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Isolation, Identification and Resistance Pattern of Microorganisms Associated with Mastitis in Buffalo

Ali AzamTalukder*
Department of Microbiology, Jahangirnagar University, Dhaka 1342, Bangladesh

Hasna Hena Rahman
Department of Microbiology, Jahangirnagar University, Dhaka 1342, Bangladesh

S. M. Jamil Mahmud
Department of Microbiology, Jahangirnagar University, Dhaka 1342, Bangladesh

Fahmida Alam
Department of Microbiology, Jahangirnagar University, Dhaka 1342, Bangladesh

Shuvra Kanti Dey
Department of Microbiology, Jahangirnagar University, Dhaka 1342, Bangladesh

Abstract/ Executive Summary:

Prevalence of subclinical mastitis along with the etiological agents and their antimicrobial susceptibility were studied around the city of Dhaka, the capital of Bangladesh. Milk samples from 30 apparently healthy buffaloes were subjected to microbiological examinations while subclinical mastitis was diagnosed based on California Mastitis Test (CTM). Seventy percent of the buffaloes examined were positive for subclinical mastitis. A total of 42 isolates of 5 different species were identified. Among them, Staphylococcus spp. was the most frequently identified bacterial species accounting for 50% of all the isolates, followed by E.coli (28.57%), Enterobacter (14.29%), Bacillus spp. (4.76%) and Proteus spp. (2.38%). Highest resistance was found against amoxicillin (21.43%) where chloramphenicol and ciprofloxac in proved to be a more successful antimicrobial agent, resistance against which was only 4.76%. Results from the present study suggest that contagious mastitis control practice along with good personal hygiene practice is required for mastitis control in the area.

Keywords: Mastitis, Buffalo, Bangladesh, Pathogen, Staphylococcus

Location: Dhaka

Start Date:

End Date:

Program Area: Pest Management

Commodity: Buffalo, Bacteria

Objectives:

To investigate the incidence of mastitis disease in buffalo prevailing around Dhaka in order to develop appropriate control and preventive measures against the disease.

Methodology:

Sample collection A total of 30-buffaloes were randomly selected from the three distinct areas of Dhaka, the capital of Bangladesh, in farm basis, in order to find out the epidemiology of clinical and sub-clinical mastitis caused by microorganisms in the study area. During sample collection, milk samples were collected by standard milk sampling technique as described previously6, while clinical and subclinical mastitis was detected using CMT. Approximately 10 ml of milk samples were collected into sterile test tube after discarding the first three milking streams. A septically collected sample were placed in ice boxes and transported to the laboratory with maintaining the appropriate storage and transport temperature and other physical conditions ensuring the maximum survival of the microorganisms in the milk sample for future analysis. Isolation and identification of bacteria from milk In the laboratory, stored samples were kept at room temperature for 30 minutes before streaking on culture plates. Collected samples were diluted when necessary. One hundred microliter of samples was streaked on various selective agar media including, MacConkey and blood agar plates for bacterial culture and isolation. Pure colonies were identified and analyzed further morphologically and biochemically for strain identification as shown below. Antimicrobial susceptibility test The antimicrobial susceptibility of the test isolates was determined in vitro by using the standardized agar-disc-diffusion method known as the Kirby Bauer method according to Clinical and Laboratory Standards Institute (CLSI) guidelines7. Total ten antibiotic discs were used in this study, namely, amoxicillin (AML), azithromycin (AZM), chloramphenicol (CHL), ciprofloxacin (CIP), ceftriaxone (CRO), erythromycin (ERM), gentamycin (GNT), levofloxacin (LEV), nitrofurantoine (NFT) and thrimethoprim (SXT).

Source of Information: Bangladesh J Microbiol, Volume 30, Number 1&2, June-December 2013, pp 1-5

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In this study, we have identified some pathogenic microorganisms responsible for mastitis in buffalo. Among them, Staphylococcus spp. isolates were most susceptible to ceftriaxone, trimethoprim, chloramphenicol, gentamycin, levofloxacin and amoxicillin; and resistant to amoxicillin, erythromycin and azithromycin. Similarly E. coli was highly resistant to ceftriaxone, trimethoprim, azithromycin and nitrofurantoine but susceptible to ciprofloxacin, azithromycin, trimethoprim, chloramphenicol and levofloxacin due to its infrequent usage. Our results are consistent with the result published recently12. However, some risk factors should be taken into account for prevention of mastitis in buffalo13. Moreover, the etiological agent of mastitis varies from region to region. Some drawbacks of our study are that we did the identification of the microorganisms on the basis of morphological and biochemical tests and the sample size in number were too small. So, for future research, sample size should be increased and additional test including WST, CMT and SCC should be done in addition to genetic identification on the basis of PCR of 16s rRNA gene of the microorganisms along with some other genetic characterization such as presence of virulence and antibiotic resistance genes should be included.

Knowledge Management: Journal