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猪人工授精新技术 （美国南达科达州立大学供稿） 施学仕 博士 美国大豆协会-国际项目 上海代表处

Training Boars for Semen Collection 训练公猪进行采精 Select a person who is patient and enjoys working with animals.选择一个有耐心并乐于与动物相处的人 Begin training during the isolation process by building a trust between you and the boar. Spend a few minutes each day scratching, rubbing and talking with the boar.训练开始于隔离过程中，这时要在操作者和公猪之间建立信任，每天花几分钟对公猪抓痒、擦拭和“对话” Boars vary in their rate of sexual development. Some 7 mo old boars are ready to be trained and others require 4 - 6 wks longer.不同公猪的性成熟速度各异，有些7月龄的公猪就可投入训练，而另一些可能要晚4～6周才行。

Training Boars (con’t)公猪训练(续) Provide a clean, dry, well lighted area free of distractions. A breeding mat helps provide footing. A boar that slips during mounting may be shy about attempting it again. 训练场地应该清洁、干燥、光照良好且无干扰。地上铺上一个防滑垫可有助于公猪站稳，公猪在爬跨时滑倒会羞于再次爬跨。 Keep the area about 8 x 10 ft, so that the boar is focused. 训练区域大小应在8×10英尺左右，以便公猪能够集中精力。 If possible, collect a previously trained boar first. The odor and sound may stimulate the “rookie” if he is located next to the collection area. 如有可能,先对先前接受了训练的公猪采精。令新公猪站在近旁,采精时的气味和声响会对新公猪产生刺激

Training Boars (con’t)公猪训练(续) Adjust the dummyso that it is equal to or slightly lower than the boar being trained.调整假母猪的高度，使其等于或略高于训练公猪的高度。 Plan for short training sessions (15 -20 min). Some boars may mount the dummy immediately but others may want to explore the pen and dummy for several session.每次训练时间应该短些(15 ~ 20分钟)。有些公猪会立即爬跨假母猪，而有些公猪在开始几次训练中只是探究猪圈和假母猪。

Training Boars (con’t)公猪训练(续) Some boars require coaxing. Crouchdown near the dummy, allow the boar to smell your hand and clothes and talk in a reassuring tone. Try pouring semen from a previous collection on the dummy.对有些公猪需要哄骗。要在假母猪旁边蹲下，让公猪嗅闻你的手和衣服，采用使公猪安定的声调召唤公猪。 Keep the boar focused on the dummy. If needed, use winged gates to keep him in front of the dummy.要使公猪集中关注假母猪。需要时可用winged gate使公猪呆在假母猪的前面。

New Technologies新技术 Boar semen extenders公猪精液稀释剂 Intra-uterine Insemination子宫内输精 Sexing of boar semen公猪精液性别化 Cloning and embryo transfer克隆和胚胎移植

增加人工授精站的有效工时 降低精液运输成本 Reasons for developing new long-term boar semen extenders研制长效公猪精液稀释剂的原因 More favorable work hours in boar stud 增加人工授精站的有效工时 Reduction or elimination of weekend collections减少或避免周末采精 Reduced transportation cost of semen 降低精液运输成本 Testing of semen for PRRS before shipping/using semen可在精液运输/应用之前测试其是否受到猪繁殖和呼吸综合征病毒污染 Longer storage life of延长精液储存时间 Fewer discards at the farm减少猪场中精液浪费

New Long-Term Extenders新的长效稀释剂 Non-published formula未发表配方 Androhep® EnduraGuard™ Shelf-life is 9 days有效期9天 Antibiotics抗生素 SafeCell Plus™ SafeCell Plus™ Liquid concentrated extender液态浓缩稀释剂 Specific blend of antibiotics与抗生素混合 Mulberry III Mulberry III Shelf-life is 7 to 14 days有效期7～14天 Antibiotic抗生素

Intra-uterine Insemination 子宫内输精 Intra-uterine “body” insemination (IUBI)子宫内输精(子宫体内输精) Deep intra-uterine “horn” insemination (DIUHI)深部子宫内输精(子宫角内输精)

Suggested beneficial aspects of intra-uterine body AI: 子宫体内输精的好处 Less back-flow减少精液回流 Fewer sperm cells per dose可减少每次输精的精子数 Smaller volume required可减少每次输精的精液量 Less time needed to inseminate可减少每次输精花费的时间 Less boars will be needed to produce sperm cells公猪的头数 Genetic cost will be lower?可降低种猪成本？

% of sows with backflow有回流母猪% Percentage of sows inseminated that have backflow with traditional AI 传统输精法的精液回流情况 Time of backflow 精液回流 No. sows evaluated 测试母猪数 No. sows with backflow 有回流母猪数 % of sows with backflow有回流母猪% During AI 输精中 120 76 63.3 0 to .5 hr after AI 输精后0～0.5 小时 112 110 98.2 .5 to 2.5 hr after AI 输精后0.5～2.5 小时 80 78 97.5 Stevernik et al., Animal Reproduction Science 54:109-119, 1998.

