Molecular characterization of lactoferrin gene as genetic marker to subclinical mastitis in water buffaloes (Bubalus bubalis)

The study characterized the lactoferrin (Lf) gene in different water buffalo breeds and determined its association with subclinical mastitis (SCM). A total of seventy five (75) and five (5) milk and blood samples, respectively were collected for the conduct of this study. Amplified Lf gene demonstrated a 2224 base pairs (bp) molecular weight. Nucleotide and amino acid sequence of Lf gene of riverine- and swamp-type water buffaloes revealed 98.83% and 98.29% identity, respectively with that of Bubalus bubalis Lf gene in the GenBank. Phylogenetic studies showed that Lf genes of both types of water buffaloes grouped with Lf gene of water buffalo sequences registered in the GenBank. Three water buffalo genotypes were documented using the restriction enzymes AluI and HaeIII. The AluI can produce three genotypes (AA, AB, BB) resulting in three cuts of 561, 217 and 123 bp. Using HaeIII, three restriction patterns were observed producing three genotypes as well with four fragment sizes of 561, 318, 112, and 70 bp. Based on the statistical analysis, no direct association has been established between the Lf genes of water buffalo with SCM using AluI and HaeIII restriction enzymes although a higher number of animals with genotype BB belongs to those with SCM group. The results of the study merit for further studies regarding the BB genotype of Lf gene as a possible resistance gene marker for SCM in water buffaloes.