Mellor et al., 2009). This species, first described by Kieffer (1910) from Puri, has since been reported in various parts of India (Prasad et al., 2009). The pupa was described by Mayer (1934) from Sumatra, whereas the egg, larva, and pupa were reported by Narladkar et al. (1991) without detailed taxonomy and illustrations based on recent terminologies. Based on adult morphology, Nandi (2014) revised the genus and recognized 11 species under the subgenus Hoffmania, including C. peregrinus. Light trap data revealing a high prevalence of this species in proximity to cattle (unpublished data) prompted us to describe all immature stages with modern terminologies based on scanning electron microscopy (SEM). This taxonomic information about the immature stages will help to precisely determine the oviposition sites, to specify larval habitats and the differentiation of larval instars, and ultimately to adopt effective control strategies against this vector species. This paper describes the ultrastructure of the egg and larval instars and offers a redescription of the pupa. Immature specimens of this species are presently recognized by the random arrangement of tubercles in two broad dorsoventral longitudinal rows of egg chorion, epipharynx combs 2 and 3 absent in larvae, and weak sclerotized band on the antennal sheath and presence of palpus tubercle in pupae.
Materials and methodsWild adult females of Culicoides peregrinus were collected by operating a downdraft UV light trap (Harsha and Mazumdar, 2015a) within a cattle shed at Adisaptagram (22°56′50″N, 88°23′08.3″E), Hooghly district, West Bengal, India. The live engorged females oviposited after 48 h onto a wet filter paper within a glass vial (6.5 × 2 cm). Thereafter, the embryonated eggs (confirmed by the presence of eye spots) were reared in two different rearing media. Some eggs were placed in petri dishes, with absorbent cotton provided as a substrate and mud broth + 2% yeast solution provided as food in an environmental test chamber at 26 °C (model CHM-10S; REMI Elektrotechnik Limited, Vasai, India), following Harsha and Mazumdar (2015b). Similarly, cattle manure (natural breeding habitat) was bought to the laboratory and a few larvae and pupae were hand-picked from the cattle manure and were subsequently treated for description, while the rest of the cattle manure was kept for the adult emergence. A few of the embryonated eggs were processed for SEM after Day et al. (1997); for differentiation of instars, 6-7 larvae were extracted every day from the rearing dishes and mounted on slides following Das Gupta and Wirth (1968). The different larval instars were differentiated based on measurements and several ratios of the head capsule. Eggs, larvae, and pupae were also studied under a scanning electron microscope (model S5 30; Hitachi, Tokyo, Japan) at the Science Instrumentation Centre at the University of Burdwan. Similarly, larvae and pupae were Abstract: Detailed descriptions of the immature stages, i.e. eggs, larvae, and pupae, of Culicoides peregrinus Kieff...