Background: Bovine mastitis is a multifactorial and is one of the most challenging disease. It can be caused by many different bacterial species, the most common of which are Staphylococcus species and Streptococcus species. The prevalence of different species varies temporally, geographically and also due to control measures adopted in herds. The present study is on the biofilm forming Streptococcus uberis causing mastitis and the effect of antibiofilm agents on the antimicrobial resistance of the microorganisms. Methods: The isolates were identified by Polymerase chain reaction (PCR). Two antibiofilm agents, resveratrol and ursolic acid (UA) each at two concentrations (30 μg/ml, 100 μg/ml) were used for the study. Biofilm formation and rate of biofilm inhibition was detected using quantitative microtiter plate (MTP) assay and biofilm gene (lux S) was detected using PCR. The isolates treated with antibiofilm agents were subjected to standard disc diffusion test with 7 antibiotics and the change in antibiotic resistance was studied. Result: The mean±SE values of inhibition rates of 29 S. uberis isolates by 30 μg/ml UA, 100 μg/ml UA, 30 μg/ml resveratrol, 100 μg/ml resveratrol were 33.96±3.17%, 57.40±2.8%, 31.35±3.12% and 46.28±3.47%, respectively. Biofilm inhibiting agents along with all antibiotics had reduced antimicrobial resistance by 1.5-2 times on in vitro antibiotic resistance testing by disc diffusion at 100 μg/ml concentration. The antibiofilm agents were found to be very effective to control antibiotic resistance of S. uberis from mastitic milk samples in vitro.
Background: Streptococcus uberis (S. uberis) is an environmental pathogen causing mastitis in Dairy cattle. It causes recurrent mastitis and reduction in milk production in livestock causing economic loss. The prevalence of S. uberis intramammary infections is due to ability of the organism to form biofilm in udder tissue. The present study is on in-vitro biofilm production, the correlation of luxS gene and the biofilm formation in S. uberis. Methods: A total of 91 mastitic milk samples were collected from cattle and buffaloes brought at Veterinary Hospitals and farms in Krishna, Guntur and West Godavari districts, Andhra Pradesh. The identification of the culture isolates was based on cultural and biochemical characteristics and confirmed by Polymerase Chain Reaction (PCR). The Streptococcus species cultures showing greyish, pinpointed colonies and/or aesculin hydrolysis on Edwards medium were further identified by various biochemical tests viz., catalase test, ninhydrin test, sodium hippurate hydrolysis test and type of haemolysis on 7% sheep blood agar. Confirmation of the isolates by PCR was followed by detection of biofilm formation using qualitative Congo red agar (CRA) method, quantitative microtiter plate (MTP) assay and biofilm gene (luxS) was detected using PCR.Conclusion: From this study it is suggestable that for biofilm study both phenotypic and genotypic methods should be taken together which can be influenced by various other factors also. MTP assay was a good choice for quantitative biofilm determination, which was giving a more accurate and understandable results. The results express that any of the isolates without luxS didn’t produce a strong biofilm and it is concluded that there may be other genes for regulation of biofilm production and/or luxS gene has a regulatory role for one or more genes related to biofilm formation in S. uberis.
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