Study of the distribution of inflammatory cells in the sow endometrium: effect of intravenous administration of adrenocorticotropin hormone

Seventeen multiparous cross-bred sows (Swedish Land-race x Swedish Yorkshire) were inseminated in their second oestrus after weaning and divided into two groups. One group (ACTH, n = 9) was given an intravenous injection of adrenocorticotropin hormone (ACTH) every 6 h commencing 4-8 h after ovulation, whereas another group (control, n = 8) was given saline solution at the same times. The sows were slaughtered 35-53 h after ovulation. Uterine samples, taken from the mesometrial side of the uterine horns immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue. The endometrium was then examined by light microscopy. There was no significant effect of the ACTH treatment on the distribution of lymphocytes and macrophages, but there was a tendency of an effect on the distribution of neutrophils (P = 0.1) in the sow endometrium.     

Influence of CRH and ACTH Administration on Endocrine Profile and Ovulation in Sows

Grouping of sows is a stressful event until the ranking is established. The purpose of this study was to simulate stress by repeated administration of porcine corticotropin releasing hormone (CRH) and adrenocorticotropic hormone (ACTH)/tetracosactide and to study its influence on endocrine profile and ovulation. Four multiparous sows were used and blood was collected every 2 h from the onset of pro-oestrus until 12 h after ovulation. The first oestrus after weaning was used to check ovulation and acclimate the sows to their environment. The second oestrus after weaning was used as control. At their third oestrus CRH (0.6 microg/kg) and at their fourth oestrus ACTH (5 microg/kg) were given every 4 h from onset of oestrus until ovulation. The total 'area under the curve' of cortisol was twofold larger in two of four sows during the CRH treatment period, and two- to fourfold larger (p < or = 0.05) during the ACTH treatment period, compared with the corresponding control period. In three sows, there was no clear effect of either CRH or ACTH on the levels of oestradiol 17beta, luteinizing hormone (LH) or on the timing of ovulation. One sow was different in all hormonal patterns and also in the timing of ovulation. In all four sows, ACTH treatment lowered the baseline level of prostaglandin F(2 alpha)-metabolite. Therefore, we conclude that stage of the oestrous cycle seems to be of importance when investigating the influence of exogenous administration of CRH/ACTH on hormonal pattern and ovulation time in the sow.     

Impact of postovulatory food deprivation on the ova transport, hormonal profiles and metabolic changes in sows.

The effect of food deprivation on ova transport, hormonal profiles and metabolic changes was studied in 20 crossbred multiparous sows during their second oestrus after weaning. To determine the time of ovulation, transrectal ultrasonographic examination was performed. The sows were divided into 2 groups, one control group (C-group), which was fed according to Swedish standards, and one experimental group (E-group). The E-group sows were deprived of food from the first morning meal after ovulation until slaughter. Blood samples were collected every second hour from about 12 h before expected ovulation in the second oestrus after weaning until slaughter and were analysed for progesterone, prostaglandin F2 alpha-metabolite, insulin, glucose, free fatty acids and triglycerides. All sows were slaughtered approximately 48 h after ovulation and the genital tract was recovered. The isthmic part of the oviduct was divided into 3 equally long segments and flushed separately with phosphate buffered saline (PBS). Uterine horns were also flushed with PBS. A significantly greater number of ova were found in the first and second part of the isthmus in the E-group (p = 0.05) while in the C-group most of the ova were found in the third part of the isthmus or the uterus (p = 0.01). The level of prostaglandin F2 alpha-metabolite was significantly higher in the E-group compared with the C-group. The concentration of progesterone increased in both groups after ovulation but there were no significant differences between the groups. The other blood parameters showed that the food-deprived sows were in a catabolic state. The 48 h period of fasting results, directly or indirectly in an delayed ova transport, which may be due to a delayed relaxation in the smooth circular muscle layer of the isthmus.     

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.     

