Fertility of deep frozen boar spermatozoa at various intervals between insemination and induced ovulation: influence of boars and thawing diluents.

The fertilizing ability of deep frozen boar spermatozoa was assessed after artificial insemination at various intervals after human chorionic gonadotropin (HCG) induced ovulation in gilts. Boar seminal plasma and OLEP were used as thawing diluent agents. The fertilization rates were similar when insemination was performed 2 or 6 hours before the time of estimated ovulation (40 hours post-HCG administration). However, fertility rates markedly declined when insemination was performed earlier than 6 hours before expected ovulation. In inseminations performed 2 hours before ovulation, the fertility rate was higher with specimens treated with boar seminal plasma diluent than with OLEP. As the difference between the time of insemination and that of ovulation increased, the difference in fertilizing ability of spermatozoa from the 2 boars used also increased. The results show that spermatozoa which have a low resistance to freezing-thawing also have a diminished fertile life in the female genital tract.     

Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin

The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal. Ejaculates from a suffolk ram were diluted with a Tris-based extender containing either 15% (v/v) egg yolk or 10% (w/v) BSA, and the diluted semen was frozen in 0.25 ml straws. A fixed-time intrauterine artificial insemination (AI) was performed 43-47 h after CIDR removal, regardless of incidence of estrus. There was no significant difference in pregnancy rates at 60 days after AI between the extenders containing egg yolk (66.7%, 20/30 animals) or BSA (65.5%, 19/29 animals). Furthermore, there were no significant difference in the lambing rates (66.7% and 62.1%) and prolificacy (1.25 and 1.56) between the two semen extenders. The present study indicates that a semi-defined semen extender containing 10% BSA produces fertility after intrauterine AI that is similar to that achieved with semen extender containing egg yolk.     

Fertility after artificial insemination using a soybean-based semen extender in sheep

The present study aimed to compare the fertility of ewes intrauterinally inseminated with frozen-thawed semen using a soybean-based semen extender (AndroMed) with those of ewes intrauterinally inseminated with frozen-thawed semen using a Tris-based extender containing either egg yolk or BSA. Suffolk ewes (n=104) were treated with an intravaginal sponge containing 40 mg fluoroprogesterone acetate (FGA) for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin to induce estrus and ovulation during the non-breeding season (July, 2007). Intrauterine insemination was carried out 40-46 h after removal of the FGA sponge (n=90), regardless of the incidence of estrus. The pregnancy rates were not significantly different among the semen extenders containing egg yolk (64.5%) or BSA (58.6%) and AndroMed extender (56.7%). The lambing rates (64.5, 55.2 and 56.7% for the semen extenders containing egg yolk, BSA and AndroMed, respectively) and prolificacy (1.59 to 1.75) were also not significantly different. The present results indicate that an egg yolk-containing semen extender can be replaced with the non-animal derived extender AndroMed, which could be used for intrauterine insemination using frozen-thawed ram semen without reducing fertility.     

Embryo collection from pigs post‐pseudopregnancy induced by estradiol dipropionate

We aimed to define whether embryo collection carried out after pseudopregnancy was of similar outcome and quality as after artificial abortion. To induce pseudopregnancy, 30 gilts or sows were given 20 mg intramuscular estradiol dipropionate (EDP) 10–11 days after the onset of estrus. Ten additional pigs were inseminated artificially at natural estrus as a control group. Prostaglandin F_2α (PGF_2α) was administered twice with a 24 hr interval beginning 15, 20, or 25 days after EDP‐treatment (n = 10 per group) or between 23 and 39 days after artificial insemination in control pigs. Following this, all pigs were given 1,000 IU equine chorionic gonadotropin and 500 IU human chorionic gonadotropin (hCG) and then inseminated. Embryos were recovered 6 or 7 days after hCG treatment and outcome was recorded. There was no significant difference in the number of normal embryos collected from the pigs with PGF_2α initiated at different time points or from the control group. Embryonic developmental stages 7 days after hCG treatment also did not differ among groups. These results indicate that the use of EDP to induce pseudopregnancy, followed by PGF_2α administration to synchronize estrus for subsequent embryo harvest, is a suitable alternative to the artificial abortion method.     

