猪精液冷冻保存技术研究进展

doi:10.16431/j.cnki.1671-7236.2024.02.020

Abstract

Abstract: Boar semen cryopreservation, as a vital technology for swine industry, is conducive to improve the utilization rate of excellent breeding boars and realize the long-term preservation of superior germplasm resources, as well as the elevation of efficiency in breeding farms.At present, various progress has been made in boar semen cryopreservation, the conception rate and the number born alive by domestic application of pig frozen semen have basically reached the level of foreign frozen semen breeding.However, due to the certain degree of damage of pig spermatozoa in frozen-thawed process which caused sperm fertility declined and the fertilization window period shortened, hence there is still a big gap in application of frozen semen.The article reviews the development process of cryopreservation technology for pig semen at home and abroad, introduces the basic characteristics of pig semen and sperm, briefly describes the mechanism of sperm damage caused by the cooling process, and analyzes and introduces the mechanism and methods of reducing freezing damage from the perspective of the composition of cryodiluents, such as adding cryoprotectants to reduce the formation of ice crystals inside and outside cells, maintaining osmotic pressure inside and outside cells, and stabilizing cell membrane structure.Adding antibacterial substances to inhibit the rapid reproduction of bacteria after thawing.Adding antioxidants to reduce reactive oxygen species levels, alleviate oxidative stress, protect sperm structure and fertilization ability, etc.Finally, the future development direction of boar semen cryopreservation technology is expected to provide reference for related research and practical applications.

Key words: boar; semen; cryopreservation

CLC Number:

S828.3

WANG Jialun, CHEN Hui, SUN Haowei, GUO Yaoyao, LIANG Guodong, MA Yunhui, GAO Lei. Advances in Boar Semen Cryopreservation Technologies[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(2): 639-648.