The Effect of Intravaginal Applied GnRH-agonist on the Time of Ovulation and Subsequent Reproductive Performance of Weaned Multiparous Sows

In order to prove the effect of 'fixed time insemination' and insemination at standing oestrus after post-weaning application of GnRH, in a Croatian large breeding unit, 502 sows were assigned to three groups and were artificially inseminated (AI) at their first post-weaning oestrus as many times as they stand, in 24-h intervals. The groups were treated as follows: group 1 (control, n = 160) were AI during their standing reflex; group 2 ['GnRH-fixed time insemination' (GnRH-FT-AI), n = 175] were AI, independent of detection of oestrus and following administration of GnRH-agonist at 96 h post-weaning; group 3 [GnRH insemination at standing oestrus (GnRH-OE-AI), n = 167] the animals were GnRH-agonist treated as group 2 and were AI at their standing reflex. Pre-trial daily average lactational feed intake, average daily feed intake from weaning to oestrus, oestrus within 6 days post-weaning (%), ovulation within 6 days post-weaning (%), weaning-to-oestrus interval (h), duration of oestrus (h), follicle size (mm), interval from oestrus to ovulation (h), subsequent day 24 pregnancy rate (%), farrowing rate (%) and total pigs born were evaluated. Pre-trial average daily lactational voluntary feed intake was 7.1 +/- 0.08 kg in group 1, 7.0 +/- 0.07 kg in group 2 and 7.1 +/- 0.17 kg in group 3 (p > 0.05). Average voluntary daily feed intake from weaning to oestrus was 5.1 +/- 0.3 kg in group 1, 5.2 +/- 0.5 kg in group 2 and 5.2 +/- 0.19 kg in group 3 (p > 0.05). Oestrus was detected within 6 days post-weaning in 134 (83.8%) in control, 164 (93.7%) in GnRH-FT-AI and 155 (92.8%) animals in GnRH-OE-AI groups (p = 0.05). Follicle size did not differ (p > 0.05) among the groups. In control 82.8%, in GnRH-FT-AI 91.5% and in GnRH-OE-AI 91.0% of the sows ovulated within 6 days post-weaning (p = 0.04), and had 80.6, 90.9 and 89.7% 24-day pregnancy rates (p = 0.16), respectively. In GnRH-FT-AI group 90.2%, in GnRH-OE-AI sows 89.7%, in control animals 79.9% farrowing rates were recorded (p = 0.17). Weaning to oestrus interval was 113.1 h in control, 114.1 h in GnRH-FT-AI and 112.6 h GnRH-OE-AI (p > 0.05). Duration of oestrus was significantly shorter in GnRH-FT-AI (44.9 h) and GnRH-OE-AI (48.1 h) animals, compared with the control (62.9 h) sows (p = 0.001). Similarly, the interval from oestrus to ovulation revealed significant (p = 0.004) differences between the groups (control 44.1 h, GnRH-OE-AI 34.1 h and GnRH-FT-AI 32.9 h). GnRH-FT-AI (12.5) and GnRH-OE-AI (12.6) sows had significantly higher (p = 0.01) number of total pigs born (n = 10.4) compared with control sows. GnRH-agonist-gel treatment to the sow shortens duration of oestrus, the interval from oestrus to ovulation, and may eliminate the need for oestrus detection in the hands of skilled personnel.     

Impact of Exogenous ACTH During Pro-Oestrus on Endocrine Profile and Oestrous Cycle Characteristics in Sows

Sows housed in freely moving groups have elevated cortisol levels until the rank order is established, which takes place within approximately 48 h. The aim of this investigation was to study the effect of repeated administration of synthetic adrenocorticotropic hormone (ACTH; Synacthen Depot), during the follicular phase (pro-oestrus) on oestrus, ovulation and endocrine parameters. Four multiparous sows were used. Follicular growth and ovulation were recorded by ultrasonography. The first oestrous cycle after weaning was used as control cycle. Onset of oestrus in the sow occurs 3-4 days after the time when plasma progesterone reaches a concentration of 8 nmol/l. The progesterone profile in the control cycle of the individual sow was used for estimation when the ACTH injections should start. In the third pro-oestrus ACTH (2.5 microg/kg) was given via an indwelling catheter every 2 h for 48 h. The sows were euthanased 4-6 days after onset of the third oestrus and the ovaries were examined. Cortisol levels were elevated during the treatment period (p < 0.05). The second cycle, in which the sows were injected with ACTH, was prolonged with 2.5 days compared with the control cycle (p < 0.05). The oestradiol pattern during oestrus was similar in the control and the treatment cycle in ovulating sows. Three sows had ovulated (fresh corpora lutea), but the ovaries contained additionally one or several luteinized follicles/cysts. In conclusion, ACTH administration during pro-oestrus caused a prolongation of the oestrous cycle and a disturbed follicular development.     

