Phenotypic, morphometric and genetic characterization of Khukhri, an indigenous chicken of Jharkhand

Introduction

In Jharkhand, indigenous poultry farming, also known as backyard poultry, is a widespread and culturally significant practice, particularly among the rural and tribal populations. Farmers raise native breeds, along with some improved varieties, using low-input, free-range systems that provide nutritional security and supplemental income. While producing fewer eggs than commercial breeds, Khukhri meat and eggs are highly prized in local markets for their flavour. The native birds are slow grower and poor layers of small sized eggs but are good foragers, efficient mother, good sitter and require minimal care to grow. Their plumage color, long neck and long shank helps in protecting themselves against predators. The native breeds of chickens are the reservoir of genomes and major genes for improvement of high yielding exotic germplasm for tropical adaptability and disease resistance. In developing countries, the major threat to genetic diversity is due to introduction of high yielding germplasm in poultry sector by indiscriminate cross breeding of local breeds with less adapted exotic germplasm to evolve highly productive breed which adversely affect the conservation of the local ecology. Keeping in view the above facts the present study has been planned with objectives to characterize all the native breeds to find out the uniqueness among them so that those traits can be exploited to design breeding and conservation strategies (Prabhakaran and Valavan, 2020) [16] . Phenotypic and genotypic Genetic characterization of chicken is done to assess the origin of birds and the geographic distribution of their diversity by using different types of molecular markers. It provides new information to guide and prioritize conservation decisions for poultry as locally adapted chicken genetic resources could become future assets in breeding programs and created new possibilities for the selection and rapid genetic improvement of chicken.

Materials and methods

The present study was conducted to survey, evaluate the phenotypic and molecular characterization of Khukhri chickens. The survey was conducted on 270 households and 540Khukhri chicken populations of Chotanagpur plateau of Jharkhand. Thirty blood samples were collected from 50 unrelated native chickens (2 ml each) from the wing veins. RAPD analysis was conducted with The 22 decamer primers (Table 1) of arbitrary sequences with high GC content (60-70%) were used. The amplifications were carried out in final volume of 25 µl containing 5 µl genomic DNA, 2.0 µl DNTPs (2.5mm), 1 µl primer (10pm), 0.25 µlof Taq polymerase and 2.5 µl of 10X reaction buffer with MgCl2. Amplifications were performed on DNA thermal cycler. Presence of band was designated as '1' and absence as '0'.

The matrix data obtained was fed to the POPGENE software (version 1.32) and the results were obtained for observed and effective number of alleles, Nei's gene diversity, Shannon's Information index, gene identity and genetic distance. Genetic diversity within each population was determined as the observed and expected number of alleles (Kimura and Crow, 1964) [11] , Nei's genetic diversity and Shanon's Information Index (Lewontin, 1972) [12] using Popgene software (Yeh et al, 1999) [21] .

Results and discussion

The overall value regarding predominant plumage colour were observed to be mix and red followed by brown, white, black and white, shank colour observed to be mostly yellow, ear lobe red, skin colour white, comb red and comb type mostly single followed by pea and rose (Table 1) Phenotypic characteristics was in accordance to the findings of Tantia et al. (2005) [20] in Kashmir Faverolla birds, Banerjee (2012) [4] in native chickens of West Bengal. The overall average body weight of male and female chicken was 1552.18±27.46 and 1182.04±14.30 g. Shank length observed to be 9.85±0.08 and 8.73±0.05 respectively for male and female (Table 1). The lesser frequencies were black (10.00%), brown (8.33%) and white (8.33%) (Table 2) Out of 22 only 16 primers were capable of generating polymorphic amplification patterns and hence, they were used in the subsequent analysis with 50 DNA samples. A total of 172 bands were generated from these primers and of these, 168 bands (97.674%) were polymorphic. It was observed that primer OPA-16 amplified maximum number (16) of polymorphic bands and PBG-5 amplified lowest number (5) of polymorphic bands. Among the 16 primers used, three-fourth (75%) were highly polymorphic. The average number of amplified bands per primer ranged from 2.2 to 7.46 and the size of the amplified bands varied between 100 to 3000 base pairs (Table2).The per cent polymorphism observed in the present study was in close agreement with the findings of Saleem (2019) who reported 96.96% polymorphism in three Indian breed of poultry. However lower percent of polymorphism was documented by Jallad et al. (2013), Baloza et al. (2013) [3] , Ibrahim et al. (2015) [8] , Abdulrazaq and Suliaman (2016) [1] , Shivaprasad et al. (2016) [19] , Fadhil et al. (2016) [6] , Gorkhail et al. (2020) [7] and Sang-Hwan Kim, (2021) [9] . However, Karabag and Balcioglu, (2010) [10] observed higher polymorphism (99.49%) than the present finding in Japanese quail.

