Distribution of Spermatozoa and Embryos in the Female Reproductive Tract after Unilateral Deep Intra Uterine Insemination in the Pig

The present study was performed to investigate the number of either the spermatozoa or the embryos in the reproductive tracts of sows after unilateral, deep, intra uterine insemination (DIUI). Two experiments were conducted, 10 sows were used in experiment I and eight sows were used in experiment II. Transrectal ultrasonography was used to examine the time when ovulation took place in relation to oestrus behaviour. The sows were inseminated with a single dose of diluted fresh semen 6-8 h prior to expected ovulation, during the second oestrus after weaning. In experimental I, five sows were inseminated by a conventional artificial insemination (AI) technique using 100 ml of diluted fresh semen, containing 3000 x 10(6) motile spermatozoa and five sows were inseminated by the DIUI technique with 5 ml of diluted fresh semen, containing 150 x 10(6) motile spermatozoa. The sows were anesthetized and ovario-hysterectomized approximately 24 h after insemination. The oviducts and the uterine horns on each side of the reproductive tracts were divided into seven segments, namely ampulla, cranial isthmus, caudal isthmus, utero-tubal junction (UTJ), cranial uterine horn, middle uterine horn and caudal uterine horn. Each segment of the reproductive tracts was flushed with Beltsville thawing solution (BTS) through the lumen. The total number of spermatozoa in the flushing from each segment were determined. In experimental II, eight sows were inseminated by the DIUI technique using 5.0 ml diluted fresh semen containing 150 x 10(6) motile spermatozoa. The sows were anesthetized 61.1 +/- 12 h after insemination (48-72 h) and the embryos were flushed from the oviduct through the proximal part of the uterine horn. It was revealed that, in experimental I, the spermatozoa were recovered from both sides of the reproductive tract in the AI-group, and from unilateral side of the reproductive tract in the DIUI-group (three sows from the left and two sows from the right sides). The number of spermatozoa recovered from the reproductive tracts was higher in the AI- than the DIUI-group (p < 0.001). In experiment II, fertilization occurred in five of eight sows (62.5%) after DIUI. The number of ova that ovulated were 16.4 +/- 2.6 per sow and the embryos numbering 11.4 +/- 2.3 per sow were recovered from both sides of the reproductive tract. In conclusion, the spermatozoa given by DIUI could be recovered from only one side of the reproductive tract of sows at approximately 24 h after DIUI via the flushing technique. However, embryos were found in both sides of the oviducts and the proximal part of the uterine horns 48-72 h after insemination, indicating that the fertilization occurred in both sides of the oviducts.     

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.     

Sperm distribution in the reproductive tract of sows after intrauterine insemination

The purpose of the present study was to compare the number of spermatozoa obtained from different parts of the oviducts and the uterine horns of sows after intrauterine insemination (IUI) and conventional artificial insemination (AI), 24 h after insemination. Twelve crossbred (Landrace x Yorkshire) multiparous sows were used in the experiment. The sows were examined for standing oestrus using a back pressure test and were examined every 4 h after standing oestrus by real-time B-mode ultrasonography to estimate the time of ovulation. The sows were allocated to two groups, group I sows (n = 6) were inseminated by a conventional AI technique with 3 x 10(9) motile spermatozoa in 100 ml of extended semen, and group II sows (n = 6) were inseminated by an IUI technique using 1 x 10(9) motile spermatozoa in 50 ml of extended semen. A single dose of AI or IUI was given using the same boar, 8-10 h before the expected time of ovulation during the second oestrus after weaning. Twenty four hours after insemination, the sows were ovario-hysterectomized. The oviducts and the uterine horns were removed and divided into seven parts, the cranial, middle and caudal uterine horns, the utero-tubal junction (UTJ), the cranial and caudal isthmus, and the ampulla. All parts of the reproductive tract were flushed and the spermatozoa were counted using a haemocytometer. The results revealed that the spermatozoa were found in both the oviducts and the uterine horns in all animals. The number of flushed spermatozoa in the UTJ of groups I and II, was 142,500 and 131,167 (p > 0.05), and in the caudal isthmus was 1411 and 1280 (p > 0.05), respectively. The proportion of spermatozoa in different parts of the reproductive tract in relation to the total number of spermatozoa within the tract was not significantly different between groups I and II (p > 0.05). It could be concluded that IUI, with a three-time reduction in the number of spermatozoa used resulted in the same number of spermatozoa to be deposited in the sperm reservoir around ovulation time.     