Postovulatory Effect of Intravenous Administration of Lipopolysaccharide (E. coli,O55:B5) on the Contractile Activity of the Oviduct,Ova Transport,Binding of Accessory Spermatozoa to the Zona Pellucida and Embryo Development in Sows

The effect of lipopolysaccharide (LPS) (E. coli, O55:B5), administered 18 h after ovulation in the second oestrus after weaning, on the contractile activity of the oviduct, ova transport, sperm binding to zona pellucida (ZP) and embryo development, was studied in 14 Swedish crossbred (Landrace Yorkshire) multiparous sows. The endotoxin group (E‐group) sows were administered with 300 ng/kg of LPS while the control group (C‐group) sows were administered with 5 ml of saline i.v. via an indwelling jugular cannula. Immediately after evidence of standing oestrus, a Millar^® pressure transducer was placed intraluminally about 3 cm into the mid‐isthmus, via laparotomy. Pressure recordings of the oviduct were collected from all conscious sows until slaughter. After slaughter, the genital tract opposite to the side with the transducer was retrieved, and three equal isthmic segments and the first third of the uterine horn part adjacent to the utero‐tubal‐junction (UTJ) were flushed separately to recover the ova. The intervals (mean±SD) from ovulation to slaughter (OS) and insemination to ovulation (IO) were not different between the E‐group (44.5±5.7 h; 13.3±6.5 h) and the C‐group (42.7±5.9 h; 14.8±4.1 h), respectively. Ova recovery rate (RR) in the E‐group (80.2±22.9%) did not differ from that in the C‐group (85.2±4.5%). The frequency distribution of ova recovered in the different segments did not significantly (p>0.05) differ between the groups. The E‐group showed higher cleavage rate than controls. A higher proportion of spermatozoa bound to the ZP was also found in the E‐group compared with controls. The isthmic intraluminal pressure slightly increased (p=0.07) 18 h after ovulation and immediately following LPS in the E‐group, compared with the C‐group. The frequencies of phasic pressure fluctuations were significantly (p<0.05) lower at 30 and 38 h after ovulation in the E‐ than in the C‐group. It can be concluded from the present study that a single i.v. administration of LPS (300 ng/kg body weight) to sows, 18 h after ovulation might be associated with changes in isthmic pressure and the frequency of phasic pressure fluctuations, increased numbers of spermatozoa attached to the ZP and an enhanced embryo development but not with ova transport rates.     

Effect of Modified Eros Centre on Some Reproductive Parameters of the Sow

The effect of a modified eros centre on weaning to oestrus interval, follicle size, ovulation and farrowing rate and total born litter size was investigated. In modified eros centre 94.4% and in group housing 79.1% of the sows (p < 0.01) expressed oestrus within 10 days post‐weaning. Weaning to oestrus interval was shorter (p < 0.001) for sows kept in modified eros centre. The interval from onset of oestrus to the time of ovulation was longer for sows in group housing (p=0.05). The time of ovulation was negatively correlated (r=?0.50) with the interval from weaning to oestrus (p=0.005). The time of ovulation after onset of oestrus was significantly (p < 0.05) shorter for sows expressing oestrus within 2–4 days of weaning, compared with the animals that expressed oestrus between days 5 and 6 post‐weaning and was shortest for sows expressing oestrus after day 6 post‐weaning. Farrowing rate was not affected by a modified eros centre. Litter size tended to be smaller in group‐housed weaned sows (p=0.10). The timing of last artificial insemination relative to time of ovulation did not affect litter size (p > 0.10). The implication of these results is that a modified eros centre may improve some of the post‐weaning oestrous parameters of the sow.     

Immunohistochemical Studies on Oestrogen Receptor Alpha (ERα) and the Proliferative Marker Ki-67 in the Sow Uterus at Oestrus and Early Pregnancy

