorship but it remains (McMurtry 1992, Chang & Kareiva 1999, Hagen unclear if this is related to facultative plant feed- et al. 1976). However, rarely have trophic level in- ing or to an increase in relative humidity from the teractions of generalist predators and their prey vegetation within the arena. Not surprisingly, been reported for unmanaged systems (Chang & survivorship increased to 95% in the presence of Kareiva 1999). One explanation for the paucity of either coleopteran diet (Table 2). studies concerning generalist predators may be When compared between prey diets, total de- related to a limited knowledge of their identity velopment time was slower with O. vitiosa larvae and life history traits, specifically in immature than with T. molitor larvae (Table 2). One expla- stages. The predaceous pentatomid P. mucrona- nation for this may pertain to the antipredatory tus, for instance, is widely distributed in south activity of the viscous coating that covers imma-

TABLE 2. DEVELOPMENTAL RATES AND SURVIVORSHIP OF PODISUS MUCRONATUS WHEN REARED UNDER DIFFERING DI- ETARY REGIMES.

Duration (days) of nymphal stages: mean (SD)

Dieta 1 2345Total Survivorship

1 1.00 (0.00) bb ———— —0.00 a 2 0.95 (0.22) ab 2.33 (0.58) ab — — — — 0.00 a 3 0.70 (0.47) a 3.65 (1.18) a 2.45 (0.76) 2.55 (0.69) 3.87 (0.69) 13.60 (1.88) a 0.95 b 4 0.90 (0.45) ab 2.70 (0.66) b 2.35 (0.59) 2.35 (1.18) 4.00 (0.56) 12.30 (1.59) b 0.95 b

P-value 0.04c 0.01 0.64 0.52 0.80 0.02 <0.00

aDiets consisting of: 1) moistened filter paper only, 2) diet 1 plus the addition of a M. quinquenervia tip, 3) diet 2 plus a single O. vitiosa larva (2-3 in- star) or 4) diet 2 plus a 2-4th T. molitor larval instar. bStudent-Newman-Keuls all pairwise multiple comparison procedure (SigmaStat 1995). cOne-way ANOVA. 350 Florida Entomologist 85(2) June 2002

ture stages of O. vitiosa (Purcell & Balciunas CELL, AND P. D. PRATT. 2000. Field colonization of 1994). Montgomery & Wheeler (2000), for in- the melaleuca snout beetle (Oxyops vitiosa) in South stance, demonstrated that larvae covered with Florida. Biological Control 19: 112-123. this coating were repellent to the red imported CHANG, G. C., AND P. KAREIVA. 1999. The case for indig- fire ant, Solenopsis invicta Buren. While P. mucr- enous generalists in biological control, pp. 103-115. In B. Hawkins and H. V. Cornell [eds.], Theoretical onatus readily attacks O. vitiosa larvae under Approaches to Biological Control. Cambridge Univ. field conditions, it is unclear whether this larval Press, London 412 pp. coating influences development of nymphs. DEBACH. 1964. Biological Control by Natural Enemies. When comparing immature developmental Cambridge University Press, Cambridge 323 pp. rates with those of other Podisus species, P. mucr- DE CLERCQ, P., AND D. DEGHEELE. 1990. Description onatus (11-18 d at 25°C) appears to have a shorter and life history of the predatory bug Podisus sagitta developmental time than P. maculiventris (25-31 (FAB.) (: Pentatomidiae). Can. Ent. 122: d at 27°C), P. placidus (33.1 d at 27°C) and P. sag- 1149-1156. itta (18-32 d at 23°C) (De Clercq & Degheele DE CLERCQ, P. 2000. Podisus online. http://allserv.rug.ac. be/~padclerc/ Last updated: May, 4 2000. 1990). DeBach (1964) suggests that a short devel- DECOURSEY, R. M., AND C. O. ESSELBAUGH. 1962. De- opmental time is a desirable characteristic of scriptions of the nymphal stages of some North insect biological control agents. Assuming differ- American Pentatomidae (Hemipera-Heteroptera). ences in developmental times are not due to prey Ann. Ent. Soc. Amer. 55: 323-342. suitability or experimental design, rates of popu- DRUMMOND, F. A., R. L. JAMES, R. A. CASAGRANDE, AND lation increase for P. mucronatus may be higher H. FAUBERT. 1984. Development and survival of Po- than those Podisus species commonly introduced disus maculiventris (Hemiptera: Pentatomidae), a for pest suppression in other agroecosystems. predator of the Colorado potato beetle (Coleoptera: These findings suggest that P. mucronatus may Chrysomelidae). Environ. Entomol. 13: 1283-1286. GENUNG, W. G. 1959. Notes on the syntomid moth be a useful biological control agent of pest co- Lymire edwardsi (Grote) and its control as a pest of leopteran and lepidopteran larvae. Ficus in south Florida. Florida Entomol. 42: 39-42. Podisus mucronatus has been observed as the GENUNG, W. G., E. GREEN, JR., AND C. WELBERG. 1964. most common predator associated with the weed Inter-relationship of stinkbugs and diseases to Ever- biological control agent, O. vitiosa. Our labora- glades soybean production. Proc. Soil Crop Sci. Soc. tory-based data suggested that P. mucronatus Florida 24: 131-137. readily feeds and completes development when HAGEN, K. S., S. BOMBOSCH, AND J. A. MCMURTRY. provided O. vitiosa prey. However, it remains un- 1976. The biology and impact of predators, pp. 93- clear if P. mucronatus regulates populations of 142. In C. B. Huuaker and P. S. Messenger [eds.], Theory and Practice of Biological Control. Academic this weed biological control agent under field con- Press, New York 788 pp. ditions, and what impact this predation has on LEGASPI, J. C., AND R. J. O’NEIL. 1993. Life history of weed suppression. Future studies will focus on Podisus maculiventris given low numbers of Epil- assessing the association, predator-prey interac- anchna varivestis as prey. Environ. Entomol. 22(5): tions and population dynamics of this native pen- 1192-1200. tatomid and the introduced herbivore. LEGASPI, J. C., AND R. J. O’NEIL. 1994. Developmental response of nymphs of Podisus maculiventris (Het- eroptera: Pentatomidae) reared with low numbers of ACKNOWLEDGMENTS prey. Environ. Entomol. 23(2): 374-380. MCMURTRY, J. A. 1992. Dynamics and potential impact The authors would like to thank J. Eger for com- of ’generalist’ phytoseiids in agroecosystems and ments on earlier versions of the manuscript. We appre- possibilities for establishment of exotic species. Exp. ciate Susan E. Halbert of the Florida Department of Appl. Acarol. 14: 371-382. Agriculture (Division of Plant Industries) for the initial MONTGOMERY, B. R., AND G. S. WHEELER. 2000. Anti- identification of P. mucronatus. We also thank Willey predatory activity of the weevil Oxyops vitiosa: a bi- Durden for assistance with the photography of P. mucr- ological control agent of Melaleuca quinquenervia. J. onatus. This is Journal Article R-08322 of the Florida Ins. Behav. 13 (6): 915-926. Agricultural Experiment Station. PURCELL AND BALCIUNAS. 1994. Life history and distri- bution of the Australian weevil Oxyops vitiosa, a REFERENCES CITED potential biological control agent for Melaleuca quin- quenervia. Ann. Entomol. Soc. Amer. 87: 867-873. ALDRICH, J. 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Liang & Jiang: First Record and a New Species of Punana 351

PUNANA SINICA NEW SPECIES AND FIRST RECORD OF THE GENUS FROM CHINA (HEMIPTERA: FULGOROIDEA: DELPHACIDAE)

AI-PING LIANG AND GUO-MEI JIANG Department of Entomology, Institute of Zoology, Chinese Academy of Sciences 19 Zhongguancun Road, Beijing 100080, P.R. China

ABSTRACT Punana sinica Liang sp. nov. (Hemiptera: Fulgoroidea: Delphacidae) is described and illus- trated from Sichuan, southwest China. This represents the first record of the genus Punana Muir from China and the fifth known species of Punana. The new taxon extends the range of the genus Punana northward considerably, previously known only from southeast Asia and south India. A key for separation of the species of Punana is included.

Key Words: Punana, new species, Delphacidae, Fulgoroidea, China

RESUMEN Se describe y se ilustra Punana sinica sp. nov. (Hemiptera: Fulgoroidea: Delphacidae) de Si- chuan, en el suroeste de China. Este representa el primer registro del género Punana Muir en China y la quinta especie de Punana conocida en el mundo. Este nuevo taxón extiende la distribución geográfica del género Punana hacia el norte considerablemente, que antes se co- nocía solamente en el suroeste de Asia y en el sur de la India.

The Delphacidae is the largest family of the The genus Punana was described by Muir Fulgoroidea, comprising more than 2000 de- (1913) for P. brunnea Muir from Borneo. Asche scribed species in approximately 300 genera and (1983) correctly separated Punana from six subfamilies worldwide (Asche 1985, 1990). Neopunana (8 species, the Caribbean) and Equa- Members of the group are predominantly mono- systatus (monotypic, Ecuador). Four species are cot-feeders and a few are major agricultural pests currently included in the genus from Borneo, on grasses, such as rice, maize, and sugarcane Philippines (Luzon, Negros), and south India (Wilson & O’Brien 1987, Wilson et al. 1994). (Muir 1913, 1916, Distant 1916, Metcalf 1943, The delphacid fauna of China remains inade- Asche 1983, 1985). Asche (1985) placed Punana quately studied. The only comprehensive treat- Muir in the tribe Ugyopini Fennah 1979 of the ment of Chinese Delphacidae was that of Kuoh et subfamily Asiracinae Motschulsky 1863. How- al. (1983) in their Delphacidae volume of the Eco- ever, Emeljanov (1995) upgraded the Ugyopini to nomic Insect Fauna of China, which deals with the subfamily rank and recognized three tribes in 123 species distributed in 47 genera, 2 tribes and the Ugyopinae, e.g., Neopunanini Emeljanov 2 subfamilies. The number of described species 1995, Eodelphacini Emeljanov 1995 and Ugyo- likely represents only a small fraction of the ac- pini. Punana Muir was placed by Emeljanov tual diversity of the whole Chinese delphacid (1995) in the tribe Eodelphacini together with Eo- fauna considering the vast territory and various delphax Kirkaldy 1901, Ostama Walker 1857, complex habitats of China. Paranda Melichar 1903, Melanesia Kirkaldy The basal delphacid taxa, the Asiracinae and 1907, Livatiella Fennah 1956 and Prolivatis Vizcayinae, of Delphacidae in the Chinese fauna Emeljanov 1995. Eodelphacini is characterized by have received very little attention. To date, only the distal part of the aedeagal shaft arched clock- six species in four genera from the Asiracinae wise (from the base curved to the left); the pres- (Asiracini, Ugyopini) and Vizcayinae are de- ence of a row of teeth on the metatarsomere II, in scribed or recorded from China, e.g., Asiraca which the marginal teeth are considerably longer choui (Yuan & Wang) (Shaanxi in central China), than all others; the presence of the sinus on the A. clavicornis (F.) (Xinjiang in northwest China), hindwings opposite CuAP; forewings without A. granulipennis (Kato) (Manchuria and Jinlin in postnodal transverse veins; the presence of a northeastern China), and Ugyops zoe Fennah well-defined bend of the membrane when the (Hainan Island in south China), all belonging in wings are folded; and the intermediate carinae of the Asiracinae, and Vizcaya longispinosa Liang mesonotum straight (Emeljanov 1995). (Yunnan in southwest China) and Neovizcaya sin- In the present paper the senior author de- ica Liang (Yunnan in southwest China) of the scribes a new species of the genus Punana Muir subfamily Vizcayinae (Fennah 1956, Asche 1985, which was recently found in Sichuan, southwest 1990, Liang 1996, 1998, 2002). China. The new species represents the first record

352 Florida Entomologist 85(2) June 2002 of Punana in China, and its discovery has broad- ened our knowledge of the morphology and bio- geography of the genus, as well as that of the primitive delphacid taxa in the Chinese fauna.

MATERIALS AND METHODS The specimen used in this study is from the In- sect Collection of the Institute of Zoology, Chinese Academy of Sciences, Beijing, China (IZCAS). Morphological terminology follows that of Kramer (1950) and Asche (1985).

DESCRIPTIVE TAXONOMY Punana Muir Punana Muir, 1913: 249. Type species: P. brun- Figs. 1-2. Punana sinica Liang sp. nov. 1. male holo- nea Muir, 1913, by original designation. type, dorsal view; 2. same, lateral view. Onkelos Distant, 1916: 137. Type species: O. annulatus Distant, 1916, by original designation and monotypy. [Synonymized by Muir, 1919: 6.] late, veins distinctly prominent and thickly Head (Figs. 1, 3, and 4) relatively broad, broadly covered with fuscous granules with long setae, a roundly produced anteriorly. Vertex (Figs. 1 and 3) more or less continuous series of transverse veins very short, broader than long, disk foveate with a before apical area. For forewing and hindwing median carina (very faint in P. sinica sp. nov.) and venation see Asche (1985: fig. 204). two sublateral carinae which meet anteriorly with Male genitalia. See description of Punana sin- the extreme apical part incised posteriorly. Frons ica Liang sp. nov. below and Asche (1985: figs. (Fig. 4) longer than broad, anterior margin trun- 393, 441, 442). cate, convex medially, lateral marginal areas de- Remarks. The above generic description was pressed, centrally finely carinate. Postclypeus (Fig. based on the specimen of the new species de- 4) more than half length of frons, convex medially, scribed below. lateral marginal areas depressed, with median Punana is distinguished from other genera in longitudinal carina. Anteclypeus (Fig. 4) very nar- the tribe Eodelphacini by the shape and length of row and convex, with median carina. Rostrum the antennae, the shape of vertex and frons, the long, reaching between hind trochanters, basal number of longitudinal carinae on mesonotum, segment relatively long, apical segment relatively the number of lateral spines on hind tibiae, the short, greater than 1/2 length of basal segment. wing venation, and the minutiae of the male gen- Antennae (Figs. 1, 3, and 4) moderately long, scape italia, as noted above. and pedicel with many sturdy bristles on surface, Punana can be distinguished from Melanesia scape shorter than pedicel, with base distinctly Kirkaldy (7 species, Borneo, Philippines and Fiji) narrow, strongly broadening toward apex, apex and Paranda Melichar (monotypic, Sri Lanka) by distinctly broad; pedicel cylindrical with base the hind tibiae having 3 lateral spines (2 in slightly flattened, with about 18 sensory fields. Melanesia and Paranda) and the antennal pedicel Pronotum (Figs. 1 and 3) centrally slightly shorter distinctly short (distinctly elongate in Melanesia than head, disk slightly sloping anteriorly, ante- and Paranda). It can be distinguished from Eodel- rior lateral areas strongly sloping laterad, hind phax Kirkaldy (2 species, Sri Lanka) by the an- margin centrally slightly arched anteriorly, with tennal scape and pedicel subcylindrical (both median longitudinal carina. Mesonotum (Figs. 1 scape and pedicel compressed and scape obliquely and 3) longer than head and pronotum combined, triangular in Eodelphax). Punana differs from with five longitudinal carinae on disk, extreme Prolivatis Emeljanov (monotypic, Vietnam) in the apex somewhat lobately produced. Legs moder- antennal pedicel short (relatively long in Proliva- ately long, hind tibiae with three lateral spines on tis) and the male parameres relatively broad outer edge, five apical spines (the outer one largest (very slender and narrow in Prolivatis). It differs and the middle one smallest) and a long apical con- from Ostama Walker [2 species, Borneo, Mentawi ical mobile spur, metatarsomere I with 5 apical Island (East Indies)] in the antennae short (very spines: four spines in a row and the fifth, middle elongate in Ostama), mesonotum with 5 carinae spine, shifted proximally from the row, metatar- (3 in Ostama) and forewings with about 12 closed somere II with 3 apical spines (outer side 2, inner apical cells (about 15 in Ostama). Punana can be side 1). Forewings (Figs. 1 and 2) relatively narrow separated from Livatiella Fennah (2 species, east- and elongate, about 3 times as long as broad, ob- ern Caroline Island) by the forewing with Sc+R liquely depressed laterally, apices broadly angu- with two branches before subapical transverse

Liang & Jiang: First Record and a New Species of Punana 353

nodal line (one branch in Livatiella) and the hind- P. brunnea Muir, 1913 (Borneo), P. negrosensis

wing with the apical cell enclosed by M3 and M4 Muir, 1916 [Philippines (Negros)], P. philippina relatively long and large (very short and small in Muir, 1916 [Philippines (Luzon)], and P. sinica sp. Livatiella). nov. [southwest China (Sichuan) (new record)]. Included species and distribution. Five spe- A key to the known species of Punana is pro- cies; P. annulata (Distant, 1916) (south India), vided below.

KEY TO SPECIES OF PUNANA MUIR

1. Antennae with three dark rings (Muir 1913); Borneo...... brunnea Muir —Antennae without dark rings ...... 2 2. Forewings with distinct stramineous suffusions, especially on basal, subapical and apical areas ...... 3 —Forewings without distinct stramineous suffusions ...... 4 3. Vertex (Figs. 1-3) shorter and broader with anterior margin shorter and more rounded; male pygofer (Figs. 5 and 6) with a large median triangular process on ventrocaudal margin; and parameres (Figs. 5, 8, and 9) dis- tinctly angulately produced laterad near midlength in ventral aspect; southwest China (Sichuan) ...... sinica Liang sp. nov. —Vertex relatively long, relatively distinctly produced anteriorly (Distant 1916: Fig. 98; Asche 1985: Figs. 11c, 161); male pygofer without distinct process on ventrocaudal margin (Asche, 1985: Fig. 393); parameres not angulately produced laterad near midlength in ventral aspect (Asche 1985: Fig. 441); south India ...... annulata (Distant) 4. Male pygofer with median process on ventrocaudal margin angular (Muir 1916); fore and middle coxae and fem- ora very light brown; clavus without fuscous spots; Philippines (Luzon)...... philippina Muir —Male pygofer with median process on ventrocaudal margin square (Muir 1916); fore and middle coxae and femora dark brown; clavus with a small fuscous spot; Philippines (Negros) ...... negrosensis Muir

Punana sinica Liang sp. nov. (Figs. 1 and 2) fuscous brown, with irregular stra- (Figs. 1-10) mineous suffusions mainly on basal, subapical and apical areas, granules on veins fuscous, setae Holotype male, CHINA: Sichuan, Wan County in granules on veins pale yellowish brown; hind- (30°8’N, 108°3’E), 1200 m, 27.v.1994 (Jin Yao) (IZ- wings pale fuliginous, veins brown. Abdomen CAS). with tergites brown and sternites ochraceous Description. Length (from apex of vertex to tip with anterior margin broadly brown. of forewings): male 5.8 mm. Structural characters as in generic description, Vertex, frons, postclypeus and anteclypeus but vertex with disk short and transversely broad, brownish, the pit on frons in front of vertex with very faint median longitudinal carina, frons darker, frons (Fig. 4) with two columns of small with a very small shallow rounded pit dorsally, rounded pale spots (one next to median longitudi- and pronotum shorter than vertex medially. nal carina and the other on lateral margin), epis- Male genitalia with pygofer (Fig. 5) in ventral tomal suture ochraceous, genae and lora view with apical 2/5 broad and nearly parallel- ochraceous with the latter suffused with fuscous. sided, narrowing to base over basal 3/5, ventrocau- Antennae ochraceous with apex of scape and dal margin broadly excavated with a large median pedicel suffused with brown, scape and pedicel triangular process. Pygofer (Fig. 6) in lateral view covered with long brownish sturdy bristles. Ros- elongate, very short anteriorly and very high pos- trum with basal segment ochraceous, apical seg- teriorly, dorsal margin very short and ventral mar- ment brown with extreme apex black. Pronotum gin very long. Parameres (Figs. 5, 6, 8, and 9) ochraceous with anterior lateral depressed areas slender and elongate, distinctly angulately pro- blackish brown. Mesonotum (Figs. 1 and 3) fus- duced laterad near midlength in ventral aspect, cous or blackish brown, with longitudinal carinae base relatively broad, tapered to apex, somewhat (excluding the median carina), posterior lateral constricted medially, apical 1/2 directed inwardly margins and extreme apex ochraceous. Thorax and covered with long hairs on surface. Connective ventrally ochraceous, pleurae with fuscous spots. (Fig. 9) relatively short, slender, parallel-sided, Legs ochraceous with dark brown or fuscous suf- apex and base expanded laterad. Aedeagal shaft fusion, femora dark brown or fuscous (hind fem- (Figs. 5, 6, and 10) nearly C-shaped with basal 1/2 ora much paler), tibiae with two broad fuscous compressed and apical 1/2 spinous; gonopore at rings basally and medially respectively (those on midlength on dorsal surface. Suspensorium (Fig. hind tibiae much paler), tarsi and claws brown 10) long, compressed, base ring-shaped, embrac- (hind tarsi and claws much paler), tips of apical ing base of aedeagus, apical part curved and broad spines on hind tibiae and tarsi black. Forewings with apex bluntly forked. Anal tube (Figs. 6 and 7) 354 Florida Entomologist 85(2) June 2002

Figs. 3-10. Punana sinica Liang sp. nov. 3. head, pronotum and mesonotum, dorsal view; 4. head, ventral view; 5. pygofer, ventral view; 6. pygofer, lateral view; 7. anal segment, caudal view; 8. genital styles, ventral view; 9. gen- ital style and connective, ventral view; 10. aedeagus, lateral view. Scale bars = 0.5 mm in Figs. 3, 4 and 0.2 mm in Figs. 5-10. short and broad, gradually widening from base to & Consumer Service, Gainesville, FL, USA) for review- apex in dorsal aspect, apex without appendages, ing the manuscript and suggesting improvements. The anal style slender, small and short. work on which this paper is based was supported by the Female. Unknown. National Natural Science Foundation of China grant Etymology. This new species is named sinica number 39925006 and the Chinese Academy of Sciences Biological Innovation Fund A2999084. after the Latin sinicus, adjective (“of China”), re- ferring to its occurrence in China. Distribution. Southwest China (Sichuan). LITERATURE CITED Remarks. This new species is externally simi- ASCHE, M. 1983. Aufgliederung der Asiracinen—Gat- lar to P. annulatus (Distant) from south India tung Punana Muir, 1913; Equasystatus gen. nov. aus (Kodaikanal), which was illustrated by Distant Equador und Neopunana gen. nov. von den Karibis- (1916) and Asche (1983, 1985), but differs from chen Inseln (Homoptera Auchenorrhyncha Fulgoro- the latter in having the antennal pedicel subcy- morpha Delphacidae). Marburger Entomol. Pub. 1(8): lindrical and in the shape of the vertex, male gen- 127-166. italia and pygofer, as noted in the above key. ASCHE, M. 1985. Zur Phylogenie der Delphacidae Leach, 1815 (Homoptera Cicadina Fulgoromorpha). Marburger Entomol. Pub. 2(1-2): 1-912. ACKNOWLEDGMENTS ASCHE, M. 1990. Vizcayinae, a new subfamily of Delpha- cidae with revision of Vizcaya Muir (Homoptera: We thank Dr. Manfred Asche, Museum für Natur- Fulgoroidea)—a significant phylogenetic link. Bishop kunde der Humboldt-Universität zu Berlin, Germany Mus. Occ. Papers 30: 154-187. and Dr. A. F. Emeljanov, Zoological Institute, Russian DISTANT, W. L. 1916. The fauna of British India, includ- Academy of Sciences, Leningrad, Russia, for several dis- ing Ceylon and Burma. Rhynchota Vol. 6 (Homoptera: cussions during the preparation of this manuscript. We Appendix). Taylor & Francis, London. viii + 248 pp. also thank Dr. Manfred Asche, Dr. Steve Wilson (Depart- EMELJANOV, A. F. 1995. On the question of the classifi- ment of Biology, Central Missouri State University, cation and phylogeny of the Delphacidae (Homop- Warrensburg, MO, USA), Dr. Lew Deitz (Department of tera, Cicadina), with reference to larval characters. Entomology, North Carolina State University, Raleigh, Entomol. Obozr. 74: 780-794 [In Russian with Eng- NC, USA), Dr. R. M. Baranowski (University of Florida, lish summary; Russian summary separately pagi- TREC, Homestead, FL, USA) and Dr. Gary Steck (Divi- nated, pp. 944-945. English translation in Entomol. sion of Plant Industry, Florida Department of Agriculture Rev. 75(9): 134-150, 1996.] Liang & Jiang: First Record and a New Species of Punana 355

FENNAH, R. G. 1956. Fulgoroidea from southern China. MUIR, F. A. G. 1913. On some new Fulgoroidea. Proc. Proc. California Acad. Sci. (4)28: 441-527. Hawaiian Entomol. Soc. 2: 237-269, pl. 6. KRAMER, S. 1950. The morphology and phylogeny of MUIR, F. A. G. 1916. Additions to the known Philippine auchenorhynchous Homoptera (Insecta). Illinois Delphacidae (Hemiptera). Philippine J. Sci. 11: 369- Biol. Monog. 20: 1-109, pls. 1-15. 385. KUOH, C.-L., J.-H. DING, L.-X. TIAN, AND C.-L. HWANG. MUIR, F. A. G. 1919. Notes on the Delphacidae in the Brit- 1983. Economic Insect Fauna of China. Fasc. 27. ish Museum collection. Canadian Entomol. 51: 6-8. Homoptera: Delphacidae. Science Press. Beijing, 166 WILSON, S. W., C. MITTER, R. F. DENNO, AND M. R. WIL- pp., 13 pls. [In Chinese, English abstract p. 137.] SON. 1994. Evolutionary patterns of host plant use LIANG, A.-P. 1996. Taxonomic changes in Chinese Lopho- by delphacid planthoppers and their relatives, pp. 7- pidae with a check list of Chinese species (Homoptera: 45. In R. F. Denno and T. J. Perfect (eds.), Planthop- Fulgoroidea). Pan-Pacific Entomol. 72: 145-151. pers: their ecology and management. New York: LIANG, A.-P. 1998. On the Eurasian planthopper genus Asi- Chapman Hall. raca Latreille (Homoptera: Auchenorrhyncha: Fulgoro- WILSON, S. W., AND L. B. O’BRIEN. 1987. A survey of morpha: Delphacidae). Reichenbachia 32: 187-196. planthopper pests of economically important plants LIANG, A.-P. 2002. New taxa of Vizcayinae (Hemiptera: (Homoptera: Fulgoroidea), pp. 343-360. In M.R. Wil- Auchenorrhyncha: Delphacidae), including a remark- son and L.R. Nault (eds.), Proc. 2nd Int. Workshop on able new genus from China. J. Nat. Hist. 36: 601-616. Leafhoppers and Planthoppers of Economic Impor- METCALF, Z. P. 1943. General Catalogue of the Hemip- tance. CAB International Institute of Entomology, tera. Fasc. IV. Fulgoroidea, Part 3. Araeopidae (Del- London. phacidae). Smith College, Northampton, MA. 552 pp.

