Low Dose Insemination in the Sow – A Review

Artificial insemination (AI) in pigs has been established for about four decades but ejaculates are still used insufficiently. Higher demand of semen for AI and new techniques that involve low sperm concentration require the optimization of insemination protocols. Based on the knowledge of the physiology of sperm transportation and events in the female genital tract prior to fertilization, new strategies are under development to minimize sperm losses. One goal is to deposit the semen into the uterine horn rather than into the proximal cervix. It was shown that the minimal number of spermatozoa necessary for surgical AI at the utero‐tubal junction (UTJ) were at least 1 × 10⁶ diluted in 0.5 ml of a special extender. Artificial insemination into the distal part of the uterine horn required about 1 × 10⁷ million sperm in 20 ml of extender. Meanwhile, first insemination devices for non‐surgical intra‐uterine AI are commercially available. Using similar sperm concentrations as for surgical AI, non‐surgical uterine insemination did not differ significantly from control inseminations in terms of pregnancy rate and litter size. With respect to the fertilizing capacities of their ejaculates, boars have to be selected more strictly for sperm quality parameters as most of the compensatory effects of sperm cells disappear in maximally extended semen samples.     

Successful Low Dose Insemination of Flow Cytometrically Sorted Ram Spermatozoa in Sheep

The fertility of ram spermatozoa that had undergone flow cytometric sorting (MoFlo SX) and cryopreservation was assessed after low-dose insemination of synchronized Merino ewes. Oestrus was synchronized with progestagen-impregnated pessaries, PMSG and GnRH treatment. Ewes (n = 360) were inseminated with 1 x 10(6), 5 x 10(6) or 15 x 10(6) motile sorted frozen-thawed (S(1), S(5), or S(15) respectively) or non-sorted frozen-thawed (C(1), C(5) or C(15) respectively) spermatozoa from three rams. An additional group of ewes were inseminated with 50 x 10(6) motile non-sorted frozen-thawed spermatozoa (C(50)) to provide a commercial dose control. The percentage of ewes lambing after insemination was similar for C(50) (24/38, 63.2%), C(15) (37/54, 68.5%), S(15) (38/57, 66.7%), S(5) (37/56, 66.1%) and S(1) (32/52, 61.5%) groups (p > 0.05), but lower for C(5) (19/48, 39.6%) and C(1) (19/55, 34.5%) treatments (p < 0.05). This study demonstrates sorted ram spermatozoa are equally fertile to non-sorted spermatozoa even when inseminated at 2% of the dose. Furthermore, at very low artificial insemination doses (1 or 5 million motile) the fertility of sorted ram spermatozoa is superior to non-sorted spermatozoa inseminated in equal numbers. These results have significance for the future commercialization of sex-preselection technology in sheep as a reduction in the minimum effective sperm number will allow a corresponding decrease in the associated cost per dose.     

奶牛性控冻精低剂量人工授精技术规范

奶牛性别控制繁育技术是继常规人工授精和胚胎移植之后,家畜繁育改良技术的第三次革命.由于性控冻精自身特点,在人工授精操作上与常规冻精输精略有不同,为了提高奶牛性控冻精受胎率和利用效率,节省成本,本规程对奶牛性控冻精低剂量深部人工授精技术环节进行了规范和要求.     

Incidence of Unilateral Fertilizations after Low Dose Deep Intrauterine Insemination in Spontaneously Ovulating Sows under Field Conditions