性控冻精对同期发情奶牛输精效果观察

为观察奶牛养殖场应用同期发情后,实施性控冻精人工授精的效果进行试验。本试验设立性控组和常规组,分别进行人工授精,在相同饲养管理条件下观察两组试验奶牛配种、妊娠、产母犊情况。结果表明:同期发情率为81.43%,而且84.38集中在36h内;试验组奶牛产母犊率达94.12%,明显高于常规组55.56%的产母犊率,说明该性控冻精在奶牛性别控制中的效果明显,值得推广。     

Studies of gel with immobilized semen by intrauterine endoscopy post-artificial insemination

An extended lifespan of spermatozoa following artificial insemination (AI) can make the timing of insemination less critical, as previously demonstrated with immobilized spermatozoa that are gradually released from an alginate gel. The purpose was to examine the in vivo dissolution of SpermVital (SV) alginate gel over time by endoscopy and secondly to assess spermatozoa quality after incubation of the gel. In vivo endoscopy showed SV gel in the uterus 3, 6, 20 and 24 hr after AI, demonstrating the potential release of spermatozoa to the uterus during this period. In utero ex vivo incubation of the semen demonstrated that high motility and viability of sperm cells was sustained following overnight incubation.     

Effect of Timing of Postovulatory Insemination Relative to Human Chorionic Gonadotropin/Buserelin Treatment With 1 Straw of Frozen-Thawed Semen on Mare Fertility

The reproductive management of mares for frozen semen artificial insemination (AI) can be costly and labor intensive. Predicting the exact time of ovulation can be challenging even when ovulation-inducing drugs are used. The main objective of this retrospective study was to determine whether there was an effect of interval between examinations to detect ovulation on likelihood of pregnancy and early embryonic loss in mares after postovulatory breeding with a single straw of frozen/thawed semen. The second objective was to determine the efficacy of two different drugs (human chorionic gonadotropin vs. buserelin) for timely induction of ovulation. The length of the interval from penultimate check to ovulation had no significant effect on pregnancy or embryo loss rates (4 hours: 34.1% and 13.3% vs. 8 hours: 26.1% and 0% and 16 hours: 34.5% and 10%, respectively) nor did the ovulation-inducing drug used, number of the cycle, or the stallion. In conclusion, there appears to be no advantage of checking mares for ovulation during the late evening and night hours when using a postovulatory AI protocol and ovulation-inducing drugs.     

Transmission of classical swine fever virus by artificial insemination.

Classical swine fever (CSF) virus was introduced into an artificial insemination centre during the CSF epizootic of 1997-1998 in the Netherlands. The risk of further spread of CSF virus via contaminated semen was recognised, but could not be assessed because scientific data on this issue were not available. An animal experiment was performed to determine whether CSF virus could be transmitted via artificial insemination with contaminated semen. Three boars were inoculated with a CSF virus field isolate and from Day 5 till Day 18 thereafter, ejaculates were collected and prepared for insemination. Ruttish sows were inseminated with the extended semen from Day 5 till Day 18 after inoculation of the boars. All the inoculated boars remained healthy throughout the experiment and developed CSF neutralising antibodies between 14 and 21 days after inoculation. Virus was isolated from several semen samples collected from 5 till 11 days after inoculation. Two out of six sows inseminated with CSF contaminated semen seroconverted after insemination. All the other sows remained seronegative. In the foetuses of both the seropositive sows, CSF virus was detected at approximately 35 days post insemination. These results demonstrate that adult boars infected with CSF virus can excrete virus with semen and can, subsequently, transmit the virus to sows and their foetuses via artificial insemination.     

Serum progesterone and estradiol-17beta concentrations, and lapaloscopic observations of the ovary in the cheetah (Acinonyxjubatus) with pregnant mare serum gonadotropin and human chorionic gonadotropin treatments.