356 Florida Entomologist 85(2) June 2002

TROLLING: A NOVEL TRAPPING METHOD FOR CHRYSOPS SPP. (DIPTERA: TABANIDAE)

RUSSELL F. MIZELL, III,1 RUSSELL F. MIZELL, IV2 AND RYAN A. MIZELL3 1University of Florida, NFREC-Monticello, Route 4, Box 4092, Monticello, FL 32344

2Metropolitan Life Insurance Co., Inc., Tampa, FL

3Undergraduate, Junior, University of Florida, Gainesville, FL 32611

ABSTRACT

Trolling, a novel trapping method, was developed and tested for deer flies, Chrysops spp., and other Tabanidae. The trap is a plastic plant container coated with Tanglefoot that is mounted upside down on a rod in an apparatus attached to a vehicle. The vehicle is then driven “trolled” to attract Tabanidae. Trap movement, color, shape, dimension and size were evaluated to improve trap catch. Response by Chrysops vittatus Wiedemann/C. pikei Whit- ney and C. macquarti Philip to the trap’s parameters are reported. The most effective trap is a 15 cm diameter pot (Lerio C-360, B-6, The Lerio Corp. Mobile, AL) painted bright blue, placed 1-2 m above the ground and moved at a speed of less than 3.13 m/sec. Response of Chrysops spp. to the trap indicated the hierarchy of behavioral stimuli to Chrysops spp. in order of importance to be height, movement, speed, dimensions, color, size, and contrast. No

known tabanid attractants tested including CO2, acetone or octenol increased trap captures and the insect repellent, DEET, N,N-diethyl-3-methylbenzamide, did not reduce trap cap- tures. Other biological information derived during the trapping experiments is reported. The trolling trap appears very valuable to detect and monitor certain Chrysops spp. and other ta- banid populations for scientific purposes. In addition the trap or modifications of it, mounted either on or near humans (hat or walking stick) or on vehicles, may be useful to reduce or eliminate attacks from Chrysops spp. or to suppress their activity for short time periods in small areas such as dooryards.

Key Words: Chrysops vittatus, C. pikei, C. macquarti, trap, Tabanidae, trolling

RESUMEN

Trolear, un novedoso método de captura, fue desarrollado y probado para moscas del genero Chrysops spp., y otros Tabanidae. La trampa consiste en un envase plástico para plantas recubierto con Tanglefoot que esta montado el revés sobre una barra en un aparato que va adherido a un vehículo. El vehículo posteriormente es manejado “troleado” para atraer Ta- banidae. El movimiento de las trampas, color, forma, dimensión y tamaño fueron evaluados para mejorar la capacidad de captura de la trampa. La respuesta por parte de Chrysops vittatus Wiedemann/C. pikei Whitney y C. macquarti Philip a los parámetros de la trampa son reportados. La trampa más efectiva es un envase de 15 cm de diámetro (Lerio C-360, B- 6, La Corp. Lerio, Mobile, AL) pintado de color azul brillante, colocado a 1-2m sobre el suelo y movida a una velocidad menor a 3.13 m/seg. La respuesta de Chrysops spp. a la trampa, indicaba la jerarquía del estimulo de comportamiento a Chrysops spp. en orden de importan- cia el cual era altura, movimiento, velocidad, dimensiones, color, tamaño, y contraste. Nin-

gún atrayente para tabanidos conocido que se haya probado incluyendo CO2 acetona o octenol, aumentó el numero de capturas en la trampas, y el repelente de insectos, DEET, N, N-dietil-3-metilbenzamida, no redujo el numero de capturas en la trampa. Otra información biológica obtenida durante los experimentos de trampeo se reportan. La trampa de troleado parece ser muy valiosa para detectar y monitorear ciertos Chrysops spp. y otras poblaciones de tabanidos para propósitos científicos. además la trampa o modificaciones de esta, coloca- das tanto sobre o cerca de humanos (sombreros o bastones para caminar) o sobre vehículos, pueden ser útiles para reducir o eliminar ataques de Chrysops spp. o para suprimir su acti- vidad por cortos periodos de tiempo en pequeñas áreas tal como patios de casa.

Detection and monitoring of tabanids many types of adult traps for monitoring taban- (Diptera: Tabanidae) began when Hansens (1947) ids, especially horse flies have been developed. demonstrated that Tabanus nigrovittatus Mac- The Manitoba fly trap was described and used by quart could be captured with black shingles Thorsteinson (1958) and Thorsteinson et al. coated with an adhesive material. Since then, (1965). Black and red spheres and two- and three-

Mizell et al.: Trolling: A Novel trap for Chrysops spp. 357

dimensional silhouettes were used by Bracken et (CO2) in silhouette traps and captured 23 species al. (1962) and Browne and Bennett (1980). Mal- of female tabanids. Wilson (1968) using sticky aise traps (Smith et al. 1965, Wilson et al. 1966, traps with CO2 on a pasture periphery signifi- Roberts 1971a,b), silhouette traps simulating cantly reduced tabanids on cattle in the pasture. cows (Wilson et al. 1966), red and black helium- Red and black helium-filled spheres tethered 1.2- filled spheres tethered 1.2-1.8 m above the ground 1.8 m above the ground captured large numbers (Snoddy 1970), modified Manning traps (Granger of Chrysops niger taylori Philip, but Snoddy

1970), canopy traps (Catts 1970, French & Kline (1970) determined that CO2 was a poor attractant

1989, Hribar et al. 1991a), sticky traps (Hansens for deer flies. Using the canopy trap with CO2, et al.1971), rigid canopy traps (Axtell et al. 1975), Catts (1970) reported capturing ca. 1000 tabanids aerial netting (Tallamy et al. 1976, Cilek and per hour in Delaware. Roberts (1971a and b) used Schreiber 1996), box traps (Wall & Doane 1980, Malaise traps and elucidated a species-specific re-

Ailes et al. 1992), panel traps (Allan & Stoffalano sponse to CO2 rates by tabanids. Roberts (1976a)

1986), two-tiered box traps (Jackson et al. 1993, compared six types of traps with and without CO2 French and Hagan 1995), and brown boards on for collection of tabanids including the Malaise, the ground and white buckets in the air (Moore et plexiglass and canopy traps. Based on the num- al. 1996) have been used to successfully monitor ber of species captured, the Stoneville Malaise many tabanid species. However, different trap (Roberts 1971a) with CO2 was the most efficient. types vary in their ability to capture deer flies Six Tabanus spp. and C. flavidus Wiedemann (e.g., Chrysops spp.) or other tabanid subgroups were captured in numbers suitable for statistical (horse flies), but can provide diverse information analysis (Roberts 1976a). French and Kline about the behavior of individual species. Tallamy (1989) investigated the response of tabanids to

et al. (1976) compared Malaise traps to aerial net- canopy traps with CO2 plus the tsetse fly attrac- ting and reported that the latter favored tant (Hall et al. 1984) 1-octen-2-ol (octenol) iso- Chrysops spp. while Malaise traps were better in- lated from cattle. Octenol alone increased canopy dicators of tabanid diversity and species even- trap catch three fold over traps without attrac-

ness. Neither trap alone should be used in tant. Octenol with CO2 enhanced trap catch in to- tabanid community studies. tal specimens and number of species captured. Trap color is important to tabanid attraction Both Chrysops spp. and Tabanus spp. were af- and landing. Black and red spheres (Bracken et al fected. Schreck et al. (1993) evaluated configura- 1962, Snoddy 1970) were found to be superior to tions of inflated vinyl beach balls in Malaise and green and yellow spheres and white dappling or canopy traps as possible insecticide-impregnated striping of black traps dramatically reduced at- visual targets. Beach balls attracted 2× and 2-5× traction of Tabanus illotus (O.S.). Browne and more flies respectively when used alone or when Bennett (1980) reported that both Chrysops spp. treated with octenol. Leprince et al. (1994) inves- and Hybomitra spp. were attracted to blue or red, tigated tabanid response in Louisiana to Jersey but were consistently not attracted to black, yel- bullocks and canopy traps baited with ammonia,

low or white. Tabanids landed in the areas octenol and CO2. Odors did not significantly in- painted the attractive color on striped traps with crease trap catch. The results of Leprince et al. alternating attractive and unattractive colors. Al- (1994) were contrary to the findings of French and lan and Stoffalano (1986) reported that T. nigro- Kline (1989) and Hribar et al. (1991a), and sug- vittatus preferred blue, black and red panel traps. gest that there may be environmental, geograph- Increased trap capture occurred when trap inten- ical and species differences in response to sity was increased or decreased against a back- attractants by tabanids. Foil and Hribar (1995) ground. Wall and Doane (1980) used box traps of tested the tsetse fly attractants acetone, and green, black or blue to trap T. nigrovittatus from octenol + 3-n-propylphenol and 4-methylphenol 1970-1979 and indicated a measurable decrease (4:1:8 ratio) in canopy traps in Louisiana. Four- in the flies over the trapping period. Moore et al. teen species or species groups of tabanids were (1996) used brown boards on the ground and captured in equal numbers to octenol and the white buckets on poles to determine emergence, mixture, response to acetone was similar to the concentrations and flight periodicity of T. abactor control. Jackson et al. (1993) and French and Philip. Male flies preferred the boards while fe- Hagan (1995) used a two-tiered box trap to collect male flies preferred buckets (Moore et al. (1996). T. nigrovittatus, C. fuliginosus Wiedemann and Hribar et al. (1991b) showed that reduction of C. atlanticus Pechuman. Octenol significantly in- ultraviolet (UV) light increased catch of certain creased the catch of C. atlanticus. tabanids. However, Hribar and Foil (1994) re- Despite the development of traps and the tested the effect of UV reflectance of canopy traps knowledge of tabanid behavior, effective monitor- on tabanids in Louisiana and concluded that UV ing and management methods, and even basic un- had little effect on most species. derstanding of the behavior of the thousands of Odors have also been used to enhance trap re- species of tabanids remain unknown. The objec- sponse. Wilson et al. (1966) used carbon dioxide tives of this investigation were to develop and test

358 Florida Entomologist 85(2) June 2002 a novel trapping method and to use the trap to 1-5 min. through an area containing deer flies. Af- characterize the behavior of selected tabanid spe- ter each replication the captured flies were re- cies. The trap described herein is most effective moved and recorded by species and trap. Trap against Chrysops spp. and the responses of three position was rotated clockwise to the next adja- species trapped in large numbers near Monti- cent position for the succeeding replication. Traps cello, Florida, C. vittatus Wiedemann, C. pikei in the right end position were returned to the op- Whitney and C. macquarti Philip, are reported. posite left-end position. Replication generally was conducted such that each trap type was posi- METHODS AND MATERIALS tioned in each test location 1-3 times. Paints used for small pyramid and square Trap development progressed over a series of shapes were Glidden Rust Master Enamel. Paints experiments in an effort to exploit the observation used for the plastic pots were TRU-TEST HI-G, of large numbers of deer flies (Chrysops spp.) high gloss enamel of the following colors, 105 Chi- chasing and landing on the side mirrors of a vehi- nese red, U-48 Royal blue (darkest), U-9 Dutch cle. Experiments were conducted during August- blue (lightest), and FL-5 Neon blue (brightest), a September, 1994, April-August, 1995, May-Au- TRU-TEST “Easy Color” fluorescent spray. Black gust, 1996, and May-August, 1998. All trap sur- was used as the natural color of the plastic pots. faces were covered with a thin layer of heated Spectral reflectance of the blue traps (Fig. 2) Tanglefoot (The Tanglefoot Co., Grand Rapids, MI was measured using a USB UV-VIS S2000 spec- 49504) applied with a small paint brush. Because trometer with a DT-1000-MINI tungsten light C. vittatus and C. pikei are not possible to sepa- source (Ocean Optics, Dunedin, FL). Percent re- rate in the field, their counts were combined. flectance was determined in comparison to a In preliminary experiments we determined white Spectralon standard. that two-dimensional black silhouette traps Traps were baited with candidate chemicals against a completely white background and using the methods of Foil and Hribar (1995). mounted on the top or on the sides of a moving ve- Data were analyzed using the SAS Proc GLM hicle would not induce tabanid species flying procedures. Means were separated by least signif- along side to land. Therefore, we developed the icant difference test when a significant F was indi- described “trolling” apparatus used for the exper- cated by analysis of variance (SAS Institute 1999). iments to mount traps on a vehicle. Experiment 1: Pyramid-shaped traps (Tedders All experiments were conducted using a 1992 & Wood 1994) ca. 30 cm in height and 15 cm wide Dodge Dakota Clubcab pickup, white with gray- at the base, made of clear plexiglass, unpainted striped side panels and black mirrors. An appara- cardboard or cardboard painted black, red, yellow, tus was mounted on the front and/or back of the or white and coated with Tanglefoot were used. truck to support the traps and enable rotation of Eighteen replicates were conducted with one pyr- traps among 7 different positions. The apparatus amid of each color mounted on both the front and (Fig. 1) consisted of a 2.5 × 5.0 cm wooden lath back of the truck. Nine replicates were conducted base 1.8 m long with 2, 30 cm long pieces nailed by driving the truck forward and 9 by driving the perpendicularly on the bottom to serve as support truck backwards for a one minute bioassay. Cap- braces. On the front of the vehicle each end of the tures were processed as described above. The data base was strapped tightly to the truck wheel wells were analyzed as a three-way analysis of variance using bungi cords. On the back of the vehicle the with direction (driving forward or backwards), po- apparatus was mounted into the standard holes in sition on the truck (front or back), and trap color the truck bed. Along the length of the base at ca. as the factors. 30 cm intervals from each end, two 10 penny nails Experiment 2: Black plant pots (15 cm) (Lerio were driven through the bottom of the wood lath C-650, B-6, The Lerio Corp., Mobile, AL) were com- ca. 2 cm apart so as to orient with their points ver- pared in six replicates to the cardboard pyramid tically through the top side. Small wooden blocks traps of plain brown cardboard, black or white. each with a top centered hole and with 2 bottom Experiment 3: Black unpainted plant pots holes to fit snugly over the nails were attached. (Lerio) of 15, 17.5, and 20 cm were compared to Metal rods 30 cm long of ca. 5 mm wire were flat squares of cardboard of 15, 17.5, and 20 cm on formed at the top into a L shape and the straight a side and painted black on both sides in eight bottom was placed into the top hole in the wooden replicates. block. A small binder clip was fastened over the Experiment 4: Trap size was tested in this two wire’s L-shaped end for trap support. Plant pot part experiment. Black Lerio pots of 5, 7.5, 10, traps were placed on the rods upside down by 12.5, 15, 17.5, and 20 cm in diameter were used. drilling 7 mm hole in the center of the bottoms. Po- In part one trap sizes from 5-15 cm were tested in sition of the traps was changed by moving the 15 replicates. In part two the trap sizes of 15, traps from rod to rod after each replicate. 17.5, and 20 cm were tested with 8 replicates. Experiments were replicated 5-15 times with Experiment 5: Red, U-48 Royal blue and black each replicate consisting of driving the vehicle for 15-cm pots were tested with 8 replicates. Two

Mizell et al.: Trolling: A Novel trap for Chrysops spp. 359

Fig. 1. The “trolling” deerfly trap: A. truck-mounted apparatus used in experimentation, B. cup version mounted on cap for personal protection and C. individual trap that can be mounted on a lawnmower, 4-wheeler or other ve- hicle to suppress deer flies from dooryards and other locations.

traps of each color were used and the pots of the 3 white and black. The striped pots were compared colors were grouped in two sets to begin the first to two solid black pots in five replicates. replicate then randomized as described above. Experiment 8: The “size” test (expt. 4) was re- Experiment 6: Plant pots (15 cm) were painted peated using pots painted FL-5 Neon blue. Pots with one of three different blue paints. In ascend- similar to those described in experiment 3 of sizes ing order of intensity (authors’ vision, Fig. 2) the 5, 10, 15, 20, and 25 cm were compared in ten rep- colors were: U-9 Dutch blue, FL-5 Neon blue, U- licates. 48 Royal blue. Ten replicates were conducted with Experiment 9: Background color under the 2 traps of each color randomized by position on trap. Cardboard fruit harvesting boxes (25 × 30 × the apparatus. 45 cm) were stapled to form a cup shape. The in- Experiment 7: Two 15 cm black pots were side of one box each was painted white, FL-5 painted with 2.5 cm alternating vertical stripes of Neon blue or covered with aluminum foil. To pro-

360 Florida Entomologist 85(2) June 2002

Fig. 2. Spectral reflectance patterns of three blue paints tested on traps for deer fly response. FL-5 Neon and U- 48 Royal blues captured significantly more deer flies than U-9 Dutch blue (see text). vide a trap background under the trap, the center Experiment 12: Attractants and repellents. of the cupped box was placed over the nails (fitted Using the methodology of Hribar and Foil (1994) into the wood for holding the rods) on the truck- for elution devices and concentration ranges, CO2 mounted apparatus which held the box in place. (2.5 l/min - open box of dry ice), acetone (500mg/ FL-5 Neon blue15 cm traps were placed on the hr), and octenol (0.5-5.0 mg/hr) were evaluated to rods as described above in the center of the back- determine the attraction and repellence to C. vit- ground boxes. Fifteen replicates were conducted tatus/C. pikei. CO2 was investigated using con- and trap position was changed after 5 replicates trolled elution rates and dry ice in a variety of were completed so that each trap was similarly containers to produce a range of low to high vol- located next to the other 2 traps for 5 replicates, umes both on and around the traps. Ten repli- i.e., three blocks in time. cates were also conducted using cannister CO2 Experiment 10: Height above ground: A eluded through a flowmeter at the rate of 2.5 l/ wooden lath of 5 × 5 cm was used to construct a min. For octenol, 3 FL-5 Neon blue 15 cm pot 3.7 m pole that fit snugly into the stake holder on traps were used with one trap containing the at- the truck bed. Another piece of lath drilled to hold tractant and 2 traps serving as controls. Traps the metal rod supporting the trap was nailed per- were separated on the apparatus by 50 cm (2 sta- pendicular to the pole at 3 heights: 150, 300 and tions) and position was randomized after each of 450 cm. A trap was also placed in the truck bed on the 11 replications. The octenol with 98% purity a rod which was at 75 cm above the ground. Traps was purchased from Sigma Chemical Co. Histo- of 15 cm diameter painted FL-5 Neon blue were logical grade acetone was purchased from Fisher mounted on the perpendicular stakes at 150, 300, Scientific and was bioassayed as described for and 450 cm and 15 replicates were conducted. octenol with 7 replicates. N,N-diethyl-3-methyl- Experiment 11: Silhouettes of 30 × 60 cm made benzamide, DEET, was tested using several elu- of blue chloroplast sheeting (BBE Graphics Ware- tion methods: vials as with octenol after Hribar house, Tampa, FL) were mounted under a trap and Foil (1994) and larger dental wicks to provide placed at 150 cm above ground on the bed of the higher concentrations. Results from the 8 repli- truck as described for the height experiment. Two cates conducted with dental wicks are reported. silhouette traps were compared for each replicate. C. macquarti: Experiments (1) with color, size The silhouettes were placed either vertically or and background were conducted in April- May, horizontally to mimic an animal body under the 1997 when this early-emerging species was ob- trap “head”. Replicates were conducted with the served in relatively high numbers. Color and size silhouettes oriented differently or the same for were investigated together using 2 Lerio pots side by side comparison of the effect on deer fly re- each, one black and the other FL-5 Neon blue, of sponse. diameter 7.5, 15, and 22.5 cm. The six pots were

Mizell et al.: Trolling: A Novel trap for Chrysops spp. 361

initially arranged on the truck apparatus with TABLE 1. RESPONSE OF CHRYSOPS VITTATUS WIEDE- the traps of similar size and color adjacent and MANN/C PIKEI WHITNEY TO PYRAMID TRAPS OF then randomized as described above for each of DIFFERENT COLORS AND A 15 CM BLACK PLANT the 12 replicates. Experiment 2 testing back- POT WHEN “TROLLED” ON A MOVING VEHICLE. ground colors was conducted as described above for C. vittatus/C. pikei with 16 replicates. Popula- Experiment 1A Experiment 2 tions of C. macquarti were much lower than C. Trap Type Mean ± SE Mean ± SE vittatus/C. pikei and also more variable in space. Thus, the numbers of flies captured by traps in Black pyramid 2.1 ± 0.33 Aa 3.5 ± 0.8 A each replicate were more variable and trap counts Tan pyramid 1.5 ± 0.29 AB 5.2 ± 0.6 AB often were zero. Therefore, the data were trans- Red pyramid 1.8 ± 0.27 B — formed before analysis of variance by taking the Clear pyramid 0.2 ± 0.07 C — square root of trap capture number +1. The un- White pyramid 0.2 ± 0.06 C 0 ± 0 C transformed means are reported. Yellow pyramid 0.2 ± 0.05 C — Black plant pot — 8.2 ± 2.3 A RESULTS aMeans in columns not followed by the same letter are significantly different as determined by LSD. Trap efficiency was ca. 95% or better as long as the Tanglefoot was fresh. Deer flies occasionally escaped from the Tanglefoot. Therefore, at the be- inefficient, but “trolling” appeared promising, we ginning and during the experiments as needed, compared other potential trap types. Significant the Tanglefoot was refreshed by rubbing the sur- differences were determined by analysis of vari- faces of two traps together. ance with F = 7.3; df = 3, 20; P = 0.0017 and LSD The seasonal occurrence and emergence dates = 3.7. Black 15 cm plant pots captured 8.2 ± 2.3 by years were: first detection of C. vittatus/C (mean ± SEM) C. vittatus/C. pikei, in comparison pikei—20 April 1995, 3 May 1996, 29 March 1997, to 5.2 ± 0.6 or less for the other pyramid traps (Ta- and 9 April 1998. The last collection date for C. ble 1). vittatus/C. pikei was 9 October 1998. We did not Experiment 3, two-dimensional squares vs evaluate the diurnal activity of specific Chrysops plant pots of 3 sizes: Shape was significantly dif- spp, but we did collect C. vittatus/C. pikei from ferent (F = 42.3; df = 1,42; P = 0.0001), size was dawn to dusk and occasionally just after dark. C. not significant (F = 2.8; df = 2,42; P = 0.074) and macquarti emerged first in Spring and was the shape x size interaction was significant (F = present only from late March to early May in 4.92; df = 2,42; P = 0.012) indicating that the 1998. C. macquarti is also much less aggressive three-dimensional 15 cm pot trap captured signif- towards humans and smaller in size than C. vit- icantly more (13.9 ± 2.2) than the 15 cm two-di- tatus/C. pikei. These data agree generally with mensional trap (1.4 ± 0.6). In this test the 15 cm Jones (1953) who reported two generations of black pot trap also captured significantly more C. vittatus in north Florida. C. vittatus/C. pikei than the 17.5 cm (7.5 ± 1.0) Experiment 1, colored pyramid traps: Colored and 20 cm black pot traps (7.3 ± 1.5), LSD = 2.34. pyramid traps mounted on the front on rear of the Experiment 4, size and color interaction: This vehicle captured deer flies inefficiently relative to experiment was conducted in two parts. In evalu- the numbers of C. vittatus/C. pikei that were fly- ating black pot traps varying in size from 5-15 cm ing around them. However, they indicated that we found significant differences (F = 3.84; df = dark colors were more attractive (F = 17.4; df = 4,70; P = 0.007) LSD = 3.44. In part two, evaluat- 5,216; P = 0.0001) (Table 1). Comparison by anal- ing traps of 15, 17.5, and 20 cm, we also found a ysis of variance of the trap catch on the front and significant size effect (F = 6.72; df = 2,21; P = rear of the vehicle driven either forwards or back- 0.006) LSD = 2.2 (Table 2). The 15 cm pot trap wards indicated no significant differences: direc- captured significantly more C. vittatus/C. pikei in tion (F = 0.92; df = 1, 216; P = 0.33) vehicle end both experiments. (F = 1.39; df = 1,216; P = 0.24) but the interaction Experiment 5, 15 cm pots of red, black and of direction x vehicle end was significant (F = blue: Blue traps captured significantly more C. 8.07; df = 1, 216; P = 0.005). The interaction was vittatus/C. pikei than the red and black traps, (F significant because the number of deer flies cap- = 21.5; df = 2,135; P = 0.0001) LSD = 1.56 (Table tured on the front (engine end) of the vehicle was 3). larger when the vehicle was driven forward or Experiment 6, 15 cm pots of 3 intensities of backward. This result may indicate that heat blue: FL-5 Neon blue and U-48 Royal blue pots from the engine or other characteristic of the ve- captured significantly more C. vittatus/C. pikei hicle front (white hood providing more contrast) than the U-9 Dutch blue traps, (F = 4.68; df = may have enhanced attraction and/or landing. 2,59; P = 0.01) LSD = 1.54 (Table 3). Experiment 2, black plant pot vs colored pyra- Experiment 7, 15 cm black pots with white midal traps: Because the pyramidal traps were stripes: Striped traps captured significantly less 362 Florida Entomologist 85(2) June 2002

TABLE 2. RESPONSE OF CHRYSOPS VITTATUS WIEDEMANN /C PIKEI WHITNEY TO BLACK AND NEON BLUE PLANT POT TRAPS OF DIFFERENT SIZES WHEN “TROLLED” ON A MOVING VEHICLE.

Trap Color Size (cm) Black Black Neon Blue Neon Blue

5.0 2.4 ± 0.7 Ba —1.9 ± 0.59 B — 7.5 3.5 ± 0.7 B — — — 10.0 4.2 ± 0.8 B — 4.5 ± 0.93 A — 12.5 4.3 ± 0.6 B — — — 15.0 8.7 ± 2.3 A 8.0 ± 1.4 A 4.3 ± 0.94 A 7.1 ± 1.2 A 17.5 — 3.5 ± 0.8 B — — 20.0 — 4.0 ± 0.5 B 3.7 ± 0.68 AB 7.4 ± 1.6 A 22.5———— 25.0 — — 3.8 ± 0.87 AB 4.9 ± 1.2 A

aMeans in columns not followed by the same letter are significantly different as determined by LSD.

C. vittatus/C. pikei 1.72 ± 0.22 (total = 34) than placed at 75 and 150 cm above ground. No deer the solid black traps (2.84 ± 0.39, total = 94) (F = flies were captured on traps placed higher. 6.33; df = 1,18; P = 0.022) (Table 3). The over- Experiment 11: Blue silhouettes of 60 × 120 cm whelming majority of the deer flies captured on had no effect on fly response in any orientation the striped trap landed on the black areas and relative to trap placement. avoided the white. Experiment 12, response to odors: Response by Experiment 8, repeat of the size test with FL- C. vittatus/C. pikei to octenol (5.1 ± 1.5) was not 5 Neon blue pot traps: Part one indicated a signif- significantly different from the unbaited control icant size effect (F = 1.59; df = 4, 45; P = 0.019) (5.2 ± 0.8) (F = 0.01; df = 1,31; P = 0.93). Similarly, LSD = 2.32 (Table 2). Part two evaluating 15, 20, no differences were found in response by C. vitta- and 25 cm traps did not indicate significant size tus/C. pikei in these bioassays to FL-5 Neon blue differences (F = 1.02; df = 2,29; P = 0.37). Trap traps baited with acetone (11.9 ± 3.0 vs 10.6 ± ± ± captures in relation to FL-5 Neon blue trap size 1.2), CO2 (5.9 1.1 vs 5.2 0.9) or DEET (control differed from the results obtained with the black -10.0 ± 2.8 vs DEET - 8.4 ± 3.1). traps in experiment 4 (Table 2). C. macquarti: Experiment 1, size and color in- Experiment 9, background contrast: A FL-5 teraction: The two-way analysis of variance indi- Neon blue trap on a white background captured cated that color (F = 14.38; df = 1,66; P = 0.0003) significantly more C. vittatus/C. pikei (16.7 ± 2.2) had a significant effect on C. macquarti response than traps with a FL-5 Neon blue background with FL-5 Neon blue trap collection (11.4 ± 1.1) (11.6 ± 1.1) which captured significantly more greater than black traps (6.5 ± 0.7). Size (10, 15, than the traps with a background of aluminum 22.5 cm) was not significant (F = 1.16, df = 2, 66; foil (6.7 ± 1.0) (F = 10.49; df = 2,42; P = 0.0002) P = 0.32) nor was the color x size interaction (F = LSD = 4.44. In the blue trap with blue back- 1.81, df = 2, 66; P = 0.17). ground treatment, some deer flies landed on the Experiment 2, background contrast: Back- background which decreased trap capture. ground color had a significant effect on trap re- Experiment 10, trap height. All C. vittatus/ sponse by C. macquarti, (F = 3.55; df = 2,45; P = C. pikei deer flies were captured on the traps 0.037) LSD = 0.57 with FL-5 Neon blue (6.81 ±

TABLE 3. RESPONSE OF CHRYSOPS VITTATUS WIEDEMANN /C PIKEI WHITNEY TO PLANT POT TRAPS OF DIFFERENT COL- ORS AND STRIPES WHEN “TROLLED” ON A MOVING VEHICLE.