Acknowledgement

AcknowledgementsThe authors are thankful to the Ranchi College of Veterinary Science, Birsa Agricultural University and ICAR-AICRP on Poultry Breeding for their support in generating records and providing facilities for the current study.

References

1. Abdulrazaq HS, Ali Suliaman NM. Polymorphism between five local chickens in Erbil using RAPD. ZANCO J Pure Appl Sci. 2016;58(1):65-69.
2. Ahmad LS, Al-Barzinji YMS. Genetic diversity among local quail using RAPD-DNA marker. Rev Cient Vet. 2019;29(3):178-185.
3. Baloza SH, Hemeda SA, Sosa GA, Shady AA. Assessment of genetic variability among Fayoumi, Rhode Island Red and their crosses using RAPD-PCR. Benha Vet Med J. 2013;25(2):232-239.
4. Banerjee S. Morphological characterization of indigenous chickens of Sikkim and West Bengal, India. Anim Genet Resour. 2012;51:57-71.
5. Dehghanzadeh H, Mirhoseini SZ, Romanov MN, Ghorbani A. Evaluation of genetic variability and distances among five Iranian native chicken populations using RAPD markers. Pak J Biol Sci. 2009;12(11):866871.
6. Fadhil IA, Dakheel MH, Hussein TH. Detection of genetic diversity through two poultry breeds using RAPD-PCR technique. J Babylon Univ Pure Appl Sci. 2016;24(9):2661-2668.
7. Gorkhail NA, Dhakal R, Sapkota S, Koirala P, Pokhrel P, Pokhrel BR, et al. Polymorphism of Sakini chicken population from different locations/altitudes of Nepal International Journal of Advanced Biochemistry Research https://www.biochemjournal.com using RAPD markers. SAARC J Agric. 2020;18(2):115-124.
8. Ibrahim AM, Sabry AM, Hassan MM, El-Hallous EI, Mohamed AA. Genetic characterization of local chicken from Taif region in Saudi Arabia using RAPD markers. Int J Biosci. 2015;6(10):142-148.
9. Kim SH. Identification of genetic association among different colors of Korean native chicken breeds through RAPD-PCR method. J Anim Health Prod. 2021;9(1):33-39.
10. Karabag K, Balcioglu MS. Genetic diversity among selected Japanese quail (Coturnix coturnix japonica) lines using RAPD markers. J Appl Anim Res. 2010;38:149-152.
11. Kimura M, Crow JF. The number of alleles that can be maintained in a finite population. Genetics. 1964;49:725-738.
12. Lewontin RC. The apportionment of human diversity. Evol Biol. 1972;6:381-398.
13. Nei M. Genetic distance between populations. Am Nat. 1972;106:283-292.
14. Nei M. Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci USA. 1973;70:33213323.
15. Olowofeso O, Wang JY, Zhang P, Dai GJ. Genetic analysis of Haimen chicken populations using decamer random markers. Asian Australas J Anim Sci. 2006;19(11):1519-1524.
16. Prabhakaran R, Valavan ES. Breeding strategies for native chicken production in India. J Entomol Zool Stud. 2020;8(3):193-199.
17. Radwan LM, Mahrous MY, El-Denary ME. Assessing genetic variability in egg production and egg quality traits of four Egyptian local strains of chickens using RAPD and SSR techniques. Egypt J Anim Prod. 2015;52(1):55-61.
18. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press; 1989.
19. Tantia MS, Ganai N, Vij PK, Vijh RK, Ahlawat SPS. Chicken breeds of India: Kashmir Faverolla. Karnal: National Bureau of Animal Genetic Resources; 2005.
20. Yeh FC, Boyle T, Rongcai Y, Ye Z, Xian JM. POPGENE version 3.1. Edmonton: University of Alberta; 1999.