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Rhabditida: Steinernematidae) Journal of Nematology 22(2): 187-199. 1990. © The Society of Nematologists 1990. Steinernema scapterisci n. sp. (Rhabditida: Steinernematidae) K. B. NGUYEN AND G. C. SMART, JR3 Abstract: Steinernema scapterisci n. sp., isolated in Uruguay from the mole cricket Scapteriseus vicinus, can be distinguished from other members in the genus by the presence of prominent cheilorhabdions, an elliptically shaped structure associated with the excretory duct, and a double-flapped epitygma in the first-generation female. The spicules of the male are pointed, tapering smoothly to a small terminus, and the shaft (calomus) is long, bearing a sheath. The gubernaculum has a long, upward- bent anterior part. The ratio of head to excretory pore divided by tail length of the third-stage juvenile is greater for S. scapterisci n. sp. than for S. carpocapsae. Steinernema scapterisci n. sp. did not hybridize with S. carpocapsae strain Breton. In laboratory tests, S. scapterisci n. sp. killed 10% or less of non-orthopteran insects, including the wax moth larva, a universal host for other species of Steinernema. Key words: biological control, entomopathogenic nematode, mole cricket parasite, morphology, new species, Neoaplectana, Scapteriscus, Steinernema scapterisci n. sp., taxonomy, light microscopy (LM), scanning electron microscopy (SEM). In 1985, several isolates of a steinerne- MATERIALS AND METHODS matid nematode collected from mole crick- Nematodes collected in Uruguay were ets in Uruguay were brought to Florida placed on moist filter paper in several vials, and cultured. The morphology and biol- a live mole cricket (Scapteriscus vicinus) was ogy of the nematode showed that it is a placed in each vial, and the vials were placed new species ofSteinernema Travassos, 1927. in an insulated container with ice and hand The genus was erected by Steinernema carried to the quarantine facility in Gaines- Travassos in 1927 (6) for the species Aplec- ville, Florida. The populations of the which Steiner (3) had de- tana kraussei nematode were increased in mole crickets scribed in 1923 from a sawfly (Cephaleia (S. vicinus and S. acletus) and later in the abietis). Steiner (4) established the genus house cricket (Acheta domesticus). These Neoaplectana in 1929 for N. glaseri which nematodes, or their progeny, were used for he described from the Japanese beetle (Po- all studies. pillia japonica). Light microscopy (LM): First-generation Wouts et al. (7) considered Neoaplectana and second-generation adult nematodes a junior synonym of Steinernema. We con- were obtained by dissecting infected mole cur with these authors and describe our crickets 2-3 and 5-7 days, respectively, af- nematode herein as Steinernema scapterisci ter they died; J3 were obtained when they n. sp., named after its host, Scapteriscus (Or- emerged from the cadavers in 7-15 days. thoptera), a genus containing mole crick- The nematodes were killed and relaxed in ets. warm water (40 C), then mounted in water on glass slides with coverglass supports. In addition, live specimens or those killed and Received for publication 3 April 1989. stained with acid fuchsin were examined 1 Florida Agricultural Experiment Station Journal Series No. 9846. A portion of a dissertation submitted by the first to confirm the presence and (or) detail of author to the University of Florida in partial fulfillment of some structures. the requirements for the Ph.D. degree. Former Graduate Student (now Biological Scientist III) Scanning electron microscopy (SEM): Speci- and Professor, Entomology and Nematology Department, In- mens prepared for SEM were placed live stitute of Food and Agricultural Sciences, University of Flor- ida, Gainesville, FL 32611-0611. in lactophenol at 43 C for 30 minutes, The first author has published previously under the names transferred to a desiccator for 2 days, re- Nguyen Ba Khuong and N. B. Khuong. The authors thank Drs. A. C. Tarjan and R. P. Esser for moved, rinsed with water, and prepared by suggestions. the method of Stone and Green (5). Spic- 187 188 Journal of Nematology, Volume 22, No. 2, April 1990 ules and gubernacula were obtained from specimens of each species were retained in first-generation males placed in a petri dish drops of blood in two dishes as controls. containing water, heat killed, and stored Comparativepathogenieity: Four species of at room temperature. After 2-3 days the insects in the order Lepidoptera--the fall bodies became softened by decay and were army worm (Spodopterafrugiperda), the vel- transferred to clean water. The posterior vet beam caterpillar (Anticarsia gemmatalis), of each nematode was opened with two the granulate cut-worm (Feltia subterrania), small needles, and the spicules and guber- and the greater wax-moth larva (GaUeria naculum were dissected out and washed meUoneUa)--were