Document Type : Research Paper
Authors
1 Graduated of master of Genetics and Animal Breeding of Sari Agricultural Sciences and Natural Resources University, Sari,
2 Professor, Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University
3 Associate Professor, , Department of Animal Science, Faculty of Animal Science and Fishery, Sari Agricultural Sciences and Natural Resources University, Sari
Abstract
The aim of this study was to identify the allelic variants of C4-A and LF genes and to investigate their associations with the nunber of milk somatic cells in Holstein cows. Mastitis is one of the most common diseases in dairy cows, that reduces milk production and imposes high costs on breeders. C4-A and lactoferrin genes are amonge the genes that affect the immune system to fight pathogens, microbial agents and mastitis. 384 blood samples were prepared from Holstein cows and DNA extraction was performed using optimized salting out method. IPLEX technique was used to determine the genotype of the samples. To identify single nucleotide polymorphisms, the marker sites of 1-rs137485678; C<G and 2-rs132741478; C<T for C4- A complement gene and 1-rs384176726; A<G and 2-rs445918028; A<T were selected for the LF gene. Genotyping at 1-rs137485678; C<G locus of C4-A gene showed two C and G alleles with frequencies of 58 and 42 %, and also three genotypes CC, CG and GG with frequencies of 31, 53 and 16%, respectively. No polymorphisms were observed at the other sites. Marker-trait analysis showed a statistically significant association (P<0.05), so that cows with CG genotype had the lowest number of somatic cells. Phase software was used to identify haplotypes. Based on the results of this research, the marker rs137485678; C <G-1 of the C4-A gene can be used as a genetic marker to improve mastitis in dairy cows.
Keywords
and Edriss MA (2016) Single nucleotide
polymorphism of the lactoferrin gene and its
association with milk production and
reproduction traits in Iranian Holstein cattle.
Journal of Livestock Science and
Technologies, 4 (1): 71-76. (In Persian)
2. Avanus K and Altinel A (2017) Comparison of
allele-specific PCR, created restriction-site PCR,
and PCR with primer-introduced restriction
analysis methods used for screening complex
vertebral malformation carriers in Holstein cattle.
Journal of Veterinary Science, 18 (4): 465-470.
3. Barlow J (2011) Mastitis therapy and
antimicrobial susceptibility: a multispecies
review with a focus on antibiotic treatment of
mastitis in dairy cattle. Journal of Mammary
Gland Biology, 16 (4): 383-407.
4. Cecchinato A, Chessa S, Ribeca C, Cipolat-
Gotet C, Bobbo T, Casellas J and Bittante G
(2015) Genetic variation and effects of
candidate-gene polymorphisms on coagulation
properties, curd firmness modeling and acidity
in milk from Brown Swiss cows. Journal of
Animal Science, 9(7): 1104-1112.
5. Cho CI, Alam TJ, Choy JG, Choi SS and Cho
KH (2015) Models for estimating genetic
parameters of milk production traits using
random regression models in Korean Holstein
cattle. Asian-Australas Journal of Animal
Science, 29 (5): 607-614.
6. Gunther J, Koczan D, Yang W, Nurnberg G,
Gunther J, Koczan D, Yang W, Nurnberg G,
Repsilber D, Schuberth H-J, Park Z, Maqbool N,
Molenaar A and Seyfert S (2009). Assessment of
the immune capacity of mammary epithelial
cells: comparison with mammary tissue after
challenge with Escherichia coli. Veterinary
Research. 40(4): 31.
7. Guerra-Junior G, Grumach AS, de Lemos-
Marini SHV, Kirschfink M, Neto AC, de
Araujo M and Mello MPD (2008) Complement
4 phenotypes and genotypes in Brazilian
patients with classical 21- hydroxylase
deficiency. Clinical and Experimental
Immunology, 155(2): 182-188.
8. Hemati Doust V, Rahimi Mianji G and Farhadi
A (2013) Association between bovine
lactoferrin gene variant and somatic cell count
in milk based on EcoRI restriction site. Iranian
Journal of Veterinary Research, Shiraz
University, 46: 62-65. (In Persian)
9. Huang J, Wang H, Wang C, Li J, Li Q, Hou M
and Zhong J (2010) Single nucleotide
polymorphisms, haplotypes and combined
genotypes of lactoferrin gene and their
associations with mastitis in Chinese Holstein
cattle. Molecular Biology Reports, 37: 477-483.
