Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India

Abstract

Buffaloes are the second largest source of milk. Mastitis is a major impediment for milk production, but not much information is available about bubaline mastitis, especially subclinical mastitis. The aim of this study was to (a) investigate the application of various tests for the diagnosis of bubaline subclinical mastitis, (b) identify the major bacteria associated with it, and (c) evaluate the antibiotic resistance pattern of the bacteria. To this end, 190 quarter milk samples were collected from 57 domesticated dairy buffaloes from organized (64 samples) and unorganized (126 samples) sectors. Of these, 48.4%, 40.0%, 45.8%, 61.1%, and 61.6% were positive for subclinical mastitis by somatic cell count, electrical conductivity, California mastitis test, bromothymol blue test, and N-acetyl glucosaminidase test, respectively. As compared to the gold standard of somatic cell count, California mastitis test performed the best. However, a combination of the two methods was found to be the best option. Microbiological evaluation, both by biochemical methods as well as by monoplex and multiplex polymerase chain reaction, revealed that coagulase-negative staphylococci were the most predominant (64.8%) bacteria, followed by streptococci (18.1%), Escherichia coli (9.8%) and Staphylococcus aureus (7.3%). Most of the pathogens were resistant to multiple antibiotics, especially to β-lactam antibiotics. We propose that California mastitis test be combined with somatic cell count for diagnosis of subclinical mastitis in domestic dairy buffaloes. Further, our results reveal high resistance of the associated bacteria to the β-lactam class of antibiotics, and a possible major role of coagulase-negative staphylococci in causing the disease in India.

Antibiotic Resistance Pattern of Mastitis Causing Escherichia Coli Toxinotypes

“…This low percentage was because of good milking practices and use of teat dips before milking. While, another study by Preethirani et al (2015) showed high percentage (9.8) of E. coli isolates from an unorganized dairy farm. These results were not in accordance to present study.…”

Insights Into the Resistome of Bovine Clinical Mastitis Microbiome, a Key Factor in Disease Complication

“…The most frequently isolated pathogens are Staphylococcus aureus, Escherichia coli, Klebsiella spp., Streptococcus spp., Mycoplasma spp., Enterobacter spp., Bacillus spp., and Corynebacterium species (Abebe et al, 2016;Gao et al, 2017;Hoque et al, 2018). Therefore, the accurate identification of pathogens that cause CM enables appropriate choices for antimicrobial treatment and preventive mastitis management (Preethirani et al, 2015;Van Boeckel et al, 2015;Cheng et al, 2019). Over the past two decades, a wide range of phenotyping and genotyping methods have been implemented to study mastitis-causing bacteria (Preethirani et al, 2015;Gao et al, 2017;Hoque et al, 2018;Cheng et al, 2019).…”

“…Therefore, the accurate identification of pathogens that cause CM enables appropriate choices for antimicrobial treatment and preventive mastitis management (Preethirani et al, 2015;Van Boeckel et al, 2015;Cheng et al, 2019). Over the past two decades, a wide range of phenotyping and genotyping methods have been implemented to study mastitis-causing bacteria (Preethirani et al, 2015;Gao et al, 2017;Hoque et al, 2018;Cheng et al, 2019). Although culture-based techniques are traditionally used to detect CM bacteria, these methods are time-consuming and have the inherent drawback of not being applicable to non-cultivable bacteria (Baron et al, 2018).…”

Antibacterial effects and resistance induction of silver and gold nanoparticles against Staphylococcus aureus‐induced mastitis and the potential toxicity in rats

“…Bovine mastitis is one of the major problems facing the dairy industry worldwide (Benić, Habrun, & Kompes, 2012;Elbehiry, Al-Dubaib, Marzouk, Osman, & Edrees, 2016;Preethirani et al, 2015). Several bacterial species are responsible for bovine mastitis infections, Staphylococcus aureus (S. aureus) being one of the most prevalent mastitis pathogens, causing the most virulent forms of bovine mastitis (El Behiry, Schlenker, Szabo, & Roesler, 2012;Yang et al, 2008).…”