In 3 adult female cheetahs, induced-superovulation treatment was conducted, by means of 200 IU of pregnant mare serum gonadotropin (PMSG) and 100 IU of human chorionic gonadotropin (hCG) 80 hr after PMSG. The administration of PMSG created a sharp increase in the estradiol-17beta concentration, resulting in 232 pg/ml 8 hr later in one specimen out of three. The hCG administration showed an increase in the progesterone concentration of 2.29 ng/ml 46 hr later. In addition, after direct observation of the ovary surface by laparoscopy, 5 follicles in the right ovary over 2 mm in diameter, and 7 corpora lutea (5 in the right ovary and 2 in the left) were found. It is assumed that ovulation can be induced with hCG after 80 hr on PMSG during a cheetah's diestrus or proestrus.     

牦牛的人工授精技术

人工授精技术是牦牛养殖的重要环节,本文就公牦牛驯养,人工授精前的准备、过程、精液品质检查、母牛发情鉴定,输精等环节做了介绍.     

Use of Direct Thaw Insemination to Establish Pregnancies with Frozen–Thawed Semen from a Standard Jack

Pregnancy rates reported after artificial insemination with frozen–thawed jack spermatozoa have been relatively low compared with those attained in other species. Cholesterol is known to influence post-thaw fertility of both jack and stallion semen, and altering the amount of cholesterol in the freezing extender may help improve the fertility of frozen–thawed jack semen samples. In this study, we report clinical work that was performed using semen samples collected from a single jack. Samples were extended in EZ Mixin OF and then slowly cooled to 5°C. Extended semen samples were centrifuged at 400 × g for 10 minutes and the supernatant was discarded. Spermatozoa were resuspended in freezing medium to a final concentration of 400 × 10⁶ cells/mL and were later frozen in liquid nitrogen vapor. Freezing extender treatments containing 2% ethylene glycol included the following: (1) 20% egg yolk (EY), (2) 5% EY, and (3) 20% EY + 60 mM hydroxypropyl-β-cyclodextrin (β-CD). For this study, a total of 28 mares aged 2 to 18 years was used over five breeding seasons (82 total cycles). Mares were administered human chorionic gonadotropin to induce ovulation when the dominant follicle was ≥35 mm in diameter. They were inseminated within 6 hours before ovulation and again within 6 hours after ovulation. Pregnancy rates obtained were as follows: (1) 6.25% (one of 15 matings) for 20% EY, (2) 46.5% (20 of 43 matings) for 5% EY, and (3) 58.5% (14 of 24 matings) for 20% EY + 60 mM β-CD. These data suggest that binding of cholesterol with β-CD enhances post-thaw fertility of jack semen samples. We conclude that acceptable pregnancy rates could be achieved with frozen–thawed jack semen samples cryopreserved in 5% EY or 20% EY + 60 mM β-CD using direct post-thaw insemination.     

Unilateral intrauterine horn insemination of fresh semen in cats

The sperm count required were investigated to obtain a conception rate of 80% by unilateral intrauterine insemination (UIUI) of fresh semen in cats. The conception rates obtained by insemination before and after ovulation were also examined. Thirty-six female cats aged 1-7 years were used in the experiments, and the number of experimental cases was 44. Seven male cats aged 2-12 years from which semen could be collected by the artificial vagina method were used. In artificial insemination, 100 iu x 2 or 250 iu of hCG was administered on days 2-4 of estrus, and sperm were introduced into the uterine horn with a greater number of ovulations (or mature follicles) 15, 20 and 30 hr after hCG administration by laparotomy. The inseminated sperm counts were 2 x 10(6) (Exp. 1). 4 x 10(6) (Exp. 2), and 8 x 10(6) (Exp. 3). As a result, ovulation was induced in 42 of 44 cases (induction rate: 95.5%) regardless of the dosage of hCG. Conception was obtained by UIUI in two of 16 animals (conception rate: 12.5%) in the Exp. 1, five of 16 animals (31.3%) in Exp. 2, and eight of 10 animals (80.0%) in Exp. 3. Regarding the relationship between the ovulation state at insemination and conception, the conception rate obtained by insemination before ovulation was clearly higher than that obtained by insemination after ovulation (p<0.05). Regarding the number of kits compared to the number of ovulations on the inseminated side, the percentages of cases in which the number of kits exceeded the number of ovulations on the inseminated side were similar in all groups inseminated with a different number of sperm. It is therefore necessary to investigate conception rates obtained by bilateral insemination to increase the fertility rate. Based on the above findings, it was shown that the sperm count required for fertilization by UIUI is 8 x 10(6).     

