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Pathogenicity of Steinernema Scapterisci to Selected Invertebrates 1

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Pathogenicity of Steinernema Scapterisci to Selected Invertebrates 1 Journal of Nematology 23(1):7-11. 1991. © The Society of Nematologists 1991. Pathogenicity of Steinernema scapterisci to Selected Invertebrates 1 K. B. NGUYEN AND G. C. SMART, JR3 Abstract: Steinernema scapterisci was more pathogenic to insects tested in the order Orthoptera than to those in the orders Lepidoptera or Hymenoptera; it was not pathogenic to earthworms. The nematode also infected and killed the mole crickets Scapteriscus acletus and S. vicinus when released four successive times at 10-day intervals in containers of soil infested with the nematode. Key words: biological control, entomopathogenic nematode, mole cricket, Scapteriscus, Steinernema scapterisci. Three species of mole crickets, Scapteris- lates of the nematode from Uruguay to the cus abbreviatus (short-winged mole cricket), quarantine laboratory in Gainesville, Flor- S. acletus (southern mole cricket), and S. ida. We chose an isolate that, in initial vicinus (tawny mole cricket), were intro- pathogenicity tests in petri dishes, caused duced accidentally into Florida from South the highest mortality (38%) of S. vicinus America around the turn of the century and S. acletus. The virulence of the isolate (9). Currently, S. vicinus is present in the was increased by serial passage (i.e., re- coastal plain regions of South Carolina peatedly exposing infective-stage juveniles through Alabama; S. acletus is present in collected from infected mole crickets to the coastal plain regions of southern North other mole crickets) through the two mole Carolina through eastern Texas (8) and in cricket species. These nematodes were used Arizona (5); and S. abbreviatus is restricted in all experiments. to Puerto Rico (9) and to three coastal The purpose of the research reported regions in Florida and one in Georgia. here was to determine the pathogenicity The three species have become the most of S. scapterisci to a spectrum of insects and important insect pests of turf and pasture an earthworm and to determine whether grasses in Florida (7). They also damage uninfected mole crickets would become in- seedlings of agronomic and vegetable crops fected and die when released in containers and ornamentals. Losses, including costs to of soil infested with S. scapterisci. control mole crickets in Florida, are about $45 million annually (4). Therefore, a mole MATERIALS AND METHODS cricket research project was established at Pathogenicity tests: Test organisms were the University of Florida in 1978 with em- adults of Orthoptera: house cricket (Acheta phasis on biological control. A search for domesticus), American cockroach (Peripla- natural enemies has extended 'to South neta americana), field cricket (GryUus sp.), America, which appears to be the native and the mole crickets (S. abbreviatus, S. acle- homeland for these mole crickets (8). Mole tus, and S. vicinus); last-instar larvae of crickets infected with a steinernematid Lepidoptera: fall armyworm (Spodoptera nematode, recently described as Steinerne- frugiperda), granulate cutworm (Feltia sub- ma scapterisci (6), were found in Brazil and terranea), greater wax moth larva (GaUeria Uruguay. In 1985, we brought several iso- meUonella), and velvet bean caterpillar (An- ticarsia gemmatalis); adults of Hymenop- Received for publication 16 May 1989. tera: honeybee (Apis mellifera); and adults Florida Agricultural Experiment Station Journal Series No. 9910. A portion of a dissertation submitted by the first of Annelida: earthworm, Allolobophora ca- author to the Graduate School of the University of Florida liginosa. The organisms (except the hon- in partial fulfillment of the requirements for the degree of Doctor of Philosophy. eybee and earthworm) were tested in petri Former Graduate Student, now Biological Scientist III, dishes (100 x 15 mm) containing two piec- and Professor, Entomology and Nematology Department, In- stitute of Food and Agricultural Sciences, University of Flor- es of filter paper (Whatman #2, 90 mm d). ida, Gainesville, FL 32611-0611. Approximately 8,000 infective-stage ju- 8 Journal of Nematology, Volume 23, No. 1, January 1991 veniles of S. scapterisci in 2 ml water were times. The controls contained earthworms added to each dish. Nematodes were used but no nematodes. within 2 weeks after they were produced. Additionally, to determine whether the Not all organisms tested nor the quantities nematode can develop in severely dam- desired were available at the same time. aged earthworms, five earthworms were cut Therefore, the tests were not conducted in half and placed in cups ½ filled with simultaneously, and the number of test or- organic soil. Eight thousand infective ju- ganisms per treatment and the number of veniles in 2 ml water were added to each replicates varied as follows: treatments with cup, and the cups were shaken well to mix GryUus sp. and Feltia subterranea, 10 spec- the