Number of Spermatozoa in the Crypts of the Sperm Reservoir at About 24 h After a Low‐Dose Intrauterine and Deep Intrauterine Insemination in Sows

The aim of this study was to investigate the number of spermatozoa in the crypts of the utero‐tubal junction (UTJ) and the oviduct of sows approximately 24 h after intrauterine insemination (IUI) and deep intrauterine insemination (DIUI) and compared with that of conventional artificial insemination (AI). Fifteen crossbred Landrace × Yorkshire (LY) multiparous sows were used in the experiment. Transrectal ultrasonography was performed every 4 h to examine the time of ovulation in relation to oestrous behaviour. The sows were inseminated with a single dose of diluted fresh semen by the AI (n = 5), IUI (n = 5) and DIUI (n = 5) at approximately 6–8 h prior to the expected time of ovulation, during the second oestrus after weaning. The sperm dose contained 3000 × 10⁶ spermatozoa in 100 ml for AI, 1,000 × 10⁶ spermatozoa in 50 ml for IUI and 150 × 10⁶ spermatozoa in 5 ml for DIUI. The sows were anaesthetized and ovario‐hysterectomized approximately 24 h after insemination. The oviducts and the proximal part of the uterine horns (1 cm) on each side of the reproductive tracts were collected. The section was divided into four parts, i.e. UTJ, caudal isthmus, cranial isthmus and ampulla. The spermatozoa in the lumen in each part were flushed several times with phosphate buffer solution. After flushing, the UTJ and all parts of the oviducts were immersed in a 10% neutral buffered formalin solution. The UTJ and each part of the oviducts were cut into four equal parts and embedded in a paraffin block. The tissue sections were transversely sectioned to a thickness of 5 μm. Every fifth serial section was mounted and stained with haematoxylin and eosin. The total number of spermatozoa from 32 sections in each parts of the tissue (16 sections from the left side and 16 sections from the right side) was determined under light microscope. The results reveal that most of the spermatozoa in the histological section were located in groups in the epithelial crypts. The means of the total number of spermatozoa in the sperm reservoir (UTJ and caudal isthmus) were 2296, 729 and 22 cells in AI, IUI and DIUI groups, respectively (p < 0.01). The spermatozoa were found on both sides of the sperm reservoir in all sows in the AI and the IUI groups. For the DIUI group, spermatozoa were not found on any side of the sperm reservoir in three out of five sows, found in unilateral side of the sperm reservoir in one sow and found in both sides of the sperm reservoir in one sow. No spermatozoa were found in the cranial isthmus, while only one spermatozoon was found in the ampulla part of a sow in the IUI group. In conclusion, DIUI resulted in a significantly lower number of spermatozoa in the sperm reservoir approximately 24 h after insemination compared with AI and IUI. Spermatozoa could be obtained from both sides of the sperm reservoir after AI and IUI but in one out of five sows inseminated by DIUI.     