补饲与犊牛断乳对产后母牦牛发情周期恢复的影响研究

为了提高当年产犊母牦牛的发情率,我们对选定的120头怀孕母牦牛进行补饲,同时在犊牛产后3个月对犊牛进行隔离断奶,补饲试验于2010年3月初开始：100头母牦牛随机分为两组,试验组Ⅰ 50头母牦牛隔离断奶后进行同期发情和定时授精处理;Ⅱ组50头母牦牛隔离断乳.不进行同期发情处理,发情母牛进行人工授精。人工授精冻精液为荷斯坦和野牦牛冻精。配种45d后进行怀孕诊断。Ⅰ组发情39头,发情率为78%（39/50）,怀孕34头妊娠率为68%（34/50）;Ⅱ组发情31头,发情率为62%（31/50）,怀孕27头,妊娠率为54%（27/50）。相同草场放牧50头适配母牦牛作为对照。对照组母牛和犊牛不进行隔离和短期断乳;对照组发情率为12%（6/50）,妊娠率为6.32%（3/50）。试验结果表明,通过对带犊的母牦牛实施隔离断奶,能够明显提高当年产犊母牦牛的发情率和妊娠率。     

人工授精技术在骡鸭生产中的应用

番鸭与家鸭为不同属的鸭种,自然交配受精率较低,平均受精率为30%～40%。本研究采用人工授精技术,对公番鸭进行隔离调教,按摩法采精,母家鸭以翻肛法输精。结果表明:公番鸭调教1周即可使用,每天采精1次,采精量可达0.8毫升/只;精子在输精后第4天活力最强,两次输精间隔以4天为宜;常温下采出的新鲜精液应在30分钟用完;每只母家鸭每次输精0.05毫升,种蛋受精率平均达71.5%。     

Some factors affecting pregnancy rate in ewes following laparoscopic artificial insemination

The fertility of 646 ewes and gimmers bred by laparoscopic artificial insemination (LAI) in the autumn of 2006 was investigated using a questionnaire and individual ewe breeding records kept for 13 commercial sheep flocks that used LAI routinely. Overall, the pregnancy rate was 66 per cent, but it was highest in ewes bred for the fourth time. Technical aspects of LAI influenced fertility: pregnancy rates were 70 per cent for ewes bred using frozen semen compared with 58 per cent when fresh semen was used (P≤0.01), and 74 per cent of ewes that travelled to an artificial insemination centre for mating conceived, compared with 62 per cent that remained on their own farm (P<0.01). Higher doses of equine chorionic gonadotrophin (>400 iu) used for oestrus synchronisation reduced pregnancy rates to only 49 per cent (P<0.001). However, the largest effect was associated with shepherds gathering, handling and treating breeding ewes four to six weeks before mating; pregnancy rates were 54 per cent among ewes where this was carried out, compared with 74 per cent for ewes not treated in this way (P<0.01).     

Fertility after different artificial insemination methods using a synthetic semen extender in sheep