nematodes with the soil. The treatment imens per replicate and 5 replicates; S. acle- was replicated five times. After 5 days, the tus, S. vicinus, and S. frugiperda, 5 specimens earthworm segments were collected, dis- per replicate and 5 replicates; G. meUoneUa sected in water, and examined for nema- and A. domesticus, 10 specimens per repli- todes. Also, two earthworms were cut in cate and 10 replicates; A. gemmatalis, 6 half and placed in a petri dish (60 x 15 specimens per replicate and 5 replicates; ram) containing organic soil and 8,000 in- and S. abbreviatus, 4 specimens per repli- fective juveniles. The treatment was rep- cate and 3 replicates. Dishes were stored licated five times. The earthworm seg- in the dark at 25 C. Numbers of dead or- ments were dissected after 5 days and ganisms were counted after 3 days, except examined for nematodes. the earthworms which were examined af- Serial infection of mole crickets in soil: Scap- ter 5 and 10 days. Controls contained the teriscus vicinus was used in one experiment test organism but no nematodes. and S. acletus in another. In both experi- The honeybee test was conducted in cy- ments, containers (11.4-liter capacity) were lindrical screen wire cages (8.5 cm d x 20 filled with soil (97.9% sand, 1.8% clay, 0.3% cm high) containing 20 adult honeybees of silt) to within 5 cm of the top. There were different ages. About 8,000 infective-stage four treatments and 14 replicates per treat- juveniles in a water-saturated cotton ball ment for S. vicinus and four treatments and were placed in a petri dish which served as 10 replicates per treatment for S. acletus. the bottom of each cage. The saturated The four treatments were 1) one live cotton was a source of water for the bees nematode-infected mole cricket per con- and assured that they came in contact with tainer, 2) two live nematode-infected mole the nematodes when they landed on the crickets per container, 3) 12,000 infective- cotton. The controls were prepared in the stage juveniles sprayed on the soil surface same way but without nematodes. All cages per container, and 4) control with mole were placed in the dark at 25 C. The treat- crickets but no nematodes. After the treat- ment was replicated five times. Dead hon- ments were applied, 10 uninfected mole eybees were collected daily for 3 days. crickets were released in each container Dead organisms were placed individually and lids were applied to prevent the mole in vials (25 ml). Two days later, the cadav- crickets from escaping. After 10 days the ers were dissected; if they contained S. scap- live and dead mole crickets were counted, terisci which had developed beyond the and the live mole crickets were removed third stage, they were considered to have from the containers. The dead crickets been killed by the nematode. were dissected; and if the nematode was Pathogenicity tests against earthworms detected, they were assumed to have been were conducted in cups (237 ml) contain- killed by it and were returned immediately ing organic soil (composted oak leaves). to the original containers. On the same Approximately 8,000 infective juveniles day, 10 other uninfected mole crickets were and 10 earthworms were added to each released into each container. The process cup. The treatment was replicated five was repeated three times for a total of four Pathogenicity of S. scapterisci: Nguyen, Smart 9 TABLE 1. Mortality of organisms exposed to Steinernema scapterisci. Treated Control Tested Died Mortality]" Tested Died (n) (n) (%) (n) (n) Orthoptera Acheta d~mesticus 100 100 100 100 4 Gryllus spp. 50 11 22 50 0 Scapteriscus abbreviatus 12 10 75 12 4 Scapteriscus acletus 25 25 100 25 3 Scapteriscus vicinus 25 25 100 25 2 Periplaneta americana 50 2 4 50 0 Lepidoptera Anticarsia gemmatalis 30 1 3 30 0 Feltia subterranea 50 5 10 50 0 Galleria mellonella 100 9 9 100 0 Spodoptera frugiperda 25 2 8 25 0 Hymenoptera Apis meUifera 1O0 16 10 1O0 7 Annelida Allolobophora caliginosa 50 0 0 50 3 ]- Percentage of mortality corrected by Abbott's formula (1) which is (X - Y)/X × 100, where X = % insects alive in the controls; Y = % insects alive in the nematode treatment. releases per container at 10-day intervals. ders Lepidoptera and Hymenoptera; it was The containers were held at 25 C + 4 C. not pathogenic to earthworms (Table 1). In dead or injured earthworms, one adult RESULTS male and four adult female nematodes were Pathogenicity tests: Steinernema scapterisci dissected from the earthworm segments in exhibited a higher degree of pathogenicity the first experiment and none in the sec- to insects in the order Orthoptera (except ond experiment. Thus, although a few in- for the Cockroach) than to those in the or- fective juveniles developed to the adult stage in dead earthworms, they did not re- TABLE 2. Mole crickets Scapteriscus vicinus and S. produce. In both experiments, however, acletus killed by Steinernema scapterisci in each of four large numbers of diplogasterids and rhab- releases.t ditids developed on the bacteria associated with the earthworm segments.
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