10. Legrand D and Mazurier J (2010) A critical
review of the roles of host lactoferrin in
immunity. Biometals, 23: 365-376.
11. Mao Y, Zhu X, Xing Sh, Zhang M, Zhang H,
Wang X, Karrow N, Yang l and Yang Zh
(2015) Polymorphisms in the promoter region
of the bovine lactoferrin gene influence milk
somatic cell score and milk production traits in
Chinese Holstein cows. Research in Veterinary
Science, 103: 107-112.
12. Miller SA, Dykeys DD and Plesky HF (1988)
A simple salting out procedure for extracting
DNA from human nucleated cells. Nucleic
Acid Research, 16: 1215-1220.
13. Musayeva K, Sederevicius A, Zelvyte R,
Monkeviciene I, Beliavska AD and Garbenyte
Z (2018) Lactoferrin and immunogl obulin
content in cow milk in relation to somatic cell
count and number of lactations. Veterinarija IR
Zootechnika, 76 (98).
14. Naserkheil M, Miraie-Ashtiani SR, Nejati-
Javaremi A, Son J and Lee D (2016) Random
regression models using Legendre polynomials
to estimate genetic parameters for Test-day
milk protein yields in Iranian Holstein dairy
cattle. Asian-Australas Journal Animal
Science, 29(12): 1682-1687. (In Persian)
15. Needs EC and Anderson M(1984) Lipid
composition of milks from cows with
experimentally induced mastitis. Journal of
Dairy Science Researche, 51: 239-249.
16. Ujita A, Alberto Negrao J, Vercesi Filho AE,
Rabelo Fernandes A and El Faro L (2019) Milk
lactoferrin and milk constituents in dairy Gyr
heifers. Livestock Science, 226: 78-92.17. Verbreke J, Poucke MV, Peelman L, Piepers S
and Vliegher SD (2014) Associations between
CXCR1 polymorphisms and pathogen-specific
incidence rate of clinical mastitis, test-day
somatic cell count, and test-day milk yield.
Journal of Dairy Science, 97(79): 27-39.
18. Viale E, Tiezzi F, Maretto M, De Marchi M,
Penasa M and Cassandro M (2017) Association
of candidate gene polymorphisms with milk
technological traits, yield, composition, and
somatic cell score in Italian Holstein-Friesian
sires. Journal of Dairy Science, 100(9): 7271-
17281.
19. Wang X, Zhong j, Gao Y, Ju Z and Huang J
(2014) A SNP in intron 8 of CD46 causes a
novel transcript associated with mastitis in
Holsteins. BMC Genomics, 630(15): 1-11.
20. Yang Y, Chung EK, Wu YL, Savelli SL,
Nagaraja HN, Zhou B, Hebert M, Jones KN,
Shu Y, Kitzmiller K, Blanchong CA, McBride
KL, Higgins GC, Rennebohm RM, Rice RR,
Hackshaw KV, Roubey RA, Grossman JM,
Tsao BP, Birmingham DJ, Rovin BH, Hebert
LA, and Yu CY (2007). Gene copy-number
variation and associated polymorphisms of
complement component C4 in human systemic
lupus erythematosus (SLE): low copy number
is a risk factor and high copy number is a
protective factor against SLE susceptibility in
European Americans. American Journal of
Human Genetics, 80: 1037-1054.
21. Yanga BY, Li AQ, Jua Z, Huanga J, Zhoua L,
Li AR, Li AJ, Shib F, Zhonga J and Wanga CH
(2011) Three novel single-nucleotide
polymorphisms of complement component 4
gene (C4A) in Chinese Holstein cattle and their
associations with milk performance traits and
CH50. G Model Veterinary Immunology and
Immunopathology, 145(1-2): 223-232
22. Yang YY, Huang JM, Ju1 ZH, Li1 QL, Zhou1
L, Li1 JB (2012) Increased expression of a
novel splice variant of the complement
component 4 (C4A) gene in mastitis-infected
dairy cattle. Genetics and Molecular Research,
11(3): 2909-2916.
23. Zabolewicz T, Brym P, Olenski K, Suchocki T,
Malewski T, Szydam J and Kaminski S (2012)
Polymorphism within TATA box of bovine
lactoferrin gene and its association with
performance traits in Holstein cattle. Journal of
Livestock Science, 149(3): 267-274.
24. Zipfel PF and Reuter M (2009) Complement
activation products C3a and C4a as
endogenous antimicrobial peptides.
International Journal of Peptide Research and
Therapeutic, 15(2): 87-95.