used to compare the rate free of debris by shaking them in water. of kill by S. scapterisci n. sp. to that of some Then they were picked up with a fine needle other species and strains of Steinernema. and placed on a SEM stub close to a hair Two pieces of Whatman No. 2 filter pa- used for a marker. Specimens were ex- per were placed in a petri dish (100 x 15 amined with a Hitachi $450 SEM. ram), and 8,000 J3 in 2 ml water and 10 Cross hybridization: Two techniques were insects were added. Controls were pre- used. In the first, a drop of blood (hemo- pared similarly, but without nematodes. lymph) from a mole cricket was placed in Treatments were replicated four times. a sterile petri dish (35 x 10 mm), and one The dishes were maintained in the dark at J3 of S. scapterisci n. sp. and one J3 of S. 25 C. After 2 days the number of dead carpocapsae strain Breton were added. The insects was determined. dish was placed in a plastic bag containing SYSTEMATICS a paper towel saturated with water, and the bag was closed, tied, and stored in the dark Steinernema scapterisci n. sp. at 25 C. The treatment was replicated 25 (Figs. 1-8) times. Holotype (male, first generation, in glycerine): In the second technique, a drop of blood Length 1,554 #m; width 131 #m; stoma was placed in each of two petri dishes and length 3.9 ~m; stoma width 6 ~m; head to 10 J3 of S. scapterisci n. sp. were placed in excretory pore 77 #m; head to nerve ring one drop and 10 J3 orS. carpocapsae strain 130 gm; head to end of esophagus 189 #m; Breton were placed in the other. The dish- testis from reflexion to terminus 416 #m; es were maintained as for the first tech- body width at anus 37.5 #m; tail length 28 nique. The treatment was replicated 10 ~m; spicule length 89 ~m; spicule width times for each species. The nematodes were 13.5 #m; gubernaculum length 65 urn; observed daily. When the sexes could be spicule width 13.5 #m; gubernaculum distinguished in the preadult stage, all males length 65 #m; gubernaculum width 7.8 #m; of S. scapterisci n. sp. were removed and mucron length 4.5 #m. placed in a new drop of blood and all males Male, first generation: Measurements of of S. carpocapsae strain Breton were placed 10 males in Table 1. Smaller than first- in a separate drop of blood. Then the males generation female. Body usually rotund, of S. scapterisci n. sp. were transferred to curved ventrally posteriorly (Fig. 4E), spi- the drop of blood containing females of S. ral or C shaped when heat-killed. Gonad carpocapsae strain Breton, and the males of monorchic, reflexed. Spicules paired, uni- S. carpocapsae strain Breton were trans- formly curved, dark brown in color; head ferred to the drop of blood containing fe- large somewhat angular (Figs. 3D; 4A, B; males ofS. scapterisci n. sp. The nematodes 5B); shaft and blade angle averages 110 were observed for mating, continuously the degrees (range 100-120); shaft appears en- first hour and then hourly for the remain- cased in a sheath (Fig. 5B); blade tapers der of the first day. On subsequent days smoothly, posterior portion thin (Figs. 4A, they were observed hourly for mating, egg 7A). In SEM cross section, spicule blade laying, or the presence of juveniles. Other has two lumina (Fig. 5C), only one aperture Steinernema scapterisci n. sp.: Nguyen, Smart 189 B A ", .... ' "4 A 20pro , ..-...,t.~.;,.;.,,. :, ~:",~.: .. C V.IP' 50 Pm == 120Pro " '~"'.; "': ~':. ~ ~- • '"~ O_ lOOPm Fig. 1. Females ofSteinernema scapterisci n. sp. A) Diagrammatic face view of first-generation female showing unevenly distributed papillae. B) Entire body of second-generation female. C) Double-flapped epiptygma on vulva of first-generation females. D) Variation in tails of first-generation females. E) Variation in tails of second-generation females. F) Anterior region of the first-generation female showing large cheilorhabdions, esophagus, nerve ring, excretory pore and duct, and elliptically shaped structure and gland cell associated with the excretory system. observed on ventral side and close to tip 5; pairs 6-7 postanal, subventral; pairs 8 (Fig. 7A). Each spicule has two internal ribs and 9 caudal, subventral; pair 10 caudal, (Figs. 4A, B; 8F), with termination points subdorsal. Pairs 1 and 6 obscure. Tail con- variable. Gubernaculum boat-shaped, an- oid, mucron present (Fig. 4B). terior part thin, ventrally curved, posterior Male, second generation: Measurements of end bifurcate (Fig. 5D). Cloacal area raised, 10 males in Table 1. Second-generation anterior flap present. Ten pairs and one male similar to that of the first generation single genital papillae present (Fig. 8A). except that its length is shorter, 1,147 t~m Pairs 1-5 preanal, subventral; single pa- vs. 1,728 ~m; and the width is about one- pilla preanal, ventral, between pairs 4 and half that of the first-generation male.
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