ABSTRACT
Identification of filarial species in dogs is clinically important because of zoonotic concerns and therapeutic implications. The present study was carried out to identify the filarial parasites causing microfilaraemia in dogs in Thrissur District, Kerala- an endemic area for human Brugian filariasis. Out of the 1600 dogs screened by wet blood film examination, 130 were positive for microfilariasis. Giemsa staining of blood smears revealed that 90 out of 130 dogs had unsheathed microfilariae, 24 had sheathed microfilariae and 16 had combined infection of sheathed and unsheathed microfilariae. Results of micrometry and histochemical staining of the sheathed microfilariae were in conformity with that of Brugia malayi. The DNA isolated from the sheathed microfilariae amplified the primers specific for the Hha 1 repeats of the B. malayi. Cloning and sequencing revealed that the amplified fragment corresponded to the 140-292 base pairs of the 320 base pair Hha1 repeat of Brugia malayi. The amplified DNA fragment also contained restriction sites for Alu 1 and Rsa 1which confirmed that the present isolate is Brugia malayi. The present study confirmed the presence of B. malayi in dogs in Kerala, India.
ABSTRACT
Brugia malayi is a filarial parasite that causes lymphatic filariasis in human beings. Kerala (India) is endemic for human Brugian filariasis. B. malayi microfilariae, similar to that causing filariasis in human beings, have been identified in dogs in Kerala. This is the first report of western blotting analysis of canine B. malayi microfilarial proteins. SDS PAGE analysis of B. malayi microfilarial protein revealed 5 major protein bands with molecular weights of 125, 80, 64, 54 and 27 kDa. Among these, the prominent bands were those having molecular weights of 64, 54 and 27 kDa. We raised polyclonal antibodies against these somatic proteins of dog microfilariae in a rabbit. The polyclonal antibodies recognized predominantly the 54 kDa and 64 kDa antigens in a Western blot analysis. Based on previous publications with B. malayi, these two protein bands appear to be important for diagnosis and for vaccine development against lymphatic filariasis.
ABSTRACT
This study was conducted to identify the aetiological agents associated with a particular type of lower leg dermatitis, locally called pododermatitis, among dairy cattle in Kerala. Skin scabs and scrapings were collected aseptically from 82 naturally occurring cases of lower leg dermatitis in cattle and were subjected to direct microscopical examination and bacterial and fungal culture. Microscopical examination of the skin scrapings with 10% potassium hydroxide revealed fungal spores in hair shafts from only two samples and did not reveal the presence of mites or other parasites. Fungal culture yielded dermatophytes from only five samples; these were identified as Trichophyton mentagrophytes in two cases, T verrucosum in one case, Epidermophyton floccosum in one case and Microsporum nanum in one case. Microscopical examination of Giemsa- and Gram-stained smears of the scab material from the lesions from 72 cases revealed characteristic Gram-positive septate branching filaments with multiple rows of spherical to ovoid cocci, with a typical 'tram-track' appearance suggestive of Dermatophilus congolensis. Culture of the scab materials on sheep blood agar in the presence of 10% carbon dioxide yielded typical beta haemolytic colonies of D. congolensis from 75 samples. The isolates were further confirmed by the macroscopic and microscopic morphology of the colonies, and biochemical test results. This study confirmed the presence of dermatophilosis caused by D. congolensis in cattle in Kerala.