A new procedure for non-surgical deep intrauterine insemination (DUI) in unrestrained sows hormonally induced to ovulate, has been reported. In comparison with standard artificial insemination (AI), with this procedure, the sperm numbers inseminated can be reduced 20-fold without reducing the reproductive performance of these hormonally treated sows. The present study evaluated, using two experiments, the reproductive performance applying 20-fold different sperm numbers per AI dose using DUI or standard AI in spontaneously ovulating sows, under field conditions. In experiment 1, AI was applied to crossbred sows at 12, 24 and 36 h after onset of spontaneous oestrus using one of the following two regimes: (i) DUI (treatment) with 0.15 x 10(9) fresh boar spermatozoa in 5 ml of Beltsville thawing solution (BTS) extender (n = 95), and (ii) standard cervical AI (control) with 2.85 x 10(9) fresh spermatozoa in 95 ml of BTS extender (n = 95). The farrowing rates of the two groups of sows were statistically similar (NS). However, a decrease (p < 0.002) in litter size and the total number of pigs born alive was observed in sows inseminated with the DUI procedure. In experiment 2, 42 post-weaned oestrus sows were inseminated following the same design described for experiment 1 during spontaneous oestrus. On day 6 after onset of oestrus, the proximal segment of the uterine horns of the sows were flushed under surgery to retrieve eventual embryos and evaluate the success of fertilization per cornua (e.g. occurrence of effective uni- vs bilateral sperm transport rendering uni- or bilateral, complete or partial fertilization). Retrieved embryos were assessed for cleavage and number of accessory spermatozoa. Although identical overall pregnancy rates were achieved in both insemination groups, the percentage of sows with partial bilateral fertilization and unilateral fertilization was markedly higher (p < 0.05) in the DUI group (35%) compared with the control (standard AI) group (5%), with a consequent lower (p < 0.001) percentage of viable early embryos after DUI. The number of accessory spermatozoa in the zona pellucida of the embryos was highly variable, but higher (p < 0.001) in control animals than in DUI-AI. No accessory spermatozoa were found in oocytes retrieved from sows depicting unilateral fertilization. In conclusion, DUI in spontaneously ovulating sows with 0.15 x 10(9) spermatozoa renders similar farrowing rates but a lower litter size compared with use of standard AI with a 20-fold higher sperm dose. The lower litter size ought to be related to a decreased distribution of spermatozoa after DUI leading to a higher incidence of partial bilateral and unilateral fertilization.     

不同稀释液和输精量对天柱黑番鸭人工授精效果的影响

天柱黑番鸭是我国4个地方番鸭品种之一,已被列为地方品种资源进行保护和利用.由于骡鸭属于属间杂交的产物,自然交配受精率很低,仅为15%～45%,因此,开展人工授精是提高番鸭受精率根本途径.     

Distribution of T cells in the Endometrium of the Mare 6 and 48 h after Insemination

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In order to study the T-cell response after the introduction of semen into the uterine cavity, the distribution of helper T cells (CD4+) and cytotoxic/suppressor T cells (CD8+) was examined immunohistochemically in endometrial biopsy specimens. Endometrial tissue samples were obtained from 19 gynaecologically healthy mares during oestrus, both before and 6 or 48 h after deposition of a single dose of neat stallion semen. An increase (p = 0.04) in the numbers of helper T cells (CD4+) was observed at 6 h after insemination; thereafter the number of CD4+ cells decreased to pre-insemination values by 48 h after insemination. No significant variations in numbers of CD8+ cells were recorded either 6 or 48 h after insemination. There seems to be an early (6 h), recruitment of helper T cells into the equine endometrium after semen deposition, which might be related to the activation of the endometritis-like reaction seen as part of the equine uterine immune defence system during oestrus.     

Fertility in Single‐ovulating and Superovulated Dairy Heifers after Insemination with Low Dose Sex‐sorted Sperm

