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Pathological Investigation and Identification of Bacteria from Bovine Uterus

Mst. Naznin Nahar
Upazila Livestock Office
Charghat, Rajshahi, Bangladesh.

Rumi Majumder
Rabies Vaccine Section, Livestock Research Institute, Mohakhali, Dhaka-1212, Bangladesh.

Md. Mokbul Hossain
Department of Pathology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

The present study was carried out to determine the occurrence of bacteria as well as pathological lesions in the uterus of slaughtered cattle. A total of 20 selected samples from 30 randomly collected samples were taken from the uteri with grossly identifiable lesions such as congested, hemorrhagic, cystic, and mucus-filled uteri. The used methods were cultural technique, staining procedure, and different types of biochemical test and histopathology. The bacteriological swab samples were collected from the lumen of the uteri in nutrient broth. The histopathological samples were collected in 10% neutral buffered neutral formalin. The collected tissues were fixed, processed, sectioned, stained, and studied with a light microscope. The mostly observed gross lesions were hemorrhagic, congested, and mucus-filled uterus with some abnormalities on the ovary. All the gross lesions were not found in all uteri. The occurrence of bacteria 22.92% Bacillus spp, 12.50% Streptococcus spp, 25.00% Staphylococcus spp, 31.11% Escherichia coli, and 10.417% Klebsiella spp. were recorded in 20 uteri of cattle. Microscopically, acute endometritis was characterized by thickening of the epithelial layer, infiltration of neutrophils in the submucosal layer, dilatation of endometrial glands, and congestion of blood vessels. In chronic endometritis, infiltration of lymphocytes, macrophages, plasma cells in the submucosal layer and proliferation of fibrous connective tissue within the myometrium. In adenomyosis, dilatation of endometrial glands and the presence of endometrial glands in the myometrium were recorded. The microscopic lesions were variable in uteri. The occurrence of acute endometritis was 33.33%, chronic endometritis 26.66%, and adenomyosis 6.66% respectively. In acute and chronic endometritis, the isolated bacteria were Bacillus spp, Streptococcus spp, Staphylococcus spp, Escherichia coli, and Klebsiella spp. while in adenomyosis cases the isolated bacteria were Bacillus spp, Staphylococcus spp, and Escherichia coli. To understand the role of these bacteria in the production of pathological lesions further studies are needed.

Keywords: Uterus, Cow, Bacteria, Histo- Pathology, Streptococcus, Staphylococcus, Escherichia coli, Klebsiella

Location: Department of Pathology, BAU, Mymensingh.

Start Date: 00-07-2009

End Date: 00-12-2009

Program Area: Animal Health and Management

Commodity: Cattle, Bacteria

Objectives:

To find out the uterine bacterial organisms associated with different types of histopathological changes in slaughtered cows.

Methodology:

Grossly identifiable lesions such as congested, hemorrhagic, cystic, and mucus-filled uteri from slaughtered cattle of different ages from kewatkhali region and district slaughterhouse of Mymensingh Sadar during July to December 2009 were used for this investigation. Aseptically swabs were collected from the lumen of uteri in nutrient broth for bacterial isolation and identification. Isolation and identification of different organisms were performed by culturing, staining, different biochemical (carbohydrate fermentation, Methyl Red and Voges-Proskauer (MR-VP), indole test) tests, and enzyme activity (catalase test) test. For histopathology, the samples were collected and fixed in 10% buffered formalin. After fixation, the samples were trimmed and again fixed for 1-2 days. Then processing of samples (processing, embedding, sectioning, etc.) was done. The processed samples were collected on slides. Staining with routine H & E stain was performed following a routine procedure. Finally different pathological changes in tissue level were studied under microscope available in the Department of Pathology. A total of 20 selected samples were collected directly from 20 uteri of cattle (at different ages) for bacteriological isolation in nutrient broth. These samples were carried in a cool box with ice to the Department of Pathology, BAU, Mymensingh. Test tubes containing samples were incubated for 72 hours at 37°C. Grossly identifiable lesions such as congestion, hemorrhage, cystic, and mucus filled uteri of collected samples from Kewatkhali and district slaughterhouse of Mymensingh Sadar were preserved for histopathology in a plastic pot containing 10% neutral buffered formalin. From the nutrient broth, 20 uteri swab samples were placed in a nutrient agar plate and incubated overnight at 37°C for the growth of the organisms. All samples were subcultured in nutrient agar, MacConkey agar, EMB agar, and Blood agar. A small amount of inoculums from the nutrient agar were spread into different culture media and incubated at 37°C for overnight. On the basis of the colony, staining characters, and biochemical tests, the organisms were isolated and identified. A microscopic study was done for all 20 uterine swabs samples after primary culture and subculture. Gram’s staining was performed as per the recommendation of Hucker and Conn; Merchant and Packer. Leishman's staining was performed for 4 samples. With the inoculation loop, a small amount of bacterial colony was taken on a clean glass slide. One drop of water was added then it was mixed properly. Later, the slide was first air-dried and then fixed with gentle heat. Biochemical (carbohydrate fermentation, Methyl Red and Voges-Proskauer (MR-VP), indole test) tests and enzyme activity (catalase test) tests were performed as described by Cowan. The collected uterine samples from slaughtered cows were selected for histopathological study. The formalin-fixed tissues were trimmed with 1.5 cm × 1 cm size. Then the tissues were processed, sectioned, and stained following standard procedure. Specific samples containing lesions from each group were used in the histopathological study. Hematoxylin & Eosin staining was performed and Photornicrography was taken at the Department of Pathology using a photomicrographic camera (Olympus PM-C 35 Model) fitted with an Olympus microscope.

Source of Information: International Journal of Natural and Social Sciences, 2020, 7(3):59-70

Article Link (Online Journal): DOI: 10.5281/zenodo.4043448

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

A total of 20 selected samples were collected directly from 20 uteri of 30 randomly collected uterine samples of cattle at different ages. Post mortem examination was performed on 20 selected uteri. The gross pathological lesions were hemorrhage, congestion, mucus-filled uterine lumen, etc. found almost in all cases. The overall occurrences of bacteria in 20 uterus were 22.92% Bacillus spp, 12.50% Streptococcus spp, 25.00% Staphylococcus spp, 31.11% Escherichia coli, and 10.417% Klebsiella spp. Microscopically, the lesions were categorized into acute endometritis characterized by thickening of the epithelial layer, infiltration of neutrophil in the submucosal layer, dilatation of endometrial glands, and slight congestion of blood vessel; chronic endometritis characterized by infiltration of lymphocytes, macrophages, plasma cells in the submucosal layer and proliferation of fibrous connective tissue within the myometrium and adenomyosis characterized by the presence of endometrial glands within the myometrium. These microscopic lesions were variable in uteri. Bacteria isolated from acute endometritis were E. coli (26.08 %), Streptococcus spp. (13.04 %), Bacillus spp. (13.04 %), Staphylococcus spp. (30.43 %) and Klebsiella spp. (17.39 %); in chronic endometritis E. coli (35.00 %), Streptococcus spp. (15.00 %), Bacillus spp. (30.00 %), Staphylococcus spp. (15.00 %) and Klebsiella spp. (5.00 %); in adenomysis E. coli (20.00 %), Bacillus spp. (40.00 %) and Staphylococcus spp. (40.00 %). Here, the same types of bacteria have been found in different pathological conditions. However, further study would be necessary to know the role of specific bacteria in developing specific lesions in the uterus.

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