References

[1] 刘刚,杨幼明.我国主要肉类市场调控政策思路研究--肉类市场调控的逻辑脉络:短期调供求、中期调预案、长期调结构[J].价格理论与实践,2020,7:40-43. LIU G,YANG Y M.Research on the thoughts of regulation and control policy for my country's main meat market-Logic context of meat market regulation:Short-term adjustment of supply and demand,mid-term adjustment plan,long-term adjustment of structure[J].Price:Theory&Practice,2020,7:40-43.(in Chinese)
[2] 邱小田,张振,王长存,等.应对非洲猪瘟疫情保护种猪基本产能--对当前种猪产业的调研[J].中国畜牧业,2019,12:39. QIU X T,ZHANG Z,WANG C C,et al.Dealing with African swine fever and protecting basic production capacity of breeding pigs[J].China Animal Industry,2019,12:39.(in Chinese)
[3] 张亮,潘红梅,郭宗义,等.猪冷冻精液应用研究进展[J].中国畜牧杂志,2023,59(8):20-30. ZHANG L,PAN H M,GUO Z Y,et al.Advances in application of boar frozen-thawed semen for artificial insemination[J].Chinese Journal of Animal Science,2023,59(8):20-30.(in Chinese)
[4] SHERMAN J K.Research on frozen human semen:Past,present,and future[J].Fertility and Sterility,1964,15:485-499.
[5] POLGE C,SMITH A U,PARKES A S.Revival of spermatozoa after vitrification and dehydration at low temperatures[J].Nature,1949,164(4172):666.
[6] HESS E A,TEAGUE H S,LUDWICK T M,et al.Swine can be bred with frozen semen[J].Ohio Farm and Home Research,1957,42(309):100.
[7] 左晓宇,任尚,吴方超,等.猪精子冷冻保存技术的研究进展[J].黑龙江动物繁殖,2022,30(1):18-25. ZUO X Y,REN S,WU F C,et al.Research progress on sperm cryopreservation technology in pigs[J].Heilongjiang Journal of Animal Reproduction,2022,30(1):18-25.(in Chinese)
[8] PURSEL V G,JOHNSON L A.Freezing of boar spermatozoa:Fertilizing capacity with concentrated semen and a new thawing procedure[J].Journal of Animal Science,1975,40(1):99-102.
[9] WESTENDORF P,RICHTER L,TREU H.Deep freezing of boar sperma.Laboratory and insemination results using the Hülsenberger paillete method[J].DTW. Deutsche Tierarztliche Wochenschrift,1975,82(7):261-267.
[10] 潘红梅,彭刚,吴玲,等.猪冷冻精液在我国的应用现状和存在的问题[J].猪业科学,2020,37(6):44-48. PAN H M,PENG G,WU L,et al.Application and problems of pig frozen semen in China[J]. Swine Industry Science,2020,37(6):44-48.(in Chinese)
[11] 杨公社,郭传甲,王爱国,等.猪生产学[M].北京:中国农业出版社,2002. YANG G S,GUO C J,WANG A G,et al.Swine Production Science[M].Beijing:China Agriculture Press,2002.(in Chinese)
[12] 张渭斌.猪细管冷冻精液技术研究[D].杨凌:西北农林科技大学,2011. ZHANG W B.Study on freezing-preservation technology on boar semen frozen in straws[D].Yangling:Northwest A&F University,2011.(in Chinese)
[13] PARKS J E,LYNCH D V.Lipid composition and thermotropic phase behavior of boar,bull,stallion,and rooster sperm membranes[J].Cryobiology,1992,29(2):255-266.
[14] CUMMINS J M,WOODALL P F.On mammalian sperm dimensions[J].Journal of Reproduction and Fertility,1985,75(1):153-175.
[15] KONDRACKI S,BONASZEWSKA D,MIELNICKA C.The effect of age on the morphometric sperm traits of domestic pigs (Sus scrofa domestica)[J].Cellular&Molecular Biology Letters,2005,10(1):3-13.
[16] GADELLA B M,EVANS J P.Membrane fusions during mammalian fertilization[J].Advances in Experimental Medicine and Biology,2011,713:65-80.
[17] 朱振东.猪精子能量代谢的调控机理研究[D].杨凌:西北农林科技大学,2020. ZHU Z D.Mechanisms underlying energy metabolism in boar sperm[D].Yangling:Northwest A&F University,2020.(in Chinese)
[18] WESTHOFF D,KAMP G.Glyceraldehyde 3-phosphate dehydrogenase is bound to the fibrous sheath of mammalian spermatozoa[J].Journal of Cell Science,1997,110(15):1821-1829.
[19] FEIDEN S,STYPA H,WOLFRUM U,et al.A novel pyruvate kinase (PK-S) from boar spermatozoa is localized at the fibrous sheath and the acrosome[J].Reproduction,2007,134(1):81-95.
