Apply traditional and molecular protocols for the detection of carrier state of visceral leishmaniasis in black Bengal goat

Abstract

Visceral leishmaniasis (VL) is the second largest parasitic killer of human in the world after malaria which is responsible for an estimated 500,000 new cases of VL in each year with 8-10% mortality. The parasite is transmissible to humans and animals by the bite of phlebotomine sand fly. The clinical manifestations are highly diverse, humans and dogs are naturally infected, and the diseases are associated with several risk factors, yet to understand. The aims of this study were to apply traditional and molecular detection tools and more emphasis was given to identify goat as a carrier of visceral leishmaniasis. To demonstrate the promastigote and amastigote phases of Leishmania in tissues, traditional impression smear staining technique was used. For the confirmation of the species of Leishmania involved specific technique like polymerase chain reaction (PCR) was applied. A total of twenty goats were investigated and samples were tested using impression smear staining, histopathology and PCR. Blood smear and impression smears were prepared from spleen, liver, bone marrow and stained with Giemsa's stain. Using Giemsa's staining out of twenty goats investigated, six (6/20) were found to contain promastigote and amastigote stages of Leishmania in their visceral organs. Histopathological examination from the liver section of suspected goats showed degeneration, necrosis and non specific fibrous connective tissue proliferation compared to non-reactive goats. There was accumulation of macrophages in lymphoid follicles of spleen in five suspected goats. A highly sensitive and specific primers were used in PCR amplification with the extracted DNA from liver and spleen of suspected six goats. Results of PCR showed that two of them were generated 145bp amplicon selective for L. donovani in their liver and spleen. Leishmaniasis has a great public health significance and the protozoa found in goats of Fulbaria Upazilla may possess threat for transmission in human and other animals, require further investigation.

Livestock infected with Leishmania spp. in southern Iran

“…To determine whether the same parasite clone is present, the Leishmania ITS1 DNA sequences of VL and CL patients in this study area should be compared to that extracted from infected dogs, rodents, sand flies and livestock. Several previous studies in Leishmania endemic regions around the world have reported Leishmania infection in livestock, consistent with present study findings, although these studies differ in terms of sample size, diagnostic method applied, types of samples, presence of heterogeneous parasite populations and immune responses to the Leishmania parasite [ 12 – 14 , 16 , 26 ]. In the present study, some samples were positive by both molecular and serological methods (51/181; 28.2%), while others were positive by only one method.…”

Passive surveillance on occurrence of deadly infectious, noninfectious and zoonotic diseases of livestock and poultry in Bangladesh and remedies

“…The important zoonotic diseases only noted in the hospital data sheet during passive surveillance was anthrax. Brucellosis (Dey et al, 2013) and Tuberculosis in dairy cattle , Leishmaniasis in goats (Labony et al, 2014) and canids and avian influenza in chickens (Bari et al, 2009) and ducks were endemic in Bangladesh and extremely zoonotic, these diseases were not included in the hospital data sheet. The passive surveillance protocols were, therefore, unable to analyze frequency distribution of the existing zoonotic diseases of livestock and poultry in Bangladesh.…”

Evaluate comparative efficacy of tuberculin test, gross and histopathology, Ziehl-Neelsen staining and polymerase chain reaction technique for the detection of tuberculosis in dairy cattle

“…Portion of lungs were collected aseptically, snap frozen and used for bacterial DNA extraction and detection of TB by using multiplex and uniplex PCR. In brief, 2 gm frozen tissues were crushed in liquid nitrogen and extracted bacterial DNA using conventional method [9]. The quality and quantity of the extracted DNA was measured by using agarose gel electrophoresis and spectrophotometry (SpectronicR Genetics TM New York, USA) (A260/A280).…”