Trap Type (15 cm) Experiment 5 Experiment 6 Experiment 7

Black 3.43 ± 0.46 Ba — 2.84 ± 0.39 A Red 3.87 ± 0.68 A — — Neon Blue 8.10 ± 0.51 B 4.20 ± 0.60 A — Royal Blue — 3.95 ± 0.62 A — Dutch Blue — 2.05 ± 0.40 B — Black & White Stripes — — 1.72 ± 0.22 B

aMeans in columns not followed by the same letter are significantly different as determined by LSD. Mizell et al.: Trolling: A Novel trap for Chrysops spp. 363

0.93) and white (6.69 ± 1.46) equal, but greater Movement of the traps was necessary to elicit than response to traps with an aluminum foil deer fly landing. Both quality and quantity of trap background (3.63 ± 0.97). movement were important. The method of trap suspension on the metal rods allowed the traps to DISCUSSION rotate and to shake back and forth during troll- ing. Trolling on the moving truck provided angu- The specimens of C. vittatus/C. pikei and C. lar displacement and velocity. A crude macquarti captured on traps in this study were determination indicated that a speed of ca. 3.16 almost all females with only a rare male. Parity of m/sec (7 mi/h) was the maximum vehicle velocity the females was not tested. Occasional samples of at which flying C. vittatus/C. pikei could land on captured flies dissected for gut contents found the traps. This figure is within the range of the about equal numbers with and without clear gut flight speed reported for tsetse flies (2.5-5m/s) contents on any date. No dissected deer flies con- (Gibson and Brady 1985). Deer flies, as do many tained evidence of blood feeding. hematophagous Diptera (Galun 1977), will follow The truck-mounted experimental apparatus a moving vehicle and will continue to circle it for containing the entire array of traps presented a a brief period after it stops moving. Occasionally, moving attraction at a distance (ca. 10-15 m = deer flies would land on traps after the truck be- maximum distance from the woods, most repli- came stationary. Manually rotating or shaking cates were closer to vegetation) to responding flies the traps after stopping the truck had little effect which then selected a single trap on which to on deer fly landing on traps. Hanging traps such land. The distance from the woods in these bioas- as blue balloons or pots swaying in the wind in says is the distance that Phelps and Vale (1976) the same area (as the stationary truck) also only suggested was the maximum from which taban- occasionally induced deer flies to land. ids could detect a moving target. The truck- No significant response by the Chrysops spp. to mounted trap design is analogous to the cluster any behavioral chemicals tested (acetone, octenol,

design of Hribar et al. (1991b) and Hribar and CO2, DEET) were observed in these experiments. Foil (1994) and provides more precise comparison Trap movement, color and size either singularly of fly treatment preference for activation and or in combination were perhaps such strong stim- landing cues than other designs that offer single uli to the Chrysops spp. tested that odorant ef- trap replicates in separate locations. The trolling fects were not detectable. For example, we were trap does not fully measure fly attraction, as at- only able to detect the effect of trap size by using traction could occur without landing and capture. black traps that are less preferred than blue. Al- The flight orientation pattern and degree of ternatively, because the traps measure landing aggression exhibited by Chrysops spp. to the behavior, the effects of attractants in our design traps during trolling was variable, and while the perhaps were undetectable. Other species of ta- causes were not measured in these experiments, banids including large horse fly species were reg- these behaviors were most likely related to the ularly observed flying near the traps or the truck. time since emergence, feeding history and the However, very few were captured which may indi- prevailing weather conditions. During periods of cate that the trolling traps provided some of the relatively warm temperatures, deer flies at the primary attractive stimuli, (i.e., movement and time of the experiments predominantly flew color), but not the secondary or final landing cues straight into the trap from any direction attack- (Gibson and Torr 1999). Nevertheless, the repel- ing the trap in a manner similar to Vespidae de- lent DEET must ultimately affect fly landing be- fensive behavior. Some deer flies oriented from havior which was measurable in these bioassays. behind the trap and followed it for a short period No repellent affect of DEET was observed. More- of time before landing. This behavioral response over, true attractants if operative would in part could be induced by increasing the vehicle speed. serve to activate and draw flies from a greater dis- Other orientation behavior included circling tance to the trap vicinity while the landing behav- around several traps and then landing. Often cap- iors would likely remain constant for the test tured deer flies tended to be in clusters on the species. As a result, if attractants were operative, traps which may have been the result of short higher numbers of flies in the test replicates us- range visual orientation to other flies that had ing attractants (in comparison to other experi- been captured previously. However, an experi- ments without chemicals) would be expected. ment placing either live or dead tabanids of dif- Higher numbers were not observed, however, the ferent sizes and species on the traps in different appropriateness of the trolling trap bioassay for places as bait prior to trolling (data not shown) determining the effects of chemicals on deer fly had no effect on deer fly landing patterns. Gener- behavior remains for future research. ally, the first arriving deer flies landed on the trap Many times during the course of the experi- positions at random, but tended to land on the top ments, C. vittatus/C. pikei apparently were elim- portions first and filled in other areas as avail- inated from local areas that had been repeatedly able. trolled. Lack of deer flies made it necessary to find 364 Florida Entomologist 85(2) June 2002 populations at new locations. During these had the lowest intensity and also reflected a low searches we trolled the traps through different level of UV. Dutch blue reflected a high percentage landscape habitat types. We found that habitat of UV and was the most intense (brightest). Color characteristics affected deer fly presence and is primary and size is secondary in the behavioral abundance as shown previously by Dale and Ax- hierarchy as indicated by the significant interac- tell (1976). C. vittatus/C. pikei was found almost tion in the behavioral response to size of the less exclusively on the edges of open areas adjoining preferred black traps (Table 2). woodlands (within 25-30 m of the borders) and in Contrast was also an important determinant narrow road corridors through forests. C. mar- of deer fly behavior. A white background provid- quarti was found further from the forest edge in ing the most contrast increased trap catch signif- more open habitats such as old fields with sparse icantly. Aluminum foil which provides high tree density. The two species were found together contrast but reflects ca. 80% UV (Allan and Stof- in road corridors through areas adjacent to habi- folano 1986) significantly decreased trap catch for tats described above. C. vittatus/C. pikei and C. macquarti and these The collective results of these experiments in- results agree with reports by Allan and Stoffolano dicate the importance of habitat context and the (1986) for T. nigrovittatus and the results of the hierarchy of stimuli determining the behaviors blue series (Exp. 6, Fig. 2) above. The addition of used in the orientation to and landing on hosts by white stripes to black traps also significantly re- C. vittatus/C. pikei, C. macquarti and presum- duced trap collection (95 C. vittatus/C. pikei on ably related Chrysops spp. Habitat characteris- the black traps vs 34 on black and white striped tics such as amount of sunlight and vegetation traps). pattern that may affect micro habitats of perches Although silhouettes have been shown to be im- where the species were found during trolling were portant cues for tabanids (Phelps & Vale 1976) in-

different and deserve further research enabled by cluding Chrysops spp. in the presence of CO2 trolling traps. Height of deer fly flight above (Browne & Bennett 1980), we found no effect when

ground as indicated by orientation to traps was traps were trolled with silhouettes without CO2. always less than 3.0 m. Roberts (1976b) captured A Malaise trap run concurrently with these 70% of tabanids in Malaise traps with openings at studies and placed along the edge of a forest with less than 1.8 m above ground and Snoddy (1970) central swampy areas (the edges and road and Schulze et al. (1975) reported similar flight through the middle of this ca. 2 ha area were used patterns by other tabanids. for trolling) indicated that the trolling trap did not C. vittatus/C. pikei and C. macquarti were catch all of the Chrysops spp. present in our trap/ strongly attracted to movement of 3-dimensional trolling locations. However, we apparently cap- objects and would rarely land on stationary tar- tured most Chrysops spp. present that generally gets. However, other horse fly species have been attack humans in the area. Diaclorus ferrugatus captured effectively using flat panels (Allan and (F.) was not captured by trolling, although this Stoffalano 1986, Moore et al. 1996). Moving tar- species also attacks humans usually around the gets may be more attractive to most tabanids lower legs. Other species regularly captured in- (Phelps and Vale 1975). Quality of movement is im- cluded C. geminata Wiedemann, C. dorso-vittatus portant to the captured Chrysops spp. such that Hine, and the C. flavidus Wiedemann complex. C. angular displacement is required to elicit landing. davisus Walker, Tabanus trimaculatus Palisot de Movement of the trap without angular displace- Beauvois, T. pumilus MacQuart, and T. fulvulus ment—stationary placement with rotation or Wiedemann were captured occasionally. Approxi- shaking—often attracted deer flies in low num- mately 30 Chrysops spp. have been captured by bers, but induced landing only in a few cases when using the Malaise trap in Monticello, Florida (Mi- the deer flies had just followed the moving vehicle. zell & Roberts 1997-1999, unpublished). Deer flies preferred the 3-dimensional traps Many humans are allergic to deer fly bites and presumably due to the increased reflectiveness of often develop painful boil-like areas in response to the surface as shown by Thorsteinson et al. (1966). tabanid bites. Even in the absence of allergic reac- Deer fly response to traps was also strongly af- tions, deer flies are a nuisance and bites are pain- fected by color and size and the interaction of color ful. Our results provide an inexpensive method to with size was evident in the results of experiments reduce attacks by certain deer fly species and to 1A, 2, 4, 6, and 8 (Table 2). Blue was preferred over suppress deer fly populations in small areas such black and red, which were preferred over traps of as dooryards. Placing a small blue trap covered clear plexiglass, yellow and white. Trap spectral with Tanglefoot such as a plastic drinking cup at- reflectance patterns indicated that intensity and tached to a hat either worn or carried on a walk- ultraviolet (UV) light may be important determi- ing stick, exploits the deer fly’s tendency to attack nants of Chrysops spp. behavior (Fig. 2). FL-5 the highest point on the target animal, and sup- Neon blue captured the highest numbers of deer presses or eliminates most deer fly species that flies and was of medium intensity, the most satu- land on the body. Because deer flies are ambush rated (pure) blue and reflected no UV. Royal blue predators, placement of traps on a lawnmower, 4- Mizell et al.: Trolling: A Novel trap for Chrysops spp. 365

wheeler, or other vehicle will enable trapping-out BROWNE, S. M., AND G. F. BENNETT. 1980. Color and in small areas such as dooryards for a few days or shape as mediators of host-seeking responses of Sim- so until new deer flies re-colonize the area. uliids and Tabanids (Diptera) in the Tantramar As with all tabanid traps developed to date, the marshes, New Brunswick, Canada. J. Med. Entomol. trolling trap has limitations. The trap catches pre- 17: 58-62. CATTS, E. P. 1970. A canopy trap for collecting Taban- dominately female Chrysops spp., but only a few idae. Mosq. News. 30: 472-474. other tabanids. The trap requires specific move- CILEK, J. E., AND E. T. SCHREIBER. 1996. Diel host-seek- ment (angular displacement) in order to attract ing behavior of Chrysops celatus (Diptera: Taban- deer flies and has the added disadvantage of re- idae) in northwest Florida. Florida Entomol. 79: 520- quiring a sticky substance to trap landing insects. 525. However, cleanup of Tanglefoot is easy with soaps DALE, W. E., AND R. C. AXTELL. 1976. Salt marsh Ta- containing d-limonene. During this study many banidae (Diptera): comparisons of abundance and other unidentified species of biting flies were cap- distribution in Spartina and Juncus habitats. J. tured on the trolling trap. As such our trolling Med. Entomol. 12: 671- 678. FOIL, L. D., AND L. J. HRIBAR. 1995. Evaluation of tsetse trap offers a novel method to detect and collect attractants as baits for horse flies and deer flies Chrysops spp. and perhaps many other Dipteran (Diptera: Tabanidae) in Louisiana. Florida Entomol. species, and may be useful to investigate a variety 78: 129-133. of scientific questions heretofore unattainable. In FRENCH, F. E., AND D. L. KLINE. 1989. 1-Octen-3-ol, an addition, the simple, economical methodology has effective attractant for Tabanidae (Diptera). J. Med. great potential for demonstration of basic insect Entomol. 26: 459-461. behavior to science students of all ages. FRENCH, F. E., AND D. V. HAGAN. 1995. Two-tiered box trap catches Chrysops atlanticus and C. fulginosus (Diptera: Tabanidae) near a Georgia salt marsh. J. ACKNOWLEDGMENTS Med. Entomol. 32: 197-200. GALUN, R. 1977. Responses of blood-sucking arthropods We especially recognize, thank and remember Dr. Ri- to vertebrate hosts, pp. 103-115. In H. H. Shorey and chard Roberts, USDA-ARS, (deceased) for identifying J. J. McKelvey [eds.]. Chemical control of insect be- the Tabanidae and for many enjoyable conversations havior, theory and application. John Wiley and Sons. about horse fly behavior and research. We thank Pat Mi- N.Y. 414 pp. zell, wife and mother, for her kind patience and encour- GIBSON, G., AND J. BRADY. 1985. ‘Anemotactic’ flight agement during the course of these studies. We thank paths of tsetse flies in relation to host odors: a pre- Stephanie Bloom, Peter Andersen, Richard Roberts, liminary video study in nature to host odor. Physiol. Frank French, Lane Foil and two anonymous reviewers Entomol. 10: 395-406. for helpful comments on an earlier draft of the manu- GIBSON, G., AND S. J. TORR. 1999. Visual and olfactory script. Special sentiment and thanks are sent to Dr. responses of haematophagous Diptera to host stim- Robert Combs, Mississippi State University, whose jo- uli. Med. and Vet. Entomol. 13: 2-23. vial humor and wit were often of great amusement and GRANGER, C. A. 1970. Trap design and colors as factors encouragement to the senior author during graduate in trapping salt marsh greenhead fly. J. Econ. Ento- school. This is the University of Florida Agricultural Ex- mol. 63: 1670-1672. periment Station Journal Series No. R-07188. HANSENS, E. J. 1947. Greenhead flies (Tabanus nigro- vittatus) like dark colors. New Jersey Agr. 29: 3-4. REFERENCES CITED HANSENS, E. J., E. M. BOSLER, AND J. W. ROBINSON. 1971. Use of traps for study and control of saltmarsh AILES, M. C., L. J. BROWN, C. CHURCH, D. P. FRENCH, greenhead flies. J. Econ. Entomol. 64: 1. AND W. GALE. 1992. Mechanical control of greenhead HRIBAR, L. J., AND L. D. FOIL. 1994. Color and UV reflec- flies (Diptera: Tabanidae) in a marsh environment. tance of canopy traps for collecting horse flies J. Med. Entomol. 29: 160-164. (Diptera: Tabanidae) in Louisiana. Bull. Soc. Vector ALLAN, S. 1984. Studies on vision and visual attraction Ecol. 19:49-52. of the salt marsh horse fly, Tabanus nigrovittatus HRIBAR, L. J., D. J. LEPRINCE, AND L. D. FOIL. 1991a. Macquart. Ph.D. Dissertation. University of Mass., Design for a canopy trap for collecting horse flies Amherst. 178 pp. (Diptera: Tabanidae). J. Am. Mosq. Contr. Assoc. 7: ALLAN, S., AND J. G. STOFFALANO, JR. 1986. The effects 657-659. of hue and intensity on visual attraction of adult Ta- HRIBAR, L. J., D. J. LEPRINCE, AND L. D. FOIL. 1991b. In- banus nigrovittatus (Diptera: Tabanidae). J. Med. creasing horse fly (Diptera: Tabanidae) catch in can- Entomol. 23: 83- 91. opy traps by reducing Ultraviolet light reflectance. J. ALLAN, S. A., J. F. DAY, AND J. D. EDMAN. 1987. Visual Med. Entomol. 28: 974-877. ecology of biting flies. Ann. Rev. Entomol. 32: 2 97- JACKSON, J. W., D. V. HAGAN, AND F. E. FRENCH. 1993. 313. Two-story French box traps for Tabanids (Insect: AXTELL, R. C., T. D. EDWARDS, AND J. C. DUKES. 1975. Diptera). J. GA. Science. 51: 24-25. Rigid canopy trap for Tabanidae (Diptera). J. GA En- JONES, C. M. 1953. Biology of Tabanidae in Florida. J. tomol. Soc. 10: 64-67. Econ. Entomol. 46:1108-1109. BRACKEN, G. K., W. M. HANEC, AND A. J. THORSTEIN- MOORE, T. R., J. E. SLOSSER, J. COCKE, JR., AND W. H. SON. 1962. The orientation behavior of horse flies NEWTON. 1996. Effect of trap design and color in and deer flies (Tabanidae: Diptera) II. The role of evaluating activity of Tabanus abactor Philip in some visual factors in the attractiveness of decoy sil- Texas rolling plains habitats. Southwest. Entomol. houettes. Canadian J. Zool. 40: 685-695. 21: 1-11. 366 Florida Entomologist 85(2) June 2002

PHELPS, R. J., AND G. A. VALE. 1976. Studies on the local for sampling a horsefly and deer fly community. En- distribution and on the methods of host location of viron. Entomol. 5: 788-792. some Rhodesian Tabanidae (Diptera). J. Entomol. TEDDERS, W. L., AND B. W. WOOD. 1994. A new tech- Soc. Southern Africa. 39: 67-81. nique for monitoring pecan weevil emergence. J. En- ROBERTS, R. H. 1971a. The seasonal appearance of Ta- tomol. Sci. 29: 18-30. banidae as determined by Malaise trap collections. THORSTEINSON. A. J. 1958. The orientation of horse flies Mosq. News. 31: 509-512. and deer flies (Diptera: Tabanidae). I. The attraction of

ROBERTS, R. H. 1971b. Effect of amount of CO2 on collec- heat to Tabanids. Entomol. Expt. et Appl. 1:1 91-196. tion of Tabanidae in Malaise traps. Mosq. News. 31: THORSTEINSON, A. J., G. K. BRACKEN, AND W. HANEC. 552-558. 1965. The orientation behavior of horse flies and ROBERTS, R. H. 1976a. The comparative efficiency of six deer flies (Tabanidae: Diptera). III. The use of traps trap types for the collection of Tabanidae (Diptera). in the study of orientation of Tabanids in the field. Mosq. News 36: 530-535. Entomol. Exp. et Appl. 8: 189-192. ROBERTS, R. H. 1976b. Altitude distribution of Taban- THORSTEINSON, A. J., G. K. BRACKEN, AND W. TOSTO- idae as determined by Malaise trap collections. WARYK. 1966. The orientation behavior of horse flies Mosq. News. 36: 518-520. and deer flies (Tabanidae: Diptera) V. The influence SAS INSTITUTE. 1996. SAS User’s Guide v. 6.12. SAS In- of the number and inclination of reflecting surfaces stitute, Cary, NC. on attractiveness to Tabanids of glossy black polyhe- SCHULZE, T. L. E. J. HANSENS, AND J. R. TROUT. 1975. dra. Can. J. Zool. 44: 275-279. Some environmental factors affecting the daily and WALL, W. J., AND O. W. DOANE, JR. 1980. Large scale seasonal movements of the salt marsh greenhead, Ta- use of box traps to study and control saltmarsh banus nigrovittatus. Environ. Entomol. 4: 965-971. greenhead flies (Diptera: Tabanidae) on Cape Cod, SMITH, G. E., S. G. BREELAND, AND E. PICKARD. 1965. Massachusetts. Environ. Entomol. 9: 371-375. The Malaise trap—a survey tool in medical entomol- WILSON, B. H. 1968. Reduction of Tabanid populations ogy. Mosq. News. 25: 398-400. on cattle with sticky traps baited with dry ice. J. SNODDY, E. L. 1970. Trapping deer flies with colored Econ. Entomol. 61: 827-829. weather balloons (Diptera: Tabanidae). J. Ga. Ento- WILSON, B. H., N. P. TUGWELL, AND E. C. BURNS. 1966. mol. Soc. 5: 207-209. Attraction of Tabanids to traps baited with dry ice TALLAMY, D. W., E. J. HANSENS, AND R. F. DENNO. 1976. under field conditions in Louisiana. J. Med. Entomol. A comparison of Malaise trapping and aerial netting 3: 149-149.

Scientific Notes 367

EFFECT OF TOMATO MOTTLE VIRUS (TOMOV) ON BEMISIA TABACI BIOTYPE B (HOMOPTERA: ALEYRODIDAE) OVIPOSITION AND ADULT SURVIVORSHIP ON HEALTHY TOMATO

C. L. MCKENZIE USDA-ARS, U.S. Horticultural Research Laboratory, 2001 South Rock Road, Ft. Pierce, FL 34945

The population dynamics of the sweetpotato wood, and an aluminum hair clip was glued to the whitefly, Bemisia tabaci (Gennadius) biotype B balsa wood and cup portion. Whiteflies were intro- (= silverleaf whitefly, Bemisia argentifolii Bel- duced through a small hole in the side of the cup. lows & Perring) are driven by seasonal climatic After a 48-h access period, adult whiteflies were conditions, natural enemies (including entomo- removed and eggs were counted. Treatments were pathogens), host-plant interactions, and IPM maintained separately at 25 ± 1°C and a photo- practices (Byrne & Bellows 1991; Coudriet et al. period of 16:8 L:D. For each experiment, 20 test 1985; Nava-Cameros et al. 2001; Tsai & Wang plants were typically used for each treatment; 1996; Yee and Toscano 1996). Physiological disor- however, the final number of replicates (= clip ders (Schuster et al. 1990, 1991) and the spread of cages) per treatment varied when leaves of test geminiviruses (Blair et al. 1995; Polston & Ander- plants died or were severed during the experi- son 1997) associated with whitefly infestations in ment. A minimum of 8 replicates was used for all Florida vegetables are becoming increasingly im- treatments. Experiments were repeated five portant limitations to grower profitability. The ef- times. There were no significant interactions be- fect of plant viruses on the reproductive potential tween experiment*treatment (F = 2.0; df = 4,80; of the vector is key to understanding geminivirus P = 0.10) or treatment*clip cage (F = 0.66; df = epidemiology and developing effective control 19,80; P = 0.85) so results were pooled over exper- measures. The objective of this study was to de- iments for mean comparison (Tukey option in termine the effect of tomato mottle virus (ToMoV) SAS GLM procedure, SAS Institute 1998). In the on whitefly oviposition and survival rates on last two experiments, progeny from cohorts used healthy tomato. in the oviposition clip cage experiments (n = 15) Adult B. tabaci biotype B were obtained from were used to include survival to adult emergence laboratory colonies maintained by the U.S. Horti- which was evaluated 30 days after egg lay to en- cultural Research Laboratory, Ft. Pierce, FL. sure that all viable whiteflies had emerged. Whiteflies used in these experiments were origi- Whiteflies infected with ToMoV deposited sig- nally obtained from Dr. Lance Osborne, Univer- nificantly more eggs (F = 19.51; df = 1,80; P < 0.01) sity of Florida, Apopka, FL and have been on healthy tomato leaves than nonviruliferous maintained on dwarf cherry tomato (Lycopersicon whiteflies (Table 1) and supports earlier findings esculentum cv. Florida Lanai) since 1996 by serial (McKenzie et al. 2002). There was no significant transfer. In 1997, a ToMoV whitefly colony was es- difference between virus-infected and nonvirulif- tablished by obtaining tomato plants infected erous whiteflies for the number of adults emerged with ToMoV from Dr. Philip Stansly, University of or the proportion of those adults surviving from Florida, Immokalee, FL and infesting with white- the egg stage. There was no significant correlation flies from the healthy colony. Whitefly biotyping between the number of eggs deposited per female was based on RAPD PCR analysis using primers and progeny survival rates on healthy tomato for developed by De Barro and Driver (1997). Nonvir- whitefly infected with or without the virus (SAS uliferous and viruliferous whitefly colonies were CORR procedure, SAS Institute 1998). housed separately in screened Plexiglass cages lo- A report by Costa et al. (1991) demonstrated cated in separate growth chambers at 25 ± 1°C that whitefly maintained on pumpkin for ~ 6 and a 16:8 L:D photoperiod. Whiteflies from the years had a higher rate of survival on virus-in- viruliferous colony were confirmed to be infected fected pumpkin compared to healthy pumpkin with ToMoV prior to infestation by PCR analysis out of six virus-plant hosts evaluated, including (Sinisterra et al., 1999). tomato. In those experiments, researchers did not In each experiment, one male and one female take host-plant adaptation time into consider- whitefly of unknown age from healthy or ToMoV- ation. For example, whitefly survival rates from infected whitefly colonies were confined in clip egg to adult on tomato were 8% on virus-infected cages attached to the terminal leaf of the 3rd fully plants and 17% on noninoculated control plants, expanded leaflet of a healthy cv. Florida Lanai but the parental generation of whiteflies used in plant. Clip cages were made from clear plastic those experiments were maintained on pumpkin. cups (PC100 30 ml cups, Jet Plastica, Harrisburg, In our experiments, high survival (Table 1) of PA) fitted with a foam seal on the bottom and an both healthy and ToMoV-infected whitefly reflect organdy window on top for ventilation. The foam this host-plant adaptation when compared to the bottom was backed with a thin square of balsa previous work by Costa et al. (1991). Plants from

368 Florida Entomologist 85(2) June 2002

TABLE 1. MEAN NUMBER OF NONVIRULIFEROUS VERSUS VIRULIFEROUS B. TABACI BIOTYPE B EGGS/FEMALE/48-HR AC- CESS, ADULTS AND PERCENT SURVIVAL ON HEALTHY TOMATO.