The present study was performed to test fertility in single‐ovulating and superovulated dairy heifers after insemination with low dose sex‐sorted sperm under field conditions. Some parameters, including the dosage, deposition site and timing, were assessed with the pregnancy rates after artificial insemination (AI). Moreover, the use of oestrus synchronization in combination with sorted sperm was evaluated. Besides that, we also improved the embryo production efficiency in superovulated dairy heifers by optimizing the timing of inseminations and repartitioning the sexed sperm dosage among multiple inseminations. The conception rate (52.8%) in heifers after low dose (2 × 10⁶) insemination with sorted sperm deep into the uterine horn did not differ (p > 0.05) from that (59.6%) of conventional AI (1 × 10⁷ non‐sorted sperm) and that of deep insemination with low dose non‐sorted sperm (57.7%). There was also no difference (p > 0.05) between conception rates after single (51.7%) and double (53.8%) deep insemination with sorted semen. Heifers inseminated with sorted sperm at synchronous oestrus had a lower pregnancy rate (48.1%) than heifers at spontaneous oestrus (53.6%), but this did not reach statistical difference (p > 0.05). The average number of transferable embryos collected in vivo from heifers inseminated with sorted sperm (4.81 ± 2.04) did not differ (p > 0.05) from that obtained from heifers after insemination with non‐sorted sperm (5.36 ± 2.74). Thus, we concluded that the pregnancy rate after deep intra‐uterine insemination with low dose sorted sperm was similar to that of non‐sorted sperm, which was either also deposited at a low dose deep intra‐uterine or into the uterine body. Sychronization of oestrus can be beneficial in combination with sorted sperm to optimize the organization and management of dairy herds. The results from superovulated heifers demonstrated that our insemination regime can be used to obtain a comparable embryo production efficiency with sorted sperm than with non‐sorted sperm.     

Uterine Insemination with a Standard AI Dose in a Sow Pool System

The effect of uterine AI with a standard dose of spermatozoa on fertility of the sow was studied in a field trial. The trial involved a sow pool system with 440 sows using AI as the primary method of breeding. Sows were twice a day checked for oestrus symptoms by back pressure test in front of a boar on days 3–6 after weaning. When in standing heat, sows were randomly allocated into either a uterine insemination group (UTER, n = 157) or standard AI group (CONT, n = 169) and bred accordingly using 3 billion spermatozoa in 80 ml of extender. In both treatment groups, insemination was repeated once if the sow was still receptive 24 h later. Using pregnancy (farrowed or not) and live‐born litter size as the outcome variables, a logistic and linear regression approach, respectively, was taken to study the effect of the following factors: treatment (UTER vs CONT), AI operator, breed, satellite herd preceding weaning, parity, weaning‐to‐oestrus interval and length of lactation. Overall, live‐born litter size was 11.3 ± 2.9, repeat breeding rate 4.2% and farrowing rate 91.2%. In the UTER group, 93.6% of inseminated sows farrowed, whereas farrowing rate for the CONT group was 88.8% (p = 0.13). Intrauterine insemination with a standard AI dose did not result in a significant improvement in the live‐born litter size (11.5 ± 2.8 for the UTER and 11.1 ± 3.0 for the CONT sows, respectively, p = 0.13). However, the preceding satellite herd had a highly significant effect on the live‐born litter size (12.4 ± 2.6; 11.1 ± 2.9; 10.8 ± 2.9 and 10.9 ± 2.9 for the four satellite herds, p < 0.01). We conclude that uterine insemination did not have a significant effect on live‐born litter size and farrowing rate and we also conclude that satellite herd appears to have a major effect on fertility in a sow pool system.     

Effect of Dose of Cloprostenol on the Interval to Ovulation in the Diestrous Mare: A Retrospective Study

Although the ovulatory effects of prostaglandins are well documented in several domestic species including horses, there has been little attention paid to the use of this ovulatory effect for clinical purposes. Mares often grow large follicles during the luteal phase that may or may not ovulate before progesterone levels decline. Clinical observations of administering prostaglandins in diestrous mares with large follicles suggest that there may be a negative correlation between follicular diameter and interval from treatment to ovulation. The objectives of this study were twofold: to investigate the cloprostenol dose rate effect on interval to ovulation and to confirm the negative correlation between follicular diameter and interval to ovulation. The hypothesis tested was that high doses of cloprostenol given in diestrus to mares with larger follicles would induce ovulation more rapidly than in mares given lower doses or with smaller follicles. To test the hypothesis, a total of 1,234 estrous cycles were induced with different doses of cloprostenol (ranging from 8.75 to 625 μg). All mares had at least one follicle of 28 mm or larger. Dominant follicles were followed by transrectal ultrasound examinations every other day until ovulation was detected. There was a significant effect of dose (P < .000) and follicular diameter (P < .000) on the interval from treatment to ovulation. The shortest mean interval (2.4 days) was observed after administration of 625 μg in mares with follicles 36 mm or larger, whereas the longest (4.9 days) occurred after 8.75 μg in follicles of 28 to 31 mm.     

