EFFECTS OF PROTEIN SUPPLEMENTS ON FERTILITY AND PHYLOGENIC INFERENCE OF FERTILITY GENES IN INDIGENOUS SHEEP OF BANGLADESH

Abstract

The study was carried out to know the effects of protein supplementations on fertility and inference of fertility genes. Three iso-caloric but different graded levels of protein containing rations were feed to three different groups of sheep. For molecular study, DNA was extracted from the whole blood sample for PCR of GDF9 and BMP15 fertility genes and purified PCR products were sequenced with the help of Sanger sequencer. Sequence alignment, pair and multi-alignment comparison of the BMP15 and GDF9 gene were performed with MEGA6 software. The semen volume, sperm counts, percentages of normal and viable sperm were higher in Treatment 2 than the other two treatments. Ewes treated with supplemented, protein concentrate reached conception at earlier age (Treatment 1, 7.4±0.09 months and Treatment 2, 6.67±0.18 months) than control (8.37± 0.15 months). Conception rate was significantly higher in treated groups than control group. The gestation length and lambing interval were significantly higher in control (155.25±4.1 days, 188.25±2.3 days, respectively) than treated groups. Lamb’s birth weights of three groups were ranged from 1.11 to 1.55 kg. The designated sequences of BMP15 gene and GDF9 gene were revealed 100% homology with the sequence of Pelibuey sheep. In addition, the sequences of BMP15 gene showed 99% similarity with the Balochi and Lohi breed of sheep and GDF9 gene asserted that sequences were 95% harmonious with the Awassi breed. The evolutionary divergences in indigenous sheep ranged from 0.00 to 3.35 for BMP15 gene and 0.00 to 0.77 for GDF9 gene with clear differences within indigenous sheep. In the phylogenetic tree nucleotide sequence of BMP15 was closely related with Pelibuey sheep and Rahmani sheep breed had strong association with GDF9. Jukes-Cantor (JC) model designed with the 3866.36 BIC score and Kimura 2-parameter model carried 6222.80 BIC score were the best fitted models for BMP15 gene and GDF9 gene, respectively. The overall nucleotide frequencies of BMP15 were 27.87% (A), 26.34% (T/U), 21.70% (C), and 24.10% (G) and GDF9 were 20.22% (A), 26.32% (T/U), 27.91% (C) and 25.54% (G), respectively. The study indicated that the influence of nutrition on reproductive performance and these genomic inceptions will be open a new era in the indigenous sheep production of Bangladesh.