Field Experiences on Post-cervical Artificial Insemination in the Sow

The present study was performed in order to evaluate the effects of post-cervical artificial insemination (post-CAI) in eastern European continental climate with multiparous sows. The sows were randomly allocated into two groups, and were AI by using CAI with 3 x 10(9) spermatozoa per dose (group 1, n = 859) or by post-CAI, using pooled semen with 1 x 10(9) spermatozoa per dose (group 2, n = 924). Wean-to-oestrus intervals, duration of oestrus, day 24 pregnancy rates, farrowing rates, and total pigs born were evaluated. Wean-to-oestrus intervals (CAI 114.3 +/- 4.1 h; post-CAI 115.2 +/- 5.2 h), duration of oestrus (CAI 64.1 +/- 4.1 h; post-CAI 65.0 +/- 5.2 h), day 24 pregnancy rates (CAI 90.2 +/- 1.7%; post-CAI 89.3 +/- 1.8%) and farrowing rates (CAI 88.1 +/- 2.3%; post-CAI 87.8 +/- 2.9%) did not differ significantly between CAI and post-CAI inseminated sows. The total number of pigs born differed significantly (p < 0.01) between the groups (CAI 12.3 +/- 1.1; post-CAI 10.2 +/- 0.9).     

Expression of Progesterone Receptor in the Utero‐tubal Junction After Intra‐uterine and Deep Intra‐uterine Insemination in Sows

The aim of this study was to investigate the expression of progesterone receptor (PR) in the utero‐tubal junction (UTJ) of sows at 24 h after intra‐uterine insemination (IUI) and deep intra‐uterine insemination (DIUI) compared with conventional artificial insemination (AI) in pigs. Fifteen multiparous sows were used: AI (n = 5), IUI (n = 5) and DIUI (n = 5). The sows were inseminated with a single dose of diluted semen during the second oestrus after weaning at 6–8 h prior to ovulation (AI: 3000 × 10⁶ spermatozoa, IUI: 1000 × 10⁶ spermatozoa and DIUI: 150 × 10⁶ spermatozoa). The UTJ was collected and subject to immunohistochemical staining using avidin‐biotin immunoperoxidase technique with mouse monoclonal antibody to PR. In the oviductal part of the UTJ, the intensity of PR in the tunica muscularis and the proportion of PR‐positive cells in the surface epithelium after DIUI were lower than AI (p < 0.05). The intensity and the proportion of PR‐positive cells between AI and IUI in all compartments of the UTJ did not differ significantly (p > 0.05). When comparing between tissue compartments, prominent staining was observed in the muscular layer of the UTJ. It could be concluded that the expression of PR in the UTJ prior to fertilization after DIUI with a reduced number of spermatozoa was lower than that after AI. This might influence sperm transportation and the fertilization process.     

采用深部输精与常规输精研究不同剂量和有效精子数对经产母猪繁殖力的影响

为探讨不同输精剂量与不同有效精子数对经产母猪繁殖力的影响,使用深部输精和常规输精作为主要配种方式。将发情母猪随机分为2组,A组为深部输精组(n=60),B组为常规输精组(n=60)。对A、B组内母猪分别用不同的输精剂量(Ⅰ:30亿/80 mL、Ⅱ:15亿/80 mL、Ⅲ:15亿/40 mL;n=20)进行输精。结果表明:与常规输精相比,深部输精组的受胎率和分娩率分别为93.33%和88.33%,略高于常规输精组的86.67%和83.33%,但差异不显著(P>0.05);但深部输精可显著提高经产母猪的产仔数和产活仔数,分别提高1.07头和1.15头。无论是深部输精还是常规输精,15亿/80 mL的输精量,均可获得与30亿/80 mL输精量一致的产仔效果,这一结果可以使优秀种公猪的利用率提高1倍,降低了最新国标《GB23238-2009》中对外种猪输精剂量的要求,建议扩大实验并推广使用。     

Within-herd Use of Boar Semen at 5°C, with a Note on Electronic Monitoring of Oestrus

