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PubMed2004

Intra-uterine insemination in farm animals and humans.

Artificial insemination (AI) is the oldest and currently most common technique in the assisted reproduction of animals and humans. The introduction of AI in farm animals was forced by sanitary reasons and the first large-scale applications with a commercial goal were performed in cattle in the late 1930s of last century. After the Second World War, cryopreservation of semen facilitated distribution and AI was mainly performed for economic reasons, especially in dairy cattle industry. In humans however, AI was initially performed in cases of physiological and psychological sexual dysfunction, but later on also in cases of infertility caused by immunological problems. Currently, the most common indications for intra-uterine insemination (IUI) in humans are unexplained infertility and male subfertility. In these cases, IUI is considered as the treatment of the first choice, before more invasive techniques such as in vitro fertilization (IVF) and intracytoplasmatic sperm injection (ICSI) are used. In contrast with humans, the quantity and quality of semen produced by farm animals is much higher and permits dilution and production of several insemination doses per ejaculate. However, with the introduction of sex-sorted semen in farm animals, the same problem of low-quality semen as in humans has arisen. In cattle, pigs and horses, conventional insemination with low numbers of sex-sorted spermatozoa results in a significant decrease in fertility. To improve the fertility rates with this semen, new insemination techniques have been developed in order to deposit spermatozoa closer to the site of fertilization. In sows and mares the advantage of utero-tubal junction (UTJ) insemination has already been proven; however, in cattle it is still under investigation. In this review, the differences and similarities in the application of AI between animals and humans are discussed and as AI in farm animals is most successful in cattle, the situation in this species is elaborated the most.

Animals↗
PubMed2005

Synchronization of ovulation and fixed time intrauterine insemination in ewes.

A novel method for oestrus-ovulation synchronization in sheep followed by fixed time insemination is presented herewith. Mature dry ewes (n = 28) of Karagouniko breed being at an unknown stage of the oestrous cycle, were used during the middle of breeding season. The treatment protocol consisted of an initial administration of a GnRH analogue followed 5 days later by a prostaglandin F2alpha injection. Thirty-six hours later a second GnRH injection was administered to synchronize ovulation, and laparoscopic intrauterine insemination was performed 12-14 h later. Three days after insemination, fertile rams were introduced into the flock twice daily and oestrus-mating detection was carried out. For progesterone (P(4)) determination, blood samples were collected on alternate days, starting 2 days before the first GnRH injection and continuing for 17 days after insemination. An additional sample was taken on the day of insemination. Pregnancy diagnosis was carried out by trans-abdominal ultrasonography. Fourteen ewes (50%) conceived at insemination and maintained pregnancy; from the remainder 14 ewes 10 became pregnant at natural service, while four, although they mated at least two to three times, failed to conceive. In response to the first GnRH, P(4) concentration increased at higher levels in ewes that conceived at AI compared with those that failed to conceive (47.54 and 22.44%, respectively; p < 0.05). Significant differences (p < 0.05) in mean P(4) concentration between pregnant and non-pregnant animals were detected 1 day before AI (0.17 +/- 0.06 and 0.26 +/- 0.14 ng/ml, respectively) on the day of AI (0.15 +/- 0.04 and 0.24 +/- 0.08 ng/ml, respectively) as well as 9 and 11 days thereafter (0.48 +/- 0.12 and 0.38 +/- 0.12 ng/ml; 0.68 +/- 0.14 and 0.50 +/- 0.18 ng/ml, respectively). These results indicate that using the proposed protocol, an acceptable conception rate can be achieved which could be further improved by modifying the time intervals between interventions.

Animals↗
PubMed2005

Artificial insemination of gilts with 1.5 billion sperms stored in different periods associated with different pre-ovulatory intervals.

