Abstract：Abstract Clinical mastitis is a kind of the most serious disease with high incidence in dairy industry, and has been plagued in the development of dairy industry. It has significant effects on milk performance and lifetime which will cause huge economic losses in milk production process. It's very important to study the related genes about mastitis for the improvement of milking traits and elongation of lifetime of cows with molecular techniques, and scientific basis would be provided for the breeding of anti-disease and improvement of economic benefits of dairy farm. In order to verify the effects of lactoferrin gene polymorphism on the production traits of Chinese Holstein (Bos taurus), SNPs of LF gene were selected by PCR and direct sequencing for 20 cows with low SCS and 20 cows with high SCS according to the sequence of 5' untranslating region (UTR) of LF gene. Finally, LF-131C>T and LF-28A>C for 866 Chinese Holstein cows were detected by flight mass spectrometry. The dairy herd improvement production recordings information including the number of test cow, ID number of test cow's father, date of test, related genealogical information, test-day milk yield, fat content, protein content, somatic cell score, lactose, total solid, milk urea nitrogen, clinical mastitis and productive life of tested cows were collected from the management system of dairy farm. The effects of LF-131C>T and LF-28A>C on milk performances and lifetime were analyzed by multi factor variance analysis and Cox regression. The results showed that the dominant genotype of LF-131C>T and LF-28A>C loci were TT and AA with the frequency of 0.813 and 0.580, respectively. The dominant genes were T and A, respectively, and their frequencies were 0.903 and 0.762, respectively. Two loci were in the state of Hardy-Weinberg Equilibrium. LF-131C>T had extremely significant effects on SCS, lactose content and 305 days milk yield (P＜0.01), the effects on test-day milk yield and protein content were close to significant level (0.05＜P＜0.1), but LF-131C>T did not have significant effects on fat content total solid and milk urea nitrogen (P>0.05). Multiple comparisons showed that individuals with genotype of LF-131CC had significantly higher 305 days milk yield and lower SCS and lactose content than that of LF-131 CT and TT. LF-28A>C extremely significantly affected test-day milk yield, fat content, protein content, lactose content, total solid and 305 days milk yield (P＜0.01), but had no significant effects on SCS and MUN (P>0.05). Individual with LF-28 CC genotype had significantly higher fat content, protein content, total solid and 305 days milk yield and lower lactose content than that of LF-131 AA and AC, they also had significantly higher test-day milk yield than that of LF-131 AA. LF-28A>C had significant effects on production lifetime (P＜0.05)，individuals with CC genotype had shorter lifetime than that of AA genotype. LF -131C>T had effects on lifetime, but the effects were not significant. Relative researches about the association between LF-131C>T and LF-28A>C and clinical had important significance for improving milk performance and prolonging production life of dairy cows. It could provide scientific basis for disease resistance breeding and economic benefit improvement of dairy cows.