The present study aimed to investigate the fertility of ewes artificially inseminated with three different methods using a synthetic semen extender, AndroMed. The three methods of artificial insemination (AI) were cervical AI with fresh-diluted or frozen-diluted semen at observed estrus, and an intrauterine AI with frozen-thawed semen. A total of 80 ewes were treated with a controlled internal drug release (CIDR) containing 0.3 g progesterone per device for 12 days. In Experiment 1 (26 Suffolk ewes), superovulation was induced with 20 mg follicle-stimulating hormone and 250 IU equine chorionic gonadotropin (eCG) two days and one day before CIDR removal, respectively, during the non-breeding season. In Experiment 2 (54 Suffolk and Suffolk crossbred ewes), an intramuscular injection of 500 IU eCG was administered one day before CIDR removal to synchronize estrus and ovulation during the breeding season. In Experiment 1, fresh-diluted or frozen-thawed semen was deposited into the cervical orifice after estrus detection, and an intrauterine AI with frozen-thawed semen was performed by laparoscopy at a fixed-time basis without estrus detection. Embryos were recovered by uterine flushing 6 days after AI, and the rates of recovered, fertilized (cleaved) ova and embryos at the morula or blastocyst stage were compared among the three AI methods. In Experiment 2, the pregnancy rates after the three AI methods were compared. In Experiment 1, the rates of recovered ova were not significantly different among the three AI methods (52.5-56.7%). The rate of fertilized ova (81.0%) by laparoscopic AI with frozen-thawed semen was significantly higher compared with cervical AI of fresh-diluted (25.5%) or frozen-thawed (3.5%) semen, but the rate of embryos at the morula or blastocyst stage (17.6%) was significantly lower than that of the cervical AI with fresh-diluted semen (69.2%). The rates of ewes yielding fertilized ova were not significantly different among the three groups (44.4, 11.1 and 62.5% for cervical AI with fresh-diluted and frozen-thawed semen and intrauterine AI with frozen-thawed semen). In Experiment 2, the pregnancy rate of ewes intrauterinally inseminated with frozen-thawed semen (72.2%) was significantly higher than those of ewes inseminated cervically with fresh-diluted (5.5%) or frozen-thawed (0.0%) semen. The present results showed that acceptable fertilization and pregnancy rates could be obtained by an intrauterine AI with frozen-thawed semen using a synthetic semen extender (AndroMed), but not sufficient by the cervical AI with either fresh or frozen semen.     

Artificial intravaginal insemination using fresh semen in cats

To clarify the sperm count required for fertilization by artificial intravaginal insemination (AIVI), twenty-nine female cats were examined. Six male cats aged 2-12 years with normal semen quality, copulation capability, and fertility were used. In AIVI, animals received administration of 250 iu hCG once or 100 iu twice on days 2-4 of estrus to induce ovulation, and were inseminated 15, 20, or 30 hr after the initial hCG administration. The success of ovulation was judged by elevation of the peripheral progesterone level after hCG administration. AIVI was investigated at three sperm counts, 20 x 10(6) (Experiment 1), 40 x 10(6) (Experiment 2), and 80 x 10(6) (Experiment 3), with semen collected by the artificial vagina method. Semen was infused in the vagina under general anesthesia by advancing a 9 cm-long nylon probe with 1.5 mm diameter connected to a 1 ml syringe in the vagina for 3-4 cm. Ovulation was induced in 43 of 45 animals (95.6%). One of 16 animals was fertilized (conception rate: 6.6%) by AIVI in Experiment 1. In Experiments 2 and 3, conception was obtained in six of 18 animals (33.3%) and seven of nine animals (77.8%), respectively, and the mean numbers of kits were 4.0 +/- 0.4 and 3.3 +/- 0.5, respectively, and the mean numbers of kits were 4.0 +/- 0.4 (SE) and 3.3 +/- 0.5, respectively, showing no significant difference. There were no differences in the time of insemination after hCG administration and the conception rate among these groups. Our findings showed that the number of sperm required for fertilization by AIVI of fresh semen in cats was 80 x 10(6).     

Effect of Insemination–Ovulation Interval and Addition of Seminal Plasma on Sow Fertility to Insemination of Cryopreserved Sperm

In swine, the use of frozen-thawed (FT) sperm for artificial insemination (AI) is limited because of poor sow fertility, possibly associated with a post-thaw capacitation-like status resulting in fewer fully viable sperm. Sow fertility to AI with FT sperm may improve with deeper deposition of sperm within the female tract, insemination very close to ovulation, or reversal of cryocapacitation by seminal plasma (SP). We performed two experiments to examine these suggestions. In experiment 1, 122 multiparous Yorkshire sows received 600 IU equine chorionic gonadotrophin at weaning and 5 mg pLH 80 h later to control time of ovulation. The predicted time of ovulation (PTO) was 38 h after pLH injection. Thereafter, sows were assigned on the basis of parity to a single AI of FT sperm at 2 h before PTO, or at 12 h before PTO, or FT sperm supplemented with 10% SP at 12 h before PTO. Control sows received fresh semen at 12 h before PTO. All semen doses were adjusted to 3 x 10(9) live cells and deposited into the cervix. Experiment 2 employed 99 multiparous crossbred sows and repeated the treatments of experiment 1 except that all FT inseminations were intrauterine. In both experiments, farrowing rates were lower (p < 0.01) following FT inseminations with no effect of time of insemination or of supplemental SP. In experiment 1, litter size was smaller following FT insemination (p < 0.05), but no effect on litter size was evident in experiment 2. Supplemental SP had no effect on litter size in either experiment. The lack of effect of either SP or timing of FT insemination on sow fertility suggests that the non-lethal sperm cryoinjury affecting fertility involves more than just cryocapacitation.     