This study evaluated the reproductive performance of gilts inseminated at three intervals before ovulation (0-12, 13-23, 24-30 h) with sperm doses (SD) stored for 0-48 and 96-120 h. A total of 218 PIC Camborough 22 gilts were inseminated once with SD of 1.5 x 10(9) sperms. Pregnant gilts (n = 166) were slaughtered 30.8 +/- 3.7 days after artificial insemination. The number of corpora lutea (CL) and total embryos (TE) was counted. Pregnancy rates (PR) were analysed by chi-square test. TE and embryonic survival (ES), obtained as the ratio between viable embryos and CL, were analysed by GLM procedure (SAS) and mean values were compared by Tukey's test. Pregnancy rate was similar among artificial insemination-ovulation (AIOV) intervals when semen was stored for 0-48 h. However, the lowest PR was observed in the 24-30 h AIOV interval with storage time (ST) of 96-120 h (p < 0.05). There was a significant effect of the interaction between ST and AIOV (p < 0.05) on TE and ES variables. Total embryos and ES did not differ (p > 0.05) among AIOV intervals in ST of 0-48 h. However, gilts inseminated at 24-30 h AIOV interval with ST of 96-120 h showed a reduction of 6.7 embryos (p < 0.05) compared with gilts in the same interval inseminated with semen stored for 0-48 h. ES for the 24-30 h AIOV interval and ST of 96-120 h was lower than that observed in the other groups (p < 0.05).

Animals↗
PubMed2005

Establishment of a pregnancy following intravaginal insemination with epididymal semen from a dog castrated due to benign prostatic hyperplasia.

Benign prostatic hyperplasia was diagnosed in an American Staffordshire Terrier of high breeding value presenting concurrent haematuria. Castration as a treatment was synchronized with the oestrus cycle of a bitch selected for insemination. After castration the cauda epididymis was flushed with Gent semen extender and collected spermatozoa were filtered and analysed by Hamilton Thorn computer assisted sperm analysis. A total of 7 ml semen containing 742 x 10(6) spermatozoa with 76.5% mean motility was used for insemination. Intravaginal insemination of the bitch was performed with an insemination catheter for dogs (Kruuse, Marslev, Denmark) on the day when plasma progesterone levels reached 9.9 ng/ml. Normal pregnancy without complications resulted in eight live-born puppies 63 days after insemination. This is the first report of a normal pregnancy and birth of puppies from a bitch inseminated with epididymal semen obtained from a dog affected by benign prostate hyperplasia.

Animals↗
PubMed2006

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.

Animals↗
PubMed1993

Gonadotrophin-releasing hormone treatment in cattle: a meta-analysis of the effects on conception at the time of insemination.

Data from 40 trials described in 27 published papers were analysed by meta-analysis (pooling of data from numerous trials into a single analysis) to evaluate the effects of injecting gonadotrophin-releasing hormone (GnRH) in cattle at the time of insemination on the risk of pregnancy. A total of 19,019 cows were involved in these studies. Analyses were stratified by trial and by the effects of dose of GnRH, type of GnRH and insemination number (first, second or third and greater). Mantel-Haenszel analysis revealed that use of GnRH at insemination significantly increased the overall risk of pregnancy by 12.5% in treated cows (P < 0.05). However, increases in risk of pregnancy were greatest (22.5%) when repeat breeders were tested. Results for use of GnRH or analogue at first service were similar, with increased risks of pregnancy in treated cows of 5.2 and 8.0%, respectively. The risk of pregnancy tended to be higher (11.1%) when the dose of GnRH was > or = 250 micrograms and when used at second insemination (9.9%). The analyses demonstrated that while treatment at insemination with GnRH and GnRH analogues increased conception rates in dairy cattle, some variation in study results was attributable to the number of inseminations after calving at which GnRH is administered. Further efforts should be made to determine characteristics of populations of cows that have good fertility responses to GnRH and to determine the mode of action of GnRH in increasing fertility.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Effect of factors associated with insemination on calving rate in dairy cows.

The results of 7569 inseminations performed on 6007 cows in 94 dairy herds were used to investigate the relative importance of time of insemination, bull and inseminator on calving rates in dairy cows. Although the time of insemination in relation to the first observation of oestrus was shown to have some effect, the effect was minimal during the first 24 hours. The maximum difference in expected calving rates between cows served with semen from groups of bulls with a history of either low or high fertility was 20 per cent, and the maximum difference in expected calving rate between cows inseminated by groups of inseminators who had consistently achieved either low or high fertility was 13 per cent.

Animals↗
PubMed1993

Increased reproductive losses in cattle infected with bovine pestivirus around the time of insemination.