[20] SHARAFI M,BORGHEI-RAD S M,HEZAVEHEI M,et al.Cryopreservation of semen in domestic animals:A review of current challenges,applications,and prospective strategies[J].Animals: An Open Access Journal from MDPI,2022,12(23):3271.
[21] JOHNSON L A,WEITZE K F,FISER P,et al.Storage of boar semen[J].Animal Reproduction Science,2000,62(1-3):143-172.
[22] QUINN P J.A lipid-phase separation model of low-temperature damage to biological membranes[J].Cryobiology,1985,22(2):128-146.
[23] QUINN P J,WILLIAMS W P.The structural role of lipids in photosynthetic membranes[J].Biochimica et Biophysica Acta,1983,737(2):223-266.
[24] DARIN-BENNETT A,WHITE I G.Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock[J].Cryobiology,1977,14(4):466-470.
[25] 谢言射,徐志谦,蔡更元,等.猪精液冷冻保存技术研究进展[J].家畜生态学报,2020,41(10):78-83. XIE Y S,XU Z Q,CAI G Y,et al.Research progress of cryopreservation of boar semen[J].Journal of Domestic Animal Ecology,2020,41(10):78-83.(in Chinese)
[26] PAPAHADJOPOULOS D,POSTE G,SCHAEFFER B E,et al.Membrane fusion and molecular segregation in phospholipid vesicles[J].Biochimica et Biophysica Acta,1974,352(1):10-28.
[27] YESTE M.Recent advances in boar sperm cryopreservation:State of the art and current perspectives[J].Reproduction in Domestic Animals,2015,50(Suppl 2):71-79.
[28] NAWROTH F,ISACHENKO V,DESSOLE S,et al.Vitrification of human spermatozoa without cryoprotectants[J].Cryo Letters,2002,23(2):93-102.
[29] MAZUR P,LEIBO S P,CHU E H.A two-factor hypothesis of freezing injury.Evidence from Chinese hamster tissue-culture cells[J].Experimental Cell Research,1972,71(2):345-355.
[30] WOLFE J,BRYANT G.Freezing,drying,and/or vitrification of membrane-solute-water systems[J].Cryobiology,1999,39(2):103-129.
[31] GARVEY C J,LENNÉ T,KOSTER K L,et al.Phospholipid membrane protection by sugar molecules during dehydration-insights into molecular mechanisms using scattering techniques[J].International Journal of Molecular Sciences,2013,14(4):8148-8163.
[32] MAZUR P.Freezing of living cells:Mechanisms and implications[J].The American Journal of Physiology,1984,247(3 Pt 1):C125-C142.
[33] 章啸君,屠平光,项云.金华猪精液冷冻保存技术研究[J].浙江畜牧兽医,2020,45(1):1-3. ZHANG X J,TU P G,XIANG Y.Research on the technology of semen cryopreservation for Jinhua pig[J].Zhejiang Journal Animal Science and Veterinary Medicine,2020,45(1):1-3.(in Chinese)
[34] MORRIS G J,GOODRICH M,ACTON E,et al.The high viscosity encountered during freezing in glycerol solutions:Effects on cryopreservation[J].Cryobiology,2006,52(3):323-334.
[35] MORRIS G J.Rapidly cooled human sperm:No evidence of intracellular ice formation[J].Human Reproduction,2006,21(8):2075-2083.
[36] MORRIS G J,FASZER K,GREEN J E,et al.Rapidly cooled horse spermatozoa:Loss of viability is due to osmotic imbalance during thawing,not intracellular ice formation[J].Theriogenology,2007,68(5):804-812.
[37] 高俊锋,郑筱峰,葛立军,等.用0.25 mL细管冷冻保存猪精液[J].农业生物技术学报,2008,2:264-269. GAO J F,ZHENG X F,GE L J,et al.Cryopreservation of boar semen using 0.25 mL straws[J]. Journal of Agricultural Biotechnology,2008,2:264-269.(in Chinese)
[38] TOMÁS-ALMENAR C,DE MERCADO E.Optimization of the thawing protocol for iberian boar sperm[J]. Animals: An Open Access Journal from MDPI,2022,12(19):2600.
[39] 甘麦邻,刘麟,赵雪,等.不同解冻程序对荣昌猪冷冻精液解冻效果的影响[J].中国畜牧杂志,2021,57(7):140-146. GAN M L,LIU L,ZHAO X,et al.Effect of different thawing procedures on the thawing Rongchang pigs frozen semen[J].Chinese Journal of Animal Science,2021,57(7):140-146.(in Chinese)
[40] RAJU R,TORRENT-BURGUÉS J,BRYANT G.Interactions of cryoprotective agents with phospholipid membranes-A Langmuir monolayer study[J].Chemistry and Physics of Lipids,2020,231:104949.