Oestrogen receptor alpha (ERalpha), the main subtype in the uterus, is involved in the regulation of uterine growth/proliferation. A relationship between ERalpha and proliferative activity has been shown in the cyclic sow uterus, but to our knowledge, no study has been carried out on early pregnant sows. Therefore, by means of immunohistochemistry and use of mouse monoclonal antibodies to ERalpha and a proliferative marker, Ki-67, the localization of these proteins was investigated in the sow uterus during early pregnancy. Eighteen crossbred multiparous sows were artificially inseminated once at 20-15 h before expected ovulation. After artificial insemination (AI), they were slaughtered at five different times: at oestrus, 5-6 h after AI (n = 4), 20-25 h after ovulation (n =4), 70 h after ovulation (n = 4), on day 11 (the first day of standing oestrus = day 1, n = 3) and on day 19 (n = 3). Immediately after slaughter, uterine samples were collected at the mesometrial side of the uteri, fixed in 10% formaldehyde and embedded in paraffin. Immunohistochemistry was performed by using mouse monoclonal antibodies to ERalpha (C-311) and Ki-67 (MM1). All sows slaughtered after ovulation were pregnant. In general, positive immunostaining for ERalpha and Ki-67 was found in the nuclei. Variations in staining intensity and proportion of positive nuclei were observed in different uterine compartments and stages of early pregnancy. The highest level of ERalpha presence in the surface epithelium and myometrium was found at oestrus (5-6 h after AI), and low levels of ERalpha in these compartments were observed as early as 20-25 h after ovulation. In the glandular epithelia, presence of ERalpha was highest at 70 h after ovulation. The largest number of ERalpha-positive cells in the stroma was observed at oestrus and early after ovulation. Low proliferation was observed, and with no significant difference in tissue compartments except in the glandular epithelium. High proliferative activity in the glandular epithelium at 70 h after ovulation indicated involvement in preparation for secretory activity and growth during pregnancy establishment. Significant positive correlation was found between the number of ERalpha-positive cells in the stroma and Ki-67-positive cells in the surface epithelium. In conclusion, the present study showed differences in immunolocalization of ERalpha and the proliferative marker Ki-67 in different tissue compartments of the sow uterus at oestrus and early pregnancy. In some uterine compartments, the patterns of ERalpha and Ki-67 immunostaining seemed to be influenced by insemination and the presence of embryos, in addition to the effects of steroid hormones.     

The effect of intravaginal applied gonadotropin-releasing hormone agonist on different oestrous parameters and reproductive performance in post weaned sows

In a Hungarian large breeding unit, 481 weaned sows were assigned to three groups and were treated as follows. Sows in Group 1 (Control, n=161) were artificially inseminated (3.01 +/- 0.4 times) during their standing reflex; sows in Group 2 (n=160) were artificially inseminated 3 times at 12-hour intervals, independent of detection of oestrus and immediately after administration of a GnRH-agonist at 96 hours postweaning; and sows in Group 3 (n=160) were artificially inseminated 3 times at 12-hour intervals, beginning at their standing reflex after administration of a GnRH-agonist. Pre-trial daily average lactational feed intake, average daily feed intake from weaning to oestrus, oestrus within 6 days of weaning (%), ovulation within 6 days of weaning (%), wean-to-oestrus interval (h), duration of oestrus (h), follicle size (mm), interval from oestrus to ovulation (h), subsequent day 24 pregnancy rate (%), farrowing rate (%) and total number of pigs born were evaluated. Pre-trial average daily voluntary lactational feed intake was 7.1 +/- 0.5 kg in Group 1, 7.2 +/- 0.4 kg in Group 2, and 7.3 +/- 0.7 kg in Group 3 (P > 0.05). Average voluntary daily feed intake from weaning-to-oestrus was 4.3 +/- 0.9 kg in Group 1, 4.2 +/- 0.8 kg in Group 2, and 4.1 +/- 0.5 kg in Group 3 (P > 0.05). Oestrus was detected within 6 days of weaning in 143 (88.8%) sows in Group 1, 143 (89.4%) sows in Group 2, and in 142 (88.8%) sows in Group 3. Follicle size did not differ (P > 0.05) among the groups. In Group 1, 83.2%, in Group 2, 90.6%, and in Group 3,91.3% of the sows ovulated within 6 days of weaning (P < 0.05), but there were no significant (P > 0.05) differences in 24 Day pregnancy rates (81.4%; 91.3%; and 92.5%). Farrowing rates were in Group 1, 84.5%, in Group 2, 91.3%, in Group 3, 91.9% (P > 0.05). Wean-to-oestrus interval was 115.5 h in Group 1, 114.9 h in Group 2, and 115.7 h in Group 3 (P > 0.05). Duration of oestrus was significantly shorter in Group 2 (41.9 h) and Group 3 (42.1 h) than in Group 1 (68.3 h) (P < 0.001). Similarly, the interval from oestrus to ovulation was significantly different (P < 0.01) between the groups (Group 1, 49.0 h Group 2, 32.0 h, and Group 3, 31.1 h). Sows in Group 2 (12.7) and Group 3 (12.6) had a significantly higher (P < 0.01) number of pigs born than sows in Group 1 (n = 10.9). The interval between oestrus and ovulation was highly and positively correlated (r = 0.83) with the duration of oestrus.     