A system was designed to allow a small swine farm in a northern latitude to use its own boars for artificial insemination (AI) conveniently. Semen was collected twice weekly for 3 day use (days 0, 1 and 2), extended in an egg yolk extender and stored at 5°C. Farm personnel were trained to manage the entire AI programme. For simplicity all semen collected was used for insemination. In the first test 47 gilts and 15 sows were inseminated with semen from four boars. One boar was subfertile with a farrowing rate of 36%. The averages for the other boars ranged from 71 to 100%. Then semen was collected from seven boars and all was used to inseminate 70 gilts and 55 sows with 3 × 10⁹ or more sperm. Overall 63% farrowed an average of 10.1 piglets per litter. Litter size for sows was 1.5 piglets larger than for gilts. There was no difference in farrowing rate when more than 3 × 10⁹ sperm were inseminated. The feasibility of initiating a complete AI programme within a small herd using herd boars was established. However, selection of the boars, use of only high quality semen, and experience with detecting oestrus was required to increase the farrowing rate. The use of various agents to protect sperm against cold shock below 15°C is worthy of further investigation. A new type of electronic probe, which measures the conductivity of cervical mucus, could be helpful if a boar is not available for conventional detection of 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.     

Artificial Insemination of Swine: Fecundity of Boar Semen Stored in Beltsville TS (BTS), Modified Modena (MM), or MR-A and Inseminated on One,Three and Four Days After Collection12

Contents: A field trial was conducted on 1,463 farms in The Netherlands to compare the fertilizing capacity of boar spermatozoa extended in Beltsville TS (BTS), Modified (MM), or MR-A, and inseminated in 2896 sows and gilts on the first, third or fourth day following collection. Semen was collected, extended, and stored at 18°C at six different AI centers and inseminated by inseminators on their regular routes. Sows inseminated with BTS and MR-A extended semen had higher farrowing rates than MM (79.3, 77.6 us 50.4, P <.0001), higher total pigs per litter (11.4, 11.1 vs 10.0, P <.0001) and higher total pigs born alive (10.7, 10.5 vs 9.4, P <. 0001). The farrowing rate of gilts inseminated with BTS extended semen was superior to MM (73.5 vs. 50.2%, P <.004), while MR-A gave farrowing rates greater than MM (64.1 vs 50.2%, P =.06). There was no difference in litter data for gilts. Farrowing rates for 1 and 4 day semen were superior to 3 day semen (73.2, 73.8 vs 60.3, P <.0001). The semen inseminated on day 4 contained 6 billion sperm per dose rather than the 3 billion per dose for 1 and 3 day semen. Based on the results of this study, BTS and MR-A are effective diluents for extension and storage of boar semen for use within the same week of collection. In addition, semen extended in either BTS of MR-A and stored and inseminated on the fourth day after collection can give fertility equal to first day insemination if the sperm per insemination dose is doubled. Inhalt: Künstliche Besamung beim Schwein: Fruchtbarkeitsergebnisse von Ebersamen nach Flussigkonservierung mit BTS, MR-A oder modifiziertem Modena-Verdünner (MM) und Besamung am ersten, dritten und vierten Tag nach Samengewinnung In einem in den Niederlanden durchgeführten Feldversuch wurden 3 verschiedene Verdünner-Medien für Ebersamen in vivo verglichen. Die Medien waren Beltsville TS (BTS), modifizierter Modena-Verdünner (MM) und das spanische MR-A-Medium. Insgesamt wurden 2896 Jung- und Altsauen in 1463 Herden mit 1, 3 oder 4 Tage altem Sperma besamt. Der Versuch wurde als “split-sample”-Versuch an 6 Eberstationen durchgeführt, der verdunnte Samen bei + 18°Cgelagert und von den Besamungsbeauftragten im Rahmen ihrer regulären Fahrten eingesetzt. BTS- und MR-A-konserviertes Sperma gab bei Altsauen gegenüber MM höhere Abferkelraten (79,3, 77,6 vs 50,4%, P < 0.0001), eine höhere totale Wurfzahl (11,4, 11,1 vs 10,0, P < 0.0001) und eine höhere Anzahl lebendgeborener Ferkel)10,7, 10,5 vs 9,4, P < 0.0001). Die mit BTS-uerdünntem Samen inseminierten Jungsauen wiesen gegenüber MM höhere Abferkelraten auf(73.5 vs 50,2%, P < 0.004). MR-A-Sperma lag ebenfalls hüher als MM-Samen (641 vs 50,2%, P=0.06). Die Wurfgröβen der Jungsauen reigten keine Unterschiede. Die Abferkelergebnisse nach Besamung mit 1 und 4 Tage altem Samen lagen höher als die des 3 rage alten Spermas (73,2, 73,8 vs 60,3%; P < 0.0001). Die am Tag 4 verwendeten Samenportionen enthielten 6 Milliarden Spermien gegenüber 3 Milliarden der nach 1 bzw. 3 Tagen inseminierten Portionen. Die Ergebnkse dieses Versuches zeigen, daβ BTS und MR-A effektive Verdünnermedien sind und die Lagerung von Eberfrischsperma zum Einsatz innerhalb der Entnahmewoche ermoglichen. Darüber hinaus können mit BTS oder MR-A verdünntes und am Tag 4 nach Gewinnung eingesetrtes Sperma Fruchtbarkeitsergebnisse erzielt werden, die ebenso gut sind wie nach Verwendung des 1-Tage-Spermas, wenn die Spermienzahl pro Besamungsdosis verdoppelt wird.     

