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Selection of Best Performing Exotic Sheep Breed (s) and Performance Evaluation of Their Crosses with Indigenous Sheep

G. K. Deb
Bio-technology Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh.

M. Y. A. Khan
Animal production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh.

M. P. Choudhury
Bio-technology Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh.

M. Ershaduzzaman
Goat and Sheep Production Research Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh.

T. N. Nahar
Bio-technology Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh.

M. S. Alam
Bio-technology Division, Bangladesh Livestock Research Institute, Savar, Dhaka, Bangladesh.

M. A. Alim
National Institute of Biotechnology, Savar, Dhaka, Bangladesh.

Abstract/ Executive Summary:

This study was conducted to evaluate the genetic relationship among indigenous sheep populations of Bangladesh (Barind, Jamuna river basin, Coastal and Garole sheep) using microsatellite marders. A total of 96 blood samples were collected from adult sheep of (i) Barind (24), (ii) Jamuna River Basin (24), (iii) Coastal (24), iv) Garole (10) and (vi) Chotanagpuri (10) sheep of India. Chotanagpuri sheep available in the Meherpur district of Bangladesh was used as a reference breed. DNA was extracted and quantified from blood samples using commercially available DNA extraction kit. DNA was quantified using a nanodrop machine. FAO recommended 13 labeled microsattelite markers were used for polymerase chain reaction (PCR). PCR product was confirmed by running on high resolution agarose gel and visualized by staining with ethidium bromide. The exact allele sizes in each primer were determined by GeneMaker V1.85 demo. Microsatellite tool kit and Dispan software package were used for calculation of allele frequency, number of alleles per locus, observed and expected heterozygosity and genetic distances (D A). The Dispan Computer program was used to calculate inter-individual genetic distances. These distance values were used to construct an UPGMA tree. Results showed that average number of polymorphic alleles per locus was 4 in HUJ616 to 12 in MAF70. Observed heterozygosity was also varied from 0.5481±0.0488 in coastal to 0.6313±0.0303 in Barind sheep population. Genetic distance between Jamuna river basin and Barind was lowest (0.0891)and between Garole and Costal was highest (0.1786). Garole and Chotonagpuri sheep has higher genetic distance from other three sheep populations. Phylogenetic dendogram showed that sheep of Jamuna river basin and barind belong to same genetic group. Whereas, coastal, garole and Nagpur sheeps were shown higher genetic distances from Jamuna river basin and coastal sheep. Considering findings of this study it may be concluded that the Barind and Jamuna river basin sheep belongs to a similar genetic group while, Garole and coastal sheep are belonging to two distinct genetic groups.

Keywords: Dendogram, Genetic distances, Heterozygosity, Native sheep

Location: Naogaon, Rajshahi and Chapainoabgonj district.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Sheep

Objectives:

To evaluate their genetic relationship using higher number of microsattelite markers including coastal sheep.

Methodology:

Based on geographic distribution, management system and breeding history, a total of 96 blood samples were collected from five sheep population of Bangladesh. The populations were Barind (from Naogaon, Rajshahi and Chapainoabgonj district), Jamuna River Basin (from Tangail, Sirajgonj and Mymensingh district), Coastal (from Noakhali district), Chotonagpuri (from Meherpur district) and Garole (from Khulna district) breeds. Samples were collected only from adult sheep of subsistent farmers and BLRI flock. Samples were collected avoiding related animals. Blood was collected in venoject tube, treated with anticoagulant and carried to Molecular Genetics Laboratory of BLRI and preserved at -200C until DNA extraction. DNA was extracted from blood samples using a commercial kit (QIAGEN DNA Mini Kit) following manufacturer instruction. DNA samples were quantified using Nanodrop 2000 spectrophotometer. 13 microsatellite primer pairs were used according to FAG recommendation. Most of primers used were independent and belonged to different chromosomes. All the microsatellite markers were forwardly labeled with a capillary based dye: 6 FAM (blue), PET (Red), VIC (Green) and NED (yellow) for the purpose of genotyping. All the selected primers showed polymorphism in current study. Thirteen microsatellite markers used in this study along with their sequences, size range and their locations were showed. PCR amplifications were performed with a programmable thermal cycler (Bio-Rad, DNA Engin, Dyad and Tetad 2, Peltier Thermal Circlers, Mexico) in a final reaction volume of 25 µL using PCR reagents from Invitrogen. Two types of PCR reaction mixture were used. The first reaction system consisted of 2µL of 50 ng of genomic DNA, 0.5 µL of each primer, 2.5µL 10X PCR buffer, 2µL dNTPs, and 0.937 U Taq DNA polymerase [TaKaRa Biotechnology (Dalian) China] and the second one consisted of 2 µI containing 50 ng genomic DNA, 1µI of each primer, 12.5µ1 premix and 8.5 µI ddH₂O. The amplification conditions were as follows: 5 min at 94°C for initial denaturing followed by 35 cycles at 94ºC for 30 s; annealing at Tm (ºC) for 30 s; extension at noc for 30 s and a final extension at 72°C for 7 min for all primers. Amplification was confirmed by gel electrophoresis of the PCR products on 2% agarose gel followed by visualization under UV. PCR products with different florescent dye were subjected to capillary electrophoresis (Life Biotech Ltd. China). The exact allele sizes in each primer were determined by GeneMaker V1.85 demo. Genotypes were assigned for each animal based on allele size data. Allele frequency, the mean number of alleles per locus, observed heterozygosity, and heterozygosity expected from Hardy-Weinberg assumptions for each locus, and Nei's D_A genetic distances were computed using the Microsatellite tool kit and Dispan software package. The Dispan Computer program was used to calculate inter-individual genetic distances, based on the proportion of shared alleles. These distance values were used to construct an unweighted pair group-method with arithmetic mean (UPGMA) phylogenetic tree using Dispan software.

Source of Information: Proceedings of the Annual Research Review Workshop 2013-2014, BLRI, Savar, Dhaka. Publication No.261, pp: 90-101, October-2015.

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Considering findings of the present study on average allele numbers, heterozygosity and standard genetic distances and dendrogram, it may be concluded that the Barind and Jamuna river basin sheep are belonging to a similar genetic group while, Garole and Coastal sheep are two distinct genetic groups.

Knowledge Management: Report/Proceedings