Petya VELEVA, Deyana GENCHEVA, Darina PAMUKOVA, Nikolina NAYDENOVA

Genetic polymorphism of alpha S1-casein in Bulgarian sheep breeds and its effect on milk composition

The genetic polymorphism of the alpha S1-casein (CSN1S1) was investigated in five sheep breeds reared in Bulgaria: Sofia (Еlin-Pelin, SEPL), Copper–Red Shumen (CRSH), Local Karnobat (LKNB), Pleven Blackhead (PLBH), and Stara Zagora (STZG) sheep with an aim to establish the possible effect of a particular genotype on ovine milk composition. Based on nucleotide variation in exon III of the CSN1S1gene, two genetic variants (A and C) and three genotypes (AA, AC, and CC) have been identified using PCR–RFLP analysis on a total of 217 unrelated ewes. The allele frequencies determined a prevalence of the allele C (0.886) over the allele A (0.114) across the studied sheep populations. The homozygous CC genotype was observed in nearly 80% of the studied ewes. The calculated values for observed (Hо = 0.548) and expected (He = 0.468) heterozygosity at CSN1S1 locus indicated a relatively high degree of genetic variability in the Sofia sheep population. The greatest Nei’s genetic distance (DA = 0.080) was found between the populations STZG and SEPL, while the closest relationship was established (DA = 0.000) between PLBH and CRSH, also between STZG and CRSH. The results of the association analysis indicated that CSN1S1 AC genotype was significantly associated (P < 0.05) with the highest percentages of the fat, protein, casein, solids-nonfat and total solids in Sofia sheep ewes. The genotype CSN1S1 CC was associated with the highest noncasein protein percentage, while the genotype AA was linked with the highest lactose percentage. The CSN1S1genotype did not show a significant effect (P > 0.05) in the Sofia sheep population in relation to the renneting time. In conclusion, the established single nucleotide polymorphism in the CSN1S1 locus could be used as a potential genetic marker for ovine milk composition traits, as well as for developing an effective conservation strategy towards traditional sheep breeds in the country

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1. Petrović MP, Selionova MI, Caro-Petrović V, Ružić-Muslić D, Maksimović N et al. The application of modern molecular techniques in animal selection. Biotechnology in Animal Husbandry 2018; 34 (4): 371-386.
2. Frajman P, Dovc P. Milk production in the post-genomic era. Acta Agriculturae Slovenica 2004; 84 (2): 109-119.
3. Ceriotti G, Chiatti F, Bolla P, Martini M, Caroli A. Genetic variability of the ovine alpha S1-casein. Italian Journal of Animal Science 2005; 4: 64-66.
4. Hristov PI, Teofanova DR, Mehandzhiyski ID, Zagorchev LI, Radoslavov GA. Significance of milk protein genes polymorphism for Bulgarian Rhodopean cattle: comparative studies. Biotechnology & Biotechnological Equipment 2013; 27 (2): 3659-3664.
5. Mroczkowski S, Korman K, Erhardt G, Piwczynski D, Borys B. Sheep milk protein polymorphism and its effect on milk performance of Polish Merino. Archiv fur Tierzucht 2004; 47 (6): 114-121.
6. Carillier-Jacquin C, Larroque H, Robert-Granié C. Including αs1 casein gene information in genomic evaluations of French dairy goats. Genetics Selection Evolution 2016; 48: 54. doi: 10.1186/s12711-016-0233-x
7. Park Y, Juarez M, Ramos M, Haenlein G. Physico-chemical characteristics of goat and sheep milk. Small Ruminant Research 2007; 68: 88-113.
8. Rampilli M, Cecchi F, Giuliotti L, Cattaneo TM. The influence of β-lactoglobulin genetic polymorphism on protein distribution and coagulation properties in milk of Massese breed ewes Milk protein polymorphism. International Dairy Federation, Brussels, Belgium; 1997. pp. 311-315.
9. Pirisi A, Piredda G, Papoff C, Di Salvo R, Pintus S et al. Effects of sheep alpha S1-casein CC, CD and DD genotypes on milk composition and cheesemaking properties. Journal of Dairy Research 1999; 66: 409-419.
10. Amigo L, Recio I, Ramos M. Genetic polymorphism of ovine milk proteins: its influence on technological properties of milk-a review. International Dairy Journal 2000; 10: 135-149.
