Holotrichia longipennis Blanch. (Coleoptera: Scarabaeidae) is a serious pest of commercial crops in Siduwa, Dhankuta, Nepal. Seven indigenous isolates of entomopathogenic nematodes (Steinernema lamjungense LMT5, S. lamjungense SS4, S. everestense DKP4, S. abbasi CS1, S. sp. KL1, Heterorhabditis indica CK2 and H. indica CK6) were used in a series of bioassays against the insect. All isolates showed an increased dispersal in response to H. longipennis. Nematodes were more attracted towards third instar larvae than to second instars. Differences in penetration and multiplication in the insect were observed amongst the seven isolates. Steinernema lamjungense LMT5, S. everestense DKP4 and S. abbasi CS1 caused greater mortality than other isolates to different developmental stages. Pupae and second instar larvae were more susceptible than third instar larvae. Significant differences were observed in LT 50 values of the isolates against different stages of H. longipennis. Three isolates (S. lamjungense LMT5, S. everestense DKP4 and S. abbasi CS1) along with a commonly used insecticide (chlorpyrifos) were tested against this insect in pot and field experiments. In pot experiments using maize and cabbage as a host crop, S. lamjungense LMT5 and S. everestense DKP4 performed better than S. abbasi CS1 and yielded a mortality comparable with chlorpyrifos. Similar results were observed in field experiments 3 weeks after nematode application. These experiments overall suggest S. lamjungense LMT5 to be a promising biocontrol agent against H. longipennis followed by S. everestense DKP4 and S. abbasi CS1.
A new species of entomopathogenic nematode, Steinernema lamjungense n. sp., was recovered from soil samples collected from Tarku, Lamjung district of Nepal, during a survey in 2007. The analysis of ITS-rDNA sequence placed S. lamjungense n. sp. in the arenarium-glatseri-karii-longicaudum group. The nematode can be separated from other described species of the group by morphological and morphometric characteristics of different stages and by characterisations and phylogeny of DNA sequences of the D2D3 domain of the LSU or ITS regions of rDNA. For infective juveniles, the new species can be recognised by medium-long body length of 832 (690-950) mu m, pharynx length of 127 (110-142) mu m, H% = 61(55-69) and E% = 79 (69-97). The lateral field pattern is 2, 6, 7, 8, 3, 2. The males have well curved, yellowish-brown spicules with a prominent velum. The spicule terminus is blunt with a longitudinal ventral notch. Spicule length is 87 (81-94) mu m and gubernaculum length is 57 (50-66) mu m in the first generation males. A tail mucron is absent in first but present in second generation males. First generation males have a lateral field with a single ridge. A post-anal swelling is not developed in first generation females but is well developed in second generation females. Three other isolates of the same species were recovered from the neighbourhood of the type locality and yet another isolate was collected from Swarketari, Syangja district
A new species of entomopathogenic nematode, Steinernema everestense n. sp., was recovered from soil samples collected from Pakhribas, Dhankuta district of Nepal, during a survey in 2007. The analysis of ITS-rDNA and D2D3 LSU sequences placed S. everestense n. sp. in the feltiae-kraussei-oregonense group. The nematode can be separated from other described species of Steinernema by morphological and morphometric characteristics and by characterisations and phylogeny inferred from DNA sequences of the D2D3 LSU or ITS regions of rDNA. For infective juveniles, the new species can be recognised by the body length of 775 (705-838) μm, pharynx 119 (110-130) μm long, H% = 61 (47-68) and E% = 78 (60-89). The lateral field pattern is 2, 8, 6, 4, 3. The males have well curved, brownish-yellow spicules with a prominent but short velum, gubernaculum plump, boat-shaped to almost straight in lateral view, cuneus long, needle-shaped, corpus with two wings in ventral view. Spicule and gubernaculum length of the first generation males is 79 (65-87) μm and 50 (39-57) μm, respectively. A tail mucron is present in both male generations. Genital papillae total 23 or 25, comprising 11 or 12 pairs and a single midventral papilla. Specimens with 23 genital papillae have eight precloacal pairs (seven pairs subventral and one pair lateral), two pairs subterminal and one pair subdorsal. The most posterior two pairs are consistently located near the tail tip. The specimens with 25 genital papillae have an extra adcloacal pair of papillae and the anteriormost two pairs are close together. The vulva is slightly protruding, mostly with equal lips, and low epiptygmata. A post-anal swelling is developed in most first and all second generation females.
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