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Subject: Veterinary Parasitology × Author: ARAVINDA, K. × Start date: 2006 × Type: Thesis ×
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    ThesisItemOpen Access
    MOLECULAR CHARACTERISATION AND IDENTIFICATION OF IMMUNOREACTIVE POLYPEPTIDES OF FILARID NEMATODES OF DOGS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCIES UNIVERSITY, BIDAR, 2019) ARAVINDA, K.; ANANDA, K. J.
    A study on the molecular characterization and identification of immunoreactive polypeptides of filarid nematodes of dogs in Mangalore, Karnataka state was conducted. Different methods were used for detection of microfilariosis in dogs. A total of 250 dogs screened during the present study, 62 % harboured microfilaria. In the present study, three methods used for detection of microfilariosis in dogs. Modified knott’s method detected 94.19 % of the positive samples, direct wet smear and giemsa’s stained blood smear examination detected 61.93 and 57.42 % positive samples. Morphological studies of microfilaria were done by using giemsa’s and methylene blue phosphate staining techniques revealed microfilariae were unsheathed with blunt head and a tapering tail and cuticle appeared striated. The length and width of microfilaria when measured by giemsa’s and modified knott’s technique were 292.2 ± 1.267 μm, 334.9 ± 4.573 μm, 10.65 ± 0.068 μm and 6.862 ± 0.133 μm respectively. The histochemical staining technique revealed acid phosphatase activity at anal pore region of microfilaria by showing red azo dye indicating the microfilaria were of D. repens. Confirmatory of identification of species was done by PCR using species specific primers showing amplification of mitochondrial cytochrome oxidase subunit 1 (COI) gene with single amplicon 350 bp indicated D. repens. The protein profile of microfilarial and adult worm antigen of D. repens was evaluated using SDS-PAGE. The protein profile of microfilarial antigen showed six polypeptides ranging between 183 kDa to 29 kDa. A total of thirteen polypeptides were identified in adult worm antigen ranging from 208 kDa to 29 kDa. The immunoreactive polypeptides detected on western blot with microfilarial and adult worm antigen of D. repens using known positive serum with anti-dog IgG conjugate at 1:1000 dilution revealed 50 kDa and 29 kDa. Key words: Molecular characterization, Immunoreactive polypeptides, filarid nematodes, dog, Mangalore.