An Abortion of Monozygotic Twins in a Warmblood Mare

Naturally occurring monozygotic twins are extremely rare in the horse. This paper describes an abortion in a mare after 260 days of pregnancy with monozygotic twins, one a fresh foal and the other a mummified foal.     

猪场低剂量深部输精技术应用研究

本文旨在研究低剂量深部输精操作及应用技术。每批受配母猪头数设计为1、5、10、15、20头,探讨每批母猪头数对内管插入阻碍率的影响;输精剂量设计为40、60 mL深部输精和80 mL常规输精,探讨不同输精剂量和输精方式对妊娠率、孕囊数和产仔率的影响;内管插入深度设计为10、12、15、18 cm,探讨不同输精深度对精液回流的影响;每组试验重复3次,每次实验母猪50头。结果表明:猪每批深部输精5~10头,内管插入阻碍率显著低于每批1头、15头和20头(P0.05);40、60 mL深部输精组与80 mL常规输精组母猪妊娠率差异不显著(P0.05);40 mL深部输精组B超检测孕囊数≤3的比例最高(29.87%),显著高于其余各组(P0.05);内管插入深度12 cm时的精液回流率显著低于内管插入10 cm和15 cm组(P0.05)。综上,每批输精5~10头、内管深入12 cm、输精剂量为60 mL更有利于猪深部输精操作和取得较好深部输精效果。     

Common Peripartum Problems in the Mare

Reproductive problems in the perinatal period can adversely affect a mare's future fertility or, worse, may be life threatening to her or her foal. A number of abnormalities can occur, including uterine torsion, uterine artery or uterine hemorrhage, retained fetal membranes, metritis, rectovaginal injuries, and necrotic vaginitis. Because hemorrhage, retained placenta, metritis, and necrotic vaginitis can happen after a normal delivery or after dystocia, all mares should be monitored closely in the first days after parturition for signs of lethargy, depression, colic, sore feet, or anorexia. Methods for diagnosing, managing, and treating these conditions and the possible complications are discussed.     

Some Aspects of Twin-gestation in the Mare

A study has been made of 44 cases of twin pregnancy in mares. The fetuses, foals, fetal membranes and blood groups of parents and foals were examined in order to determine the incidence of chorio-vascular anastomosis, blood chimaerism and freemartinism.

All of the twins proved to be dizygotic and 33 of the 34 pregnancies were bicornual. Fusion of the chorion was seen in 14 of 22 placentas and macroscopic choriovascular bridges were observed in five of these, although blood chimaerism was diagnosed in 11 of 25 cases (44 per cent) examined. This would indicate a high incidence of microchoriovascular anastomosis. The fetal female genital organs of four heterosexual twins were found to be anatomically and histologically normal at autopsy despite the existence of blood chimaerism. Five other sexvalley mature mares born as heterosexual twins with chimaerism all had normal genital organs and estrus cycles and one of these has since become pregnant. The important interspecies differences with respect to the incidence and degree of choriovascular anastomosis and freemartinism are stressed and suggestions for a plausible explanation were advanced.

     

Effect of Artificial Insemination Protocol and Dose of Frozen/Thawed Stallion Semen on Pregnancy Results in Mares

Deep intra‐uterine insemination is commonly accepted as a routine procedure for artificial insemination in horses. The motives and principles of deep insemination are well described, but the equipment used may differ. In this trial, the efficiency of two different insemination pipettes for deep intra‐uterine insemination in the mare was compared with insemination into the uterine body using commercially available frozen–thawed semen of two stallions of proven fertility. These inseminations were performed using two different doses. The semi‐flexible Minitube pipette was compared with a newly designed insemination device with a more flexible telescopic insemination catheter (Ghent device). The semi‐flexible Minitube pipette performed better than the newly designed insemination device with respect to pregnancy outcome (p = 0.008). The superiority of deep horn insemination over uterine body insemination was reflected by the better pregnancy rates obtained after deep insemination using the same low doses (30.6% better pregnancy rates) (p = 0.0123).     

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.