11. Othman OE, El-Fiky SA, Hassan NA, Mahfouz ER, Balabel EA. Genetic variations of β-and K-casein genes in Egyptian sheep breeds. Journal of Applied Biosciences 2013; 64 (1): 4858-4866.
12. Othman OE, El-Fiky SA, Hassan NA, Mahfouz ER, Balabel EA. Genetic polymorphism detection of two α-Casein genes in three Egyptian sheep breeds. Journal of Genetic Engineering and Biotechnology 2013; 11 (2): 129-134.
13. Clement P, Agboola S, Bencini R. A study of polymorphism in milk proteins from local and imported dairy sheep in Australia by capillary electrophoresis. LWT-Food Science and Technology 2006; 39: 63-69.
14. Chessa S, Rignanese D, Berbenni M, Ceriotti G, Martini M et al. New genetic polymorphisms within β- and αS2-caseins. Small Ruminant Research 2010; 88: 84-88.
15. Ferranti P, Malorni A, Nitti G, Laezza P, Pizzano R et al. Primary structure of ovine alpha S1-caseins: localization of phosphorylation sites and characterization of genetic variants A, C and D. Journal of Dairy Research 1995; 62 (2): 281-296.
16. Chianese L, Caira S, Garro G, Addeo F. Primary structure of ovine deleted variant αs1-CN E. In: Proceedings of 5th International Symposium on the Challenge to Sheep and Goats milk sectors. Alghero/Sardinia, Italy; 2007. p. 70.
17. Giambra IJ, Brandt H, Erhardt G. Milk protein variants are highly associated with milk performance traits in East Friesian Dairy and Lacaune sheep. Small Ruminant Research 2014; 121 (2-3): 382-394.
18. Moioli B, Pilla F, Tripaldi C. Detection of milk protein genetic polymorphisms in order to improve dairy traits in sheep and goats: a review Small Ruminant Research 1998; 27: 185-195.
19. Scintu M, Piredda G. Typicity and biodiversity of goat and sheep milk products. Small Ruminant Research 2007; 68: 221-231.
20. Barillet F. Genetic improvement for dairy production in sheep and goats. Small Ruminant Research 2007; 70: 60-75.
21. Pilla F, Bevilacqua C, Leroux C, Fraghi A, Martin P. Genotyping of alpha-S1 casein in sheep. Animal Genetics 1998; 29: 472-473.
22. Corral J, Padilla J, Izquierdo M. Associations between milk protein genetic polymorphisms and milk production traits in Merino sheep breed. Livestock Science 2010; 129: 73-79.
23. Barbano DM, Dellavalle ME. Rapid method for determination of milk casein content by infrared analysis. Journal of Dairy Science 1987; 70: 1524-1528.
24. Chomakov H, Velev S, Dimitrov T, Iliev T, Miteva C et al. Milk and Milk Products. Univers; 2000. pp. 84-85 (in Bulgarian).
25. Excoffier L, Lischer HE. Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Molecular Ecology Resources 2010; 10 (3): 564- 567.
26. Yeh F, Yong R. POPGENE version 1.31 (02.04. 2011). Microsoft based Freeware for Population Genetic Analysis. University of Alberta, Edmonton, Canada; 1999.
27. Labate JA. Software for population genetic analyses of molecular marker data. Crop Science 2000; 40 (6): 1521-1528.
28. Nei M. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 1978; 89 (3): 583-590.
29. Guo SW, Thompson EA. Performing the exact test of Hardy– Weinberg proportion for multiple alleles. Biometrics 1992; 361- 372.
30. SPSS Statistics 17.0.0 WinWrap Basic, Copyright 1993-2007 Polar Engineering and Consulting.
31. Russo V, Fontanesi L, Scotti E, Beretti F, Davoli R et al. Single nucleotide polymorphisms in several porcine cathepsin genes are associated with growth, carcass, and production traits in Italian Large White pigs. Journal of Animal Science 2008; 86 (12): 3300-3314.
32. Stuber CW, Edwards MD, Wendel JF. Molecular-marker facilitated investigations of quantitative trait loci in maize. II Factors influencing yield and its component traits. Crop Science 1987; 27: 639-648.
33. Executive Agency for Selection and Reproduction in Animal Breeding. Livestock Breeds in the Republic of Bulgaria Executive Agency for Selection and Reproduction in Animal Breeding, Catalogue, 5th ed. In: Nikolov V (editor), Sofia, Bulgaria; 2017. pp. 80-81.
