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The unique ability of modern proteomics techniques offers a great opportunity for studying simultaneously all proteins in
a sample. The present study was undertaken to examine proteome changes in milk in response to the severity of mastitis
vis-?-vis normal milk in Murrah buffaloes. Milk was collected by hand milking and mastitis was detected by California Mastitis
Test (CMT), followed by milk culture to identify micro flora. Samples collected from 4 normal and 13 CMT positive samples
from organised herd at the Central Institute for Research on Buffaloes, Hisar (India) were classified as normal (4), CMT positive
no abnormality detected in culture (NAD; 2), subclinical (5; Streptococci infection), subclinical mixed (2; Streptococci and
Staphylocci infection) and clinical (4; Streptococci) infection. Fat removed samples were subjected to reduction of pH (~ 4.0)
for casein removal, followed by sample preparation for two dimensional gel electrophoresis (2 DE). Samples (equal volume
of proteins from individual samples) were pooled to make single sample for all proteomic studies. Isoelectric focussing (IEF)
was accomplished using broad range (pH 3-10; 7cm) dry IPG strips at 5750 Volt- hour (Vh) followed by a 12.5% SDS-PAGE.
The commisse blue stained gels so obtained were densitometrically analysed by ImageMaster 2D Platinum 7.0 software (GE
Healthcare, USA) to visualize % volume changes in proteins embedded in spots of different categories. Taking normal milk as
reference, a total of 17 spots ? 8 from subclinical, 5 from subclinical mixed, 3 from clinical and 1 from NAD were earmarked from
a total of 110 matched spots which showed a change of ≥ 5 times. These differential exhibitions in protein profile of each category
suggest that the udder immune system behave differently according to stage and type of infection. Mass spectrometric analysis
of the proteins in differentially expressed spots can help in identifying these proteins which can further our understanding of the
host immune response to mastitis.
S K Phulia is currently working as senior scientist in the Buffalo Physiology and Reproduction division of Central Institute for Research on Buffaloes
(ICAR), Hisar, Haryana. He has earlier worked in the CIRG, Makhdoom. He has completed his master?s degree in Animal physiology from National
Dairy Research Institute, Karnal and Ph.D. from CCSHAU, Hisar. He has done extensive research in lactation physiology, ultrasonography of
mammary gland and reproductive systems in buffaloes. He is presently working in the area of milk and mammary gland proteomics and embryo
transfer technology in buffaloes.
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