Peri-oestrus hormone profiles and follicle growth in lactating sows with oestrus induced by intermittent suckling.

This study describes follicle dynamics, endocrine profiles in multiparous sows with lactational oestrus compared with conventionally weaned sows (C). Lactational oestrus was induced by Intermittent Suckling (IS) with separation of sows and piglets for either 12 consecutive hours per day (IS12, n = 14) or twice per day for 6 h per occasion (IS6, n = 13) from day 14 of lactation onwards. Control sows (n = 23) were weaned at day 21 of lactation. Pre-ovulatory follicles (> or =6 mm) were observed in 100% of IS12, 92% of IS6 and 26% of C sows before day 21 of lactation and in the remaining 74% C sows within 7 days after weaning. All sows with pre-ovulatory follicles showed oestrus, but not all sows showed ovulation. Four IS6 sows and one IS12 sow developed cystic follicles of which two IS6 sows partially ovulated. Follicle growth, ovulation rate and time of ovulation were similar. E(2) levels tended to be higher in IS sows (p = 0.06), the pre-ovulatory LH surge tended to be lower in IS12 (5.1 +/- 1.7 ng/ml) than in C sows (8.4 +/- 5.0 ng/ml; p = 0.08) and P(4) levels were lower in IS12 and IS6 than in C sows (at 75 h after ovulation: 8.8 +/- 2.4 ng/ml vs 7.0 +/- 1.4 ng/ml vs 17.1 +/- 4.4 ng/ml; p < 0.01). In conclusion, sows with lactational oestrus induced by IS are similar to weaned sows in the timing of oestrus, early follicle development and ovulation rates, but the pre-ovulatory LH surge and post-ovulatory P(4) increase are lower.     

Effects of ACTH injections during estrus on concentrations and patterns of progesterone, estradiol, LH, and inhibin alpha and time of ovulation in the sow

This study investigated whether injections of ACTH for 48 h, from the onset of the second standing estrus after weaning, had any impact on time of ovulation and patterns of progesterone, estradiol, luteinizing hormone (LH), and inhibin alpha. The studied sows (n=15) were fitted with jugular vein catheters and randomly divided into a control (C group) and an ACTH group. From the onset of standing estrus, the sows were injected (NaCl or synthetic ACTH, 5 microg/kg) every 4h; blood samples were collected immediately before and 45 min after each injection. Ovulation was monitored using ultrasonography. The ACTH-group sows stopped displaying signs of standing estrus sooner after ovulation in their second estrus, but no impact was found on time of ovulation. There were no significant differences in the intervals between LH peak, estradiol peak, and the onset of standing estrus between the C and ACTH groups. The cortisol and progesterone concentrations were significantly elevated (p<0.001) in samples taken 45 min after ACTH injection. There were minor differences in estradiol and LH concentrations between the groups. Overall inhibin alpha concentrations were significantly higher during the treatment period in the ACTH than in the C group, but there were no significant differences between samples taken either 45 min or 4h after injection. In conclusion, injections of synthetic ACTH during estrus in the sow apparently disturb the duration of signs of standing estrus and the hormonal pattern of progesterone, and possibly of inhibin alpha, estradiol and LH.     

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.     

