doi:10.5455/vetworld.2013.479-481 Prolificacy in Raighar goats is independent of FecB gene

Aim: The Research was undertaken to find the association between FecB and high prolificacy in Raighar goats. 
 
Materials and Methods: DNA was extracted from blood, collected from does (n=101) with history of high prolificacy. 
Further tetra-primer amplification refractory mutation system (T-ARMS) PCR and agarose gel electrophoresis were followed 
to screen the mutation. 
 
Results: Raighar goats were found to be wild homozygotes suggesting absence of FecB mutation. 
 
Conclusion: Prolificacy in case of Raighar goats is not due to the mutation at FecB locus. It is thought to search for other 
genes or loci in goat fecundity.


Introduction
Odisha and are well known for their high prolificacy with frequent twinning and triplet kidding except first Goat farming is relatively easy and profitable as kidding which is usually single type.goats are well adapted to diverse environments.In The aim of current research was to find out the commercial farming the benefit is mainly dependent contribution of an established ovine fecundity gene upon the litter size.In mammals, multiple litter size is like FecB towards high prolificacy of Raighar goat by due to multiple ovulations.Though twinning is common analyzing its polymorphism.in goats but less frequent is occurrence of quadruplet, quintuplet, and sextuplet kids.As the heritability of

Materials and Methods
litter size is very low, selective breeding as a route will Blood samples (n=101) were collected in sterile be a slow process for developing the reproductive vacutainer containing K -EDTA from Raighar does 3 performance of low prolific goat breeds [1].In with history of multiple births over different regions of alternative, introduction of a prolificacy gene into non-Nabarangpur district of Odisha as per the guidelines of prolific goat breeds having other desired traits may IAEC (Institutional Animal Ethics Committe) and effectively increase their reproductive performances transported to the laboratory under refrigeration.The [1,2].
genomic DNA was isolated from white blood cells Genetic mechanism of caprine prolificacy using standard phenol-chloroform protocol [10] and remains to be fully explored [3].Relationship between dissolved in TE buffer before storing at -20ºC for ovulation rate and litter size was initially studied in further use.For detection of FecB point mutation, sheep and it was reported that a set of genes were regulating Tetra-primer amplification refractory mutation system this complex phenomenon known as Fecundity (Fec) Polymerase Chain Reaction (T-ARMS-PCR) was genes [4,5].Inheritance of twinning and triplet followed [7].Two sets of primers were used; forward tendency is similar in both sheep and goats [3].Fec outer primer (5'-GTCGCTATGGGGAAGTTTGGA genes belong to transforming growth factor â (TGF â) TGGGAA-3'), reverse outer primer (5'-CCCGTCC CT super family [6,7] and among Fec genes BMPR1B, TTGATATCTGCAGCAATG-3'), inner forward (Bone morphogenetic protein receptor 1B, also known primer for A allele (5'-GCTGGTTCC GAGAGACA G as Booroola or FecB or Activin like Kinase 6) present AAATATAGCA-3') and inner reverse primer for G on 6th chromosome of sheep is the first to be identified allele (5'-ATGTTTTCA TGCCTCAT CAACA CC GA contributing towards increase in ovulation rate [8] CC-3').These primers were previously used for invesensuing greater prolificacy in Merino sheep [4,9].tigation of FecB mutation in Black bengal goat [9,11].The Raigarh Goats are popular meat animals in PCR was done with reaction mixture of 50 µl containing primers 0.5 µl each; Template DNA 2.0 µl; dNTPs (10mM) 1.0 µl; 10X Taq polymerase buffer 5 µl; 25mM MgCl 3 µl; Taq DNA polymerase 2 (Promega, USA, 5U/µl) 0.5 µl and rest volume was the monomorphic status of Raighar goat with respect to phoretogram is presented in Fig. 1.From the figure it the gene.FecB mutation was also not observed in Boer, was found that all the tested animals represented Haimen, Huanyhuai, Nubi and Matou goat breeds amplification of common outer PCR product that was having the litter size varied from 1.4 to 2.7 [3].This of the amplicon size 1100 bp.This product size defined suggests there may be some other genes contributing the wild homozygote genotype.There was absolutely toward caprine fecundity other than BMPR1B.no inner band of any product size to describe the mutant Similarly Hu, the most famous high prolific sheep in genotypes.All goats were found to be wild homozygote China is also wild homozygous for BMPR1B [19].It suggesting absence of FecB mutation.Amplicon size validates involvement of different ovine fecundity of 136 bp would have suggested the presence homozygous genes in different sheep breeds viz BMP15 and GDF9 mutant genotype whereas; concurrent bands of 1100 contributing to greater prolificacy in small tailed Han and 136 bp proposed genotype of heterozygous mutant ewes [20] and Brazilian SI sheep [21] respectively.type.
The prolificacy in Raighar goats may not be due to this established ovine fecundity gene.From above Discussion findings it is clear that there may be other genes or loci The careful regulation of the number of eggs shed in goat fecundity.Recent findings suggest the involveand hence the litter size is crucial to successful ment of INHA gene [22] and growth hormone gene reproduction in all species of animals [5].High [23] in increase litter size in goat-breeds.prolificacy in some sheep breed like Kendrapada of

Conclusion
India is due to mutation in BMPR1B gene [12].Number of mutations in BMPR1B gene is directly proportional FecB gene has no affinity for greater prolificacy to litter size and ovulation rate [7,13].This increase in in Raighar goats of Odisha.The high prolificacy, which ovulation rate of FecB carriers is associated with a was evident from the collected history during the precocious maturation of many of antral follicles that course of sample collection is may be due to some other may undergo ovulation with a smaller size [7,13].
genes which are yet to be explored or may be due to In goat breeds there is no such report of FecB gene some other factors.being responsible for high prolificacy except in Black Authors' contribution bengal goat.Prolificacy in Black bengal is due to FecB TKP carried out sample collection, experimental mutation of BMPR1B gene [11].In this context, execution, preparation of draft and revision of polymorphism study of FecB may offer a route to manuscript.PC Bisoi and SD participated in explore the fecundity in Raighar goats as it was the only conception and designing of experiment, trouble established ovine fecundity gene responsible for shooting and final analysis of result.AM helped in fecundity in one goat breed.
Sample collection and draft preparation, SP assisted in For fecundity gene analysis, restriction fragment experimental execution and interpretation of findings, length polymorphism [14], allele specific oligonucleotide doi:10.5455/vetworld.2013.479-481adjusted with nuclease free water.Touchdown cycling melting typing [15], primer extension assay [16] and Tconditions were followed for amplification [11].The ARMS-PCR [7] are some of the single nucleotide PCR amplified product was analysed by 2% agarose polymorphism (SNP) typing techniques commonly gel along with DNA molecular weight marker.used.T-ARMS-PCR is a simple, rapid and economical method for SNP scoring [11, 17] so followed in the Results current study.It combines the principle of ARMS and Three possible genotypes due to A and G alleles of the tetra-primer PCR method which can amplify both BMPRIB gene are AA (wild type), GG (Homozygous wild-type and mutant alleles as well as the control mutant) and AG (Heterozygous mutant).For screening fragment in a single-tube PCR reaction [18].FecB of genotypes in Raighar goats (n=101), FecB polymorpolymorphism studied through T-ARMS PCR reported phism was studied through T-ARMS-PCR and electro-