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Characterization and Antibiogram of Staphylococcus Aureus Isolated from Mastitic Milk

N. Parveen
Veterinary Surgeon
District Veterinary Hospital, Gaibandha, Bangladesh.

M. M. Kamal
Veterinary Surgeon
Upazilla Livestock Office, Palashbari, Gaibandha, Bangladesh.

S. Saha
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

K. A. Choudhury
Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

Bacteriological examination of milk samples collected from 60 clinically mastitis affected milch cows, was performed during the period from July 1999 to March 2000, of which 21 (35.0%) had Staphylococcus aureus infection. The chromogenic characters of the coagulase positive Staphylococcus showed nearly equal distribution of golden, yellow and white strains and, therefore, were not regarded as a satisfactory criterion to indicate their coagulase status. Of biochemical tests anaerobically mannitol fermentation and phosphatase activity proved to be of value in the identification of Staph. aureus. In vitro drug sensitivity pattern depicted most sensitivity of the organism to gentamicin, chloramphenicol and erythromycin (100% each) while oxacillin, neomycin, amoxycillin, streptomycin and ampicillin inhibited the growth of 19 (90.48%), 17 (80.95%), 16 (76.19%), 15 (71.43%) and 14 (66.67%) strains of Staph. aureus respectively. Resistance to penicillin-G (57.14%) and tetracycline (42.86%) was comparatively high. It was evident that gentamicin, chloramphenicol and erythromycin can be better value in the treatment of Staph. aureus infections.

Keywords: Characterization, Antibiogram, Staphylococcus aureus, Mastitic, Milk

Location: Bangladesh Agricultural University

Start Date: 00-07-1999

End Date: 00-03-2000

Program Area: Pest Management

Commodity: Milk

Objectives:

1. To analyze the physiological characteristics of the isolates and

2. To know their antibiotic sensitivity pattern against a range of antimicrobial agents.

Methodology:

A total of 60 mastitic milk samples were collected during the period from July 1999 to March 2000 from Bangladesh Agricultural University dairy farm and cases of mastitis brought to the Veterinary Clinic at the Bangladesh Agricultural University Campus, Mymensingh. Standard laboratory aseptic procedures were adopted during and after sampling. Planting on nutrient agar and 5% sheep blood agar made isolation of Staphylococcus aureus in pure culture from the collected samples. The inoculated plates were incubated aerobically 37C°C for 24 hours for growth. The isolates were identified on the basis of morphology, cultural characteristics, biochemical properties and principally on coagulase positivity. For the coagulase test rabbit plasma was used in this study. Diluted 2.5ml rabbit plasma (1:5 with sterile physiological saline) was mixed separately in two different test tubes containing an equal volume of 24 hours old Staphylococcal broth culture (or saline suspension of organisms from a solid medium) and incubated at 37°C for 4 hours. The tubes were examined after 2 to 4 hours for detecting the presence of clots of plasma. The negative tubes were left at room temperature for overnight and then re- examined. To perform the test an amount of 3ml of 3% hydrogen peroxide solution was poured into a test tube. A confluent growth of test organisms was immersed into the solution by a sterile wooden stick or glass rod. The production of gas bubbles was constituted a positive reaction. All the coagulase positive Staphylococcal strains were further tested for pigment production, haemolytic activity, anaerobic mannitol fermentation and phosphatase activity. Test for pigment production was carried out by growing the organisms on 8% NaCI agar plates and incubating them aerobically for 24 hours at 37°C followed by 48 hours at room temperature. Haemolysis due to haemolysin production was observed on 5% sheep blood agar plates. Anaerobic fermentation of mannitol was undertaken on Mannitol salt agar plates. The anaerobiosis was produced into a previously sterilized glass jar by placing a lighted-smokeless candle near the top of the jar which was made air tight by putting the cover on the jar with spreading vaseline at the both mouth edges. The production of phosphatase was examined by growing the organism on phenolphthalein phosphate agar indicator medium. In-vitro antibiotic sensitivity pattern of the coagulase Staphylococcal isolates was studied against 10 antimicrobials using commercially available biodiscs (HiMedia Laboratories Pvt. Ltd., Mumbai, India) by standard disc diffusion technique on nutrient agar and blood agar plates.

Source of Information: Bangladesh Vet. J (2001), 35 (1 - 2) : 29 - 33

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From the present investigation it is therefore evident that gentamicin, chloramphenicol and erythromycin can be used to treat the infections of Straphaureus origin. Further studies with larger number of strains from different geographical areas would be necessary before concluding the organism resistance to specific antimicrobial compound.

Knowledge Management: Journal