Influence of post-ovulatory insemination on sperm distribution,pregnancy and the infiltration by cells of the immune system,and the distribution of CD2, CD4, CD8 and MHC class II expressing cells in the sow endometrium

This study investigates the distribution of leucocytes, CD2+, CD4+, CD8+ lymphocyte subpopulations and MHC class II expressing cells in the sow endometrium following post-ovulatory insemination in relation to clinical findings and pregnancy outcome. Crossbred multiparous sows were inseminated once either at 15-20 h after ovulation [experiment 1, slaughtered at 20-25 h (5-6 h after artificial insemination (AI), group 1-A, n = 4), at 70 h after ovulation (group 1-B, n = 4), on day 11 (group 1-C, n = 4, first day of standing oestrus = day 1) or on day 19 (group 1-D, n = 4)] or 30 h after ovulation [experiment 2, slaughtered at 5-6 h after AI (group 2-A, n = 4) or on day 19 (group 2-D, n = 3)]. The uterine horns were flushed to control for the presence of spermatozoa and neutrophils and/or for recovery of oocytes and/or embryos. Mesometrial uterine samples were plastic embedded and stained. Cryofixed uterine samples were analysed by immunohistochemistry using mAbs to lymphocyte subpopulations and MHC class II molecules. Light microscopy was used to examine surface (SE) and glandular epithelia (GE), and connective tissue layers, both subepithelially (SL) and glandular (GL). In experiment 1, group 1-A, only one sow had spermatozoa in the utero-tubal junction (UTJ). Marked/moderated numbers of neutrophils and spermatozoa were observed in the flushings of two sows. In group 1-B, altogether 23 of 48 oocytes were cleaved. Day 11 (1-C), embryos with small diameter were observed. Day 19 (1-D), no embryos were found but small pieces of foetal membrane were observed in one of the sows. In group 1-A, large numbers of neutrophils were found within the SE and SL but with high individual variation. For T lymphocyte subpopulations, in the SE, most CD2+ cells were found in group 1-A. For both SE and GE in all groups, the number of CD8+ cells was significantly larger than that of CD4+ cells. In experiment 2, group 2-A, no sow had spermatozoa in the UTJ or in the uterine flushings. At day 19, no sow was pregnant. In group 2-A, large numbers of neutrophils were found within the SE and SL but with high individual variation. At day 19, high E2 levels showed a hormonal prooestrous stage but the endometrial neutrophil infiltration normally expected at pro-oestrus was absent. In conclusion, post-ovulatory insemination (about 18 h after ovulation) resulted in impaired spermatozoa transport within the uterus and embryonic degeneration. In sows post-ovulatory inseminated at a later stage (30 h after ovulation), no sow was pregnant. In both experiments, disturbed immune cell patterns were observed in some individuals.     

Effect of transportation and relocation in post-weaning anoestrous primiparous sows

The object of this investigation was to study the clinical and endocrine responses to transportation and relocation in 8 post-weaning anoestrous sows. They had been anoestrous for at least 24 days after weaning before transportation/relocation was performed. Laparoscopy, performed at the beginning of the experiment, revealed that the ovaries contained many follicles (less than or equal to 6 mm in diameter), but no corpora lutea. Blood samples, taken before and after transportation/relocation, showed that LH activity was low at the beginning of the experiment and increased after transportation/relocation in the majority of the sows. Peripheral plasma concentrations of of oestradiol-17 beta increased 1-4 days after transportation/relocation in 6 out of 8 sows which was followed by oestrus and ovulation. Progesterone concentrations were also below the practical detection limit until the end of oestrus. This study has demonstrated that a change in environment by transportation and relocation can induce oestrus by increasing the LH activity in post-weaning anoestrous sows.     

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.     

Early embryo survival and development in sows with lactational ovulation.

During lactation, daily separation of sow and piglets, intermittent suckling (IS), can induce lactational oestrus and ovulation. This study examined effects of IS on subsequent early embryo survival and development. Multiparous Topigs40 sows were separated from their piglets for either 12 consecutive hours per day (IS12, n = 13) or two times for 6 h per day (IS6, n = 10) from day 14 of lactation onwards until 23 days after ovulation. Control sows (C, n = 17) were weaned at day 21 of lactation. Oestrus was shown in all treatments within 5 days after the start of treatment. Sows were inseminated each day of oestrus and slaughtered at D23 after ovulation. Intermittent suckling did not significantly affect pregnancy rates of sows (75% IS12 vs 78% IS6 vs 94% C; p > 0.10). Embryo survival was not significantly affected by IS (IS12: 57%; IS6: 51%; p > 0.10) although it seemed to be lower than in C sows (70%). Some parameters of embryo, placental and uterine development were affected by IS, especially in the IS6 group. IS6 embryos had shorter placentas (17.5 +/- 1.2 cm; p < 0.05) than C (20.3 +/- 1.4 cm) and IS12 sows (20.9 +/- 0.7 cm) were smaller and less developed than C sows (p < 0.05). In conclusion, embryo survival does not seem significantly affected by IS, although numerical differences were great. Embryo development, however, was negatively affected in IS6 sows possibly due to a combination of high milk production, stress and lactational effects on uterine development.     

