Abstract
Bovine milk is vital for infant nutrition and is a major component of the human diet. Bovine mastitis is a common inflammatory disease of mammary gland in cattle. It alters the immune profile of the animal and lowers the quality and yield of milk causing huge economic losses to dairy industry. The incidence of sub-clinical mastitis (SCM) is higher (25–65% worldwide) than clinical mastitis (CM) (>5%), and frequently progresses to clinical stage due to lack of sensitive and specific detection method. We used quantitative proteomics to identify changes in milk during sub-clinical mastitis, which may be potential biomarkers for developing rapid, non-invasive, sensitive detection methods. We performed comparative proteome analysis of the bovine milk, collected from the Indian hybrid cow Karan Fries. The differential proteome in the milk of Indian crossbred cows during sub-acute and clinical intramammary gland infection has not been investigated to date. Using high-resolution mass spectrometry-based quantitative proteomics of the bovine whey proteins, we identified a total of 1459 and 1358 proteins in biological replicates, out of which 220 and 157 proteins were differentially expressed between normal and infected samples. A total of 82 proteins were up-regulated and 27 proteins were down-regulated, having fold changes of ≥2 and ≤0.8 respectively. Among these proteins, overexpression of CHI3L1, LBP, GSN, GCLC, C4 and PIGR proteins was positively correlated with the events that elicit host defence system, triggering production of cytokines and inflammatory molecules. The appearance of these potential biomarkers in milk may be used to segregate affected cattle from the normal herd and may support mitigation measures for prevention of SCM and CM.
Highlights
Bovine milk is vital for infant nutrition and is a major component of the human diet
Mastitis is an inflammation of the mammary gland which is broadly classified into sub-clinical and clinical mastitis on the basis of severity of infection
somatic cell counts (SCC) and California Mastitis Test (CMT) are typically used as an udder health index and as an inflammatory indicator to diagnose mastitis, because it represents the number of neutrophils in milk
Summary
Bovine milk is vital for infant nutrition and is a major component of the human diet. Bovine mastitis is a common inflammatory disease of mammary gland in cattle It alters the immune profile of the animal and lowers the quality and yield of milk causing huge economic losses to dairy industry. We used quantitative proteomics to identify changes in milk during sub-clinical mastitis, which may be potential biomarkers for developing rapid, non-invasive, sensitive detection methods. Advances in mass spectrometry-based quantitative proteomics techniques, such as two-dimensional gel electrophoresis (2D-GE)[8], label-free[9] and labelled[10] approaches, have been used to identify and quantify several host-specific milk proteins during mastitis which may be potential biomarker candidates for diagnosis of sub-clinical mastitis. To identify protein expression changes during sub-clinical and clinical infection, we used a Tandem Mass Tag TMT-based quantitative proteomic approach to identify potential biomarkers for early detection of sub-clinical mastitis
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