WF colony Egg ± SE1 Adult ± SE2 % Survival ± SE3

Healthy 8.80 ± 0.70 a 7.28 ± 1.90 a 70.56 ± 14.11 a ToMoV 14.69 ± 1.11 b 11.09 ± 2.16 a 77.44 ± 6.37 a

Means within a column followed by the same letter are not significantly different, P < 0.05, using Tukey studentized range test. 1 F = 19.51; df = 1,80; P = < 0.01. 2F = 3.16; df = 1,11; P = 0.10. 3Proportion of whitefly surviving from egg to adult stage; F = 0.44; df = 1,8; P = 0.53. the virus treatment exhibited characteristic To- COUDRIET, D. L., N. PRABHAKER, A. N. KISHABA, AND MoV symptoms 30 days after clip cages were re- D. E. MEYERDIRK. 1985. Variation in developmental moved. This suggests adaptation to the host plant rate on different hosts and overwintering of the and virus by the vector could override any ad- sweetpotato whitefly, Bemisia tabaci (Homoptera: verse effect the virus had on host plant physiol- Aleyrodidae). Environ. Entomol. 14: 516-519. CARDOZA, Y. J., H. J. MCAUSLANE, AND S. E. WEBB. ogy. McKenzie et al. (2002) found healthy plants 2000. Effect of leaf age and silverleaf symptoms on infested with ToMoV-infected whiteflies consis- oviposition site selection and development of Bemi- tently had 2.5-fold more eggs and 4.5-fold more sia argentifolii (Homoptera: Aleyrodidae) on zuc- nymphs than plants with nonviruliferous white- chini. Environ. Entomol. 29:220-225. flies (P < 0.05) 56 days after infestation with the DE BARRO, P. J., AND F. DRIVER. 1997. Use of RAPD same number of whitefly. In the present study, PCR to distinguish the B biotype from other biotypes whiteflies were well adapted to the host plant, ei- of Bemisia tabaci (Gennadius) (Hemiptera: Aley- ther with or without ToMoV. rodidae). Aust. J. Entomol. 36: 149-152. MCKENZIE, C. L., R. G. SHATTERS, JR., H. DOOSTDAR, S. D. LEE, M. INBAR, AND R. T. MAYER. (2002). Effect of SUMMARY geminivirus infection and Bemisia infestation on ac- cumulation of pathogenesis-related proteins in to- Whiteflies carrying ToMoV deposited signifi- mato. Arch Insect Biochem and Physiol. 49: 203-214. cantly more eggs than nonviruliferous whiteflies POLSTON, J. E., AND P. K. ANDERSON. 1997. The emer- when provided a healthy tomato host. Insect ad- gence of whitefly-transmitted geminiviruses in to- aptation to the host-plant is a critical factor that mato in the Western Hemisphere. Plant Disease 81: 1358-1369. should be considered on a host-by-host basis SAS INSTITUTE, INC. 1998. SAS/STAT® User’s guide when evaluating insect biology and vector-host- version 7. Cary, NC, USA: SAS Institute, Inc. plant interactions for polyphagous insect species. SCHUSTER, D. J., T. F. MUELLER, J. B. KRING, AND J. F. Mention of a trademark or proprietary product PRICE. 1990. Relationship of the sweetpotato white- does not constitute a guarantee or warranty of the fly to a new tomato fruit disorder in Florida. HortSci. product by the U.S. Department of Agriculture 25: 1618-1620. and does not imply its approval to the exclusion of SCHUSTER, D. J., J. B. KRING, AND J. F. PRICE. 1991. As- other products that may also be suitable. sociation of the sweetpotato whitefly with a silver- leaf disorder of squash. HortSci. 26: 155-156. SINISTERRA, X. H., J. E. POLSTON, A. M. ABOUZID, AND REFERENCES CITED E. HIEBERT. 1999. Tobacco plants transformed with a modified coat protein of tomato mottle begomovi- BLAIR, M. W., M. J. BASSETT, A. M. ABOUZID, E. HIE- rus show resistance to virus infection. Phytopath. BERT, J. E. POLSTON, R. T. MCMILLAN, JR., W. GRAVES, 89: 701-706. AND M. LAMBERTS. 1995. Occurrence of bean golden TSAI, J. H., AND K. WANG. 1996. Development and re- mosaic virus in Florida. Plant Disease 79: 529-533. production of Bemisia argentifolii (Homoptera: Aley- BYRNE, D. N., AND T. S. BELLOWS. 1991. Whitefly biol- rodidae) on five host plants. Environ. Entomol. 25: ogy. Ann. Rev. Entomol. 36:431-457. 810-816. COSTA, H. S., J. K. BROWN, AND D. N. BYRNE. 1991. Life YEE, W. L., AND N. C. TOSCANO. 1996. Ovipositional history traits of the whitefly, Bemisia tabaci (Homop- preference and development of Bemisia argentifolii tera: Aleyrodidae) on six virus-infected or healthy (Homoptera: Aleyrodidae) in relation to alfalfa. J. plant species. Environ. Entomol. 20: 1102-1107. Econ. Entomol. 89: 870-876.

Scientific Notes 369

BOT (DIPTERA: CUTEREBRIDAE) INFESTATION OF NEST-BOUND INFANT EASTERN GRAY SQUIRRELS

F. SLANSKY1 AND L. R. KENYON2 1Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611-0620

2Nutkin’s Nest Wildlife Rehabilitation Center, 14107 NW 61st Lane, Gainesville, FL 32653-2570

Cuterebra bot fly larvae are obligate, subcuta- tion of infestation of nest-bound infants by larvae neous parasites of rodents (mice, rats, tree squir- of this bot fly. rels, chipmunks, etc.) and lagomorphs (rabbits As part of a wildlife rehabilitation program in and hares) in the Americas (Sabrosky 1986). northcentral (Alachua Co.) Florida, over the last Infestation is revealed by lumps (referred to as 18 years we have encountered 10-15 cases of infant warbles) on an animal caused by the presence of S. carolinensis infested by bot fly larvae (out of sev- second and third instar larvae under its skin eral hundred infants brought in for rehabilitation), (Cogley 1991, Slansky & Kenyon 2000, 2001). Ev- including a record high of five in the summer, 2000 idence from a limited number of studies indicates squirrel breeding season (no cases were reported that Cuterebra flies do not oviposit directly on in 2001). This rarity contrasts with the situation in their hosts but rather on foliage, twigs or other adult squirrels, where the incidence of Cuterebra substrates, often in the vicinity of an animal’s parasitization often exceeds 50% (unpubl. obs.). In- nest (Catts 1967, Baird 1974, 1997). A potential festations of the infants typically consisted of one host thus risks exposure to newly hatched Cutere- or two, and rarely three, larvae per individual (Fig. bra larvae when leaving its nest and moving 1). All of these animals would have been nest- about in its habitat (Catts 1982). bound at the time of parasitization; most were Once the tiny, infective-stage larvae transfer young enough to have not yet opened their eyes to a potential host, they enter an orifice or wound (i.e., they were less than five weeks of age) and and begin their approximately week-long journey even those with opened eyes were too young to through the animal’s body, eventually settling un- have begun exploring outside the nest. Juvenile der its skin (Gingrich 1981). They then use their S. carolinensis typically do not begin extra-nest ac- pointed mouth hooks to cut through the host’s tivity before about eight weeks of age (unpubl. obs.). hide to create a warble pore through which they In that exposure to Cuterebra larvae is be- respire and excrete fluid. A larva typically re- lieved to occur when an animal moves about out- mains in its warble, which increases in size as the side of its nest, infestation of nest-bound infants larva grows, until completing development in seems highly unusual. We are aware of only a few about 3 to 10 weeks, depending on the species of anecdotal reports of Cuterebra-infested infant an- Cuterebra and host (Baird 1975, Jacobson et al. imals (e.g., a 6-day-old rabbit and a litter of nurs- 1978). The mature larva exits through the warble ing wood rats; Dalmat 1943). Kittens and pore and falls to the ground, where it burrows puppies, both of which are incidental hosts, can into the soil to pupate (Catts 1982). also be parasitized (Hall 1925, Rosenthal 1975, Over the past several years, we have been McKenzie et al. 1978). How infestation of nest- studying the relationship between Cuterebra bound animals occurs has not been determined. emasculator Fitch and its tree squirrel hosts. This Assuming that female Cuterebra did not deposit bot fly parasitizes eastern gray (Sciurus carolin- eggs directly on these infants, it is possible that a ensis Gmelin) and fox (S. niger L.) squirrels, as mother animal that had acquired infective-stage well as eastern chipmunks (Tamias striatus (L.)), larvae while out foraging inadvertently brought throughout the eastern and midwestern regions these back to her nest and some transferred to of North America, from southern Canada to Flor- her nursing offspring. Another possibility is that ida (Sabrosky 1986); American red (Tamiasciurus larvae from eggs laid on nest material entered the hudsonicus (Erxleben)) and flying (Glaucomys nest and parasitized the animals within. It is not spp.) squirrels seem to be rarely infested (Dorney known where C. emasculator females oviposit. 1965, Slansky & Kenyon 2000). Cuterebra emas- Parasitism by Cuterebra typically has been culator is apparently univoltine throughout its studied in juvenile, subadult and adult animals range, spending some ten months underground in (Bennett 1955, 1972b, 1973, Jacobson et al. 1981, the pupal stage (Bennett 1972a). Infested ani- Manrique-Saide et al. 2000). Individuals in these mals are observed from mid-late July to the end of age classes typically exit their nests on a daily ba- October (Bennett 1955, Jacobson et al. 1981, sis to forage in the habitat and thus risk exposure Slansky & Kenyon 2000). While there are many to infective-stage larvae; it is also these individu- published records of C. emasculator infesting tree als that are sampled through the commonly used squirrels and chipmunks active outside the nest, techniques of trapping, hunting and collection of to our knowledge this note is the first documenta- roadkills. However, as documented here, seden-

370 Florida Entomologist 85(2) June 2002

Fig. 1. Upper panel: Nest-bound infant gray squirrel (4-5 weeks old, eyes not yet opened) with a moderate-sized bot fly warble on its side behind its right front leg; the warble contained a second instar larva. Lower left panel: Sec- ond instar C. emasculator larva removed from its warble; bands of dark spines are visible (‘head’ end at left; length = 10.0 mm; original photomicrograph, 25×). Lower right panel: Ventral view of anterior portion of second instar bot fly larva showing paired mouth hooks (toward top of image) and bands of dark, posterior-pointing spines (the inter- nal cephaloskeleton to which the mouth hooks are attached is faintly visible through the translucent cuticle; orig- inal photomicrograph, 60×).

tary, nest-bound infants may also become in- with their small size and the high energy and fested. Therefore, when evaluating the effects of nutrient demands of their rapid growth, it is parasites on their host populations (Grenfell & likely that infants will be deleteriously affected Gulland 1995, Milton 1996), it would seem to be by parasites to a greater extent than adults with important to study not just animals moving about a comparable intensity of infestation. Thus, when in their habitat, but nest-bound individuals as only older, mobile animals are studied, the impact well, even though locating nests to sample the in- of parasites on the host population could be sig- fants will typically be more difficult in a field set- nificantly underestimated, especially if the inci- ting than trapping mobile animals. Associated dence of infant infestation is substantial.

Scientific Notes 371

We thank Lance Durden, Heidi Bissell, and COGLEY, T. P. 1991. Warble development by the rodent Shelley L. Miller for reviewing this manuscript, bot Cuterebra fontinella (Diptera: Cuterebridae) in and three anonymous reviewers for their helpful the deer mouse. Veterinary Parasitol. 38: 276-288. comments. This research was supported by the DALMAT, H. T. 1943. A contribution to the knowledge of Florida Agricultural Experiment Station, and ap- the rodent warble flies (Cuterebridae). J. Parasitol. 29: 311-318. proved for publication as Journal Series No. R- DORNEY, R. S. 1965. Incidence of botfly larvae (Cutere- 08126. bra emasculator) in the chipmunk (Tamias striatus) and red squirrel (Tamiasciurus hudsonicus) in SUMMARY northern Wisconsin. J. Parasitol. 51: 893-894. GINGRICH, R. E. 1981. Migratory kinetics of Cuterebra Although infestation of tree squirrels and chip- fontinella (Diptera: Cuterebridae) in the white- munks active outside the nest (i.e., juveniles, sub- footed mouse, Peromyscus leucopus. J. Parasitol. 67: adults and adults) by larvae of the bot fly Cuterebra 398-402. emasculator has been well documented, this is ap- GRENFELL, B. T., AND F. M. D. GULLAND. 1995. Intro- parently the first report of nest-bound infant duction: Ecological impact of parasitism on wildlife host populations. Parasitology 111: s3-s14. eastern gray squirrels (Sciurus carolinensis) par- HALL, M. C. 1925. The occurrence of cuterebrid larvae in asitized by this species. dogs and cats, and the possible modes of infection. J. Econ. Entomol. 18: 331-335. REFERENCES CITED JACOBSON, H. A., B. S. MCGINNES, AND E. P. CATTS. 1978. Bot fly myiasis of the cottontail rabbit, Sylvil- BAIRD, C. R. 1974. Field behavior and seasonal activity agus floridanus mallurus in Virginia with some biol- of the rodent bot fly, Cuterebra tenebrosa, in central ogy of the parasite, Cuterebra buccata. J. Wildlife Washington (Diptera: Cuterebridae). Great Basin Diseases 14: 56-66. Naturalist 34: 247-253. JACOBSON, H. A., M. S. HETRICK, AND D. C. GUYNN. BAIRD, C. R. 1975. Larval development of the rodent bot 1981. Prevalence of Cuterebra emasculator in squir- fly, Cuterebra tenebrosa, in bushy-tailed wood rats rels in Mississippi. J. Wildlife Diseases 17: 79-87. and its relationship to pupal diapause. Canadian J. MANRIQUE-SAIDE, P., S. HERNANDEZ-BETANCOURT, AND Zool. 53: 1788-1798. M. T. QUINTERO. 2000. First record of Cuterebra sp. BAIRD, C. R. 1997. Bionomics of Cuterebra austeni (Diptera: Cuterebridae) infection in Ototylomys (Diptera: Cuterebridae) and its association with phyllotis (Rodentia: Muridae). Florida Entomol. 83: Neotoma albigula (Rodentia: Cricetidae) in the south- 487-488. western United States. J. Med. Entomol. 34: 690-695. MCKENZIE, B. E., D. LYLES, AND J. A. CLINKSCALES. BENNETT, G. F. 1955. Studies on Cuterebra emasculator 1978. Intracerebral migration of Cuterebra larva in a Fitch 1856 (Diptera: Cuterebridae) and a discussion kitten. JAVMA 172: 173-175. of the status of the genus Cephenemyia Ltr. 1818. MILTON, K. 1996. Effects of bot fly (Alouattamyia baeri) Canadian J. Zool. 33: 75-98. parasitism on a free-ranging howler monkey (Alou- BENNETT, G. F. 1972a. Observations on the pupal and atta palliata) population in Panama. J. Zool., London adult stages of Cuterebra emasculator Fitch (Diptera: 239: 39-63. Cuterebridae). Canadian J. Zool. 50: 1367-1372. ROSENTHAL, J. J. 1975. Cuterebra infestation of the con- BENNETT, G. F. 1972b. Further studies on the chipmunk junctiva in a puppy. Veterinary Medicine/Small Ani- warble, Cuterebra emasculator (Diptera: Cuterebri- mal Clinician 70: 462-463. dae). Canadian J. Zool. 50: 861-864. SABROSKY, C. W. 1986. North American Species of Cute- BENNETT, G. F. 1973. Some effects of Cuterebra emascu- rebra, the Rabbit and Rodent Bot Flies (Diptera: lator Fitch (Diptera: Cuterebridae) on the blood and Cuterebridae). Entomol. Soc. Amer. Thomas Say activity of its host, the eastern chipmunk. J. Wildlife Foundation Monograph, College Park, MD. Diseases 9: 85-93. SLANSKY, F., AND L. R. KENYON. 2000. Lumpy squir- CATTS, E. P. 1967. Biology of a California rodent bot fly rels—bugged by bot flies. Wildlife Rehab. Today Cuterebra latifrons Coquillett. J. Med. Entomol. 4: 11(3): 24-31. 87-101. SLANSKY, F., AND L. R. KENYON. 2001. Warbles of the CATTS, E. P. 1982. Biology of New World bot flies: tree squirrel bot fly. http://botfly.ifas.ufl.edu/cutrwrb/ Cuterebridae. Annu. Rev. Entomol. 27: 313-338. cutrwrb1.htm.

372 Florida Entomologist 85(2) June 2002

PREDATION BY BLUEGILL (LEPOMIS MACROCHIRUS) ON LARVAL (DIPTERA) IN RELATION TO MIDGE STANDING CROP IN TWO CENTRAL FLORIDA LAKES

RICHARD J. LOBINSKE1, CHARLES E. CICHRA2 AND ARSHAD ALI1 1University of Florida, IFAS, Mid-Florida Research and Education Center 2725 Binion Road, Apopka, FL 32703-8504

2University of Florida, IFAS, Department of Fisheries and Aquatic Sciences 7922 NW 71st Street Gainesville, FL 32653-3071

Large swarms of non-biting midges (Diptera: were most numerous in gut contents of fish from Chironomidae) emanating from some central both lakes (Table 1), comprising 55.9-62.8% of Florida lakes can cause severe nuisance and eco- total consumed midge larvae from Lake Dora and nomic problems for businesses, residents, and vis- 4.8-48.1% from Lake Yale. Geoldichironomus spp. itors within the dispersal range of these insects larvae were the next most common in the gut con- (Ali 1995). Midges are also a cause of allergies to tents of fish from Lake Dora (0.0-27.5% of total humans (Cranston 1995). Because of these prob- larval chironomids). Other midge larvae consumed lems, a systematic research program on the bio- by fish from Lake Dora included Chironomus nomics and management possibilities of midge crassicaudatus, Glyptotendipes paripes, Crypto- populations in central Florida has continued for chironomus spp., Pseudochironomus spp. and the past two decades (Ali 1996). As a part of this Tanypodinae. Seasonal mean number of total program, a preliminary investigation of fish pre- midge larvae in fish gut contents ranged from 4.7 dation on chironomid midge larvae for the biolog- to 44.0. Midge larvae were present in the gut con- ical control perspective of midges was conducted. tents of all fish from Lake Dora, except those col- Midge predatory fish (bluegill, Lepomis macrochi- lected in December 2000, when 40% of collected rus) were collected from two lakes on four occa- fish had empty guts. This was likely due to low sions to elucidate any relationships between water temperatures reducing feeding activity, as consumption of midge larvae by these fish and the suggested for bluegill during winter months by associated larval composition and distributions in Gilinsky (1984). Pseudochironomus spp. larvae the lakes. Information concerning fish predation were the second most prevalent midge larvae in on midge larvae, species or habitat specific, would the fish gut contents of Lake Yale, forming up to be useful in devising new control strategies. 46.2% of total midge larvae, followed by C. crassi- Fish were collected (May 1999, July, Septem- caudatus (collected only during May 1999), ber and December 2000) from Lakes Dora and G. paripes, Cryptochironomus spp., and Tanypod- Yale (Lake County, Florida) by electrofishing un- inae. Seasonal mean number of larvae per fish in der permit from Florida Fish and Wildlife Conser- Lake Yale ranged between 1.0 and 18.9. Bluegill vation Commission. Collections were made between feeding on midge larvae in Lake Yale was also re- 0830 and 1200 h and up to twenty fish were col- duced during December 2000, though only one lected from near-shore areas. Fish were identified fish had an empty gut. Other food items identified and killed immediately, maintained on ice while from fish in these two lakes included immature transported to the laboratory, and stored at -10°C Insecta (Odonata, Ephemeroptera and Trichop- until examined. For examination, each fish was tera), Crustacea (Decapoda, Amphipoda and Ostra- thawed and the foregut was dissected (Bowen coda), Nematoda, Oligochaeta, Gastropoda, and 1996), and the contents transferred to 4-dram some unidentifiable material. These food items vials containing 70% ethanol. Gut contents were numerically were only a small part of total gut examined under variable magnification of a dis- contents in most fish examined (data not shown). secting microscope and enumerated. Chironomi- To estimate relative selective feeding by blue- dae head capsules and associated fragments were gill on examined chironomid larvae, percent com- wet mounted on slides and examined at 400× position of chironomid larvae in gut contents of magnification using a phase-contrast microscope collected fish was compared to overall percent and identified to lowest possible taxonomic level composition of chironomid larvae in study lakes using the keys of Epler (1995). Only head cap- and percent composition of midge larvae in the sules with sufficient morphological features re- nearshore areas with firm sediments representa- maining for identification were counted for gut tive of the areas from where the fish were caught, content enumeration, other fragments were used collected concurrently and reported by Lobinske to improve identification where possible. (2001) (Fig. 1). In Lake Dora, Tanytarsini were Five to 20 fish were successfully collected per most common, exhibiting similar percent compo- sampling occasion (Table 1). Midge larvae of the sitions in fish gut contents in the entire lake as tribe Tanytarsini (>90% Cladotanytarsus spp.) well as in nearshore areas. During July, Septem-

Scientific Notes 373

TABLE 1. GUT CONTENTS OF BLUEGILL (LEPOMIS MACROCHIRUS) COLLECTED ON FOUR OCCASIONS (MAY 1999-DECEM- BER 2000) FROM LAKES DORA AND YALE (LAKE CO., FL). MEAN ± SD NUMBER OF MIDGE LARVAE PER FISH AND PERCENT COMPOSITION OF TOTAL MIDGE LARVAE IN PARENTHESES.

Taxa May 1999 July 2000 September 2000 December 2000

Lake Dora No. fish collected 10 20 20 20 Chironomus crassicaudatus 0 ± 0 (0.0) 0.1 ± 0.3 (0.3) 0 ± 0 (0.0) 0.8 ± 2.6 (16.1) Glyptotendipes paripes 0.4 ± 0.7 (0.1) 1.8 ± 2.5 (6.0) 0.2 ± 0.7 (0.5) 0.3 ± 1.1 (5.4) Cryptochironomus spp. 0 ± 0 (0.0) 2.5 ± 3.4 (8.2) 4.4 ± 7.3 (10.4) 0.3 ± 1.1 (6.5) Pseudochironomus spp. 0 ± 0 (0.0) 0 ± 0 (0.0) 0.5 ± 1.5 (1.2) 0 ± 0 (0.0) Geoldichironomus spp. 0 ± 0 (0.0) 8.3 ± 11.0 (27.5) 10.5 ± 13.1 (24.7) 0.7 ± 1.7 (14.0) Tanytarsini 25.1 ± 18.2 (57.0) 16.9 ± 17.5 (56.3) 26.7 ± 22.1 (62.8) 2.6 ± 6.3 (55.9) Tanypodinae 0.7 ± 1.3 (1.6) 0.5 ± 1.1 (1.5) 0.1 ± 0.3 (0.2) 0 ± 0 (0.0) Unidentified/other midges* 17.8 ± 7.8 (40.5) 0 ± 0 (0.0) 0.1 ± 0.4 (0.2) 0.1 ± 0.4 (2.1)

Total Chironomidae 44.0 ± 21.0 30.0 ± 31.6 42.5 ± 34.2 4.7 ± 12.0

Lake Yale No. fish collected 5 7 15 10 Chironomus crassicaudatus 6.2 ± 13.9 (75.6) 0 ± 0 (0.0) 0 ± 0 (0.0) 0 ± 0 (0.0) Glyptotendipes paripes 0.2 ± 0.4 (2.4) 1.7 ± 4.1 (9.0) 0 ± 0 (0.0) 0.2 ± 0.4 (20.0) Cryptochironomus spp. 0 ± 0 (0.0) 1.7 ± 3.1 (9.0) 0.5 ± 1.3 (18.5) 0.2 ± 0.4 (20.0) Pseudochironomus spp. 0 ± 0 (0.0) 8.6 ± 9.6 (45.5) 0.6 ± 1.6 (22.2) 0.2 ± 0.4 (20.0) Tanytarsini 0.4 ± 0.9 (4.8) 6.0 ± 6.4 (31.7) 1.3 ± 1.3 (48.1) 0.3 ± 0.5 (30.0) Tanypodinae 1.4 ± 3.1 (17.1) 0.4 ± 1.1 (2.1) 0 ± 0 (0.0) 0.1 ± 0.3 (10.0) Unidentified/other midges* 0 ± 0 (0.0) 0.5 ± 1.1 (2.6) 0.3 ± 0.8 (11.1) 0 ± 0 (0.0)

Total Chironomidae 8.2 ± 13.5 18.9 ± 17.2 2.7 ± 3.0 1.0 ± 0.9

*Larvae could not be identified due to damage to head capsules.

ber and December 2000, Geoldichironomus spp. (Lobinske 2001) and thus were not in the immedi- larvae in gut contents represented a much larger ate grazing area of the collected fish. Mean per- percentage of total chironomids when compared cent composition of G. paripes larvae in all gut to their percentage composition in the entire lake contents and their overall percent composition in and nearshore populations. In Lake Dora, no sig- lake total larval population was significantly dif- nificant differences (t-test) were noted between ferent (t = 2.938, P = 0.026, n = 4), but there was percent composition of individual midge taxa in no significant difference noted with percent com- gut contents and the prevailing populations in position in firm sediment areas (t = 0.440, P = the nearshore or entire lake area. In Lake Yale, 0.676, n = 4). No significant differences were C. crassicaudatus was the most common chirono- noted among any other chironomid taxa collected mid in gut contents during May 1999, but was from Lake Yale. only a minor component (<20%) of overall midge Based on the data, bluegill (L. macrochirus) community in the lake during that time. During seem to be indiscriminate feeders on midge larvae July 2000, Pseudochironomus spp. and Tanytar- inhabiting nearshore areas of Lakes Dora and sini comprised a similar percentage of nearshore Yale. However, Gilinsky (1984), Rieradevall et al. chironomids and in the gut contents of collected (1995), and Wolfram-Wais et al. (1999) reported fish. In the remaining periods of the study in Lake selectivity of fish predation on chironomids. The Yale, Tanytarsini were the most common midge significant difference between G. paripes in Lake larvae in fish gut contents, as well as in the firm Yale benthos and fish gut contents indicates blue- sediments. Glyptotendipes paripes was the most gill collected during this study were possibly fo- common midge in Lake Yale during July and Sep- cusing their feeding efforts in sandy, nearshore tember 2000 and composed a large proportion of areas, a behavior similar to the preference of total midge larvae during December 2000, but midge predatory fish foraging in restored, sand comprised a smaller component of the gut con- bottom areas of Lake Tohopekaliga reported by tents. This was probably because G. paripes lar- Butler et al. (1992). Rieradevall et al. (1995), But- vae were aggregated in deeper areas of the lake ler et al. (1992) and Gilinsky (1984) reported that 374 Florida Entomologist 85(2) June 2002

Fig. 1. Composite percent composition of total midge larvae of various midge taxa in bluegill (Lepomis macro- chirus) gut contents and prevailing field populations in Lakes Dora and Yale (Lake County, Florida) May 1999-De- cember 2000. Chironomus crassicaudatus (Cc), Glyptotendipes paripes (Gp), Cryptochironomus (Cryp), Pseudochironomus (Pseu), Geoldichironomus (Geol), Tanytarsini (Tant), Tanypodinae (Tanp), and unidentified/ other chironomid taxa (Oth).

fish predation significantly lowered chironomid SUMMARY larval density in various studied habitats. How- ever, Batzer et al. (2000) reported that fish preda- Compositions of midge larvae consumed by the tion in a marsh weedbed did not significantly predaceous fish, bluegill (Lepomis macrochirus) reduce chironomid densities when compared to were examined in relation to associated standing fish-excluding enclosures because of increased crop of midge larvae in two central Florida lakes. predation by other macroinvertebrates. Rasmus- Overall, these preliminary results indicate that sen (1990) reported midge larvae were a major bluegill may be indiscriminate predators on part of whitefish diet in Hjarbaek Fjord, Den- midge larvae in shallow, nearshore areas and dis- mark, but whitefish only consumed 2-5% of total play limited predation on larvae aggregated in chironomid productivity. Further work is needed deeper areas of the lakes. to refine these preliminary results, enumerate ac- tual standing crop of bluegill in Florida lakes to REFERENCES CITED estimate the total consumption of midge larvae in relation to the chironomid standing crop and to ALI, A. 1995. Nuisance, economic impact, and possibili- determine if bluegill are a major source of midge ties for control, pp. 339-364. In P. D. Armitage, P. S. reduction. Cranston, and L. C. V. Pinder (eds.), The Chironomi- dae: the biology and ecology of non-biting midges. Gratitude is expressed to the Florida Fish and Chapman and Hall, London, UK. Wildlife Conservation Commission for necessary ALI, A. 1996. Pestiferous Chironomidae and their man- permits. This is Florida Agricultural Experiment agement, pp. 487-513. In D. Rosen, F. D. Bennett, and Station Journal Series No. R-08369. J. L. Capinera (eds.), Pest management in the sub- Scientific Notes 375

tropics: integrated pest management—A Florida LOBINSKE, R. J. 2001. Ecological studies of larval Glyp- Perspective. Intercept, UK. totendipes paripes (Chironomidae: Diptera) in se- BATZER, D. P., C. R. PUSATERI, AND R. VETTER. 2000. Im- lected central Florida lakes for creating an pacts of fish predation on marsh invertebrates: di- exploratory temporal and spatial model of nuisance rect and indirect effects. Wetlands 20: 307-312. populations. Ph. D. Dissertation. University of Flor- BOWEN, S. H. 1996. Quantitative description of the diet, ida, Gainesville. 161 pp. pp. 513-532. In B. R. Murphy and D. W. Willis (eds.), RASMUSSEN, K. 1990. Some positive and negative ef- Fisheries techniques. 2nd Edition. American Fisher- fects of stocking whitefish on the ecosystem redevel- ies Society. Bethesda, MD. opment of Hjarbaek Fjord, Denmark. Hydrobiologia BUTLER, R. S., E. J. MOYER, M. W. HULON, AND V. P. 200/201: 593-602. WILLIAMS. 1992. Littoral zone invertebrates communi- RIERADEVALL, M., E. GARCIA-BERTHOU, AND N. PRAT. ties as affected by a habitat restoration project on Lake 1995. Chironomids in the diet of fish in Lake Tohopekaliga, Florida. J. Freshwat. Ecol. 7: 317-328. Banyoles (Catalonia, Spain), pp. 335-340. In P. S. CRANSTON, P. S. 1995. Medical significance, pp. 365-384. Cranston (ed.), Chironomids: from genes to ecosys- In P. D Armitage, P. S. Cranston, and L. C. V. Pinder tems. CSIRO. Canberra, Australia. (eds.), The Chironomidae: the biology and ecology of WOLFRAM-WAIS, A., G. WOLFRAM, B. AUER, E. MIKSCHI, non-biting midges. Chapman and Hall, London, UK. AND A. HAIN. 1999. Feeding habits of two introduced EPLER, J. H. 1995. Identification manual for the larval fish species (Lepomis gibbosus, Pseudoorasbora Chironomidae (Diptera) of Florida. Revised Edition. parva) in Neusiedler See (Austria), with special ref- Florida Department of Environmental Protection. erence to chironomid larvae (Diptera: Chironomi- Tallahassee, FL. dae). Hydrobiologia 408/409: 123-129. GILINSKY, E. 1984. The role of fish predation and spatial heterogeneity in determining benthic community structure. Ecology 65: 455-468.