王洪梅, 孔振兴, 王长法, 等. 奶牛乳铁蛋白基因5′侧翼区遗传多态性及其与乳腺炎关联性分析. 遗传. 2009, 31(4): 393-399. Wang H M, Kong Z X, Wang C F, et al. Genetic polymorphism in 5′-flanking region of the lactoferrin gene and its associations with mastitis in Chinese Holstein cows. Gene. 2009, 31(4): 393-399. 赵佳强. 中国荷斯坦奶牛生产寿命、体型线性性状分子标记的研究[D]. 华中农业大学, 2013. Zhao J Q. Research on molecular markers and productive life, linear type traits of Holstein cattle in China. Huazhong Agricultural University, 2013. 储明星, 石万海, 邝霞,等. 奶牛乳房炎与经济性状之间关系的研究进展[J]. 中国畜牧杂志, 2002, 38(5):44-46. Chu M X, Shi W H, Kuang X, et al. Research Progress on the relationship between mastitis and economic traits in dairy cows [J]. Chinese Journal of Animal Science, 2002, 38(5):44-46. Archer, Simon C. Influence of somatic cell count in heifers on lifetime milk yield and disease management[D]. University of Nottingham, 2013. Boettcher P, Dekkers J, Warnick L, et al. Genetic analysis of clinical lameness in dairy cattle. Journal of Dairy Science, 1998, 81(4):1148-1156. Boujenane I, Aimani J E, By K. Effects of clinical mastitis on reproductive and milk performance of Holstein cows in Morocco. Tropical Animal Health & Production, 2015, 47(1):207-11. Campbell A E, Wilkinson-White L, Mackay J P, et al. Analysis of disease-causing GATA1 mutations in murine gene complementation systems. Blood, 2013, 121 (26): 5218–5227. Cartharius K, Frech K, Grote K, Klocke B, Haltmeier M, Klingenhoff A, Frisch M, Bayerlein M, Werner T. MatInspector and beyond: promoter analysis based on transcription factor binding sites. Bioinformatics, 2005, 21(13):2933-42. Daly M, Ross P, Giblin L, et al. Polymorphisms within the lactoferrin gene promoter in various cattle breeds. Animal Biotechnology, 2006, 17(1):33-42 Elhaig M M, Selim A. Molecular and bacteriological investigation of subclinical mastitis caused by Staphylococcus aureus and Streptococcus agalactiae in domestic bovids from Ismailia, Egypt. Tropical Animal Health & Production, 2014, 47(2):271-6 Fujiwara Y, Browne C P, Cunniff K, et al. Arrested development of embryonic red cell precursors in mouse embryos lacking transcription factor GATA-1. Proceedings of the National Academy of Sciences of the United States of America ， 1996， 93 (22): 12355–12358. Khatib H, Heifetz E, Dekkers J C. Association of the protease inhibitor gene with production traits in Holstein dairy cattle. Journal of Dairy Science, 2005, 88(3):1208-13. Khatib H, Schutzkus V, Chang Y M, et al. Pattern of expression of the uterine milk protein gene and its association with productive life in dairy cattle. Journal of Dairy Science, 2007, 90(5):2427-33. O’Halloran F, Bahar B, Buckley F, et al. Characterisation of single nucleotide polymorphisms identified in the bovine lactoferrin gene sequences across a range of dairy cow breeds. Biochimie, 2009，91(1):68–75 Omata Y, Satake M, Maeda R, et al. Reduction of the infectivety of Toxoplasma gondi and Eimeria stiedai sporozoites by treatment with bovine lactoferrin. journal of veterinary medical science. 2001, 63(2): 187-190. Pourzal F, Sharifiyazdi H and Mirzaei A. Assessment of a single-nucleotide polymorphism in specific protein 1 (SP1)-binding site of the lactoferrin promoter region as potential genetic markers for uterine infection in dairy cows. Comparative Clinical Pathology. 2013，DOI 10.1007/s00580-013-1781-x Seyfert H M, Tuekoriez A, Interthal H, et al. Structure of the bovine lactoferrin-encoding gene and its Promoter. Gene. 1994, 143: 265-269. Verboort B, Berry S. L. California Dairy Herd Improvement Association[J]．Dairy Tales，1995 Wakabayashi H, Kurokawa M, Shin K, et al. Oral lactoferrin prevents body weight 1oss and inerease cytoline responses during herpes simplex virus type 1 infeetion of mice. Bioscience, Biotechnology, and Biochemistry. 2004, 68(3): 537-544. Zabolewicz T, Barcewicz M, Brym P, et al. Association of polymorphism within LTF gene promoter with lactoferrin concentration in milk of Holstein cows. Polish Journal of Veterinary Sciences. 2014, 17(4):633-41. Zheng J, Ather J L, Sonstegard T S, et al. Characterization of the infection-responsive bovine lactoferrin promoter. Gene, 2005，353(1):107–117.