Fertility Results of Artificial Inseminations Performed with Liquid Boar Semen Stored in X-CellTM vs BTS Extender

The objective of the present field study was to compare the fertility results for boar semen diluted in X-cell stored up to 4-5 days before artificial insemination (AI) with semen diluted in Beltsville thawing solution (BTS) used for AI following 2-3 days of storage (where the first day being the collection day). A total number of 2601 double inseminations in Norwegian herds were included in this two-trial study. All the boars used in the study were mature cross-bred Norwegian Landrace x Duroc (LD), which were routinely used for AI in Norway. The inseminated gilts and sows were Norwegian Landrace x Yorkshire (LY). The AI doses contained 2.5 billion spermatozoa, and consisted of a mixture of semen from three, occasionally four, boars (i.e. heterospermic semen). Fertility was measured in terms of the likelihood of farrowing and subsequent litter size. The fertility of the semen in both of the extenders was satisfactory and no significant differences were found either in semen stored 4-5 days in X-cell compared with 2-3 days in BTS or in semen stored 2-3 days in X-cell compared with 2-3 days in BTS. The storage capability findings for the long-term extender X-cell could significantly simplify the practical issues of semen production and the distribution of AI doses containing 2.5 billion spermatozoa. However, in pig production systems where all semen is used within 2-3 days, the short-term extender BTS is as good as the more expensive extender X-cell.     

Assessing in vivo Fertilizing Capacity of Liquid-Preserved Boar Semen According to the 'Hanover Gilt Model'

The goal of this study was to determine the ability of the Hanover gilt model to assess in vivo fertilizing capacity of preserved sperm and to consider whether any modifications to this model were needed. This model evaluates the fertilizing capacity of semen based on the fertilization rate, the rate of normal embryos and the accessory sperm count of 3–5‐day embryos. Its distinguishing characteristics are the use of one‐time insemination of sperm in reduced numbers, of spontaneously ovulating gilts and of ovulation detection through ultrasound examination of ovaries. Reduced sperm numbers allow for an accurate evaluation of the fertilizing potential of different semen treatments, thereby avoiding the compensatory effect of doses calibrated to maximize fertility. The model's usefulness was assessed in a trial run designed to compare the fertilizing capacity of liquid boar semen diluted into two different extenders. The diluent, the boar and the backflow, had no significant effect on any of the parameters studied. Gilts inseminated less than 24 h before ovulation had a significantly higher (p < 0.01) fertilization rate and accessory sperm cell count (p < 0.05) than those inseminated more than 24 h before ovulation. Very good/good embryos from homogeneous litters (only very good/good embryos were present) had a significantly higher (p < 0.01) accessory sperm count than those from heterogeneous litters (at least one embryo was of a different quality and/or oocytes were present). Both very good/good and degenerated/retarded embryos from heterogeneous litters had low accessory sperm numbers. This suggests that accessory sperm count is significantly related to the quality of the litter, but not to the quality of the embryo within gilts. It can be concluded that the Hanover gilt model is sensitive enough to show fertility differences (in this study, those associated with in vivo ageing of semen), while using relatively few gilts and little time.     