34. Dimitrov Ts, Dimitrova I. Preservation of Livestock genetic resources in Bulgaria, In: Boyazoglu J and Chupin D, (editors). Animal Genetic Resources Information bulletin; 1994. pp. 41- 59.
35. Stankov I, Slavov R, Panayotov D, Pamukova D. State and perspective for development of sheep breeding in Republic of Bulgaria. Sheep breeding in Bulgaria and in Europe, In: Proceedings of scientific conferences, Kirilov A (editor), Sofia; 2007. pp. 23-32, (In Bulgarian).
36. Dimitrova I, Stantcheva N, Georgieva S, Nakev G, Genova K et al. Sheep (Ovis aries L.) as object of genome research. Science & Technologies. Animal studies & Veterinary medicine 2013; 3 (5): 43-50 (in Bulgarian with an abstract in English).
37. Kolev A, Nedelchev D, Dimov G, Popova M, Stoyanov S. Sofia (Elin Pelin) local sheep breed-state and tendencies. Agricultural Science 2011; 3 (6): 25-30 (in Bulgarian with an abstract in English).
38. Levéziel H, Méténier L, Guérin G, Cullen P, Provot C et al. Restriction fragment length polymorphism of ovine casein genes: Close linkage between the αS1-, αS2-, β- and κ-casein loci and κ-casein loci. Animal Genetics 1991; 22: 1-10.
39. Di Gregorio P, Rando A, Pieragostini E, Masina P. DNA polymorphism at the casein loci in sheep. Animal Genetics 1991; 22: 21-30.
40. Ordás JG, Rando A, Senese C, Masina P. DNA polymorphisms of casein genes in Spanish dairy sheep. Small Ruminant Research 1997; 26: 9-13.
41. Erhardt G. Evidence for a third allele at the b-lactoglobulin (bLg) locus of sheep milk and its occurrence in different breeds. Animal Genetics 1989; 20: 197-204.
42. Teneva A. Genetic polymorphism of proteins in the milk of Local sheep breeds and strains. Genetics and selection 1993; 26 (4): 301-305 (in Bulgarian with an abstract in English).
43. Hristova D. Genetic polymorphism of alpha S1-casein gene in Bulgarian sheep breeds. Agricultural Science and Technology 2011; 3 (1): 8-12.
44. Gencheva D, Georgieva S. Genetic diversity and population structure of Bulgarian autochthonous sheep breeds base on nucleotide variation in Alpha S1-casein gene. Bulgarian Journal of Agricultural Science 2019; (Suppl. 3): 95-102.
45. Ramos A, Matos C, Russo-Almeida P, Bettencourt C, Matos J et al. Candidate genes for milk production traits in Portuguese dairy sheep. Small Ruminant Research 2009; 82: 117-121.
46. Rustempasic A, Dokso A, Zecevic E, Hodzic A, Bandzo K et al. Polymorphism of CSN1S1 gene in Pramenka breed sheep in Bosnia and Herzegovina. In: Proceedings of the 24th International Scientific-Expert-Conference of Agriculture and Food Industry, Sarajevo, Bosnia and Herzegovina, 2013 Faculty of Agriculture and Food Sciences, University of Sarajevo; 2013. pp. 91-94.
47. Kevorkian S, Manea M, Gavrila M, Rebedea M, Georgescu S et al. Sequencing of exon three of αs1- casein sheep and goad gene. Archiva Zootechnica 2009; 12 (3): 87-92.
48. Piredda G, Papoff CM, Sanna SR, Campus RL. Influenza del genotipo della αS1-caseina ovina sulle caratteristiche chimicofisiche e lattodinamografiche del latte. Scienza e Técnica Lattiero-Casearia 1993; (44): 135-143 (in Italian).
49. Serrano Moyano B, Garzón Sigler AI, Garro G, Chianese L, Martínezn Hens. Variabilidad genética de caseínas en la raza ovina Merina. Archivos de Zootecnia 1999; 48: 197-206 (in Spanish).
50. Martini M, Salari F, Scolozzi C, Cecchi F, Ceriotti G et al. Relationship between milk genetic polymorphism and physicochemical and nutritional quality of sheep milk. In: 14th International Congress of Fe. Me. SP Rum, Lugo, Santiago de Compostela, Spain; 2006.
51. Martini M, Salari F, Scolozzi C, Cecchi F, Ceriotti G et al. Relationship between milk genetic polymorphism and phisico-chemical and nutritional quality of sheep milk. Small Ruminant Research 2008; 74: 194-201.