ISOLATION OF LACTOFERRIN IN MILK OF CROSSBRED AND INDIGENOUS BREEDS OF CATTLE AND EVALUATION OF ITS ANTIMICROBIAL ACTIVITY
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Date
2021-12-03
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PVNRTVU, HYDERABAD
Abstract
Lactoferrin (Lf) is a multifunctional iron binding glycoprotein, which exerts a broad-spectrum primary defense activity against bacteria, fungi, protozoa and viruses. Lf from bovine milk (BLf) can be mainly used in medicinal activity because of its easy availability. In the present study isolation, quantification and evaluation of the antimicrobial activity of isolated BLf from the milk samples of HF cross bred and indigenous breeds of cattle i.e. Sahiwal and Poda thurpu was done and the above parameters were compared among the mentioned breeds.
Milk samples were collected from exotic cross bred like HF cross bred and native breeds like Sahiwal and Poda thurpu breeds of cattle. Chemical analysis was performed and physico-chemical characteristics like pH, specific gravity, fat, SNF, lactose and protein were compared between the three breeds. It was found from the above analysis that Poda thurpu breed of cattle showed highest fat, SNF, Lactose and protein content significantly (P < 0.05) when compared to other two breeds whereas pH and specific gravity were not significant among the three breeds. The BLf concentration was also measured using ELISA. The concentration of BLf was also significantly (P < 0.05) highest in Poda thurpu breed of cattle when compared to HF cross bred and native breed like Sahiwal. The BLf protein was then purified using the
cation-exchange chromatography method. The eluted fractions were then subjected to SDS-PAGE along with a standard protein marker to confirm the molecular weight of isolated protein. The eluted fractions of all the three breeds showed the band of 80 kDa when compared to standard marker.
Finally, the antimicrobial activity was studied using broth microdilution and disc diffusion method. BLf isolated from milk of indigenous breeds showed higher anti bacterial activity as evidenced by their lower MIC values, compared to that of HF cross bred. Further highest zone of inhibition was obtained for isolated BLf from the milk of Poda thurpu followed by Sahiwal and HF cross bred against selected gram positive and gram negative bacteria.
From this study, it was concluded that Bovine lactoferrin (BLf) isolated from Poda thurpu breeds of cattle showed the more potent antimicrobial activity followed by Sahiwal cow milk against selected gram positive and gram negative bacteria. The antimicrobial activity was least in HF cross bred cow milk when compared to indigenous cow breeds such as Sahiwal and Poda thurpu