Management and Genetic Factors Affecting Fertility in Sows

Contents The purpose of this article is to summarize the current knowledge on genetic and management methods for improving fertility in sows. Fertilization rate, litter size and the interval between weaning and oestrus are traits that can be monitored on farms. These traits are heritable but can also be influenced by management. Age at puberty and the interval between weaning and oestrus are genetically linked ( r _g = 0.3) and a shorter weaning-to-oestrus interval is related to a longer duration of oestrus. Consequently, selection may be used to simplify detection of oestrus but because of variation in duration of oestrus between lines/breeds, farm-specific insemination strategies are needed. The direct boar effect on fertilization is small, probably due to the superabundance of sperm cells that is available in each dose of semen. Consequently, the use of a large number of sperm cells per dose is the best management tool for avoiding fertilization problems in sows. Health status of the boars, both for artificial insemination and natural mating, is of importance in order to produce a large number of sperm cells/doses of good quality per ejaculate. It is concluded that factors that influence fertility in sows, particularly prior to and during fertilization, are mainly related to oestrus and the optimal timing of insemination. Selection for adequate symptoms of oestrus, together with farm-specific insemination strategies are needed.     

The Post‐Cervical Insemination does not Impair the Reproductive Performance of Primiparous Sows

The study evaluated the reproductive performance of primiparous sows submitted to post‐cervical insemination (PCAI) compared with cervical artificial insemination (CAI). Difficulty with catheter introduction, the occurrence of bleeding or semen backflow during insemination, and volume and sperm cell backflow up to 60 min after insemination were also evaluated. Sows were homogenously distributed, according to body weight loss in lactation, lactation length, weaned piglets, weaning‐to‐oestrus interval and total born in previous farrowing, in two treatments: PCAI (n = 165) with 1.5 × 10⁹ sperm cells in 45 ml (2.4 ± 0.04 doses per sow) and CAI (n = 165) with 3 × 10⁹ sperm cells in 90 ml (2.5 ± 0.04 doses per sow). During PCAI, sows were inseminated in the absence of boars. Transabdominal real‐time ultrasonography was performed at oestrus onset, immediately before the first insemination and at 24 h after last insemination. There was no difference (P > 0.05) between treatments in farrowing rate (91.5% × 89.1%) and litter size (12.5 × 11.9 piglets born, respectively for PCAI and CAI sows). Successful passage of the intrauterine catheter in all the inseminations was possible in 86.8% (165/190) of sows initially allocated to PCAI treatment. Difficulty of introducing the catheter in at least one insemination did not affect the reproductive performance of PCAI sows (P > 0.05). Bleeding during insemination did not affect (P > 0.05) the farrowing rate in both treatments, but litter size was reduced in CAI and PCAI sows (P ≤ 0.06). Percentage of spermatozoa present in backflow within 1 h after insemination was greater in CAI than PCAI sows (P < 0.01). More than 85% of primiparous sows can be successfully post‐cervical inseminated with doses containing 1.5 × 10⁹ sperm cells in the absence of the boar during insemination without impairing the reproductive performance.     

Boar sperm production: Use in artificial insemination — A review

The production of spermatozoa increases with boar age from puberty to adulthood. Increasing collection frequency to more than once or twice a week increases the quantity of spermatozoa recovered per boar. Possible improvement of quality by collecting more often remains open to discussion. Sperm production is mainly a result of testis size, which varies with individuals and families. Selection on female ovulation rate leads to more rapid testis growth of males in the families selected. There is a correlation between pregnancy rate obtained after insemination with a given board, small size (direct boar effect) and embryonic survival rate at 25 days. Keeping boars at a high temperature lowers the quantity of spermatozoa recovered, pregnancy rate and embryonic survival in the sows inseminated. Fresh sperm may be stored for 2 days without decreasing its fertilizing ability (dose of 3 × 10⁹ spz. per AI). It may be efficiently used on day 3 of storage (double dose). The preparation and use of deep-frozen sperm is simple. Efficacious use is highly correlated with a good knowledge of sow oestrus. The number of sows which can be inseminated with one deep-frozen ejaculate is limited to 8–10 due to the number of motile spermatozoa after thawing.     