376 Florida Entomologist 85(2) June 2002

SUPPRESSION OF CITRUS ROOT WEEVIL EGG HATCH BY DIFLUBENZURON FOLIAR RESIDUES

R. C. BULLOCK AND R. R. PELOSI University of Florida, IFAS, Indian River Research and Education Center 2199 South Rock Road, Ft. Pierce, FL 34945

Previous studies with diflubenzuron (AI of lated on the twig by removing other leaves to Micromite®, Dimilin® [Cromton Corporation, facilitate caging of the oviposition site. Nine hun- Greenwich, CT 06831] and Thompson-Hayward dred and fifty liters of aqueous and 0.5% oil emul- TH-6040) revealed inhibition of embryogenesis in sion sprays (FC 435-66) containing 568 g of eggs of several citrus root weevils fed on treated Micromite 25W were prepared. These materials foliage and provided folded wax paper strips as were applied as foliar sprays to “run-off” to 3 trees oviposition sites (Schroeder et al. 1976; Love- per treatment. Three unsprayed trees served as strand & Beavers 1980). In addition, Schroeder et controls. al. (1976) found that foliar sprays of diflubenzu- After spray application, the leaves were al- ron applied at rates of 56.6 g and 113.2 g (AI)/378 lowed to dry. The paired leaves on 4 terminals liters of water with a high-pressure handgun were paper-clipped together and each terminal sprayer reduced egg hatch for 10 days. An aerial inserted into a 5.1 cm diameter, 15.2 cm long cy- application delivering 283 g (AI) + the extender lindrical mesh cage on the tree. Two male and 4 Pinolene® in 45 l/A was effective for at least 26 female field-collected D. abbreviatus adults were days against Diaprepes abbreviatus (L.). Loves- placed in each cage with a sprig of tender foliage trand and Beavers (1980) also obtained signifi- as food. Fresh, tender foliage was provided as food cant reduction of egg hatch of D. abbreviatus, every other day. Adults were removed from the Pachnaeus litus (Germar) and Artipus floridanus cages on the 7th day post-treatment. Cages were Horn for 28 days after treatment with handgun- left on the tree an additional 7 days to protect the applied foliar sprays equivalent in rates to those egg masses from predation. Cages and leaf-pairs used by Schroeder et al. in 1976. were collected on the 14th day and brought to the By eliminating wax paper strips as oviposition lab where egg masses were exposed and exam- sites, Schroeder (1996) limited the effect of di- ined under a stereomicroscope to determine their flubenzuron to residues present on the leaf sur- viability. Eggs with embryos, empty eggs and ne- faces used for oviposition. This indicated that onates were counted. This same procedure was direct egg mass contact with diflubenzuron was repeated with fresh weevils on the 7th, 14th, 21st an additional mode of action. The two rates eval- and 28th day after spraying to provide data on the uated by Schroeder (1976, 1996), 142 g and 284 g activity of residues on egg hatch over time. The (AI)/984 l of a 1% oil emulsion, were the same test was terminated after 5 weeks. Rainfall was used by Lovestrand and Beavers’ 1980 rate, recorded during the test (Table 1). which were equivalent to 1× and 2× the 568 g rate There were 116, 1267 and 4054 neonates recov- of Micromite 25W (142 g AI) recommended for ered from leaves harboring eggs treated with Micro- rust mite control (Knapp 2001). mite 25W and oil emulsion, aqueous mixture of Schroeder (1996) harvested field-sprayed Micromite 25W, and untreated control, respec- leaves, paper-clipped them together to form an tively. This is an average of 98% and 70% reduction oviposition site, and placed treated and untreated in egg hatch for all time periods with significant leaf-pairs in cages containing lab-reared oviposit- differences between treatments (Table 2). ing female adult D. abbreviatus. The 1× and 2× rates resulted in reductions of 77% and 86% in egg hatch for all time periods with no significant TABLE 1. RAINFALL RECORDED (IN CM) DURING MICRO- differences between rates. MITE 25W TRIALS AT FT. PIERCE, FL. Our research, conducted at Ft. Pierce, FL, and reported here, compares the effect of residues on 1996 1997 egg hatch reduction from aqueous and 0.5% oil emulsion sprays containing 568 g rate of Micro- Treatment applied: 30 May 21 May mite 25W. The 1996 Test. A total of twenty shoots bearing 21-31 May 0 1.39 mature flush were selected and tagged on nine 6- 31 May 2.21 0 year-old ‘Ruby Red’ grapefruit trees. On each 1-8 Jun 2.54 2.57 twig, 4 distal leaves that could be paired and held 9-16 Jun 2.18 13.18 together with a paper clip were selected. These 17-24 Jun 7.54 3.66 provided juxtaposed plant surfaces that were suitable for oviposition. The 2 leaf-pairs were iso- Gauge checked 0800 h daily.

Scientific Notes 377

TABLE 2. PERCENT REDUCTION IN HATCH OF D. ABBREVIATUS EGGS LAID ON MICROMITE 25W-TREATED FOLIAGE.

% Reduction in egg hatch at indicated days post treatmentz Rate Average Treatment per 950 l +7 +14 +21 +28 +35 % reduction

Micromite 25W+ 568 g FC 435-66 Oil 4.75 l 99 c 100 c 98 c 94 b 99 c 98 c Micromite 25W 568 g 55 b 74 b 72 b 75 b 71 b 70 b Untreated 0 2 a 14 a 3 a 6 a 4 a 6 a

zPercent separation within columns by Duncan’s Multiple Range Test, 1% level.

TABLE 3. PERCENT REDUCTION IN HATCH OF P. LITUS EGGS LAID ON MICROMITE 25W-TREATED FOLIAGE.

% Reduction in egg hatch at indicated days post treatmentz Rate Average Treatment per 950 l +7 +14 +21 +28 % reduction

Micromite 25W+ 568 g FC 435-66 Oil 4.75 l 70 b 100 b 50 b 100 b 80 b Micromite 25W 568 g 25 a 39 ab 59 b 78 ab 50 a Untreated 0 2 a 56 a 2 a 28 a 22 a

zPercent separation within columns by Duncan’s Multiple Range Test, 5% level.

Schroeder (1996) found no difference between SUMMARY two Micromite 25W treatments, viz., 568 and 1136 g of Micromite 25W in 1% oil emulsion Residues of the 568 g Micromite 25W spray sprays. However, we found that the addition of rate were less effective in reducing egg hatch of 0.5% oil emulsion to the Micromite 25W treat- Pachnaeus litus than the same rate of Micromite ment significantly reduced egg hatch compared to 25W fed by Lovestrand and Beavers (1980) to the the aqueous spray. same insect, but the addition of oil in our studies The 1997 Test. Materials and methods in this improved the effectiveness of the foliar residues experiment differed in 3 respects from the 1996 in the absence of feeding. test: (1) the weevil was P. litus, (2) 17-year-old Furthermore, Micromite 25W in the 0.5% oil “Navel” orange trees were used, and (3) the test emulsion was equal in efficacy to Micromite 25W in was terminated after 28 days. 1% oil emulsion (Schroeder 1996) in reducing Dia- The results of the test reveal that there were prepes abbreviatus egg hatch and would provide 110, 747 and 1167 neonates found on leaves the grower equivalent performance for less cost. treated with Micromite 25W oil emulsion, aque- There is no reduction in oviposition attempts ous and no spray, respectively. This is an average or hatchability because of oil residue (Schroeder of 80% and 50% reduction for all time periods & Green 1983). with significant differences among treatments (Table 3). REFERENCES CITED While Lovestrand and Beavers (1980) compared KNAPP, J. L. 2001. Florida Citrus Pest Management 568 and 1136 g of Micromite 25W in aqueous Guide: 1999. Fla. Coop. Ext. Service SP-43. sprays and found that egg hatch was significantly LOVESTRAND, S. A., AND J. B. BEAVERS. 1980. Effect of reduced by both treatments, we found that 568 g diflubenzuron on four species of weevils attacking in 0.5% oil emulsion treatment was equivalent in citrus in Florida. Florida Entomol. 63(1): 112-115. its impact on the weevils to their low rate of 568 SCHROEDER, W. J., J. B. BEAVERS, R. A. SUTTON, AND g. Our aqueous spray residue, however, per- A. G. SELHIME. 1976. Ovicidal effect of Thompson- formed poorly compared to their similar aqueous Hayward TH-6040 in Diaprepes abbreviatus on cit- treatment, perhaps partly due to the rainfall that rus in Florida. J. Econ. Entomol. 69(6): 780-82. occurred during our test (Table 1). However, the SCHROEDER, W. J., AND D. S. GREEN. 1983. Diaprepes abbreviatus (Coleoptera: Curculionidae): oil sprays oil component of our emulsion spray apparently as a regulatory treatment, affect on eggs treatment. improved residue retention and contributed to its J. Econ. Entomol. 76(6):1395-1396. acceptable performance. SCHROEDER, W. J. 1996. Diflubenzuron residue: reduc- Florida Agricultural Experiment Station Jour- tion of Diaprepes abbreviatus (Coleoptera: Curcu- nal Series No. R-07067. lionidae) neonates. Florida Entomol. 79(3): 462-3.

378 Florida Entomologist 85(2) June 2002

CALLING BEHAVIOR OF TWO DIFFERENT FIELD POPULATIONS OF PHYLLOCNISTIS CITRELLA (LEPIDOPTERA: GRACILLARIIDAE): EFFECT OF AGE AND PHOTOPERIOD

J. A. JACAS1 AND J. E. PEÑA2 1Institut Valencià d’Investigacions Agràries, Ctra. Montcada a Nàquera km 5, E-46113-Montcada, Spain Present address: Universitat Jaume I, Departament de Ciències Experimentals, Campus del Riu Sec E-12071-Castelló de la Plana, Spain

2University of Florida, Tropical Research and Education Center, Homestead, FL 33031

Phyllocnistis citrella Stainton (Lepidoptera: ing were subjected both to G-test (Sokal & Rohlf Gracillariidae), is an oligophagous leafminer of 1969) and two-way ANOVA for unbalanced de- Rutaceae, especially of Citrus spp. (Jacas et al. signs (SAS Institute 1985). Onset calling times 1997), occurring in all citrus-growing areas were also analyzed using a two-way ANOVA. worldwide. Pheromone traps have been used suc- About 10,000 observations of moth activity were cessfully in Japan (Ando et al. 1985, Ujiye 1990) recorded. but unsuccessfully in China (Tongyuan et al. Table 1 shows the influence of age and photo- 1989), Italy (Ortu 1996), Spain and Florida (pers. period on the percentage of calling females. Call- obs.), and Turkey (Uygun, N., Univ. Çukurova, ing patterns for moths subjected to the same Adana, Turkey, pers. comm.). Negative results photoperiod regime at the two locations were sig- could be attributed to the use of a non-specific at- nificantly different (G = 24.71; df = 6; P < 0.005). tractant, and to a poor understanding of the be- Nevertheless, the age pattern exhibited by fe- havior of the pest. The objective of this study was males exposed to L16:D8 at IVIA did not signifi- to determine patterns of calling of two popula- cantly differ from that exhibited by those exposed tions of P. citrella when exposed to three different to L8:D16 at TREC (G = 7.59; d.f. = 6; P = 0.2245). photoperiods as a first step toward determination Photoperiod (F = 15.56; df = 2, 38; P < 0.0001) and of a specific sex pheromone. age (F = 2.67; df = 6, 38; P = 0.0352) affected the A first set of experiments was carried out dur- proportion of females calling, whereas the geo- ing June-July 1998 at the Institut Valencià d’In- graphical/seasonal origin did not (F = 0.70; df = 1, vestigacions Agràries (IVIA) Spain (0.4°W long., 38; P = 0.4102). Calling activity of females ex- 39.6°N lat., 33 m alt). A second set was performed posed to L12:D12 photoperiod was the highest, es- at the Tropical Research and Education Center pecially for age 1 females. Moths aged 4 to 7 called (TREC) Florida (80.2°W long., 25.3°N lat., 1 m more than those aged 2 and 3, with 1-day old fe- alt.), during October-November 1998. Pupae of males being the less active ones. Females exposed P. citrella were gathered from citrus orchards. to a photoperiod opposite to the one occurring Female pupae (Garrido & Jacas 1996) were when they were collected in the field barely re- placed in 12 cm-diameter petri dishes containing sponded during the whole experimental period. 2% agar and filter paper and held in a cabinet at Mean onset calling times (MOCT) are shown 25 ± 1°C, under a photoperiod matching local one in Figure 1. Both geographical/seasonal origin (F at that moment. Emerging adults were trans- = 781.26; df = 1, 374; P < 0.0001) and photoperiod ferred daily to analogous petri dishes (max. 6 (F = 31.06; df = 2, 374; P < 0.0001) affected the moths per dish) and fed droplets of honey plus MOCT, but age did not (F = 1.35; df = 6, 374; P = pollen. These were placed in a cabinet at 25 ± 1°C, 0.2337). Moths studied during the first set of as- and exposed to one of the three photoperiods: says called significantly earlier than those tested L12:D12, L16:D8, and L8:D16. At 60 min inter- during the second set, and, similarly, the longer vals during scotophase, and using a portable red the photophase experienced by a moth, the sooner light, moth activity (quiescence, locomotion, or it called. The MOCT for females collected at IVIA calling) was observed in the cabinet and recorded. was 8.62 ± 1.29 h (N = 130; mean ± SE) when ex- Calling females opened and vibrated their wings posed to L12:D12 and 8.11 ± 0.93 h (N = 90) when and rhythmically protruded and retracted the exposed to L16:D8, whereas for those collected at ovipositor. Observations were classified per scoto- TREC, it was 11.67 ± 0.58 h (N = 71) when ex- phase hour and sorted by photoperiod, age, and posed to L8:D16, and 10.65 ± 0.61 h (N = 84) when geographical/seasonal origin. Age was expressed exposed to L12:D12. Calling occurred mostly to- as the scotophase number (1 to 7), starting with ward the end of the scotophase. Non-calling fe- the first complete scotophase after emergence. males (8L:16D for the experiments at IVIA and The onset of calling was determined as the time 16L:8D at TREC) were usually very active toward (hours after lights off) halfway between the first the end of the scotophase. time calling was observed and the previous obser- Phyllocnistis citrella has been considered a vation. Differences in percentages of female call- crepuscular and dawn-active moth. Our results

Scientific Notes 379 IVIA AT

MEASURED

OUT

CITRELLA 24 36 4229 67 13 5017 50 5 33 17 CARRIED .

P

9405956677017 OF WAS

ASSAYS

FEMALES

OF

SET

VIRGIN

FIRST

CALLING THE

). OF D

:16 L ; 8 D :12 L PERCENTAGE

; 12 D THE

:8 L ON (16 : 1-7) 30. INSECTS

WAS

PHOTOPERIODS THE

BY

FEMALES

OF

DIFFERENT 3 EXPERIENCED

NUMBER

UNDER NITIAL (1-16) OFF SCOTOPHASES

Site 1: IVIASite 1: TREC Site 2: TREC. I OF

AT

LIGHTS

WAS

NUMBER ( AFTER

ONE

AGE

OF

SECOND

INTERVALS

16L:8D 12L:12D 8L:16D 16L:8D 12L:12D 8L:16D H THE

1- NFLUENCE AND AT 1234567 12345 6 7 1234567 1234567 123 4 567 1234567 1. I ABLE 1 2 3 4 52 66222 738 49 10 915710 911101113163610 6 30 42 54 69 63 66 21 33 33 49 52 25 1 8 13 11 2 3 6 14 14 15 16 8 10 10111213 47 44 84 21 68 48 75 48 100 52 6 60 75 5 75 26 60 11 43 19 33 1 2 13 11 18 53 56 29 53 45 60 86 50 67 57 100 57 75 4 71 31 17 20 9 43 53 33 40 67 60 33 T

380 Florida Entomologist 85(2) June 2002

Fig. 1. Influence of age (1-7) on the onset of calling (in h since start of scotophase) of virgin females of P. citrella under three different photoperiods (16l:8d; 12l:12d; 8l:16d). The first set of assays was carried out at ivia (i-) and the second one was at TREC (T-). initial number of females was 30.

show a marked dawn sexual activity. Virgin fe- Age and photoperiod are two of the many fac- males modified their calling patterns in response tors affecting the occurrence of calling behavior of to photoperiod with some limitations. Females female insects (Howse et al. 1998). Further labo- could not adapt to the photoperiods completely ratory experimentation and field testing should opposite to the ones they had experienced in the take our results into account. field. Furthermore, for a given location, the This research was partially supported by a MOCT under different photoperiods extraordi- grant of the Conselleria de Cultura, Educació i narily matched current natural scotophases (9 h Ciència de la Generalitat Valenciana, Spain, to at IVIA and 14.5 h at TREC). Therefore, it is pre- J. E. P., and a fellowship under the OECD Co-op- sumed that differences observed at the two loca- erative Research Program: Biological Resource tions were due to seasonal changes rather than to Management for Sustainable Agricultural Sys- geographical differences between populations. tems to J. A. J. The ability of virgin females of many Lepidoptera Florida Agricultural Experiment Station Jour- to modify the periodicity of their calling in re- nal Series No. R-08436. sponse to different environmental changes, in- cluding day length (Haynes & Birch 1984), has SUMMARY been interpreted as an adaptation to reduce the impact of climatic fluctuations on mating success The effect of age and photoperiod on the calling (Delisle & McNeil 1987). behavior of Phyllocnistis citrella was studied. Onset calling time was not affected by moth P. citrella is sexually active at dawn. Activation is age, although age increased the proportion of fe- presumably caused by the cumulative time males calling (1 < 2 = 3 < 4 = 5 = 6 = 7 days). At a elapsed since sunset. Onset calling time was not given temperature, virgin females of numerous affected by the age of moths, but the proportion of crepuscular and nocturnal species of Lepidoptera females calling increased with age. advance the onset time of calling on successive nights (Delisle 1992). This has been interpreted REFERENCES CITED as an adaptation increasing the probability of older females to attract a mate before younger ANDO, T., K. Y. TAGUCHI, M. UCHIYAMA, T. UJIYE, AND moths initiate their calling (Swier et al. 1977). H. KUROKO. 1985. (7Z-11Z)-7,11-hexadecadienal sex

Scientific Notes 381

attractant of the citrus leafminer moth. Phyllocnis- stocks and citrus-related species for resistance to tis citrella Stainton (Lepidoptera: Phyllocnistidae). Phyllocnistis citrella (Lepidoptera: Gracillariidae). Agricultural and Biological Chemistry, Tokyo 49: Crop Protection 16: 701-705. 3633-3653. ORTU, S. 1996. Field observations on sexual attraction DELISLE, J., AND J. N. MCNEIL. 1987. Calling behaviour of Phyllocnistis citrella Stainton: (7Z, 11Z)-7,11- and pheromone titre of the true armyworm Pseuda- hexadecadienal. Proc. XX Intl. Cong. Entomol. p. 496, letia unipuncta (Haw.) (Lepidoptera: Noctuidae) under Firenze, Italy, Aug. 25-31. different temperature and photoperiodic conditions. SAS INSTITUTE. 1985. SAS User’s guide: Basics. SAS In- J. Insect Physiol. 33: 315-324. stitute, Cary, NC. DELISLE, J. 1992. Age related changes in the calling be- SOKAL, R. R., AND F. J. ROHLF. 1969. Biometry. The haviour and the attractiveness of obliquebanded lea- Principles and Practice of Statistics in Biological Re- froller virgin females, Choristoneura rosaceana, under search. W.H. Freeman and Company, San Francisco. different constant and fluctuating temperatures. En- 776 pp. tomologia Experimentalis et Applicata 63: 55-62. SWIER, S. R., R. W. RINGS, AND G. J. MUSICK. 1977. Age- GARRIDO, A., AND J. A. JACAS. 1996. Differences in mor- related calling behavior of the black cutworm Agrotis phology of male and female pupae of Phyllocnistis ci- ipsilon. Ann. Entomol. Soc. Amer. 70: 919-924. trella Stainton (Lepidoptera: Gracillariidae). Florida TONGYUAN, D., X. JINJUN, W. ZESHUA, AND K. FANLEI. Entomol. 79: 603-606. 1989. 7Z, Z11-hexadecadienal: sex attractant of HAYNES, K. F., AND M. C. BIRCH. 1984. The periodicity Phyllocnistis wampella Liu et Zeng. Kunchong Zhi- of pheromone release and male responsiveness in shi 3: 147-149 (In Chinese). the artichoke plume moth, Platyptilia carduidac- UJIYE, T. 1990. Studies on the utilization of a sex at- tyla. Physiol. Entomol. 12: 1597-1600. tratctant of the citrus leafminer moth, Phyllocnistis HOWSE, P. E., I. D. R. STEVENS, AND O. T. JONES. 1998. citrella Stainton (Lepidoptera: Phyllocnistidae). I. Insect pheromones and their use in pest manage- Analysis of seasonal population trends and some be- ment. Chapman & Hall, London, UK, 369 pp. havioral characteristics of the male moths by the use JACAS, J. A., A. GARRIDO, C. MARGAIX, J. FORNER, A. AL- of synthetic sex attractant traps in the field. Bulletin CAIDE, AND J. A. PINA. 1997. Screening of citrus root- of the Fruit Tree Research Station 18: 19-46.

382 Florida Entomologist 85(2) June 2002

SYNONYMY AND VALID NAME OF THE FAMILIES VIETNAMELLIDAE AND AUSTREMERELLIDAE (EPHEMEROPTERA: EPHEMERELLOIDEA)

MICHAEL D. HUBBARD Entomology, Florida A&M University, Tallahassee, Florida 32307, USA

Tshernova (1972) established the genus Viet- name at the time Austremerellidae was estab- namella for a new species of Asian Ephemerel- lished. lidae, Vietnamella thani. Allen (1980), with little Therefore the valid name for this family of discussion, placed Vietnamella as a subgenus of Ephemerelloidea is Vietnamellidae Allen, and Cincticostella Allen, 1971, in the tribe Ephemer- Austremerellidae McCafferty and Wang is a jun- ellini. Later, Allen (1984), with somewhat more ior synonym. discussion of the nymphal characters, in particu- lar the nymphal gills, admitted that the place- SUMMARY ment of Vietnamella as a subgenus of Cincticostella had been incorrect. He established a new subtribe Vietnamellidae Allen is shown to be a senior of the Ephemerellini, Vietnamellae, for Vietna- synonym of Austremerellidae McCafferty and mella. Wang. Edmunds and Murvosh (1995) transferred the genus Vietnamella to the related subfamily Telo- REFERENCES CITED ganodinae and retained the tribe name Vietnam- ellini for it. ALLEN, R. K. 1971. New Asian Ephemerella with notes McCafferty and Wang (1997) presented evi- (Ephemeroptera: Ephemerellidae). Canadian Ento- dence that Vietnamella was closely related to Aus- mologist 103: 512-528. tremerella Riek, 1963, and, apparently over- ALLEN, R. K. 1980. Geographic distribution and reclas- looking the existence of the subfamily name Viet- sification of the subfamily Ephemerellinae (Ephe- namellinae, established a new subfamily, Aus- meroptera: Ephemerellidae), pp. 71-91. In J. F. tremerellinae, in the family Teloganodidae for the Flannagan and K. E. Marshall (eds.), Advances in Ephemeroptera Biology. Plenum, New York. two genera. Then, McCafferty and Wang (2000) ALLEN, R. K. 1984. A new classification of the subfamily proceeded to raise the taxon to a family, which Ephemerellinae and the description of a new genus. they called Austremerellidae. Pan-Pacific Entomologist 60: 245-247. According to the International Code of Zoolog- EDMUNDS, G. F., JR., AND C. H. MURVOSH. 1995. Sys- ical Nomenclature (ICZN 1999: Article 36.1) es- tematic changes in certain Ephemeroptera studied tablishment of Vietnamellae (Allen 1984) had the by R. K. Allen. Pan-Pacific Entomologist 71: 157-160. effect of also establishing names for all other [ICZN] INTERNATIONAL COMMISSION ON ZOOLOGICAL ranks in the family group (coordinate taxa: e.g., NOMENCLATURE. 1999. International Code of Zoolog- family, subfamily, tribe). This means that the ical Nomenclature. Fourth Edition. International Trust for Zoological Nomenclature, London. 306 p. name Vietnamellidae became available and valid MCCAFFERTY, W. P., AND T.-Q. WANG. 1997. Phyloge- at that time. netic systematics of the family Teloganodidae (Ephe- In determining the valid name of a zoological meroptera: Pannota). Ann. Cape Province Museum taxon, the “Principle of Priority” applies (ICZN (Natural History) 19: 387-437. 1999: Article 23). It states that “The valid name of MCCAFFERTY, W. P., AND T.-Q. WANG. 2000. Phyloge- a taxon is the oldest available name applied to netic systematics of the major lineages of pannote it . . .” Although the Code does make exceptions mayflies (Ephemeroptera: Pannota). Trans. Amer. for family names in prevailing usage (IC2N 1999: Entomol. Soc. 126: 9-101. Article 35.5), Article 23.9.1 describes prevailing RIEK, E. F. 1963. An Australian mayfly of the family Ephemerellidae (Ephemeroptera). J. Entomol. Soc. usage to require, among other things, that the se- Queensland 2: 48-50. nior synonym has not been used as a valid name TSHERNOVA, O. A. 1972. Some new species of mayflies after 1899. This is clearly not the case here. Viet- from Asia (Ephemeroptera, Heptageniidae, Ephe- namellae (and its coordinate taxa) was estab- merellidae). Entomologicheskoe Obozrenie 51: 604 lished in 1984 and was an available and valid 614. (In Russian)

Scientific Notes 383

TUNNEL ARCHITECTURES OF THREE SPECIES OF MOLE CRICKETS (ORTHOPTERA: GRYLLOTALPIDAE)

RICK L. BRANDENBURG1, YULU XIA1 AND A. S. SCHOEMAN2 1North Carolina State University, Department of Entomology, Raleigh, NC 27695-7613

2Department of Zoology and Entomology, University of Pretoria, Pretoria 0002, Republic of South Africa

