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Isolation and Identification of Bacteria from Tracheas and Lungs of Buffaloes in Dinajpur

Md. Zulfekar Ali
Nourish Poultry Diseases Diagnostic Laboratory, Unit-2, Bogra, Bangladesh

Shirin Sultana
Department Pathology and Parasitology, Hajee Mohammad Danesh Science and Technology University (HSTU), Basherhat, Dinajpur – 5200, Bangladesh

Abstract/ Executive Summary:

The aim of the present study was the isolation and identification of bacteria from trachea and lung of slaughtered buffaloes. A total of 160 swab samples were collected from tracheas (80) and lungs (80) freshly slaughtered buffeloes from the slaughter houses of Dinajpur Sadar, Dinajpur, Bangladesh, during the period from June 2011 to February 2012. Among them, 9 tracheas (out of 80 samples) and 30 lungs (out of 80 samples) showed visible lesions. Bacterial isolates were identified based on their growth, morphological and biochemical characteristics. The most frequent (5%) bacterial isolate in tracheas was Staphylococcus spp. Pasteurella haemolytica and Staphylococcus spp. were isolated from different types of lung lesions in a frequency of 5% and 16.25%, respectively. P. haemolytica showed β- haemolysis on blood agar and did not ferment lactose and dulcitol whereas dextrose, manitol and maltose were fermented with the production of acid. This investigation would be helpful for veterinarian for the detection, treatment and control of tracheal and lung diseases of buffaloes.

Keywords: Staphylococcus; Pasteurella; buffaloes

Location: Nourish Poultry Diseases Diagnostic Laboratory, Unit-2, Bogra, Bangladesh

Start Date: 00-06-2011

End Date: 00-02-2012

Program Area: Food Safety and Security

Commodity: Buffalo

Objectives:

1. The investigation has been undertaken for the isolation and identification of the bacteria commonly available in the tracheas and lungs of buffaloes in Bangladesh.

Methodology:

Collection of samples. Samples of trachea and lungs were collected from buffaloes after slaughtering from slaughterhouses of Dinajpur Sadar, Dinajpur, Bangladesh during the period from June 2011 to February 2012. A total of 160 sterile swabs, of which 80 from mucosal surface of trachea and 80 from inner core of lung were collected, immediately after opening the thoracic cavity to avoid contamination. All the samples were collected in nutrient broth and brought to the Microbiology Laboratory of the Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh for isolation and identification of pathogenic bacteria. Isolation of Bacteria. Samples were inoculated in nutrient broth and incubated at 37 ºC for 24 hours. From the broth, subcultures were also made on blood agar, MacConkey agar, eosine methylene blue (EMB) agar and nutrient agar, and incubated overnight at 37 ºC. Identification of Pathogenic Bacteria. The representative bacteria were stained using modified Gram's staining and Leishman's stain according to the standard method. Modified Gram’s staining. Ammonium oxalate-crystal violet was applied on the bacterial smear for 1 to 2 minutes, washed with water and Lugol’s iodine solution was then applied on slide for 1 to 2 minutes. Then iodine was applied on slide following decolorization with a few drops of acetone for 5-15 seconds. After washing with water the smear was counter stained with 0.5% safranine for 1 to 2, then washed, air dried and examined. Leishman's staining. Neat Leishman's stain was added on the bacterial smear for 3 minutes. The stain was diluted gently with double volume of tap water. After 20 minutes excess stain was washed off and air dried. Acid fast staining (Ziehl-Neelsen's method). Carbol fuschin was applied on heat fixed bacterial smear and heated the underside of slide until steaming (without boiling) for 5-7 minutes and washed thoroughly under running tap water. Acid alcohol was applied for 1 minute, followed by counter staining with methylene blue for 1 minute, then washed and examined. Biochemical Tests. Five basic sugars such as dextrose, dulcitol, lactose, maltose and menthol were used for sugar fermentation test. Besides these, other biochemical tests such as triple sugar iron (TSI) agar test, Voges-Proskuer (VP) tests and indole tests were also performed in this study.

Source of Information: Stamford Journal of Microbiology, December 2012. Vol. 2, Issue 1; ISSN: 2074-5346

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In this study, P. haemolytica was isolated as the most prevalent bacteria in the congested stage of pneumonia. So, it was speculated that P. haemolytica might be the cause of pneumonia in congested stage as found previously. The etiological agent of tuberculosis in buffaloes is Mycobacterium bovis that causes nodular lesion in lungs though, no bacteria were found in section of lung nodules as observed through acid fast staining. In conclusion, this study on trachea and lung samples collected from buffaloes has focused the different diseases and the detection of pathogenic bacteria in those organs. These findings would be helpful to diagnose the etiological agents and their prevalence in respiratory diseases of buffaloes. Furthermore, our findings will also help the field veterinarian to treat and control the respiratory diseases of buffaloes.

Knowledge Management: Journal