[41] LOVELOCK J E.The protective action of neutral solutes against haemolysis by freezing and thawing[J].The Biochemical Journal,1954,56(2):265-270.
[42] HOLT W V.Basic aspects of frozen storage of semen[J].Animal Reproduction Science,2000,62(1-3):3-22.
[43] HAMMERSTEDT R H,GRAHAM J K.Cryopreservation of poultry sperm:The enigma of glycerol[J].Cryobiology,1992,29(1):26-38.
[44] SI W,ZHENG P,LI Y,et al.Effect of glycerol and dimethyl sulfoxide on cryopreservation of rhesus monkey (Macaca mulatta) sperm[J].American Journal of Primatology,2004,62(4):301-306.
[45] ALVARENGA M A,PAPA F O,LANDIM-ALVARENGA F C,et al.Amides as cryoprotectants for freezing stallion semen:A review[J].Animal Reproduction Science,2005,89(1-4):105-113.
[46] PINHO R O,LIMA D M,SHIOMI H H,et al.Effect of different cryo-protectants on the viability of frozen/thawed semen from boars of the Piau breed[J]. Animal Reproduction Science,2014,146(3-4):187-192.
[47] KIM S,LEE Y J,JI D B,et al.Evaluation of different cryoprotectants (CPAs) in boar semen cryopreservation[J].The Journal of Veterinary Medical Science,2011,73(7):961-963.
[48] 马玲琴,贾永宏,马国际,等.鸵鸟卵黄对猪精子冷冻后质量的影响[J].畜牧兽医杂志,2012,31(2):29-31. MA L Q,JIA Y H,MA G J,et al.Effect of ostrich egg yolk on boar spermatozoa quality after freezing-thawing[J].Journal of Animal Science and Veterinary Medicine,2012,31(2):29-31.(in Chinese)
[49] MOUSSA M,MARINET V,TRIMECHE A,et al.Low density lipoproteins extracted from hen egg yolk by an easy method:Cryoprotective effect on frozen-thawed bull semen[J].Theriogenology,2002,57(6):1695-1706.
[50] 吕瑞凯,胡建宏,王红,等.5种禽类卵黄低密度脂蛋白对猪精子冷冻效果的影响[J].西北农业学报,2011,20(9):5-10. LYU R K,HU J H,WANG H,et al.Comparative study of LDL extracted from five avian species on the cryopreservation of boar sperm[J].Acta Agriculturae Boreali-Occidentalis Sinica,2011,20(9):5-10.(in Chinese)
[51] AMIRAT L,TAINTURIER D,JEANNEAU L,et al.Bull semen in vitro fertility after cryopreservation using egg yolk LDL:A comparison with Optidyl,a commercial egg yolk extender[J].Theriogenology,2004,61(5):895-907.
[52] 张树山,戴建军,吴彩凤,等.谷氨酰胺与鸡卵黄低密度脂蛋白提高猪精液冷冻效果的研究[J].上海农业学报,2013,29(2):5-8. ZHANG S S,DAI J J,WU C F,et al.Effect of glutamine and low-density lipoproteins of hen's egg yolk on cryopreservation of boar semen[J].Acta Agriculturae Shanghai,2013,29(2):5-8.(in Chinese)
[53] 潘彩霞.精液稀释液冷冻保护剂卵黄替代物的研究进展[J].黑龙江动物繁殖,2020,28(6):33-36. PAN C X.Research progress of egg yolk substitutes as cryoprotectants for semen diluents[J].Heilongjiang Journal of Animal Reproduction,2020,28(6):33-36.(in Chinese)
[54] 汪俊跃,张树山,戴建军,等.高压均质(HPH)大豆卵磷脂替代卵黄在猪精液冷冻保存效果的研究[J].中国兽医科学,2020,50(11):1461-1468. WANG J Y,ZHANG S S,DAI J J,et al.High pressure homogeneity (HPH) soybean lecithin-based extender as an alternative for boar sperm freezing[J].Chinese Veterinary Science,2020,50(11):1461-1468.(in Chinese)
[55] CROWE L M,CROWE J H,RUDOLPH A,et al.Preservation of freeze-dried liposomes by trehalose[J]. Archives of Biochemistry and Biophysics,1985,242(1):240-247.
[56] CROWE J H,CROWE L M,CHAPMAN D.Preservation of membranes in anhydrobiotic organisms:The role of trehalose[J].Science,1984,223(4637):701-703.
[57] AHMAD E,NASEER Z,AKSOY M,et al.Trehalose enhances osmotic tolerance and suppresses lysophosphatidylcholine-induced acrosome reaction in ram spermatozoon[J].Andrologia,2015,47(7):786-792.
[58] IQBAL S,ANDRABI SMH,RIAZ A,et al.Trehalose improves semen antioxidant enzymes activity,post-thaw quality,and fertility in Nili Ravi buffaloes (Bubalus bubalis)[J].Theriogenology,2016,85(5):954-959.
