34-Title: Proteomic analysis of milk in mastitic, subclinical mastitic and healthy Sahiwal cows
Authors: Pawanjit Singh, R Nigam, Vijay Pandey and Ambika Sharma
Source: Ruminant Science (2024)-13(1):175-180.

Abstract

How to cite this manuscript: Singh Pawanjit, Nigam R, Pandey Vijay and Sharma Ambika (2024). Proteomic analysis of milk in mastitic, subclinical mastitic and healthy Sahiwal cows. Ruminant Science 13(1):175-180.
Abstract
The present study was conducted on milk samples collected from lactating Sahiwal cows housed at the Livestock Farm Complex of the institute. The experimental animals were categorized into three groups: apparently healthy, subclinical mastitis (SCM), and clinical mastitis (CM). Classification was based on a comprehensive screening process that included Somatic Cell Count (SCC), the California Mastitis Test (CMT), and the Electrical Conductivity Test (ECT). SDS-PAGE analysis of pooled milk samples revealed distinct protein profiles across the groups. Milk from healthy cows exhibited 23 protein bands, whereas both subclinical and clinical mastitic milk samples showed 27 protein bands. Notably, four additional protein bands, corresponding to molecular weights of 14, 56, 160, and 168 kDa, were observed exclusively in the mastitic milk samples. Further protein profiling was performed using 2-D gel electrophoresis on whey proteins isolated from healthy cows as well as cows with SCM and CM. Isoelectric focusing (IEF) followed by 1-D gel analysis highlighted the expression of a broad set of proteins. However, three differentially expressed protein spots were identified specifically in the CM group. These protein spots were excised, digested, and subjected to MALDI-TOF mass spectrometry analysis. The MALDI analysis identified the following proteins-DNA-dependent protein kinase catalytic subunit, Rho-specific guanine nucleotide exchange factor (RhoGEF), Acyl-CoA-binding domain-containing protein 6 isoform X4 (Bubalus bubalis), Ankyrin repeat domain-containing protein (Homo sapiens), Chordin-like protein 1 and Kielin/chordin-like protein (KCP). These findings offer valuable insights into the protein expression changes associated with mastitis in Sahiwal cows and provide potential biomarkers for disease detection and management.
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