A study on the number of recovered spermatozoa in the uterine horns and oviducts of gilts,after fractionated or non-fractionated insemination

The objective of this study was to compare the number of recovered spermatozoa, in different parts of the uterine horn and oviduct in gilts, after insemination with fractionated (experiment) and non-fractionated (control) liquid stored semen. The number of spermatozoa and volume of backflow was also investigated. Twenty three cross-bred gilts were used in the study. They were divided into 2 groups, a control group (non-fractionated liquid stored semen, n=10) which were inseminated with 100 ml of liquid stored semen containing 3,000 million spermatozoa per dose and an experimental group (fractionated liquid stored semen, n=10) which were inseminated with 50 ml of liquid stored semen, with 3,000 million spermatozoa per dose and followed by another 50 ml of semen dilutor (Beltsville Thawing Solution, BTS). Thereafter, backflow semen was collected and measured every 15 min for a period of 1 hr. Three or 12 hr after insemination, 5 gilts from each group had the uterus, the horn of the uterus, the oviducts and the ovaries removed under general anaesthesia. The horn of uterus and the oviducts were seperated by ligation into 6 segments. All 6 segments were flushed with BTS to collect all spermatozoa within the segment. Recovered spermatozoa were counted, using a haemocytometer and the volume recorded. It was seen that the percentage of spermatozoa in the backflow semen in the experimental group was less than in the control group. The difference was not significant in the gilts that were operated on 3 hr after insemination, the mean number of spermatozoa in the uterine horn and the utero-tubular junction (UTJ) was more in the experimental than in the control group, but less in the isthmus and the ampulla of the oviduct. The gilts which were operated on 12 hr after insemination, had relativity more ovulating gilts in the control group than in the experimental group (3 of 4 gilts compare to 3 of 5 gilts). The control group had more spermatozoa in the oviduct than the experimental group, but less in UTJ and in the horn of the uterus. Again the difference was not significant. It can be concluded that fractionated (experimental) or non-fractionated (control) insemination of semen with the same number of spermatozoa provides no significant difference in the number of spermatozoa either in the horn of the uterus, the UTJ or the oviduct of gilts.     

METHOD OF SEMEN COLLECTION AND ARTIFICIAL INSEMINATION IN SNAKES

This study focuses on the method for sperm extraction and artificial insemination in snakes. Ten adult healthy snakes (4.6) were included in the study (Pantherophis guttatus 1.3; Hydrodynastes gigas 1.1; Corallus hortulanus 1.1; and Sanzinia madagascariesis 1.1). Massage of the ventral aspect of the caudal third of the male snake body for 2 to 3 minutes was performed successfully for the semen collection. Both female and male P. guttatus and H. gigas were placed in brumation, with the female snakes being assessed for ovarian activity after emerging from dormancy. Only females showing ultrasonographic evidence of vitellogenic follicles were included in the study. S. madagascariensis and C. hortulanus were maintained at the same temperature through the year, and the ovarian activity was assessed ultrasonographically prior to artificially inseminating the animal. With the aid of a rigid endoscope, fresh semen was delivered through the cloaca and into the females’ oviducts using a catheter connected to the syringe. The technique failed in female S. madagascariensis and H. gigas. Two female P. guttatus laid eggs 2 months after artificial insemination, with hatchlings emerging following 2 months of development within the eggs. The third female corn snake did not lay an egg. The female C. hortulans produced eggs 4 months after insemination. The manual massage method of sperm collection in male snakes and endoscopy-assisted insemination in female snakes may be useful for conservation programs.