[59] 张树山,李青旺,李刚,等.海藻糖、蔗糖和乳糖对猪精液冷冻保护效果的影响[J].西北农林科技大学学报(自然科学版),2006,6:41-45. ZHANG S S,LI Q W,LI G,et al.Effect of trehalose,sucrose and lactoseon on cryopreservation of boar sperm[J].Journal of Northwest A&F University(Natural Science Edition),2006,6:41-45.(in Chinese)
[60] 张伟.二甲基乙酰胺、海藻糖与低密度脂蛋白对猪精液冷冻保存效果的研究[D].长春:吉林大学,2012. ZHANG W.Effects of supplementing boar semen cryopreservation media with dimethylacetamide,trehalose and low denisity lipoprotein on post-thawed spermatozoa quality[D].Changchun:Jilin University,2012.(in Chinese)
[61] PEZO F,ZAMBRANO F,URIBE P,et al.Oxidative and nitrosative stress in frozen-thawed pig spermatozoa.Ⅱ:Effect of the addition of saccharides to freezing medium on sperm function[J].Cryobiology,2020,97:5-11.
[62] ALTHOUSE G C,LU K G.Bacteriospermia in extended porcine semen[J].Theriogenology,2005,63(2):573-584.
[63] MOREIRA S S J,SANTOS C S,CASTELO T S,et al.Investigating the need for antibiotic supplementation to the extender used for semen cryopreservation in collared peccaries[J].Frontiers in Veterinary Science,2022,9:954921.
[64] LUTHER A M,NGUYEN T Q,VERSPOHL J,et al.Antimicrobially active semen extenders allow the reduction of antibiotic use in pig insemination[J].Antibiotics (Basel, Switzerland),2021,10(11):1319.
[65] YAKANDAWALA N,GAWANDE P V,LOVETRI K,et al.Effect of ovotransferrin,protamine sulfate and EDTA combination on biofilm formation by catheter-associated bacteria[J].Journal of Applied Microbiology,2007,102(3):722-727.
[66] NASREEN S,AWAN M A,UL-HUSNA A,et al.Honey as an alternative to antibiotics for cryopreservation of Nili-Ravi buffalo bull spermatozoa[J]. Biopreservation and Biobanking,2020,18(1):25-32.
[67] GAVAZZA M,MARMUNTI M,COMPAGNONI M,et al.Antioxidant effect of lutein protects against oxidative damage to porcine spermatozoa[J].Revista Veterinaria,2019,30(1):7.
[68] ROCA J,RODRÍGUEZ M J,GIL M A,et al.Survival and in vitro fertility of boar spermatozoa frozen in the presence of superoxide dismutase and/or catalase[J].Journal of Andrology,2005,26(1):15-24.
[69] CHEN Y,WANG K,ZHANG D,et al.GPx6 is involved in the in vitro induced capacitation and acrosome reaction in porcine sperm[J].Theriogenology,2020,156:107-115.
[70] AITKEN R J.Free radicals,lipid peroxidation and sperm function[J].Reproduction, Fertility, and Development,1995,7(4):659-668.
[71] CEROLINI S,MALDJIAN A,SURAI P,et al.Viability,susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage[J].Animal Reproduction Science,2000,58(1-2):99-111.
[72] PEZO F,ZAMBRANO F,URIBE P,et al.Oxidative and nitrosative stress in frozen-thawed pig spermatozoa.Ⅰ:Protective effect of melatonin and butylhydroxytoluene on sperm function[J].Research in Veterinary Science,2021,136:143-150.
[73] BANG S,TANGA B M,FANG X,et al.Cryopreservation of pig semen using a quercetin-supplemented freezing extender[J].Life (Basel, Switzerland),2022,12(8):1155.
[74] 刘浩域,郭建华,罗兰,等.虾青素和平衡时间对猪精子冷冻保存效果的影响[J].中国畜牧杂志,2022,58(8):243-247. LIU H Y,GUO J H,LUO L,et al.Effect of astaxanthin and equilibration time on the effect of cryopreservation of porcine spermatozoa[J].Chinese Journal of Animal Science,2022,58(8):243-247.(in Chinese)
[75] ERDENEBAYAR O,刘定邦,杨丽,等.熊果酸对猪精液冷冻保存效果的影响[J].家畜生态学报,2022,43(10):49-53. ERDENEBAYAR O,LIU D B,YANG L,et al.The effect of ursolic acid on cryopreservation of boar spermatozoa[J].Journal of Domestic Animal Ecology,2022,43(10):49-53.(in Chinese)
[76] 邢之宁,曹丁壬,章晓炜,等.冷冻稀释液和抗氧化剂对金华猪精液冷冻效果的影响[J].黑龙江畜牧兽医,2023,4:41-44. XING Z N,CAO D R,ZHANG X Y,et al.Effects of diluent and antioxidants on semen freezing of Jinhua pigs[J].Heilongjiang Animal Science and Veterinary Medicine,2023,4:41-44.(in Chinese)