Effects of breeding at the second oestrus or after post-weaning hormonal treatment with altrenogest on subsequent reproductive performance of primiparous sows

This study investigated the effects of breeding at the second oestrus after weaning or after feeding an orally active progestagen (altrenogest) on the subsequent reproductive performance of primiparous sows. After 3 weeks of lactation, 663 weaned sows of two genotypes were allocated into three groups: G1--breeding at the first oestrus after weaning; G2--breeding at the second oestrus after weaning and G3--treatment with altrenogest for 5 days after weaning and breeding at the first oestrus after the end of the treatment. Body weight at breeding was lower in G1 and G3 than in G2 sows (p < 0.05). The interval to show oestrus was similar for G1 and G2 groups (p > 0.05) but higher (p < 0.05) than that observed in G3 group. Within genotype A, percentages of females in oestrus within 10 days were not different (p > 0.05) among groups, whereas in genotype B, more G1 and G2 sows (p < 0.05) showed oestrus than G3 sows. In both genotypes, lower farrowing rates were observed in G3 than in G1 and G2 sows (p < 0.05) and a greater litter size (p < 0.05) was observed in G2 sows. In genotype A, the number of total born piglets was similar for G1 and G3 groups (p > 0.05), whereas in genotype B, G1 sows had a greater litter size than G3 sows (p < 0.05). Body weight at weaning and at breeding was similar (p > 0.05) between farrowed and non-farrowed sows in all groups. Reproductive performance is not improved in primiparous sows treated with altrenogest during 5 days after weaning. The reproductive performance of genotype B sows is compromised in Control and Altrenogest-treated sows but not in those bred at the second oestrus after mating. Breeding at the second oestrus after weaning allows primiparous sows to gain weight between weaning and service, and increases their farrowing rate and subsequent litter size.     

Follicular Development and Ovulation in Sows: Effect of hCG and GnRH Treatment

Follicular growth, chronology of ovulation and embryo morphology were compared in sows ovulating spontaneously and sows, in which the ovulation was attempted induced by hCG or GnRH.Indwelling catheters were placed on day 1 (weaning = day 0) in the ear veins of 18 sows, which were then randomly divided into 3 groups: a control group (N = 6), a group (N = 6) given 750 iu hCG (Physex®) im 76h after weaning (hCG group) and a group (N = 6) given 500 µg GnRH (Fertagyl®) im 76h (N = 3) or 100h after weaning (N = 3) (GnRH group). Follicular diameter and time of ovulation were monitored by ultrasonography every 4h from day 3 until ovulation or development of cysts by means of a sector scanner fitted with a 5.0/7.5 MHz multiangle probe. Heat detection was performed every 8h from day 3 until ovulation. On day 13, the sows were slaughtered, the number of corpora luteae (CL) was counted, and embryos were flushed from the uteri. The control group showed clear heat symptoms, and on day 3, the follicles were typically 3–7 mm and grew up to 7–10 mm over 2 days, where they remained for approximately 24h until ovulation took place 41h ± 9h after first sign of standing heat. The hCG group exhibited no signs of heat, and the follicles only reached 5–8 mm in diameter at time of ovulation, which occurred 40h ± lh after hCG-injection. The GnRH group exhibited inconsistent signs of heat, and the follicles reached a maximum size of 7–12 mm in diameter where they remained for more than 24h. Only 2 sows in this group ovulated within 84–92h after the GnRH injection, and development of bursa cysts and cystic follicles was a common finding. The average number of CL was 18.2 ±5.7 per sow (N = 16, range: 3–27) with no significant difference between the groups. Total embryo recovery was 79 ± 13 % with no significant difference between groups. The embryo diversity calculated as standard deviation of the maximum diameter was higher in the hCG group as compared with the control group.It is concluded that (1) transrectal ultrasonography can be used in sows for accurate assessment of follicular growth and ovulation; (2) the use of hCG results in lack of heat symptoms and reduced follicle size at the time of ovulation when injected 76h after weaning; (3) administration of a single injection of GnRH, if given before the first signs of heat, results in inconsistent heat symptoms and no or late ovulations.     

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.