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Prevalence of Salmonella Infection in Naturally Infected Layer Birds in Bangladesh

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

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

A. K. Saha
Department of Pathology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

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

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

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

Abstract/ Executive Summary:

The seroprevalence, cultural prevalence and pathological study of Salmonella infections in chickens of selected layer farms of Birgonj Upazila (Sub-district), Dinajpur were determined. A total of 175 blood samples were tested randomly by locally prepared Salmonella coloured antigen for seroprevalence study. Out of 96 cloacal swabs, 80 samples from live birds (36 from seropositive and 44 from seronegative) and 16 samples from dead birds were collected to determine the cultural prevalence of Salmonella organisms. Postmortem examination was done in 16 dead birds. Using whole blood agglutination test (WBA) with locally prepared Salmonella Pullorum coloured antigen, the overall seropositive prevalence was 46.2%. The seroprevalence decreased with age of birds. The cultural prevalence in seropositive was 33.3% and in seronegative 22.7%. In dead birds, the cultural prevalence using cloacal swab was 25%. A total 26 Salmonella were isolated, 27% Salmonella Pullorum, 58% Salmonella Gallinarum and 15% paratyphoid group of Salmonella. Isolation rate of Salmonella from cloacal swabs was significantly higher in seropositive than seronegative group. Grossly, the livers were friable, with bronze discolouration and necrotic foci, there was severe congestion in the lung, congested haemorrhagic egg follicles with stalk formation and enlarged discoloured spleen. Microscopically, there was focal necrosis and degeneration with leukocytic infiltration in liver, congestion and pneumonic lesions in the lung and various degrees of catarrhal to haemorrhagic enteritis in the intestine. In the egg follicles, congestion and haemorrhage with leukocytic infiltration and enlarged spleen with white necrotic foci were detected. In future, isolated Salmonella organisms may be used for vaccine production, serotyping and antibiotic sensitivity test.

Keywords: Salmonella infection, Layer birds , Pathological study

Location: Birgonj Upazila under Dinajpur District

Start Date: 00-11-2009

End Date: 00-04-2010

Program Area: Animal Health and Management

Commodity: Chicken

Objectives:

To determine the seroprevalence of Salmonella infection using locally prepared Salmonella. Pullorum coloured antigen in layer birds at Dinajpur district in Bangladesh.
To determine correlation between cultural prevalence and seroprevalence.
To study the pathological lesions of organs of Salmonella-infected birds.

Methodology:

The study was conducted from November 2009 to April 2010. Blood samples were collected from chickens of 8 layer farms, which contained 30,500 birds and had no history of using Salmonella vaccine. At the rate of 0.8% (conventional method of seroprevalence determination), 175 birds were tested randomly. Sterile disposable syringe was used for collection of 3 ml blood from wing vein under aseptic condition. Birds were divided into two groups, grower (9-20 weeks) and layer (21-80 weeks). One drop of rapidly collected blood and one drop of Salmonella coloured antigen were taken in glass slide and mixed well with toothpick and rocking. In positive cases granules observed within two minutes and graded +++ highly positive response, ++ moderate, + lower positive response and – without any reaction ; Muktaruzzaman et al., 2010). However, all the reactions were graded together for considering as positive case. Isolation and identification of Salmonella organisms A total 80 cloacal swabs (36 from seropositive; 44 from seronegative birds) were collected from the same flock at the same time as the seroprevalence study. Postmortem examinations of 16 dead birds were performed from 8 poultry farms. All the cloacal swabs were collected in test tubes containing 10 ml tetrathionate broth. Iodineiodide solution (200 μl) was added in each test tube just before the swab was mixed in tetrathionate broth. For isolation and identification of Salmonella organisms, the samples were stored in icepack with sunlight-protected black box and carried to the laboratory. Tetrathionate broth (TTB), nutrient broth (NB), Salmonella-Shigella (SS) agar, brilliant green agar (BGA), blood agar (BA), Mac Conkey agar, eosin methylene blue (EMB) agar, triple sugar iron (TSI) agar, nutrient agar (NA), bacteriological peptone, methyl red-Voges Proskauer (MR-VP) medium, Gram’s stain and motility test were used for the isolation and characterization of organisms (Merchant and Packer, 1967; OIE Manual, 2004). Maintenance of stock culture To preserve the isolated organisms for further studies, the organisms from pure culture were inoculated into tubes containing nutrient agar slant and incubated at 37°C for 24 hours. After the growth of organisms, the tubes were sealed with light liquid paraffin and kept at 4°C. Gross pathology During the seroprevalence study, necropsy of dead birds was done routinely. Gross tissue changes at necropsy were recorded and representative tissue samples were preserved in 10% neutral buffered formalin for histopathological studies. Histopathology The tissues were trimmed to 1.5 × 1 cm size, then kept in running tap water overnight to wash out formalin. Dehydration was done in ascending grades of alcohol, 50, 70, 80, 95% and three changes of absolute alcohol for one hour in each. Sections were cleaned in two changes of chloroform, one and half an hours for each. Tissues were embedded in two changes of melted paraffin wax at 56°C, one and half an hours for each. Blocks were sectioned at 5μm thickness. The sections were allowed to spread on warm water bath (45°C) and taken on grease-free glass slides. A small amount of gelatin was added to the water bath for better adhesion of the section to the slide. The slides were air-dried and kept in cool place, and stained routinely (Luna, 1968). Photomicrography Photomicrography was taken using compound microscope with “Sony” digital camera.

Source of Information: The Bangladesh Veterinarian (2011) 28(1) : 8 – 18

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The presence of antibody was not directly related with the presence of organisms in the digestive tract. Seroprevalence, culture and pathological examination should be performed simultaneously for confirmation of Salmonella infections in a poultry farm. Further studies with serotyping, vaccine production and antibiotic sensitivity determination with isolated Salmonella from poultry may be performed in the near future

Knowledge Management: Journal