Relationship between sperm quality traits and field-fertility of porcine semen

An investigation involving seven boars, active in artificial insemination, and 1,350 multiparous sows was conducted at a private farm and aimed at examining the relationship between sperm quality traits and boar fertility in terms of farrowing rate and litter size. This experiment was done for 6 months. The semen samples were evaluated for subjective sperm motility and concentration. Ejaculates with at least 1 × 10⁸ sperm/mL and 70% sperm progressive motility were extended with a commercial medium to 30 × 10⁶ sperm/mL and used for artificial insemination (AI). AI dose was 100 mL semen containing 3 × 10⁹ spermatozoa. Aliquots of diluted semen were assessed for live morphologically normal spermatozoa (LMNS, eosin-nigrosin stain exclusion assay) and sperm chromatin instability (SCI, acridine orange assay). Farrowing rates according to different boar sperm varied (p < 0.001) from 59.3 to 88.92%. The mean values of LMNS (47.2~76.5%) and SCI (0.16~4.67%) differed significantly among boars. LMNS (r = 0.79, p < 0.05) and SCI (r = -0.90, p < 0.02) accounted for 62.2 and 81.7% of the variability in farrowing rates, respectively. After the combination of sperm traits, the relationship between percentage of LMNS with stable chromatin structure and farrowing rate was significant (r = 0.86, p < 0.05). The number of live piglets per parturition was not significantly correlated with sperm quality attributes. In conclusion, boar fertility after AI with freshly diluted semen can be predicted based on the evaluation of sperm morphology and chromatin integrity.     

Use of a novel double uterine deposition artificial insemination technique using low concentrations of sperm in pigs

Currently, the three most important non-surgical artificial insemination systems used in pigs are the conventional, the post-cervical (IUI), and the deep-intrauterine (DIUI) methods. In this study, a new system, termed double uterine deposition insemination (DUDI), which combines aspects of both IUI and DIUI, was evaluated. This method used a thinner, shorter and more flexible catheter than those normally used for DIUI and resulted in the deposition of semen post-cervically, approximately half-way along the uterine horn, thus potentially by-passing the threat of 'unilateral' insemination or pregnancy when using sperm of low concentration. The experiment was carried out over 8 weeks on a group of 166 sows, which were divided into seven groups, inseminated with semen of varying concentration, using the conventional system (control group) or by DUDI. There were no significant differences in fertility at day 35 post-insemination between the controls and the various DUDI sub-groups. Only sows inseminated with 500 million viable spermatozoa in a total of 30 mL of fluid using the DUDI system demonstrated decreased total litter sizes when compared to conventional insemination (P<0.001). While conventional insemination normally uses 2.5-3.5 billion sperm, the findings of this study suggest that DUDI can be used under 'field' conditions with sperm concentrations as low as 750 million spermatozoa in 50-30 mL without any detrimental effect on fertility or litter size. DUDI may provide a viable, robust alternative to IUI and DIUI, and has the potential to become incorporated into on-farm insemination systems.     

