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Investigating Infectious and Zoonotic Diseases of Street Dogs in the Residential Area of Bangladesh Agricultural University

T. K. Kasfi
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

S. S. Labony
Department of Parasitology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

M. G. A. Chowdhury
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

U. K. Rima
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

M. Z. Hossain
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

S. J. H. Chowdhury
Bangladesh Agricultural Research Council (BARC), Farmgate, Dhaka, Bangladesh.

M. M. Hossain
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

M. A. H. N. A. Khan
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

Dogs are common carnivor and scavenger at Bangladesh Agricultural University (BAU) campus. Dogs may carry many infectious and zoonotic diseases and could play a role in transmitting those diseases in other carnivors and human as well. At day time cats, dogs and golden jackals are found to eat in a same dustbin at BAU campus. So it is possible to share diseases of each other and there are possibilities of crossing species barrier. This study was, therefore, aimed at specific investigation of the occurrence of leishmaniasis, canine distemper (CD), infectious canine hepatitis (ICH) and avian influenza (AI) of dogs at BAU campus. This study was carried out during the period from January to June-2012. A total of 10 apparently healthy dogs were euthanatized and postmortem examination were carried out. Impression smears were prepared from spleen, liver, bone marrow and stained with Giemsa’s stain. Histopathological studies were conducted using routine procedures. For molecular characterization, polymerase chain reaction (PCR) and Reverse Transcriptase polymerase chain reaction (RT-PCR) were adopted for the detection of genomic fragment of visceral leishmaniasis (VL), canine distemper (CD), infectious canine hepatitis (ICH) and avian influenza (AI, viral matrix protein gene). Results of this study showed the presence of leishmania organism in the impression smears of eight dogs. Using PCR amplification technique, Leishmania donovani specific genomic DNA (145bp) was detected in two dogs. PCR amplification of genomic DNA of ICH ( 411bp) and RT-PCR amplification of genomic RNA of CD (287bp) and matrix protein gene of AI (245bp) were not detected in any of the dogs of BAU campus. Future work is needed to explore the presence of snad flies (vector of VL) in the investigating areas and establish role of street dogs of BAU campus towards disseminating leishmaniasis in other animals and human beings.

Keywords: Street dogs, Visceral leishmaniasis, Canine diseases, Avian influenza, Giemsa’s staining, PCR

Location: Department of Pathology, Faculty of Veterinary Science, BAU, Mymensingh

Start Date: 01-01-2012

End Date: 30-06-2012

Program Area: Animal Health and Management

Commodity: Diseases

Objectives:

To adopt protocols for the detection of selected genomic fragments of VL, ICH, CD and AI matrix protein genes and designed strategies to prevent its future dessimination.

Methodology:

The investigation was carried out in the Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University (BAU), Mymensingh, Bangladesh. A total of 10 apparently healthy dogs were caught by the dog catchers of district public health corporation, Mymensingh, euthanatized by intravenous injection of saturated solution of magnesium sulfate and brought for investigation to the Department of Pathology, BAU during the period from January-June, 2012. Systemic dissection and investigation was carried out and liver, lymph node, kidney, spleen, heart, lungs, bone marrow, intestine and skin were collected. During necropsy liver, lungs, spleen, intestine and skeletal muscles from all dogs were collected and fixed in 10% buffered neutral formalin for histopathological studies. Formalin fixed tissue samples were trimmed, processed, sectioned and stained with Hematoxylin and Eosin staining. A low and high power microscopic examination were carried out to observe the changes related to specific infections. DNA for the detection of genomic fragments of Leishmania and ICH. Portion of liver and spleen of street dogs were also collected and extracted genomic. DNA using manufacturer instruction (2012). Spectophotometry and agarose gel ectrophoresis were done for confirmation of the purity and concentration of DNA. Primer sets from published literature were used to identify causal agent of VL and viruses of ICH. PCR reactions were adopted for the detection of genomic fragments of L. donovani and ICH viruses. PCR was carried out in a final reaction volume of 25μl in the thin walled PCR tubes. The condition of PCR amplifications were denaturation for 60sec at 94°C, annealing for 90sec at 61°C and extension for 60sec at 72°C followed by a final extension for 10min at 72°C. The PCR reactions were held at 4°C and finally the reaction was terminated by adding 3µl 50mM EDTA. The PCR products were analyzed by electrophoresis in 1.5% agarose gel, stained with ethidium bromide and examined under UV light using an image documentation system. RNA from the host tissues including viral RNA was extracted from liver and spleen of street dogs using commercially available RNA extraction Kit and used the protocol as described by the manufacturer. The purity and concentration of RNA was determined by using spectophotometry and agar gel electrophoresis. One step RT-PCR was carried out with a total of 25μl of reaction volume. The RT-PCR protocols were adopted and carried out in a Master Cycler. The thermal profile used for CD Viruses comprised a reverse transcription for 30min at 50°C. The subsequent PCR were carried out with an initial denaturation for 4min at 94°C followed by 40 cycles of amplification reaction. The condition of PCR amplifications were denaturation for 60sec at 94°C, primer annealing for 2min at 59.5°C and extension for 3min at 72°C followed by a final extension for 10min at 72°C. The RT-PCR reactions were held at 10°C and the reaction was terminated by adding 3µl 50mM EDTA. The RT-PCR products were analyzed by electrophoresed in 1.5% agarose gel, stained with ethidium bromide and examined under UV light using an image documentation system.

Source of Information: Bangl. J. Vet. Med. (2015). 13 (1): 57-63, ISSN: 1729-7893, 2308-0922

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

This study provides evidence that Leishmania like protozoa was present in the internal organs of eight street dogs of BAU campus. Results of PCR (145bp amplicon) showed the presence of L. donovani in the liver of two dogs. Canine distemper, infectious canine hepatitis and avian influenza viral genomes were not detected in any of the dogs of BAU campus. It needs to test large number of sample using wider primers pair to confirm the presence of other species of Leishmania and detection of vector available at BAU campus.

Knowledge Management: Journal