Advances in Boar Semen Cryopreservation Technologies

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	SUN Qingyi, LI Mengqi, ZHAO Wei, ZHANG Wentao, ZHANG Xiaosheng, ZHANG Jinlong, ZHANG Hongjun, FANG Xiaohuan, TAO Chenyu, XIA Wei, LI Junjie. Establishment and Cryopreservation of Chengde Hornless Goat Ear Fibroblast Cell Line [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 183-192.
[2]	XU Shiyuan, ZHANG Qiuxue, ZHAO Lingjun, DONG Zhihao, LIU Songqi, YUAN Kaimin, WU Kaihui, WANG Dong. Research Progress on the Function of ART3 and Its Relationship with Spermatogenesis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3221-3228.
[3]	LEI Mingkai, BAN Bin, YIN Weiqi, ZHANG Junmei, SHI Panfeng, WANG Li, ZHOU Yang, ZHANG Chunxiang, LI Bibo. Effects of Coated Folic Acid and Coated Taurine on Semen Quality, Serum and Seminal Plasma Antioxidant Capacity and Hormone Level in Heat Stressed Rams [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2755-2765.
[4]	LI Zhixing, QING Yuhong, LI Zhiyuan, CAO Shaoqi, GAO Xinmei, ZENG Weibin, WANG Jing, HU Guangdong. Effects of Leonurine on Cryopreservation of Sheep Semen [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2801-2810.
[5]	LYU Chunrong, LIANG Jiachong, SHAO Qingyong, WU Guoquan, ZHANG Yumei, QUAN Guobo. Effects of Sodium Pyruvate on the Quality of Frozen Goat Sperm [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2363-2369.
[6]	LIU Jinyu, ZHANG Yang, MA Xumei, ZHOU Qian, ZHOU Yang, ZHANG Chunxiang, SHI Lei, REN Youshe. Effects of Glutathione on Cryopreservation of Sheep Semen [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2388-2394.
[7]	SONG Chenglei, ZHANG Zhe, WEI Yilin, YANG Songbai, DOU Yaqing, LI Chenlei, LIU Yingke, LI Xinjian, YANG Feng, LI Xiuling, WANG Kejun, QIAO Ruimin, HAN Xuelei. Effects of Different Breeds,Seasons and Month Ages on Semen Quality of Boars [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1938-1946.
[8]	DU Zhaohui, GE Zihao, YOU Junyi, LIU Xiaowei, LIANG Guodong, SUN Shiduo, HU Jianhong, LI Xiao. Effects of Clostridium butyricum and Sodium Butyricum on Reproductive System Injury Induced by High Fat Diet in Male Mice [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1603-1616.
[9]	ZHANG Xiaoke, LIAO Weili, LI Tingting, LI Yao, LI Jiaqi, HUANG Xiang, ZHANG Hao. Effects of Semen Collectors and Environmental Parameters in Pigpen on Semen Quality of Duroc Boars [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 220-229.
[10]	WANG Hechuan, ZHANG Han, LI Tianfeng, WANG Qian, ZHANG Qun, GUO Minghui, SUN Chao, ZHOU Yanzhi, LI Yuan, LI Yanbing, WEI Guosheng, LI Jingchun. Effects of Different Concentrations of Taurine on the Preservation of Boar Semen Under Negative Pressure at Room Temperature [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3484-3491.
[11]	LIU Gang, ZHU Fangxian, ZHANG Xiaowei, GU Feng, LIU Lijun, SU Rui, MENG Fei, XU Haitao, SUN Feizhou. Evaluation of Embryo and Sperm Cryopreservation in Hu Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3492-3499.
[12]	XU Xiaoling, BAI Jiahua, XIAO Linli, QIN Yusheng, SONG Yuqing, LIU Yan. Effects of Compound Additives on the Quality and Conception Rate of Dairy Cows Thawed Sex Controlled Frozen Semen [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2654-2660.
[13]	YUAN Chongshan, WANG Jun, LYU Wenfa. Research Progress on Effects of Polyunsaturated Fatty Acids on Animal Semen Quality [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1422-1429.
[14]	LIANG Jiachong, LYU Chunrong, HONG Qionghua, WU Guoquan, QUAN Guobo. Analysis of Sperm Differential Transcriptome Expression Before and After Freezing in Yunnan Semi-fine Wool Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 973-981.
[15]	LUO Lan, CUI Chao, DONG Ruilan, LIU Haoyu, LIU Xiaobin, YU Guanghui. Effects of Different Concentrations of Rutin and Freezing Rates on Cryopreservation of Boar Semen [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 982-991.