No. sows evaluated测试母猪数 Total volume lost, %损失量% Proportion of total volume (80 mL) that was lost during and after traditional Ai 传统输精法总输精量80毫升中 输精中和输精后的损失比例 Time of backflow回流时间 No. sows evaluated测试母猪数 Total volume lost, %损失量% Average平均 Range范围 During AI输精中 76 7% 1 to 56% 0 to .5 hr after AI 输精后0～0.5 小时 110 31% 3 to 76% .5 to 2.5 hr after AI 输精后0.5～2.5 小时 78 36% 1 to 94% Stevernik et al., Animal Reproduction Science 54:109-119, 1998.

子宫体 深部Goldenpig输精管 8英寸 (20.32毫米) Goldenpig输精管 Uterine body DeepGoldenpig 8 inches 8英寸 (20.32毫米) Goldenpig输精管 Goldenpig IMV International

Uterus is hostile to sperm 子宫不利于精子生存 新方法将精子置于子宫体内 Intra-uterine body placement of sperm cells Phagocytosis细胞吞噬作用 Massive influx of polymorphonuclear Leucocytes at 30 minutes to 3 hours after spermatozoa enter uterine horn精子进入子宫角后30分钟到3小时，多形核白细胞大量出现 Traditional placement of sperm cells传统方法输精法将精子置于此处

Deep Intra-uterine Horn Insemination 深部子宫角内输精 Surgical procedure外科手术法 Not practical for commercial use 不适合用于商业目的 Fiber optic endoscope光纤内镜 Flexible catheter柔软导管 Used on non-sedated estrous sows 用于非呆立发情的母猪 Practical for commercial use 适合用于商业目的

深部子宫角内输精 Deep Intra-uterine “Horn” Artificial Insemination FirFlex® Catheter (Patented by University of Murcia) FirFlex®输精管(专利权Murcia大学) Marketed in Europe by MAGAPOR (Spain, Portugal, Hungary) 由MAGAPOR公司在欧洲销售 (西班牙，葡萄牙，匈牙利)

DNA differences between X- & Y-chromosome-bearingspermatozoa X精子和Y精子之间的差异 Vole田鼠– 12.5% Chinchilla南美栗鼠 – 7.5% Sheep绵羊– 4.2% Dog狗– 3.9% Cattle牛– 3.8% Elk]麋鹿 – 3.8% Horse马– 3.7% Pigs猪– 3.6% Elephant象– 3.4% Camel骆驼– 3.3% Rabbit兔– 3.0% Human人– 2.8% Possum负鼠– 2.3% Turkey火鸡– 0.0% L. A. Johnson. Animal Reproduction Science 60-61:93-107, 2000. L. A. Johnson. Reproduction, Fertility, and Development 7:893-903, 1995.

Current problems with sexing boar semen:公猪精液性别化中的问题 Sorting process is too slow性别鉴定过程太慢 6 million per hour for X每小时6百万个X精子 6 million per hour for Y每小时6百万个Y精子 11 million per hour when sorting for only X仅仅选择X精子可达每小时1100万 Decreased reproductive performance 降低繁殖性能

Stimulating/Priming Uterus 刺激/敏化子宫 Adding oxytocin to each dose of semen在每份精液中加入催产素 Adding prostaglandin to each dose of semen在每份精液中加入前列腺素 “Inserting” MR-A Predil prior to insemination输精前加入人工合成的精浆制品MR-A Predil Mating gilts with sterile boar用不育的公猪与小母猪交配 Inseminating gilts with dead semen用死精给小母猪输精