The Effect of Post-ovulatory Insemination on the Subsequent Embryonic Loss, Oestrous Cycle Length and Vaginal Discharge in Sows

The aim of present study was to study the effect of post-ovulatory insemination on the subsequent embryonic loss, oestrous cycle length and vaginal discharge in sows. Ten Large White multiparous sows were divided into two groups. Group A sows were inseminated once at 15 h after ovulation. Thereafter, they were ovariohysterectomized on day 11 (n = 5, first day of standing oestrus = day 1) and flushed for recovery of embryos. Group B sows were also inseminated once at 15 h after ovulation. They were further observed for return to oestrus and vaginal discharge (n = 5) after insemination. The endometrium tissues were biopsied from sows with vaginal discharge, embedded with paraffin, stained with haematoxylin and eosin and examined under light microscope. Only two embryos were observed in one of four sows from group A. All embryos had a spherical shape but differed in size (range 1-2 mm). In group B, only one sow had a regular return to oestrus (i.e. on day 23) and another sow had an irregular return to oestrus (i.e. on day 27). The other two sows in this group had shown vaginal discharge on days 20 and 38 after standing oestrus. For the number of leucocytes in the endometrium of sows with vaginal discharge, a large number of lymphocytes and plasma cells were observed in the connective tissue of the subepithelial layer. In conclusion, post-ovulatory insemination resulted in early embryonic loss, a subsequent prolonged oestrus interval and also vaginal discharge (i.e. endometritis) in sows.     

Motility Characteristics and Fertilizing Capacity of Boar Spermatozoa Stained with Hoechst 33342

Flow cytometry sorting of spermatozoa using fluorescence dye Hoechst 33342 is the only effective sex selection methodology validated in numerous laboratories. This study was carried out to determine the effect of Hoechst 33342 on the motility and fertility of stained boar spermatozoa. Experiment 1 evaluated motility parameters (percentage of motile spermatozoa, velocity, angularity and oscillation) of boar spermatozoa stained with Hoechst 33342 by a computer‐aided sperm analysis (CASA) instrument. Spermatozoa (30 million/ml) were divided into five treatment groups and stained during 1 h at 35°C with 9, 18, 27, 60 and 90 μM of H33342. There were no differences in sperm motility patterns nor percentages of motile spermatozoa incubated in the presence of 9, 18 or 27 μM. Percentage of motile spermatozoa and motility parameters decreased significantly (p < 0.05) at 60 μM of Hoechst 33342. Spermatozoa were immotile at concentration of 90 μM. In experiment 2, pregnancy rates, farrowing rates and litter size from sows (n = 275) artificially inseminated (AI) with either Hoechst 33342 stained (27 μM) or unstained (control) spermatozoa were determined. Sows inseminated with stained spermatozoa had no significant lower pregnancy rate (88.33%) as compared with controls (90.32%). Staining neither affected farrowing rates (85.0 vs 87.7%) nor total number of piglets born (10.56 ± 0.32 vs 10.47 ± 0.24, stained and controls, respectively). No phenotypical abnormalities were registered among the newborn piglets. The data suggest that incubating spermatozoa with Hoechst 33342 at levels required for X‐ and Y‐bearing chromosome sperm sorting, does not impair sperm viability or their fertility after AI.     

The effect of Oestrophan Spofa (synthetic analog of prostaglandin F2 alpha) added to the insemination dose on pregnancy and fertility in sows

Laboratory trials were conducted to study the effect of various concentrations of cloprostenol on the motility and morphological changes of the acrosomes of boar spermatozoa. As found, a cloprostenol concentration of 250 ng per ml of semen to 2500 ng per ml of diluted semen has no adverse effect on the motility of spermatozoa and on the morphological changes of their acrosomes. The concentration of 5000 ng of cloprostenol (in the Oestrophan Spofa product) per 1 ml of diluted semen negatively influences the motility of spermatozoa. An insemination dose of 100 ml of diluted sperm treated with 500 ng of cloprostenol was used for the artificial insemination of 152 sows; 166 sows of the same farm inseminated with untreated semen were used as controls. No gilts were included in the trial. Out of the 152 test sows, 113 conceived after the first insemination, i.e. 74.35%, and the average litter size was 10.04 piglets. In the control group, 125 sows delivered their litters, i.e. 75.30% of the total number, the average litter size being 9.96 piglets. Comparing the reproduction parameters of the experimental and control groups it can be said that the treatment of an insemination dose with 500 ng of cloprostenol immediately before insemination had no influence on the pregnancy rate and on the size of litters.