A study on milk proteome and biochemical analysis of milk and blood of mastitic and healthy Sahiwal cows

Abstract

The study was performed at Department of Veterinary Biochemistry, College of Veterinary newlineSciences and Animal Husbandry, Uttar Pradesh Pandit Deen Dayal Upadhayaya Pashu Chikitsa Vigyan newlineVishwavidyalaya Evam Go- Anusandhan Sansthan, (DUVASU), Mathura, Uttar Pradesh, India. The newlinestudy was carried out on lactating Sahiwal cows maintained at ILFC of the College of Veterinary newlineScience, Mathura. Milk samples used in this study were of control/apparently healthy group, newlinesubclinical mastitis and clinical mastitis group. newlineThe animals were classified into three stages of lactation namely, Early lactating group (up to newline100 days), Mid lactating (from 101 to 200 days) and late lactating (from 201 to 280 days). Now these newlinegroups were further divided in to three subgroups, namely control/healthy group subclinical mastitis newlineand clinical mastitis group on the basis of screening by using somatic cell count (SCC) , California newlinemastitic test (CMT) and Electrical conductivity test (ECT) on the milk samples. newlineBiochemical analysis of milk samples revealed significant increase in Sodium, copper, MDA, newlineLactose, LDH, AST and ALP and decrease in Magnesium, calcium , phosphorus, potassium, iron, zinc, newlineTotal protein and Albumin compared to healthy animals. newlineMilk of healthy animal showed significant decrease in Sodium and LDH whereas Subclinical newlinemastitic animals revealed significant fall in copper and LDH and rise in copper levels in Mid and Late newlinelactation compared to early lactation. newlineBiochemical study of serum of healthy, SCM and CM Cows showed significant increase in newlineSodium, potassium, Total protein, Globulin LDH and AST while significant decrease in calcium, newlinecopper, albumin, A/G ratio, FFA, MDA and Cholesterol. newlineThe stage of lactation found to affect biochemical profile in both serum and milk and revealed newlinesignificant increase in Calcium, phosphorus, copper, glucose, cholesterol, LDH and AST and decrease newlinein Total protein in Mid and Late lactation compared to early lactation in healthy animals. Similarly newlinecows having SCM showed significant increase in calcium, phosphorus, sodium, glucose and newlinecholesterol and decrease in Total protein and MDA in Mid and Late lactation. newlineBacterial isolation and multiplex PCR performed on milk samples of SCM/CM revealed newlineinfection of Staphylococcus, Pseudomonas and E. coli in the studied cows. newlineSDS PAGE analysis of pooled milk samples healthy milk revealed 23 protein bands where as newlineboth subclinical and clinical mastitic milk 27 protein bands. The 4 protein bands which were not found newlinein the healthy milk revealed 14, 56, 160 and 168 kDa molecular weight proteins. newline2-D gel electrophoresis was carried out by using isolated whey proteins of healthy SCM and newlineCM sahiwal cows. IEF and I-D gel analysis revealed expression of large set of proteins but on gel newlineanalysis revealed 3 differentially expressed protein spots in CM group of proteins. Those spots were newlineidentified, digested and sent for MALDI analysis. MALDI analysis revealed following important newlineproteins. newlinea) DNA-dependent protein kinase catalytic subunit, protein. newlineb) Rho-specific guanine nucleotide exchange factor (RhoGEF) newlinec) acyl-CoA-binding domain-containing protein 6 isoform X4 [Bubalus bubalis] newlined) Ankyrin repeat domain-containing protein [Homo sapiens] newlinee) Chordin-like protein 1 newlinef) kielin/chordin-like (KCP) protein