The southern mole cricket, Scapteriscus vicinus similar product used in South Africa. This and Giglio-Tos, and the tawny mole cricket, S. borellii other similar products are widely available at Scudder, damage turfgrass in southeastern local hardware and automobile repair stores. United States. The two species are univoltine in Approximately of the recommended amount of most of their range. They also have similar life cy- hardener was added to the fiberglass resin (about cles and morphology. However, southern mole 1 ml hardener/100 ml resin). The fiberglass resin cricket is primarily carnivorous, whereas tawny hardens quickly after adding hardener, therefore, mole cricket is herbivorous (Taylor 1979, Ulagaraj the whole procedure must be done quickly. The fi- 1975, Matheny 1981). The African mole cricket, berglass resin container was covered and shaken Gryllotalpa africana Palisot de Beauvois, is a after adding hardener. The contents were then world-wide pest (Sithole 1986). It damages plants poured immediately into the tunnel entrance in a including wheat, maize, rice, sorghum, millet, bar- steady stream. The excavation of the castings ley, oats, potatoes, cassava, groundnuts, straw- started 1-2 h after pouring. The fiberglass resin in berries, turnips, tobacco, and vegetables in Africa, one can (1 l) usually filled two to three mole Asia, and Europe. It also causes severe damage to cricket tunnels. We used a large screwdriver to turfgrass on golf courses in South Africa and Asia clear away the grass roots surrounding the tunnel (Brandenburg, unpubl. data). Tsedeke (1979) re- entrance and to determine the direction of the ported that surface tunneling behavior, which is casting before starting to dig the cast. Finding partly determined by feeding preference, is differ- other entrance(s) of the tunnel helps to judge di- ent between the two species in the U.S. We there- rection the tunnel casting. There are at least two fore speculate that tunnel architectures of three entrances for tawny and African mole cricket tun- species are also different judging from the differ- nels. The soil on tunnel casts was washed away ences in their feeding behavior and damage. This with water following excavation. study used fiberglass resins to compare tunnel We made over 100 castings and excavations architecture of three species of mole crickets in during 3 years. Tunnels of tawny mole crickets two locations were almost always (90%) in the shape of “Y” with Tawny and southern mole cricket tunnel cast- two entrances for each tunnel (Fig. 1A, B, C). Vari- ings were made on the driving range of Oyster ations were occasionally observed in the tunnel ar- Bay Golf Course, Brunswick County, NC, during chitecture. There might be two parallel “Y”s 1998 to 2000. The turfgrass on the driving range linking together to form a tunnel, or, two entrances was hybrid bermudagrass, Cynodon dactylon (L.) observed at each end of a tunnel. The length of Pers. in sandy loam soil. African mole cricket tun- most tawny mole cricket tunnels ranged from 50 to nel castings were made in typical heavy clay soil 70 cm. Tunnels of African mole crickets also typi- at Silver Lakes Golf and Country Club, Pretoria, cally showed “Y” shape (Fig. 1G, H, I). The length of South Africa. The turfgrass on the fairway was African mole cricket tunnels ranged from 10 cm to Kikuyu grass, Pennisetum clandestinum Hochst. 23 cm. This was much shorter than that observed ex Chiov. in tawny mole crickets. The tunnels of southern We located mole cricket tunnel entrances by mole crickets were more likely in a reversed “Y” hand, and cleaned foreign matter, debris, and soil shape with only one surface entrance (Fig. 1D, E, from the area around the entrance. We then used F). The tunnels often branched within 10 cm deep a soapy water flush (Short & Koehler 1979) as an of the soil surface. The tunnels were usually much irritant to flush the mole cricket from the tunnel shorter than those of tawny mole cricket. for species identification. The soapy water flushing The difference in tunnel architecture probably also helped to find other entrances to the tunnel relates to the behavioral difference of the three and make the soil around the entrance firm. Areas species. Southern mole crickets are carnivorous. without turf were avoided because the tunnels are They seek prey throughout the soil. Our observa- often blocked by loose soil during the flushing. tions and research by Tsedeke (1979) suggested We have previously reported that fiberglass that southern mole crickets were much more ac- resin is the best material for mole cricket tunnel tive in tunneling than tawny and African mole casting (Brandenburg et al. 2001). Bondo® fiber- crickets. This may be why southern mole cricket glass resin and hardener (Dynatron/Bondo Corp., tunnels were almost always branched down into Atlanta, GA), was used in the U.S. study and a the soil rather than near the soil surface. In con-

384 Florida Entomologist 85(2) June 2002

Fig. 1. Tunnel Castings of the Tawny (A, B, C), Southern (D, E, F), and African (G, H, I) Mole Cricket. trast, tawny and African mole crickets feed on harder, clay soil in South Africa may also be re- roots near the soil surface and their activity is lated to the shorter tunnel structure for the Afri- around the branches of the “Y”. They may face fre- can mole cricket. quent threats from predators (Hudson et al. 1988) This research was funded in part by the and need more than one entrance in a tunnel. The United States Golf Association, Green Section Re-

Scientific Notes 385 search. We would like to thank Jack Bacheler, (Orthoptera: Gryllotalpidae) Using Three Materials Ken Sorensen, and Mike Waldvogel in Depart- with an Emphasis on Fiberglass Resin. J. Kansas ment of Entomology, North Carolina State Uni- Entomol. Soc. (In press). versity, for reviewing the draft of this manuscript. HUDSON, W. G., J. H. FRANK, AND J. L. CASTNER. 1988. Biological control of mole crickets (Orthoptera: Gryl- lotalpidae) in Florida. Ann. Entomol. Soc. Amer. 34: SUMMARY 192-198. MATHENY, E. L., JR. 1981. Contrasting feeding habits of The fiberglass resin used to determine tunnel pest mole cricket species. J. Econ. Entomol. 74: 444- architecture of three species of mole crickets was 445. effective in providing an accurate and durable SHORT, D. E., AND P. G. KOEHLER. 1979. A sampling record of mole cricket activity. The tunnel castings technique for mole crickets and other pests in turf- of the tawny mole cricket, Scapteriscus vicinus grass and pasture. Florida Entomol. 1979. 62 (3): 282-283. Scudder, and the African mole crickets, Gryllo- SITHOLE, S. Z. 1986. Mole Cricket. Zimbabwe Agric. J. talpa africana Palisot de Beauvois, almost always 83(1): 21-22. exhibit a “Y” shape upper tunnel structure. The TAYLOR, T. R. 1979. Crop contents of two species of mole tunnel castings of the southern mole cricket, crickets, Scapteriscus acletus and S. vicinus (Ortho- Scapteriscus borellii Giglio-Tos, are typically an ptera: Gryllotalpidae). Ibid. 62: 278-279. inverted “Y” shape. Tunnels of tawny mole crick- TSEDEKE, A. 1979. Plant material consumption and sub- ets typically go deeper into the soil and are usu- terranean movement of mole crickets (Orthoptera: ally more complex than the other two species. Gryllotalpidae: Scapteriscus) as determined by ra- dioisotope techniques, with notes on materials for laboratory feeding. M.S. thesis. University of Flor- REFERENCES CITED ida, Gainesville. 72 pp. ULAGARAJ, S. M. 1975. Food habits of mole crickets BRANDENBURG, R. L., Y. XIA, AND M. G. VILLANI. 2001. (Orthoptera: Gryllotalpidae: Scapteriscus). J. Geor- Determining Tunnel Structure of Mole Crickets gia Entomol. Soc. 10: 229-231.

386 Florida Entomologist 85(2) June 2002

ESTABLISHMENT OF A CHINESE STRAIN OF COTESIA RUBECULA (HYMENOPTERA: BRACONIDAE) IN THE NORTHEASTERN UNITED STATES

R. G. VAN DRIESCHE AND C. NUNN Department of Entomology, University of Massachusetts, Amherst, MA, 01003

Cotesia rubecula (Marshall) is a braconid par- We subsequently reared this strain both in the asitoid of Pieris spp. larvae that is relatively spe- laboratory and from field-collected larvae be- cific to Pieris rapae (L.) (Lepidoptera: Pieridae), a tween 1988 and 1993 and made 12 other releases pest of cabbage and related cole crops. The estab- in Massachusetts, three in Connecticut, and one lishment of this parasitoid in the eastern United in Rhode Island, for 17 release locations in total States to help suppress this garden pest has been (Fig. 1, two sets of MA sites overlap on map). long sought. Efforts to establish it in North Amer- Same-year recoveries of the parasitoid were made ica have a complex history. A self-introduced pop- at seven of these sites, and recoveries were made ulation of uncertain origin was discovered on after one or more years at seven other sites. Vancouver Island in British Columbia in 1963 Among the seven sites at which recoveries were (Wilkinson 1966), and the range of this popula- made in subsequent years, we observed the para- tion has extended as far south as Oregon (Biever sitoid at three sites one year after release and at 1992). This strain was later released in Missouri, single sites 2, 3, 5, and 8 years after last release. New Jersey, South Carolina, and Ontario (near Recovery efforts varied in different years and not Ottawa) (Puttler et al. 1970; Williamson 1971, all sites were visited yearly. 1972). This strain appears not to have established To assess spread away from release sites, we in Missouri (Parker & Pinnell 1972), but may periodically collected groups of P. rapae larvae have established in Ontario (Corrigan 1982). Poor from non-release locations. We have recovered establishment of this strain was attributed to an C. rubecula from 13 non-release sites, from just improperly timed diapause induction response north of Hartford, Connecticut to Craftsbury, Ver- (Nealis 1985). mont (north of St. Johnsbury) (Fig. 1). Recoveries A second population, from the former Yugo- have been made both along the Connecticut River slovia, was released in Missouri in the mid 1980s, Valley and in various locations in the Litchfield and subsequently released in Virginia and On- Hills in Connecticut, the Berkshire Hills in Mas- tario. In 1988, the Yugoslavian strain was recov- sachusetts, and the Champlain Valley of Vermont. ered in Virginia, but this population later appeared Towns in which recoveries have been made either to have died out, perhaps due to high level of at non-release sites or, if a release site, one or hyperparasitism (McDonald & Kok 1991). In more years after the release include Winsor and 1993, C. rubecula, of uncertain origin, was found Falls Village, Connecticut; Williamstown, Lanes- to be the dominant parasitoid in Quebec, in farm- boro, Westhampton, Northampton, Amherst, Had- ing areas near Montreal (about 160 km east of ley, Deerfield, Northfield, and Barre, Massachu- Ottawa) (Godin & Boivin 1998). setts; and Stamford, Rockingham, Hartland, South In 1988, a population of C. rubecula was col- Royalton, Plainfield, Burlington, and Craftsbury, lected by David Reed of the USDA in Shenyang, Vermont (Fig. 1), all of which indicate extensive China (42 north latitude, 123 east longitude), for range expansion in both agricultural valleys and release in the eastern United States. This location adjacent forested hill country. Recoveries through- matched the intended release location in Massa- out Vermont bring the known range of C. rubecula chusetts in latitude, and both locations have conti- near the Canadian border. Godin and Boivin nental type climates. Parasitized host larvae (1998)’s report of recovery of C. rubecula of uncer- (P. rapae) were shipped to the USDA quarantine tain origin in southern Quebec, seen in the light laboratory in Newark, Delaware. Adult parasitoids of the data presented here, may be a further were allowed to emerge and, following confirmation northward extension of the Chinese population, of species identity, 99 female and 49 male C. rubec- rather than an eastward extension of releases ula adults from this shipment were shipped to the from near Ottawa. This is uncertain, as no molec- senior author in Amherst, Massachusetts in July of ular markers have been identified to separate 1988 and all were released in field cages in a pesti- these populations. cide-free, 0.1 ha collard plot in Deerfield, Massa- Because establishment of C. rubecula has been chusetts (42 n. l.). That C. rubecula was not present associated with declines in density of the other in- at this site before the release (through spread, per- troduced P. rapae parasitoid, Cotesia glomerata haps from some distant source) is demonstrated by (L.), in Oregon and Washington (Biever 1992), we the absence of C. rubecula in large numbers of also counted numbers of P. rapae larvae and Cote- hosts collected at this location and dissected for sia parasitoid cocoons (as single cocoons for the parasitism rates in a population dynamics study I solitary species C. rubecula and as cocoon groups ran in 1985 and 1986 (Van Driesche 1988). for the gregarious species C. glomerata) on entire

Scientific Notes 387

Fig. 1. Locations of release and recovery sites for Cotesia rubecula (Chinese strain) in the northeastern United States. Solid circles with cross hatches are release sites at which recoveries were made one or more years after re- lease; solid circles without cross hatches are non-release sites where recoveries were made; hollow circles with cross hatches are release sites at which recoveries were made only in the year of release; hollow circles without cross hatches are release sites at which no recoveries were made; crosses (in Canada) indicate closest sites with known releases or recoveries of other strains of C. rubecula. collard plant in two years (1985, 1986) before the tor was the time period (pre- or post-release of release of C. rubecula (in 1988) in our Deerfield, C. rubecula). Data from two pre-release years Massachusetts plot and for three years (1990, were available (1985 and 1986), as well as three 1991, 1992) after the parasitoid’s establishment. post-release years (1990, 1991, and 1992). Data The size and management of this 0.1 ha collard from the year of release (1988) and the following plot was maintained in a consistent manner from year (1989) were not included in order to allow for 1985 to 1992. For each week of the growing sea- population interactions to reach a stable end son (May through September) in these years, we point before analysis. We then used a χ2 test on examined 20-100 whole collard plants (187 sam- these data to determine if the percentage of ple occasions, with an average of 58 plants per plants with C. glomerata cocoons on them varied date). Numbers of groups of C. glomerata cocoons between the pre and post release periods. Of 4098 were greatest in July, August and September. For plants examined in these months in 1985 or 1986, all samples in these three months in five years, 16% (661) bore live C. glomerata cocoons, com- we classified each plant sampled into a 2 × 2 ma- pared to only 3% (82) of 2708 plants examined in trix. One factor was whether or not the plants had July-September of 1990, 1991 or 1992, a signifi- C. glomerata cocoons on them (+/-). The other fac- cant difference (χ2 = 288.7, df = 1, P < 0.005). Total

388 Florida Entomologist 85(2) June 2002

numbers of larvae on sampled plants in the prer- REFERENCES CITED elease period (9980) were either the same or lower (2.45 larvae per plant) than in the post re- BIEVER, K. D. 1992. Distribution and occurrence of Cote- lease period (8683 larvae on 2708 plants, or 3.21 sia rubecula (Hymenoptera: Braconidae), a parasite per plant) and therefore the decline in the propor- of Artogeia rapae in Washington and Oregon. J. tion of plants bearing C. glomerata cocoons in the Econ. Entomol. 85: 739-742. post release period cannot be explained as being CORRIGAN, J. E. 1982. Cotesia (Apanteles) rubecula [Hy- menoptera: Braconidae] recovered in Ottawa, On- due to a decrease in the number of larvae per tario ten years after its release. Proc. Entomol. Soc. plant available for parasitization. Furthermore, Ontario 113: 71. in the 1990-1992 period, C. rubecula accounted GODIN, C., AND G. BOIVIN. 1998. Occurrence of Cotesia for over half of all Cotesia parasitism of P. rapae rubecula (Hymenoptera: Braconidae) in Quebec, 30 larvae in the Deerfield, MA, release plot (74% [n = years after its introduction in North America. Cana- 175], 91% [n = 87], and 49% [n = 76], in 1990, dian Entomol. 130: 733-734. 1991, and 1992 respectively, with percentage be- MCDONALD, R. C., AND L. T. KOK. 1992. Colonization ing based cocoons of each parasitoid species seen and hyperparasitism of Cotesia rubecula (Hymen.: on sampled plants). These data suggest that Braconidae), a newly introduced parasite of Pieris rapae, in Virginia. Entomophaga 37: 223-228. C. glomerata declined in density in the study area NEALIS, V. 1985. Diapause and the seasonal ecology of following establishment of C. rubecula. However, the introduced parasite, Cotesia (Apanteles) rubec- regionally C. glomerata remains common in New ula (Hymenoptera: Braconidae). Canadian Entomol. England and we cannot say if it has declined at 117: 333-342. that larger spatial scale. PARKER, F. D., AND R. E. PINNELL. 1972. Further stud- We conclude that C. rubecula has become an ies of the biological control of Pieris rapae using sup- important parasitoid of P. rapae in parts of New plemental host and parasite releases. Environ. England since its establishment in 1988 and we Entomol. 1: 150-157. suspect that its range is still increasing and PUTTLER, B., F. D. PARKER, R. E. PINNELL, AND S. E. THEWKE. 1970. Introduction of Apanteles rubecula should be examined in other states in the region. Marsh. and other parasites of Pieris rapae in British Potential effects of this new parastioid on related Columbia. J. Econ. Entomol. 63: 304-305. native Pieris butterflies have been examined VAN DRIESCHE, R. G. 1988. Survivorship patterns of Pi- (Benson et al. unpubl.). eris rapae (Lep.: Pieridae) larvae in Massachusetts kale with special reference to mortality due to Cote- sia glomerata (Hymen.: Braconidae). Bull. Entomol. SUMMARY Res. 78: 199-208. A population of Cotesia rubecula, collected from WILKINSON, A. T. S. 1966. Apanteles rubecula Marsh. near Beijing, China and released in Massachusetts and other parasites of Pieris rapae in British Colum- in 1988, has established and spread throughout bia. J. Econ. Entomol. 59: 1012-1013. WILLIAMSON, G. D. 1971. Insect liberation in Canada. much of New England. It has become a common Parasites and predators 1970. Agric. Canada Liber- parasitoid of Pieris rapae in agriculture fields and ation Bull. 34. is also found in meadow habitats. Cotesia glomer- WILLIAMSON, G. D. 1972. Insect liberation in Canada. ata appears to have declined in abundance follow- Parasites and predators 1971. Agric. Canada Liber- ing establishment of C. rubecula. ation Bull. 35.

Scientific Notes 389

FIRST REPORT OF ANASTREPHA COMPRESSA IN MEXICO AND NEW RECORDS FOR OTHER ANASTREPHA SPECIES IN THE YUCATAN PENINSULA (DIPTERA: TEPHRITIDAE)

V. HERNÁNDEZ-ORTIZ1, P. MANRIQUE-SAIDE2, H. DELFÍN-GONZÁLEZ2 AND L. NOVELO-RINCÓN2 1Instituto de Ecología, A.C. Departamento de Entomología. Km 2.5 antigua carretera a Coatepec, Apdo Postal 63, 91000, Xalapa, Veracruz, México

2UADY. Facultad de Medicina Veterinaria y Zootecnia. Departamento de Zoología, Apdo Postal 4-116 Itzimná. Mérida, Yucatán, México

Tephritid flies (Diptera: Tephritidae) are pana, El Cermeño, and Balboa) (Stone 1942), and known as “true fruit flies” due to the close relation- updated records include material from Venezuela ship between their immature stages and their wild (Norrbom et al. 1998). This is the first report from and domesticated host plants. They are the most Mexico which extends its distribution to the north important dipteran pests of agriculture world- of Central America. wide (Christenson & Foote 1960) and include 481 Material examined. MEXICO. CAMPECHE, genera and 4352 species (Norrbom et al. 1998). Alfredo Bonfil, 20-I-1997, McPhail-trap (4 , 1 Anastrepha is the most economically important IEXA). and diverse genus of fruit flies in the Americas, with 197 species distributed throughout tropical Anastrepha fraterculus (Wiedemann, 1830) and subtropical areas (Norrbom et al. 2000). To date, 32 species are known to occur in Mexico, and Its known distribution includes the United seven of them have been reported for the Yucatan States (Texas), South and Central America, and Peninsula (YP). This includes the Mexican states Trinidad (also introduced to Galapagos Is.) (Stone of Campeche, Quintana Roo and Yucatan. Anas- 1942; Norrbom et al. 1998). Records for Mexico in- trepha species reported for each state are: clude Aguascalientes, Campeche, Chiapas, Nuevo Campeche (Anastrepha fraterculus, A. hamata, León, Oaxaca, Tamaulipas, Veracruz, Yucatan, A. limae, A. obliqua, A. serpentina); Quintana Roo and Zacatecas (Arana et al. 1992; Hernández-Or- (A. hamata, A. ludens, A. obliqua, A. serpentina); tiz 1992).Here we report the first record for Quin- and Yucatan (A. fraterculus, A. ludens, A. serpen- tana Roo state. tina, A. striata) (Hernández-Ortiz 1992). Material examined: MEXICO. CAMPECHE, This work provides new locality records for Tenabo, Tinún, 10-VII-1997, Herrera (1 IEXA); Anastrepha species already reported for YP, first QUINTANA ROO, Felipe Carrillo Puerto, Chunhu- records of three species in YP (A. ampliata, A. pal- hub, 5-12-VIII-1997, Xool (11 , 128 IEXA); lens and A. spatulata), and the first record of YUCATAN, Dzilam Reserve, Rancho San Salvador, A. compressa for Mexico. Material examined is 30-XI-1992, light-trap, Delfín & Manrique (1 deposited at the Colección de Insectos, Instituto CER); Ibid. 30-XI-1992, butterfly-trap (1 CER); de Ecología A.C. Xalapa, Veracruz (IEXA), and Colonia Yucatan, Kalah Dzonot, 21-22-IX-1993, Colección Entomológica Regional of the Univer- butterfly-trap, Delfín & Manrique (2 CER). sidad Autónoma de Yucatán (CER). Anastrepha ludens (Loew, 1873) Anastrepha ampliata Hernández, 1990 This species ranges from southern USA This species has only been reported in the (Texas) to Central America (Foote 1967). It has Mexican state of Chiapas, and Guatemala been reported for 25 Mexican states including (Hernández-Ortiz 1990, 1992); new record for YP. Quintana Roo and Yucatan (Huerta et al. 1987; Material examined. MEXICO. CAMPECHE, Arana et al. 1992; Hernández-Ortiz 1992). Here Calkini, Concepción, 5-VIII-1997. Huchin (1 we report the first record for Campeche state. IEXA); QUINTANA ROO, Chunhuhub, McPhail- Material examined. MEXICO. CAMPECHE, trap, 12-VIII-1997, Xool-Cetz (8 IEXA); La Libertad, 26-VI-1993, net, Manrique (1 YUCATAN, Dzilam Reserve, Rancho San Salva- CER); QUINTANA ROO, Felipe Carrillo Puerto, dor, 30-IX-1992, light-trap, Delfín & Manrique (2 Chunhuhub, 26-XII-1997, Xool (1 , 1 IEXA). CER), Ibid. butterfly-trap (1 CER), Ibid. 30- XI-1992, net (2 CER). Anastrepha obliqua (Macquart, 1843)

Anastrepha compressa Stone, 1942 This species has a wide distribution in the New World, having been recorded from the USA This species was described on the basis of ma- (Florida, Texas) to South America and the Carib- terial from several localities in Panama (La Cam- bean Islands (Stone 1942). In Mexico its range in-

390 Florida Entomologist 85(2) June 2002 cludes both coasts and other central states (18 northern limit in USA (Texas) and a southern dis- states), including Campeche and Quintana Roo tribution in many countries in Central and South within YP (Hernández-Ortiz 1992). America (Stone 1942; Hernández-Ortiz & Aluja Material examined. MEXICO. CAMPECHE, 1993) Here we report the first record for Quintana Palizada, Rancho Santa Isabel, 23-IX-1997, Ca- Roo state. brales (1 IEXA), Palizada, Rancho Alamilla, 7- Material examined. MEXICO. QUINTANA X-1997, Cabrales (1 IEXA), Cd. del Carmen, ROO, Felipe Carrillo Puerto, Chunhuhub, 26-XII- Matamoros [no date], Dominguez (1 IEXA). 1997, Xool (1 , 1 IEXA), Felipe Carrillo Puerto, Emiliano Zapata, 15-X-1997, Xool (3 , 2 IEXA). Anastrepha pallens Coquillett, 1904 At this time, a total of 11 Anastrepha species are known to be present in the states of the This species has a known distribution in the Yucatan Peninsula. The current status of our Mexican states of Chiapas, Coahuila, Guerrero, knowledge of these species and the most complete Jalisco, Nayarit, Nuevo Leon, Oaxaca, Sinaloa, list of Anastrepha in those Mexican states is as Sonora, Tamaulipas and Veracruz (Hernández- follows: Campeche, 9 species; Quintana Roo, 7 Ortiz 1992), with its northern limits in USA species; Yucatan, 6 species. (Texas) and southern limits in El Salvador and The species A. ludens, A. serpentina and Honduras (Norrbom 1998). Here we report a new A. obliqua are economically important for fruit record for YP. The reported hosts for this species crops in Mexico. At the present time, there are no are Sideroxylon celastrinum (Kunth) T. D. Pen- reports indicating that they represent an agricul- nington (as Bumelia angustifolia) and S. lanugi- tural problem for YP states. However, the pres- nosa Michx. Baker et al. (1944) also reported as ence of species detrimental to national and host B. spiniflora A.DC. in Mexico, probably a regional agriculture, indicates the need for a per- synonymy of S. celastrinum (see Norrbom 1998). manent surveillance campaign to evaluate popu- Material examined. MEXICO. YUCATAN, Ria lation and damage levels throughout the region. Lagartos Reserve, El Cuyo, 8-II-1995, light-trap, Delfín & Manrique (1 CER). SUMMARY

Anastrepha serpentina (Wiedemann, 1830) We make an updated report for the Anastrepha This species has a wide distribution in the species that occur in the Yucatan Peninsula. For New World, from USA (Texas) to South America the first time A. compressa Stone is recorded from and Trinidad (Foote 1967). In Mexico it occurs in Mexico, and the occurrence of three other species 16 states along both coasts and in other central for this region is documented: A. ampliata Hernán- states, including the YP (Hernández-Ortiz 1992). dez, A. pallens Coquillett, and A. spatulata Stone. Material examined. MEXICO. QUINTANA Information about localities and collection dates ROO, Vallehermoso, 19-20-VII-93, butterfly-trap, of voucher specimens are provided. Delfín & Manrique (1 CER); YUCATAN, Dz- ilam Reserve, Rancho San Salvador, 30-XI-1992, REFERENCES CITED light-trap, Delfín & Manrique (3 CER); Ibid. butterfly-trap (1 CER). ARANA, P. J., P. T. VERA, AND J. CACHON. 1992. La mosca mexicana de la fruta del sur del estado de Yucatán. Experiencias en desarrollo sostenible, 1-33. Anastrepha spatulata Stone, 1942 BAKER, A. C., W. E. STONE, C. C. PLUMMER, AND M. This species has a known distribution in the MCPHAIL. 1944. A review of the studies on the Mex- Mexican states of Baja California Sur, Chiapas, ican fruitfly and related Mexican species. U.S. Dept. Agric. Misc. Publ. 531: 1-155. Guerrero, Jalisco, Morelos, Nayarit, Oaxaca, Si- CHRISTENSON, L. D., AND R. H. Foote. 1960. Biology of naloa, Sonora, Tamaulipas and Veracruz, with its fruit flies. Annu. Rev. Ent. 5: 171-192. northern limit in USA (Texas) and southern limit FOOTE, R. H. 1967. Family Tephritidae (Trypetidae, in Costa Rica and Panama (Foote 1967; Hernán- Trupaneidae), Fascicle 57, pp. 1-91. In N. Papavero dez-Ortiz 1992). Here we report it for the first [ed.], A catalogue of the Diptera of the Americas time for YP. South of the United States. Museo de Zoologia, Uni- Material examined. MEXICO. CAMPECHE, versidade de Sâo Paulo, Brazil. Hecelchacán, Blanca Flor, 20-III-1997, Dzul (1 HERNÁNDEZ-ORTIZ, V. 1990. Lista preliminar de especies IEXA). mexicanas del género Anastrepha (Diptera: Tephriti- dae) con descripción de nuevas especies, registros y si- nonimias. Folia Entomol. Mexicana. 80: 227- 244. Anastrepha striata Schiner, 1868 HERNÁNDEZ-ORTIZ, V. 1992. El género Anastrepha Schiner en México (Diptera: Tephritidae). Taxonomía, dis- This species has a known distribution in the tribución y sus plantas huéspedes. Instituto de Mexican states of Aguascalientes, Colima, Chia- Ecología Publ. 33. Xalapa, Veracruz, Mexico. 162 pp. pas, Guerrero, Jalisco, Mexico, Morelos, Nayarit, HERNÁNDEZ-ORTIZ, V., AND M. ALUJA. 1993. Listado de Oaxaca, Sinaloa, Veracruz and Yucatan, with its especies del género neotropical Anastrepha (Diptera: Scientific Notes 391

Tephritidae), con notas sobre su distribución y plan- bases of names, pp. 65-251. In F. C. Thompson [ed.], tas hospederas. Folia Entomol. Mexicana 88: 89-105. Fruit fly expert identification system and systematic HUERTA, P. R., N. S. RODRÍGUEZ, AND M. G. SILLER. information database. Backhuys Publ., Leiden, 1987. Distribución geográfica de las moscas de la Netherlands. fruta del género Anastrepha Schiner en México, pp. NORRBOM, A. L., R. A. ZUCCHI, AND V. HERNÁNDEZ-OR- 128-146. In INIFAP [ed.], Primer Informe sobre Mos- TIZ. 2000. Phylogeny of the genera Anastrepha and cas de la Fruta en Mango. INIFAP, Veracruz, México. Toxotrypana (Trypetinae: Toxotrypanini) based on NORRBOM, A. L. 1998. A revision of the Anastrepha dac- morphology, pp. 299-342. In M. Aluja and A. L. Norr- iformis species group (Diptera: Tephritidae). Proc. bom [eds.], Fruit flies (Tephritidae): phylogeny and Entomol. Soc. Washington 100: 160-192. evolution of behavior. CRC Press, USA. NORRBOM, A. L., L. E. CARROLL, F. C. THOMPSON, I. M. STONE, A. 1942. The fruit flies of the genus Anastrepha. WHITE, AND A. FREIDBERG. 1998. Systematic data- U.S. Dept. Agric. Misc. Publ. 439: 1-112.