(BTS semen extenderBTS精液稀释剂) 分娩率(%) Farrowing rate and number of piglets born alive per litter when oxytocin was injected prior to inseminating with “old” sperm cells.用老精子输精前注射催产素时母猪的分娩率和窝产仔数 (BTS semen extenderBTS精液稀释剂) Treatment处理 Farrowing rate, % 分娩率(%) Litter size 窝产仔数 Sperm cells >72 hours of age大于72时龄的精子 68.2 a (n = 55) 9.4 a Sperm cells >72 hours of age + oxytocin 大于72时龄的精子+催产素 85.2 b (n = 59) 10.1 b Difference差异 17.0 0.7 ab (P < .05) W. L. Flowers, NSCU

Adding Lutalyse to a dose of semen increased farrowing rate and litter size 在精液中加入律胎素可提高分娩率和窝产仔数 Item项目 Lutalyse 律胎素 Control 对照 Difference差异 Number of sow & gilts母猪头数 1575 1552 23 Farrowing rate, %分娩率(%) 83.80 80.61 3.19 Litter size窝产仔数 10.15 10.08 .07 FI 851 813 38 .5 to 1.0 mL Lutalyse (PGF2a) per dose of semen每份精液加入0.5～1.0毫升律胎素(PGF2a，前列腺素)

合成精浆MR-A Predil可提高分小母猪的娩率和窝产仔数(三个研究综述) MR-A Predil increased farrowing rate and litter size of gilts [Summary of three studies] 合成精浆MR-A Predil可提高分小母猪的娩率和窝产仔数(三个研究综述) Item项目 Predil 合成精浆 Control 对照 Difference 差异 Number of gilts 小母猪数 296 324 28 Farrowing rate, %分娩率% 86.80 82.07 +4.73 Litter size窝产仔数 10.28 10.04 +.24

Mating gilts at puberty with a vasectomizedboar increased farrowing rate 用输精管切除的公猪与初情期小母猪交配可提高分娩率 Study 研究 Gilts mated to sterile boar at puberty用不育公猪与初情期小母猪交配 Gilts were not mated不与小母猪交配 Difference 差异 1 (n = 265) 87.0 81.0 6.0 2 (n = 192) 88.0 85.0 3.0 Note: Gilts were mated to fertile boar at second estrus.用第二情期小母猪与有生育力公猪交配

Mating gilts at puberty with a vasectomized boar increased litter size 用输精管切除的公猪与初情期小母猪交配可提高窝产仔数 Study研究 Gilts mated to sterile boar at puberty用不育公猪与初情期小母猪交配 Gilts were not mated不与小母猪交配 Difference 差异 1 (n = 265) 9.9 9.5 0.4 2 (n = 192) 10.4 9.6 0.8 3 (n = 98) 10.9 9.3 1.6 4 (n = 34) 1.4 Note: Gilts were mated to fertile boar at second estrus.用第二情期小母猪与有生育力公猪交配

Pregnancy Diagnosis妊娠诊断 Blood tests (23 to 30 days)验血(23～30天) Progesterone (RIA procedure)孕酮(放射免疫法) Estrone sulphate硫酸雌酮 Estrone glucuronide葡(萄)糖苷酸雌酮 Estradiol glucuronide]葡(萄)糖苷酸雌二醇 Fecal tests (20 to 21 days)粪检(20～21天) Estrone (ELISA procedure)雌酮(ELISA)法 Urine tests (23 to 30 days)尿检(23～30天) Conjugated estrone (RIA) 共轭雌酮(放射免疫法) Estrone sulphate (immunoenzymatic)硫酸雌酮(免疫酶法) Ultra-sound techniques超声波技术

Synchronization of Estrus同期发情 PGF2a – can be effective in shortening the luteal phase after d 12 of the estrous cycle. Usually 2 i.m. injections of 10 mg/ea 12 h apart.前列腺素PGF2a可有效缩短发情周期12天后的黄体期。通常注射二次，每次10毫克，肌肉注射；二次注射间隔12小时 Matrix – a progestagen? (altrenogest) fed to cycling sows and gilts at 15 mg/d for 14 d. Upon removal of matrix, estrus occurs 6 days later. Costs ~ $1.50/h/d.孕激素(烯丙孕素，为人工合成的孕激素)饲喂给发情的母猪和小母猪，每天喂15毫克，喂14天。停喂后6天即发情。每天每头1.50美元 Weaning – Weaned sows exhibit estrus by d 3 -7 after weaning. This may be combined with administration of PG 600 (~$5.00/dose).断奶——断奶母猪会在断奶后3～7天发情。可在断奶的同时应用PG600(每剂5.00美元)