392 Florida Entomologist 85(2) June 2002

SCYPHOPHORUS ACUPUNCTATUS (COLEOPTERA: CURCULIONIDAE) ATTACKING POLIANTHES TUBEROSA (LILIALES: AGAVACEAE) IN MORELOS, MEXICO

MARIO CAMINO LAVIN, VICTOR R. CASTREJON GOMEZ, RODOLFO FIGUEROA BRITO, LUCILA ALDANA LLANOS AND MA. ELENA VALDES ESTRADA Departamento de Interacciones Planta-Insecto, Centro de Desarrollo de Productos Bióticos del Instituto Politécnico Nacional, COFAA, Carretera Yautepec-Jojutla km 8.5, A.P. 24, 62730 San Isidro, Yautepec, Morelos, Mexico [email protected]

Tuberose, Polianthes tuberosa L. (Liliales: had the highest incidence of damage (69%). Emil- Agavaceae), is grown as an ornamental plant and iano Zapata and Tepalcingo demonstrated 47 and as a source of a fragrant essence for perfumes 37% damage, respectively (Table 1). (Gonzatti 1981, Watson & Dallwitz 1992). It has This paper is the first formal report of S. acu- been grown commercially in the state of Morelos, punctatus attacking tuberose. Vaurie (1971), in Mexico, for 60 years. Currently there is grown ap- her revision of Scyphophorus, did not mention proximately 300 cultivated ha, which generates tuberose as a host plant of S. acupunctatus. Dampf US $6,300,000 per year (Uribe 2000). However, in mentioned the weevil as “acapiche del nardo” recent years, producers have noticed severe dam- (Anonymous 1930). Vaurie (1971) mentions that age caused by an insect that they call black weevil this insect is distributed from the southern USA to but unidentified scientifically. The objective of the north of South America, the Caribbean (Cuba, this work was to identify the pest and to assess Hispaniola, and Jamaica), East Africa, Hawaii, damage. Java, and Australia. In Mexico it has been re- The study was carried out in the central Mexi- ported attacking several economically important can state of Morelos, between 18°22’19” and plants of the family Agavaceae (McGregor & 19°07’10” north and between 99°30’8” and Gutiérrez 1983). In the Yucatán peninsula it has 19°07’10” west. Mean annual temperature is been mentioned as causing damage up to 50% in 20°C and the annual rainfall ranges from 900 to the cultivation of henequen (Agave fourcroydes 1100 mm (García 1981). Samples were collected Lem.) (Ramírez-Choza 1979). In the states of from March to September 2000 in the main areas Hidalgo, Tlaxcala, and México, it has been re- of tuberose production to include: the municipali- ported causing damage of 30% to the cultivated ties of (1) Tepalcingo, (2) Emiliano Zapata, and (3) plant “maguey pulquero” (Agave salmiana Salm- Coatlán del Río. Dyck ssp. crassispina (Trel.) Gentry and var. culta) In the first two municipalities, 100 bulbs of (Ruvalcaba 1983). In the state of Jalisco, it caused P. tuberosa were collected, whereas in the third, damage of 10% to “agave tequilero” (Agave tequi- 120 plants were collected. A 1-ha plot in each mu- lana Weber var. azul) (Valenzuela 1994). In the nicipality was selected, and samples were taken municipalities of Tepalcingo and Coatlán del Río, from the center and four sides of the plot. In the cultivation of A. tequilana began three years ago, laboratory, bulbs and whole plants were exam- whereas in Emiliano Zapata there are wild agaves, ined for insects. The insect larvae and adults were predominantly A. angustifolia Haw., in the hills deposited in 70% ethanol and processed for iden- near cultivated P. tuberosa. These two agaves may tification (see acknowledgments). serve as refuge to S. acupunctatus during chemical The weevil was identified as Scyphophorus acu- applications or when tuberose is not in the field, so punctatus Gyllenhal (Coleoptera: Curculionidae) control is complicated (Uribe 2000). Infestations in (Muñíz 2000, Marín 2000, Nápoles & Equihua Agave tequilana var. azul have been reported as 2000). Most damage is caused by weevil larvae from 4 to 24 weevils per plant (Solis et al. 1999). mining the bulb. The samples from Coatlán del Río The present survey showed 6-10 larvae per in-

TABLE 1. INFESTATION OF S. ACUPUNCTATUS IN P. TUBEROSA IN MORELOS, MEXICO (MARCH TO SEPTEMBER 2000).

Bulbs Plants Locality and date sampled/infested sampled/infested Adults/plant Percent infested

Tepalcingo 6 March 100/36 — — 36 Emiliano Zapata 7 April 100/47 — — 47 Tepalcingo 30 August 45/14 — — 40 Tepalcingo 6 September 120/42 — — 35 Coatlán del Río 5 September — 120/83 4-36 69

Scientific Notes 393

fested bulb and 4-36 adults per infested plant. GARCIA, E. 1981. Modificaciones al sistema de clasifi- S. acupunctatus is a severe pest of tuberose. Our cación climática de Koppen. Instituto de Geografía, work continue on S. acupunctatus life cycle, popu- Universidad nacional Autónoma del Estado de Méx- lation dynamics, identification and evaluation of ico. Offset Larios; México, D.F., Mexico. feeding attractants. GONZATTI, C. 1981. Flora Taxonómica Mexicana II. Ce- neti; Guadalajara, México (see pp. 87-88). We thank Dr. J. H. Frank (Entomology and Μαριν, J. A. 2000. Personal communication. INIFAP- Nematology Department, University of Florida) SAGAR, Celaya, Guanajuato, México. for guidance and corrections to a manuscript MCGREGOR, R., AND O. GUTIERREZ. 1983. Guía de insec- draft. This report was supported financially by tos nocivos para la agricultura en México. Alhambra; Fundación Produce Morelos, A. C. (project num- México. 166 pp. ber 4-I/A18/2000). We also thank Unión de Pro- MUÑIZ, R. V. 2000. Personal Communication. Sanidad ductores de Nardo del Estado de Morelos, through Vegetal, INIFAP-SAGAR, México. Mr. Rodolfo Uribe Landa. We thank R. Muñíz, NAPOLES, R. J., AND A. EQUIHUA. 2000. Personal Com- A. Marin and Dr. J. Nápoles and Dr. A. Equihua munication. Colegio de Posgraduados, Instituto de Fitosanidad, Montecillo, Estado de México, México. for identification of the insects. The authors of RAMIREZ-CHOZA, J. L. 1979. Metodología para el control this paper are scholarship awardees of Comisión del max del henequén Scyphophorus acupunctatus de Operación y Fomento de Actividades Académi- bajo condiciones de campo como resultados de tres cas del Instituto Politécnico Nacional. años de estudio. Fol. Entomol. Mexicana 42: 62-63. RUVALCABA, J. M. 1983. El maguey manso. Universidad Autónoma Chapingo; México. 122 pp. SUMMARY SOLIS, A. J., H. H. GONZALEZ, F. F. MENDOZA, S. M. CA- BRERA, H. E. CALDERON, A. VALLE DE LA PAZ, M. A. Scypophorus acupunctatus is reported attack- EQUIHUA, L. J. VAZQUEZ, AND M. A. GARZA. 1999. In- ing Polianthes tuberosa in Morelos, Mexico. All sectos asociados con Agave tequilana var. azul, en growth stages (eggs, larvae, pupae and adults) of cinco localidades de Jalisco, México. Memorias this insect were collected. The average percent in- XXXIV Congreso Nacional de Entomología, Aguas- festation was 51%. calientes, México, 23-26 May 1999: 455-457. This paper is in memory of Dr. Mario Camino URIBE, L. R. 2000. Personal communication. Presidente Lavín. de la Unión de Productores de nardo del Estado de Morelos. Coatlán del Río, Morelos, México. VALENZUELA, Z. A. G. 1994. El agave tequilero. Litteris; REFERENCES CITED México, D.F., México (see pp. 121-137). VAURIE, P. 1971. Review of Scyphophorus (Curculion- ANONYMOUS. 1930. Principales plagas y enfermedades idae: Rhynchophorinae). Coleopts. Bull. 25: 1-8. de los cultivos en la República Mexicana, incluyendo WATSON, L., AND M. J. DALLWITZ. 1992. The families of las más importantes de los Estados Unidos de flowering plants: descriptions, illustrations, identifi- Norteamérica. Oficina Federal para la Defensa Agrí- cation, and information retrieval. [Online] Available: cola, México. 195, 159, 220. http://biodiversity.uno.edu/delta/ (4 November 2000).

394 Florida Entomologist 85(2) June 2002

A NEW RECORD OF A MOTH ATTACKING SAPODILLA, WITH DESCRIPTIONS OF FEMALE GENITALIA AND THE LAST INSTAR LARVA

RUBEN IRUEGAS1, BENIGNO GOMEZ2, LEOPOLDO CRUZ-LOPEZ2, EDI A. MALO2 AND JULIO C. ROJAS2 1Rudyard Kipling No. 4800 casa 18, Jardines de la Patria, Zapopan, Jalisco, Mexico, CP 45030 2Departamento de Entomología Tropical, El Colegio de la Frontera Sur, Ap. Postal 36 Tapachula, Chiapas, México, CP 30700

ABSTRACT This paper reports a new record of a moth species, Zamagiria dixolophella Dyar (: ) attacking sapodilla (Manilkara zapota van Royen) in Southern Mexico. This is the first report of this species in Mexico. The female genitalia and the last instar larva are described and illustrated. Key Words: Pyralidae, Zamagiria dixolophella, Sapodilla, Manilkara zapota, Mexico

RESUMEN Un nuevo registro de una especie de palomilla atacando al chicozapote (Manilkara zapota van Royen) en el sur de Mexico es reportado. La palomilla fue identificada con Zamagiria dixolophella Dyar de la familia Pyralidae, subfamilia Phycitinae. Esta especie es reportada por primera vez para Mexico. La genitalia femenina y el ultimo estadio larvario es descrito e ilustrado.

The sapodilla (Manilkara zapota van Royen) is of Tapachula and Suchiate of the Chiapas state, a fruit tree native of the south of Mexico and Cen- Mexico. The pupae from tender young shoots and tral America, and is found from the Yucatan pen- fruits were taken to the laboratory and placed in insula to Costa Rica. Sapodilla is grown screen cages (20 × 20 cm) for adult emergence at commercially in India, the Philippines, Sri 25 ± 1°C, 30-50% RH and a photoperiod 16: 8 (L: Lanka, Malaysia, Mexico, Venezuela, Guatemala, D) h. After adult emergence, the insects were and other Central American countries (Mickel- killed and mounted for identification. Larvae bart 1996). The entomofauna associated with sa- were killed immediately after collection. They podilla trees has been reported from Venezuela, were fixed in Pampel liquid and preserved in 70% Puerto Rico and India (Rubio-Espina 1968, Bu- ethyl alcohol. For morphological studies the lar- tani 1975, Sandhu & Sran 1980, Medina-Goud et vae were cleared with lactic acid. The terminology al. 1987). For example, in India, over 25 species of used to describe the setae and other characters is insects have been recorded causing damage to sa- adopted from Stehr (1987). podilla (Butani 1975, Sandhu & Sran 1980), but the main insect pests are a galechid caterpillar RESULTS (Anarsia achrasella) and the pyralid caterpillar, Nephopteryx engraphella. The former species is a The moth species was identified as Zamagiria bud borer, while the latter feeds on the leaves, dixolophella Dyar (Pyralidae, Subfamily Phyciti- flower buds and young fruits. Unfortunately, the nae). This species represents the first report in entomofauna associated with sapodilla in its area Mexico. The original description of this species of original distribution has not been studied. In was based on males collected in Corazal, Panama, Mexico and Central America, the main insect pest in 1914 (Heinrich 1956). The female has not been of this tree seems to be the fruit flies of the genera described and is very close to Zamagiria pogery- Anastrepha, particularly A. serpentina (Norrbom thus Dyar (Neunzing & Dow 1993, Heinrich & Kim 1988). For several years the sapodilla pro- 1956). However, the female genitalia of Z. dixol- ducers of the Soconusco region of Chiapas state, ophella has some differences: the ductus bursae is Mexico have been reporting a bud borer moth, short and narrow, while in Z. pogerythus it is which has been causing problems in their planta- wide, as wide as the bursae. The ductus seminalis tions. This work was undertaken to identify the of Z. dixolophella is located in the apical part of moth as a first step towards further studies on the bursae, whereas in Z. pogerythus it is located in biology, ecology and control of this species. the basal part of the bursae (Figs. 1 A and B).

MATERIALS AND METHODS Description of Adults The biological material was collected from sev- In general appearance this insect is a small eral sapodilla farms located in the municipalities gray moth, resting with the wings folded along

Scientific Notes 395

Fig. 2. Mature larva of Z. dixolophella, lateral view.

matal seta S1 is situated between stemmata 2 and 3. Stemmatal seta S2 is near the opening of the stemmatal semicircle and placed in front of st- emmata 4. The length that divides S2 from the st- emmata 1 is close to 2/3 of the distance between this same seta and stemmata 4. S3 is placed out- side of the stemmatal semicircle. The length that separates S3 of the stemmata 6 is similar to the distance between S3 and S2. Pore Sa sited be- tween stemmata 6 and S3. The length that sepa- rates the pore Sa from stemmata 6 is the twice that of the distance from S3. The substemmatal seta SS1 is situated near to the mandible and sep- arated from it by a quarter of the length that ex- ists between this seta and stemmata 5. SS2 is placed below stemmatas 5 and 6 and with the Fig. 1. A) Female genitalia of Z. Pogerythrus (re- same distance separating both stemmata. The drawn from Heinrich 1956). B) Female genitalia of Z. distance that separates SS2 of SS1 is the twice dixolophella. that of the distance between SS2 and stemmata 5. SS3 is placed below SS2 and forming a straight line between them and stemmata 5. The distance the body. The forewings are dark smoky gray, that separates SS2 and SS3 is the same as that with raised scales, colored brown and dark gray. between SS2 and stemmata 5. Pore Ssa is situ- The hindwings are gray transparent, with the ated inside of substemmatal area. The length that costal area covered with gray scales. Male and fe- divides Ssa from SS1 is the twice of the distance male wing span is 19 mm. Males may be distin- between the same pore and SS2. The anterior guished by a bushy, forward-projecting labial palp seta A1 is placed in front of stemmata 2 and 3, an- and the presence of an aigrette of the maxillary terior seta A2 is situated directly above A1. The palps, which are reddish in color. Male antennae length that separates A1 from A2 is similar to have a basal sinus covered by strongly developed that which divides A1 from stemmata 3. gray scale tufts. Female genitalia (Fig. 1B): signum developed as a longitudinal, elongate, narrow and sinuous form, armed with strong spines extending the length of the bursa. The bursa has a plate armed with strong spines near the junction with the duc- tus bursae. The ductus seminalis extends from the apical part of the bursa and it is slightly scle- rotized. Ductus bursae are short and narrowed.

Description of Full Grown Larva

The total length of the last instar larva 15-16 mm and it is pinkish in color. The head is notable for the presence of dark spots located on the dor- sal and lateral surfaces of the frontal area (Fig. 2). The stemmatal area is a semicircle and formed by six stemmata (Fig. 3). Stemmata 3-5 are located in straight line and higher with respect to the other stemmata with the border sclerotized. Stem- Fig. 3. Larval head of Z. dixolophella, lateral view.

396 Florida Entomologist 85(2) June 2002

from A3-A6. The internal face of prolegs measures approximately 0.51-0.82 mm. The pinaculum of SD1 seta of A1-A8 has a circular aspect. Spiracle A8 is very large in comparison to the other ab- dominal spiracles and clearly directed towards the dorsocaudal position. Segment A9 with SV group bisetose and L group trisetose. In segment A10, the V setae are separated from each other by about half of the distance that they are separated from the V setae of segment A9. The V setae of A10 are longer or equal to twice the length of the V setae of A9.

DISCUSSION

Ten species have been described in the genus Zamagiria. The related Z. pogerythrus is distrib- uted from Campeche state, Mexico to Chejel, Fig. 4. Larval head of Z. dixolophella, frontal view. Guatemala. The best known species of this genus is Zamagiria laidion (Zeller), whose larvae also feed on the leaves and flowers of M. zapota, as well as on Manilkara emarginata (Sapotaceae) Pores Aa are placed above of A2. The length and Eriobotyra japonica (Rosaceae). This moth that divides Aa from the ecdysial line is twice that has been collected from the United States (Flor- which separates A1 from the ecdysial line. The ida), Guatemala, Panama, Colombia, Bolivia, frontal setae F1 is situated near the adfrontal lat- Venezuela and Brazil. Another species of this gen- eral suture. Pores Fa are situated in the middle era that feeds upon Sapotaceae is Zamagiria fra- part of the front. The distance between adfrontal terna, which has been reported in Cuba attacking setae AF2 with respect to F1 is twice that which Bumelia microcarpa (Heinrich 1956). separates AF1 and AF2. Lateral setae L1 is Oviposition of Z. dixolophella occurs on the placed anterior to A3 and the separation between buds of sapodilla. Larvae feed upon the ovaries both is the half of the length between L1 and st- and the petals of the flowers but frequently bore emmata 3. The Postdorsal pore Pb is separated by into tender young shoots. Also, the larvae are the same distance that exists between Pb1 and sometimes found inside sapodilla fruit. Prelimi- Pb2 setae. The clypeus has a pair of medium setae nary observations have shown that insect infesta- C1 and a pair of lateral setae C2, the latter sited tion persists almost throughout the year, in the intersection of the ecdysial line and the pro- however, the highest populations are found dur- jection of the adfrontal lateral suture and the di- ing the peak of flowering. viding line between clypeus and the anteclypeus. The labrum is incised with three pairs of medium labial setae and two pairs of lateral setae. The ACKNOWLEDGMENTS mandibles bear the characteristic tridentate structure with two basal setae. The intermediate We thank Armando Virgen and Alejandro del Mazo tooth projects beyond teeth, the oral surface con- for their help in collecting the biological material. Also cave. The submentum carries two medium setae we thank Rodulfo Muñoz Campero (Altamira), Esteban and a bilobed posterior border, recurved towards Moises (Rancho el Nayar) and Abenamar Gonzalez the dorsum. A well developed spinneret is (Rancho Cazanares) for allowing us to collect material from their farm. present. The thoraxic segment T1 with L group is bise- tose. Segment T2 bears a sclerotized very dark pi- REFERENCES CITED naculum and evident on the base of the SD1 seta. Segment T3 has a pinaculum on the SD1 seta but BUTANI, D. K. 1975. Insect pests of fruit crops and their is less conspicuous than in T2. The thoraxic spir- control, sapota. Pesticides 9: 37-39. acle is semicircular and larger than the abdomi- HEINRICH, C. 1956. American Moths of the subfamily nal spiracles, except for the spiracle on segment Phycitinae. United States National Museum Bulle- tin 207. Smithsonian Institution, Washington, D.C. A8, which is the same size and form as the tho- NEUNZIG, H. H., AND L. C. DOW. 1993. The Phycitinae of raxic spiracle. Belize (Lepidoptera: Pyralidae) Technical Bulletin The prolegs are well developed in the abdomi- 304. North Carolina Agricultural Communications. nal segments A3-A6 and A10 (caudal). Each pro- North Carolina State University. Raleigh, NC. leg carries many crochets. The crochets are MEDINA-GOUD, S., F. GALLARDO-COVAS, E. ABREU, AND arranged in an uniserial biordinal semicircle R. INGLES. 1987. The insects of nispero (Manilkara

Scientific Notes 397

zapota (L.) P. van Rogen) in Puerto Rico. J. Agric. RUBIO-ESPINA, E. 1968. Estudio preliminar de los insec- Univ. P.R. 71: 129-132. tos perjudiciales a los árboles de níspero (Achras MICKELBART, M. V. 1996. Sapodilla: a potential crop for zapota Linneus) en el estado Zulia, Venezuela. Revta. subtropical climates, pp. 439-446. In J. Janick [ed.], Facul. Agronomía 1: 1-24. Progress in new crops. ASHS Press, Alexandria, VA. SANDHU, G. S., AND C. S. SRAN. 1980. New records of NORRBOM, A. L., AND K. C. KIM. 1988. A list of the re- Lepidoptera on sapota. FAO Plant Protection Bulle- ported host plants of the species of Anastrepha tin 28: 43-44. (Diptera: Tephritidae). U.S. Dept. Agric. APHIS- STEHR, F. W. 1987. Inmature insects. Volume I. Kendall/ PPQ.81.52. Hunt Publishing Co. Dubuque, IA.

398 Florida Entomologist 85(2) June 2002

BOOK REVIEWS

ALUJA, M., AND A. NORRBOM. 2000. Fruit Flies (Tephritidae): Phylogeny and Evolution of Behavior. CRC Press; Boca Raton. 944 p. ISBN 0-8493-1275-2. Hardback. $169.95.

This book addresses the evolution and behav- Sections III through VI address the behavior ior of tephritid fruit flies, many of which are of and phylogenetic relationships of the Subfamilies, significant economic importance worldwide. The Blepharoneurinae and Phytalmiinae, Trypetinae, volume is the result of a symposium on the evolu- Dacinae, and Tephritinae. The chapters within tion of fruit fly behavior in honor of Drs. Ronald these sections use various approaches (primarily Prokopy and D. Elmo Hardy, internationally rec- cladistics, morphological characters, allozyme vari- ognized experts in the field and pioneers of the ation, and in at least one case, mitochondrial DNA) study of tephritid fruit fly evolution and behavior. to address the phylogenetic relationships among The volume represents a compilation of sympo- members of the respective Subfamilies. As with the sium papers arranged into thirty three chapters phylogenetic studies of many organisms, several as- and eight sections. The chapters range from broad sumptions have to be made regarding convergent topics such the history of the study of tephritid and divergent evolution of a variety of morphologi- fruit flies to empirical studies on the use of mito- cal (and even allozyme) traits. In some cases con- chondrial ribosomal DNA in the study of fruit fly trary results are obtained when comparisons are phylogeny. Nevertheless, they are appropriately made among some of the papers. Consequently, organized into Sections that reflect the thought- there seems to be a need for the use of existing high fulness and care the editors took to present this throughput DNA sequencing technologies to begin eclectic collection in a reasonably coherent and to study tephritid fruit fly genomes. Indeed, other logical format. Consequently, the editors achieved than the few mitochondrial DNA studies presented their stated objective that this book, though a in this book, there appears to be no significant dis- “hybrid” of wide- ranging topics, should serve as a cussion in any of the chapters on the potential use general reference to specialists in various aspects of DNA sequencing and analysis of specific te- of fruit fly evolutionary biology and behavior. phritid genes to help resolve some phylogenetic Section I provides reviews of the phylogeny of questions. For example, it would be of tremendous the Superfamily Tephritoidea (Chapter 1 by Kor- value to analyze those genes involved in/related to neyev), the behavior of fly relatives (tephritoids) sexual selection, mating, courtship behavior, and of the Tephritidae (Chapter 2 by Sivinski), and pheromone production. This book represents the the history of studies on tephritids including latest in the field of fruit fly phylogeny and behavior many of the World’s most important Genera of but provides evidence for the need of complemen- economic importance such as Anastrepha, Bactro- tary studies using state-of-the-art technology to cera, Ceratitis, and Rhagoletis (Chapter 3 by help resolve questions on tephritid fruit fly phylog- Diaz-Fleischer and Aluja). Together, these three eny and the evolution of mating systems. chapters provide an excellent overview of the Section VII contains an interesting and diverse three major themes in the book; phylogeny and collection of papers addressing the genetics of evolution, behavior, and aspects of fruit fly phero- populations, evolution of feeding behavior, sexual mones and attractants. Admittedly, Chapter one, selection and life history traits, and oviposition by virtue of the topic, presents a challenge to the behavior. A discussion of Drosophila speciation in non-taxonomist, especially one unfamiliar with Hawaii raises interesting issues that may also be tephritid systematics. Nevertheless, as the edi- addressed in tephritid fruit fly systems. Section tors indicate, the book is intended primarily for VIII, the Glossary, provides definitions and expla- specialists. Even to the non-taxonomist however, nations of various terms used throughout the a perusal of the chapter allows for a better appre- book. These are especially valuable in navigating ciation of the breadth of the Tephritoidea, far be- the papers on tephritid taxonomy/phylogeny. yond our narrow view of the tephritids that are of Overall, I found this book to be an excellent agricultural importance. The Glossary will be an and comprehensive compilation of research by invaluable tool for readers wishing to better un- experts in the field of tephritid phylogeny and derstand the systematics and indeed, other spe- behavior. I found few errors in the book. Each cialty areas addressed throughout the book. For chapter is clearly written and with the aid of the specialists and non-specialists alike, the histori- glossary and other outside references, chapters cal review of fruit fly research (Diaz-Fleischer and on taxonomy/phylogeny can be maneuvered to Aluja) and discussion of tephritoid flies (Sivinski) some degree by non-specialists. The graphics are is refreshing and informative. They provide a satisfactory and the color photographs excellent. solid background for students and researchers The font size of the nucleotide sequences and cla- alike, who wish to study novel tephritid and re- distic tables made reading somewhat difficult. lated fly species. This is an excellent book for the specialist and for

Book Reviews 399 those seeking new tephritid species for research. Drs. Handler and MacCoombs and colleagues in The major gap I found was the absence of nucle- Greece) that could help resolve questions on otide sequences of tephritids (some of which are tephritid fruit fly phylogeny and behavior. available in existing databases) that could help Pauline O. Lawrence address phylogenetic questions. The book could Department of Entomology and Nematology have benefited from a section on the role of cur- University of Florida rent molecular technology (such as that used by Gainesville, FL 32611-0620

400 Florida Entomologist 85(2) June 2002

SCHAEFER, C. W., AND A. R. PANIZZI (eds.). 2000. Heteroptera of Economic Importance. CRC Press; Boca Raton. (20+) 828 p. ISBN 0-8493-0695-7. Hardback. $94.95.

This book is a comprehensive and up-to-date Many true bugs are important predators of in- review of worldwide literature on Heteroptera sect pests and mites. De Clercq (Pentatomidae: from the point of view of their economic impor- Asopinae, Ch. 32) stated that the present litera- tance, whether in managed or natural systems. ture review has demonstrated the potential value The book’s most important contribution is as a of several pentatomid predators for the manage- resource, not just for those involved in the man- ment of a wide array of agricultural insect pests. agement of agricultural pests, but for anyone This information could be extended to other Het- interested in true bugs. Its organization is so eroptera families as well, such as Nabidae (Ch. practical as to invite the reader to simply read it 27), on which there is little information in gen- for edification and enjoyment or to find informa- eral, especially for the tropics. Berytid predators tion with immediate applicability. This book rep- (Ch. 31) have also received little attention, but resents a clear guide for future research in are potentially important. Mirids (Ch. 28) that heteropteran biology, and its reviewed references feed on delphacid planthopper eggs have been and the massive bibliography make it a necessary used successfully in classical biological control reference book in entomological libraries. programs. Other zoophagous heteropterans, mostly Currently, Heteroptera, a suborder of the insect aquatic and semiaquatic, are economically impor- order Hemiptera, include about 37,000 described tant natural enemies because they feed, in part, species, many of which feed on plants; thousands on blood-sucking Diptera, especially mosquito more await description or discovery, according to larvae and pupae (Ch. 21 to 25). Biological control Schaefer and Panizzi (Introduction, Ch.1). Little is programs have to take into account that some of known about the basic biology and ecology of most these predacious species are cannibalistic, some true bugs. This book succeeds in thoroughly sum- may damage crops, or may feed on beneficial marizing and assessing present knowledge of the arthropods such as pollinators and spiders. true bugs and presents it in the form of current The authors in this book do not restrict their re- reviews and references. The authors reviewed views to Heteroptera associated with agricultural thousands of articles and books, although not all crops, but also those associated with ornamentals, references are listed; nonetheless, the number of plants not cultivated for major commercial benefit references is impressive. For example, Sweet listed such as sycamore trees (Tingidae, Ch. 4) and royal 925 references (Lygaeoidea, Ch.6). The references palms (Thaumastocoridae. Ch. 5), and natural sys- include information on taxonomy and control of tems, where the intrinsic value of Heteroptera has true bugs, as well information on natural history, not been quantified. behavior, morphology, embryology, endocrinology, This book is also an excellent resource for be- and ecology, etc. Some of these references guide ginning entomologists. There is a general descrip- the interested reader to identification keys, but tion for each of the families included and a review which are not included in this book. of their classification and their feeding behavior. The scope of this book is beyond that of only Homeowners will also find it useful because it North America; for example, research is reviewed contains details about true bugs that occasionally from India, China, Brazil, Thailand, Poland, and become nuisances in or around homes, such as Ukraine. An insect or a plant that is considered a boxelder bugs (Rhopalidae, Ch. 9) which move pest or a beneficial species in the United States towards or into houses in large numbers in the may not be seen in the same way in other parts of autumn. Even the general public will find it easy the world. The editors are to be praised for allow- to read and use because it is devoid of much jar- ing different points of view. gon. Apparently, it has very few errors in spelling; What makes true bugs economically impor- the only one noticed was in the title of Ch. 8. It is tant? Many feed on plants, some of which trans- well organized and indexed. The book would have mit plant-pathogenic viruses. An important benefitted, though, by including some photographs example is Eurygaster integriceps Puton, a seri- of the most important pest or beneficial species. ous pest of wheat (Scutelleridae, Ch.14). Because This book is an important resource for health heteropterans feed in a unique way, with pierc- professionals since it contains information on oc- ing-sucking mouthparts, Hori (Ch. 2) explained, casional bites (Ch. 19) by non-blood-feeders, their stylets bypass many of the plants’ defenses which do not transmit pathogens, and on Chagas’ against biters and chewers, which also protects disease, of great medical and social importance in them from many pesticides. Some plant-feeding the Western Hemisphere, that is transmitted to bugs are helpful for the control of pest plants; for people by infected triatomine bugs (Reduviidae, example, Zulubius acaciaphagus Schaffner (Aly- Ch. 18). Cimicids (Ch. 17), or bed bugs, also feed didae, Ch.10) has helped to reduce the seed bank on human blood; studies on whether they can be of Acacia cyclops, a weed introduced from Austra- vectors for diseases such as hepatitis B, HIV, and lia to South Africa. yellow fever are reviewed.

Book Reviews 401

Entomologists and biological control special- book of the same style will be written. As a worker ists will find this book useful, but for heteropter- on Lygaeoidea, this book helps me to focus my in- ists, it is an essential book. The writers state the terests, guides my research and inspires me to strengths of previous and current research and write. I highly recommend this book. recommend research goals for the future. They J. Brambila emphasize the value that basic biological infor- Florida State Collection mation has with respect to applicable work for the of Arthropods control of heteropteran pests and the use of bene- Division of Plant Industry ficial species. Also, the value that good systematic Florida Department of Agriculture research has with respect to identification is dis- and Consumer Services cussed by the authors. Not all heteropteran fami- 1911 S.W. 34 Street lies are covered, for which I hope that a second Gainesville, FL 32608

402 Florida Entomologist 85(2) June 2002

HOWARD, F. W., D. MOORE, R. M. GIBLIN-DAVIS, AND R. G. ABAD. 2001. Insects on palms. CABI Pub- lishing; Wallingford, Oxon, UK, xiv + 400 pp. ISBN 0-85199-326-5. Hardback. $120.00. [Sales in the USA handled by Oxford Univ. Press, New York]

This book seems to be the first on the subject I have few criticisms. The book (p. 3) claims and thus fills a void. It deals extensively and in- that insect biogeography “lacks standardized tensively with the phytophagous insects and mites place-names.” If by this is meant that insect bio- that feed on or in palms, and with their natural en- geography uses names for biogeographical re- emies including biological control agents. As such, gions that have evolved from some proposed in it serves as a textbook for anyone wishing to pro- the mid-19th century, that is true. But, in making tect palms from damage by insects and mites, or this claim, the book equates the outmoded expres- from insect-transmitted diseases, or to encourage sion “Ethiopian Region” with all of Africa, when pollination by insects. But, it is more than that be- for over two decades sub-Saharan Africa with cause it reviews the behavior and ecology of the in- Madagascar has been called the “Afrotropical Re- sect and mite fauna of palms and makes the gion” (Crosskey & White 1977). The northern tier literature available to anyone interested in the ac- of African countries has long been attributed to ademic subject of insect/plant relationships. the Palearctic Region. It asserts (p. 94) that pres- It is not a multi-authored book with four edi- ence of a mite, Pyemotes ventricosus, detected in tors, but a book written by four authors. It is sub- Fiji in 1921, was due to “inadvertent introduc- divided into only eight chapters; this was possible tion.” This makes three assumptions: 1, that the only through good integration. F. W. Howard is mite is not native so must be adventive; 2, that it the major contributor to it. His opening chapter is was not introduced (deliberately) so it must be an “The animal class Insecta and the plant family immigrant; and 3, that it did not arrive by natural Palmae” which has a useful 29-page condensation means so must have arrived as a hitchhiker in for the uninitiated (e.g., most entomologists) some sort of cargo transported by humans. Evi- about the biology, cultivation and use of palms. dence for those assumptions is not presented. The There are four pages of introduction for the unini- weevil Metamasius mosieri, which is stated (p. 279) tiated about insects. Detailed information about to be associated with palms in tropical America, the insects is, of course, in the other chapters. in fact feeds on bromeliads as an adult and larva The other chapters are: 2, Defoliators of palms; (Larson et al. 2001). Anyone who expects to use 3, Sap-feeders on palms; 4, Insects of palm flow- the book as a source of describer (author) names ers and fruits; 5, Borers of palms; 6, Population for the scientific names of palms and their associ- regulation of palm pests; 7, Principles of insect ated insects and mites will be disappointed—they pest control on palms; and 8, Field techniques for are not given. The book’s rather high price ($120) studies of palm insects. The chapters are followed will surely deter some purchasers who are inter- by a 47-page section of integrated References, and ested but uncommitted. But, it is carefully edited this by a 20-page integrated Index. Tables in the and largely free of the typographical and gram- text summarize a lot of information usefully. Ta- matical errors, and bloated bureaucratese expres- ble 3.3 is a list of species of Derbidae (Hemiptera: sions that are all too common in many technical Auchenorrhyncha) reported on palms, with host books. This makes it a pleasure to read. palms, distributions and literature references. J. H. Frank Table 4.4 lists, by palm genus, arthropods associ- Entomology & Nematology Dept. ated with pollination of palms. It summarizes ar- University of Florida thropod behavior, but how can it write “larvae Gainesville, FL 32611-0630 probably breed in the flowers” when breeding is a function restricted to adults? Boxes in the text de- velop items of especial interest that would other- REFERENCES CITED wise destroy the flow, such as Box 2.2, the story of classical biological control of a coconut leaf-miner CROSSKEY, R. W., AND G. B. WHITE. 1977. The Afrotropi- in Fiji, and Box 5.2, an account of red-ring dis- cal Region. A recommended term in zoogeography. ease. The book’s numerous black and white photo- J. Nat. Hist. 11: 541-544. graphs, including electron micrographs, serve LARSON, B. C., J. H. FRANK, AND O. R. CREEL. 2001. admirably. It even has 16 plates of color photo- Florida bromeliad weevil. (Available: http://creatures. graphs, most of which have been reproduced well. ifas.ufl.edu/orn/M_mosieri.htm)

Book Reviews 403

MARCONDES, C. B. 2001. Entomologia médica e veterinária. Atheneu; São Paulo, Brazil, xvi + 432 pp. Paperback, 21 × 28 cm (8.25 × 11 inches). ISBN 85-7379-319-8. Reais 78.00 (US$38.50 by mail from the publisher, Editora Atheneu: [email protected]).

Reviews of three related books were published Most of the illustrations are black-and-white in Florida Entomologist in 2000 (volume 83). The drawings. Some are black-and-white photo- books were by Goddard 2000, Infectious diseases graphs. About a dozen of the photographs, mainly and arthropods (83: 384), Méndez 1999, Insectos of objects shown in transmitted-light microscopy, y otros artrópodos de importancia médica y veter- such as fleas and lice and cimicids, are rendered inaria (83: 503-504), and Service 2000, Medical in color, scattered on the text pages which are all entomology for students (83: 384-386). This new of the same matte paper stock. Most of those pho- book by Carlos Brisola Marcondes differs from tographs would have been clear enough in black- the others in three main aspects. First, it is of and-white. The chapters are followed by a 32- larger format and also has more pages, so con- page combined bibliography, a 4-page glossary, tains much more information. Second, its infor- and a 28-page combined index. The references in mation is very nearly restricted to hematopha- the text are numerical, so they waste no space in gous (blood-sucking) arthropods, with exception referring to the bibliography. only by inclusion of some non-biting flies and The principal author’s introduction (p. xiii-xiv) mites. Third, it is in Portuguese and focuses on states that this is the first general book on arthro- Brazil, with relevance to other parts of South pods of medical importance with especial refer- America. ence to Brazil. The preface (p. xi) by David Pereira The core chapters are 3, flebotomíneos—phle- Neves states that it is especially important for botomine psychodids; 4, simulídeos—simuliids; 5, students and health professionals. If it is designed ceratopogonídeos—ceratopogonids; 6, culicídeos— for students, then its objective must be to present, culicids; 7, tabanídeos—tabanids; 8, moscas— at moderate cost, which it does, the information Musca, Muscina, Stomoxys, Haematobia, Derma- that Brazilian teachers of medical and veterinary tobia, Oestrus, Cochliomyia, Gasterophilus, Wohl- entomology deem is most important for their stu- fahrtia, Hypoderma, and other flies; 9, pulgas— dents to learn. If it is designed for health profes- fleas; 10, piolhos—lice (mammalian- and avian- sionals, then it may be more comprehensive, to lice, Phthiraptera, formerly Anoplura and Mallo- serve as a reference for all aspects of medical and phaga); 11, hemípteros—triatomine reduviids veterinary entomology, and may sell for a higher and cimicids; and 12, ácaros—ticks and mites. price. That it has much more information, per- Each of these chapters includes information on haps twice as much, as do the earlier-reviewed classification, distribution, structure, behavior, books [paragraph 1 above, of which one (Service life cycle, methods for rearing, trapping, and con- 2000) was designed expressly for students] but trol, and brief information on the disease organ- yet is not fully comprehensive suggests that objec- isms transmitted to people and other vertebrates. tives were not initially fully defined. It is not fully Some chapters were written by contributing au- comprehensive because it does not deal with thors: 4 by Márcia Itiberê da Cunha, 6 by Ana stinging and otherwise toxic non-hematophagous Leuch Lozovei, 7 by Rosângela Bassi, 9 and 10 by arthropods of medical and veterinary importance Pedro Marcos Linardi, and 12 by Nicolau Maués [compare with Méndez (1999) cited in paragraph Serra-Freire. The most remarkable chapters are 1 above]. It does not include medical and veteri- 10 (lice, 56 pages) and 12 (ticks and mites, 53 nary methods for diagnosis and treatment of dis- pages) which expand the information presented eases [compare with Goddard (1999) cited in to a much more detailed level than in the three paragraph 1 above], although such subjects fall books mentioned in the first paragraph above; outside the scope of entomology. Perhaps some ad- each is longer than the chapter on mosquitoes. venturous publisher will see the need for compan- The ancillary chapters are 1, a 6-page intro- ion books: (a) medical and veterinary entomology, duction to morphology and physiology of hema- (b) diagnosis and treatment of diseases caused or tophagous arthropods; 2, a 6-page introduction to transmitted to vertebrate animals by arthropods, hematophagy; 13, a 10-page review of methods of and (c) diagnosis and treatment of diseases collection and preparation; 14, a 16-page review caused or transmitted to humans by arthropods. of the principles of control of these arthropods; 15, This new book is a very worthy first entry, which a 16-page summary of species concepts, phyloge- should serve for many years into the future. It will netics, and cytogenetic methods for distinguish- be interesting to see what changes the author de- ing species; and 16, a 10-page account of remote cides to make at its second edition. sensing for monitoring populations of pest in- J. H. Frank sects. Chapters 15 (by Jean-Pierre Dujardin) and Entomology & Nematology Dept. 16 (by Raquel Gleiser) were translated into Portu- University of Florida guese from Spanish. Gainesville, FL 32611-0630

404 Florida Entomologist 85(2) June 2002

CASTNER, J. L. 2000. Photographic atlas of entomology and guide to insect identification. Feline Press; Gainesville, FL. xii + 172 p. ISBN 0-9625150-4-3. Spiral binding. $35.00.

Insects comprise the greatest component of an- Thirty insect orders and over 175 families are imals on earth. To taxonomically (i.e., keys and/or treated in this book, including groups in a few descriptions) and photographically cover the vast related arthropod classes, namely Chilopoda, array of species in one publication would indeed Diplopoda, Crustacea, and Arachnida. Easy to be a daunting task, and virtually no publication read text of key diagnostic characters accompa- has ever done so. There have, however, been nu- nies 670 color images of insects, related groups, merous publications that cover what is a fraction and important anatomical features. Specimens of the sum total of insect species, each doing so in have been photographed alive and/or on pins, and varying degrees of detail and format. Lepidop- in most images a neutral background of gray has tera, especially butterflies, and some Coleoptera been inserted to allow subjects to stand out. In- groups, often get enough attention to be covered sect taxa are arranged roughly in phylogenetic or- by detailed illustration or photograph for identifi- der, and each section begins with a list of cation purposes, often to the great appreciation of important characters for the order, and in most students taking insect identification courses. orders coverage includes profiles of the more com- Other groups, especially groups of little economic monly encountered families. Characters for each importance, do not receive such attention. A family are listed and corresponding photographs plethora of entomological jargon must be under- are included, most of which are a common repre- stood before one can follow keys or descriptions sentative species in the family. In some instances, and accurately identify an insect to any level, and detailed photographs of important anatomical this challenges instructors restricted by time features are included. A key to the commonly en- when training students. Students can get caught countered families is also included for major in- up in the quagmire of terms presented in the lit- sect orders, and the taxa covered in the guide erature, often becoming skeptical that keys or de- generally represent groups common throughout scriptions are useful, and begin to doubt their the United States. Spiral binding allows the book ability to identify the insect under the microscope to lie flat while users are working with the guide; before them. This can ultimately lead to a general something larger books have difficulty doing turn-off to the discipline of insect taxonomy and without the binding eventually breaking. further taxonomic training in academic institu- Despite bringing together over 25 years worth tions, a trend that cannot persist at the student of images to compile this photographic atlas of in- level (or departmental level) if we are to continue sects, justly representing the millions of subjects to accurately identify and classify this extremely that comprise this extremely diverse group photo- diverse and important group of organisms. graphically is an enormous task. To cover enough Illustrations or photographs of insect anatomy forms for even an introductory course in insect or insects themselves that accompany challeng- identification is difficult. Major insect orders, ing keys or long descriptions aid not only students namely Coleoptera, Diptera, Lepidoptera, and but instructors in guiding them through keys or Hymenoptera, fall well short in photographic cov- helping them comprehend the characters pre- erage. With scores of known families in each of sented in descriptions. Accurate, and equally im- these major orders, this atlas treats 24, 20, 17, portant, enjoyable insect identification should be and 21 families respectively. There are only a few the goal of any introductory level taxonomic text, representative species pictured for each family, and James Castner’s Photographic Atlas of Ento- and users will not be able to readily identify fam- mology and Guide to Insect Identification at- ilies diverse in form by using the photographs tempts to do just that. alone. In addition, the keys included in each sec- Introductory sections include brief discussions tion treat the same number of families, which on the use of dichotomous keys and information may confuse users when they turn up one of the on taxonomic terminology, classification, and no- numerous families not covered in the key or fam- menclature. Sections on external insect anatomy ily profiles. A key to insect orders is also lacking, and insect development precede treatments of forcing students to match unknown specimens to insect orders and major classes of arthropods. a photograph for order determination. In other Photographs of several important anatomical lesser orders, family treatment is omitted alto- features and developmental stages are here gether, namely in groups such as Plecoptera, Pso- included. Most of the terms and photographs are coptera, and Trichoptera. devoted to adult insects, but some coverage of Distributional, life history, and in- immature forms is included. Castner also refers formation for families or orders is also lacking, al- the user to a few introductory entomology text- though it can help the student better identify the books and resources for courses devoted to insect specimen in question. Families that occupy one or a identification, and a glossary of terms found few types of habitat may be easier to separate from throughout the guide is also included. others that do not occupy those areas; moreover

Book Reviews 405 students who seek these families for collections dating at first, complete keys benefit students (which are often required in insect identification seeking a good taxonomic basis. This photographic courses) may not know where to look. The user will atlas, however, is indeed on the right track to cov- also find some of the higher classification out of ering the identification of forms in this enormous date, including placing Collembola in Insecta and group of organisms in an easy, enjoyable fash- the use of Cryptocerata and Gymnocerata as sub- ion—notions that are often forgotten when one is orders of Hemiptera and Raphidiodea and Mega- introducing students to the challenging world of loptera as suborders of Neuroptera. insect taxonomy. Note: paper quality enables Castner’s Photographic Atlas is a good supple- pages and print to hold up well to water or alcohol mentary text to introductory college level ento- (ethyl or isopropyl) spills if attended to in a timely mology courses in taxonomy, but it cannot replace manner—another challenge novice taxonomists traditionally used texts such as Borror, Triple- may face! horn, and Johnson’s An Introduction to the Study J. C. Dunford and L. S. Long of Insects. While the photographs are of high qual- Entomology and Nematology Dept. ity, some detailed characters are better recog- University of Florida nized though line drawings, and although intimi- Gainesville, FL 32611-0630

406 Florida Entomologist 85(2) June 2002

CAPINERA, J. L., C. W. SCHERER, AND J. M. SQUITIER. 2001. Grasshoppers of Florida. University Press of Florida; Gainesville. 143 p. ISBN 0-8130-2426-9. Paper. $34.95.

Regional insect identification guides can be very directly to that page, rather than flipping through useful and I have several in my personal library. the book or locating the page in the index. Thirdly, They vary in their completeness and level of sci- when referring to a particular anatomical feature entific information as well as ease of use. It is dif- in the key, such as shape of the cerci, it would ficult to write an insect identification guide that have been useful to have a reference in the key to contains accurate descriptions, is complete for the the figure number illustrating that feature. geographical scope of the work, and is also use- The introduction of this book is a wealth of in- able by the amateur entomologist as well as a spe- formation, concisely written, on the anatomy, life cialist. Capinera et al., however, have written history, ecological significance, pest status, cir- such a book. cumstances that cause outbreaks, and habitats of The goal of this book is to illustrate and describe grasshoppers. For example, figures 5-7 illustrate the 70 species of grasshoppers known to occur in the importance of grasshoppers in ecosystems by Florida of which 25% (18 species) are found only in showing the composition of the diets (including Florida and no where else in the world! To aid in grasshoppers and other invertebrates) of three identification this book has a bracket key that sep- bird species. Even a bird with primarily a vege- arates species or groups of similar species, excel- tarian diet as adults, such as the chipping spar- lent color photographs (taken by the senior author) row, utilize insects almost exclusively for nestling of the adults of 61 of the 70 Florida grasshoppers, food. One criticism of this figure is that the color as well as informative descriptions, pointing out of the proportion of a group in a diet (i.e. blue for the distinguishing characteristics. Also included grasshoppers in figure 5) should have been pre- are distribution maps for not only Florida, but in- served for figures 6 and 7 (where grasshoppers clusive of the known range of each species. The pho- are indicated by orange). A section on the large tos are particularly noteworthy because they are proportion of endemic species in Florida has an not from pinned specimens, but rather the insect interesting discussion on how this may have oc- has been photographed on leaf material or flowers curred including the fact that habitats have been in a natural resting pose. Also, care was taken to in- fragmented into “biological islands” with fluctua- dicate the sex of the individual in the photo (except tions of the ocean levels. On page 12 there is a list for the Ridgebacked sand grasshopper, p.72 and the of the 18 species unique to Florida as well as those Glassywinged toothpick grasshopper, p. 125). In whose range is mostly restricted to Florida. For species that are sexually dimorphic, photos of both me, the inclusion of the page number for the spe- sexes are included. For the banded-winged grass- cies description and/or photo next to the species hoppers, a photo is included of a pinned specimen name would have been handy. with the hind wing spread to show the color pattern The habitat photos nicely illustrate where used in identification. grasshoppers may be found. The descriptions of I used the key to try to identify some of the these areas as well as their suitability to support grasshopper species from our student teaching a large diversity of grasshopper species or not is a collection. I found the characters used obvious very useful addition. The section on collecting and and easy to see with the unaided eye or with a preserving grasshoppers also contains informa- hand lens. The illustrations in the introduction on tion on how to make some of your own collecting grasshopper anatomy clearly describe and label equipment (killing jar, spreading board). A list of the features used in the key and in the subse- suppliers where one could purchase this equip- quent written descriptions. Additionally, there is ment would have been useful. a glossary in the back for definitions of anatomi- The price of this book may be a deterrent to the cal and biological terms. Several things would average insect enthusiast. I think it is worth the have made this key combined with the photos and cost. The authors’ enthusiasm for grasshoppers descriptions even more useful. First, there is no and knowledge of the species is imparted in the discussion on how to use the key. Although I am writing. It will make even the casual reader ex- familiar with this type of key structure, and knew cited about observing these interesting insects. “where to go”, I don’t think all the potential users Wendy L. Meyer of this book would intuitively know this. Secondly, Tropical Research and Education Center it would have been nice to have the page referring University of Florida/ IFAS to the photo and species description next to the 18905 SW 280th St. species name in the key so the user could go Homestead, FL 33031

Sustaining Members 407

2002 FLORIDA ENTOMOLOGICAL SOCIETY SUSTAINING MEMBERS

Aventis Crop Florahome Pest Control, Inc. Attn: Eddie Ingram Attn: Richard R. Stout 1209 Hickory Lane 2689 Sunset Point Rd. Auburn, AL 36830 Clearwater, FL 34619-1500

Barcinas, Joe Florida Sugar Cane League 16120 Kraneria Ave. Attn: John W. Dunckelman Riverside, CA 92504 P.O. Box 1208 Clewiston, FL 33440 Bayer Corp. Attn: John Paige FMC Corporation 1614 Yorksire Trail Attn: Geri Cashion Lakeland, FL 33809 2948 Landmark Way Palm Harbor, FL 34684 Bayer Corp. Attn: Roy Morris II Foothill Argicultural Research 5690 58th Ave. Harry Griffiths Vero Beach, FL 32967 550 Foothill Pkwy. Corona, CA 92882 Best Termite & Pest Control Inc. Attn: Frank A Mongiovi Helena Chemical Company 8120 N. Armenia Ave. Attn: Bill Salley Tampa, FL 33604 P.O. Box 5115 Tampa, FL 33675 Cypress Sales & Marketing, Inc. Attn: Raymond J. Meyers, Jr. Lemont Entomology Services 630 Brookfield Loop Attn: Byron C. Lemont Lake Mary, FL 32746 2535 NW 182 St. Newberry, FL 32669 Dow Agrosciences Attn: Dr. Ellen Thoms Massimino, John 3225 S. MacDill Ave. #129-258 1561 Dorset Dr. Tampa, FL 33629-8171 Mount Dora, FL 32757

Dow Agrosciences Monsanto Attn: Dr. Joseph Eger Attn: Clair G. Erickson 4880 Bay Heron Pl. P.O. Box 7 Tampa, FL 33616 Tangerine, FL 32777

Dupont Ag. Products Rhone Poulenc Ag. Co. Attn: Bob Williams Attn: J. Malone Rosemond 10918 Bullrush Terrace 2812 Park Ave. Bradenton, FL 34202 Tifton, GA 31794

Eden Bio Science Syngentia Crop Pro. Attn: Henry Yonce Attn: J. Scott Ferguson 1092 Glenwood Trail 7145 58th Ave. Deland, FL 32720-2130 Vero Beach, FL 32967

E. O. Painter Printing Company T&L Consulting Service Attn: S. Dick Johnston Attn: Vern E. Toblan P.O. Box 877 161 Chrone Road DeLeon Springs, FL 32130 Nottingham, PA 19362

408 Florida Entomologist 85(2) June 2002

Taylor Pest Management Valent USA Corp. Attn: James B. Taylor Attn: John Altom 851 N. E. Jensen Beach Blvd. 3700 NW 91 St. Bldg. C, Suite 300 Jensen Beach, FL 34957 Gainesville, FL 32606

Terminex International Walt Disney World Attn: Norman Goldenberg Attn: Jerry A Hagedorn 860 Ridge Lane Blvd. P.O. Box 10000 Memphis, TN 38120-761 Lake Buena Vista, FL 32830

The Scotts Co. Wright Pest Control Attn: Wayne Mixon Attn: M. L. Wright P.O. Box 2187 P.O. Box 2185 Apopka, FL 32704 Winter Haven, FL 33880

Thermo Triology Corp. Yoder Brothers Attn: Adam Muckenfuss Attn: Nancy Recheigl 328 SE Ridge Ln. 11601 Erie Rd. Stuart, FL 34994 Parrish, FL 34219

Uniroyal Chemical Attn: Keith H. Griffith 5211 Fawnway Ct. Orlando, FL 32819-3823

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