Unmated heifers seronegative to bovine pestivirus were used to investigate the effects on conception and embryo-fetal survival of pestivirus infection around the time of artificial insemination. The reproductive performances of three groups were compared; the control group did not become infected during pregnancy, group 1 heifers were infected by contact with a persistently infected cow and calf four days after insemination and group 2 heifers were infected intranasally nine days before insemination. Conception rates and embryo-fetal survival were monitored by serial serum progesterone assays, transrectal ultrasonography and manual palpation of the uterus. The conception rates (determined 20 days after insemination) of 60 per cent (nine of 15) and 44 per cent (eight of 18) for groups 1 and 2 were lower than the 79 per cent (11 of 14) achieved by the control group. The group 1 heifers subsequently experienced significant embryo-fetal loss, resulting in a pregnancy rate (determined 77 days after insemination) of 33 per cent (five of 15), significantly lower than the control group's 79 per cent (11 of 14). The pregnancy rate of the group 2 heifers (39 per cent, seven of 18) was also significantly lower than that of the controls, largely as a result of the group's poor conception rate. All the heifers diagnosed pregnant 275 days after insemination were induced to calve. No persistently infected calves were born.

Animals↗
PubMed1992

Pregnancy by means of tubal insemination and subsequent spontaneous pregnancy in rabbits.

The purpose of the study was to determine whether or not physical stimulation by tubal insemination had any unfavourable influences upon the tubal fimbria. Tubal insemination was carried out on 14 rabbits and subsequent pregnancy results were monitored. After a period of 69-123 days following tubal insemination, 10 of the rabbits were mated spontaneously and these rabbits were then monitored for pregnancy. Newborn were obtained, with normal gestation periods in six out of the 14 rabbits, following tubal insemination and all 10 of the rabbits that were mated spontaneously with males following tubal insemination subsequently delivered. It is concluded that physical stimulation by tubal insemination does not produce adhesive changes on the tubal fimbria.

Animals↗
PubMed2003

Optimal timing for canine artificial insemination with frozen semen and parentage testing by microsatellite markers in superfecundency.

Parentage testing was performed in sixteen litters by canine artificial inseminations with frozen semen from different sires on Days 5 and 7 after the LH surge. It became apparent that only 25% of dams had superfecundation, but 43.8% of dams were whelped after insemination only on Day 5 after the LH surge and 31.3% of dams after insemination only on Day 7. Of the total 87 puppies, 46% were born after insemination on Day 5 after the LH surge and 54% after insemination on Day 7. This result strongly suggested that canine artificial insemination with frozen semen could be sufficiently successful also on Days 5 and 7 after the LH surge.

Animals↗
PubMed1989

Relationship between the number of sperms and the rate of implantation in bitches inseminated into unilateral uterine horn.

Studies were made on the number of sperms required for fertilization of ova ovulated from the bilateral ovaries in bitches. The experiment was carried out with 44 female beagles, which were divided into four groups of seven to eleven bitches each. The four groups were inseminated with 40 X 10(6) sperms/0.2 ml, 20 X 10(6) sperms/0.1 ml, 10 X 10(6) sperms/0.1 ml, and 3-5 X 10(6) sperms/0.1 ml, respectively. All the bitches were laparotomized at an optimum time for mating and inseminated at the tip of the horn of an unilateral uterus. In the four groups conception took place at a rate of 7/7, 8/8, 10/11, and 2/7, respectively. The number of puppies exceeded the number of ova ovulated on the inseminated side at a rate of 5/7, 6/8, 0/10, and 0/2 in the four groups, respectively. Therefore, it was clarified that when more than 20 X 10(6) sperms/0.1 ml were inseminated in the ipsilateral uterine horn, ova ovulated on the contralateral side were fertilized also. Then a unilateral ovary was ovariectomized in 11 bitches, which were inseminated with 10 X 10(6) sperms/0.1 ml in the ipsilateral uterine horn. As a result, only one bitch became conceptive and gave birth to only one puppy. From these results, it was considered that 10 X 10(6) sperms/0.1 ml may be most adequate for the ova on the side of insemination alone to be fertilized.

Animals↗
PubMed1984

Intrauterine insemination enhances fertility of frozen semen in superovulated ewes.

Superovulated ewes were inseminated with fresh or frozen semen in a factorial experiment which compared two techniques of artificial insemination; i.e. conventional cervical deposition and intrauterine deposition at laparoscopy. Similar fertilization rates resulted from insemination with fresh semen at cervical (81% of ova from 11/11 ewes) and intrauterine (83% of ova from 10/12 ewes) sites. These results approached those observed in a naturally-mated group (95% of ova from 5/5 ewes). In ewes inseminated with frozen semen, fertilization rate was markedly reduced (P less than 0.05) after cervical insemination (11% of ova from 3/11 ewes) and partly restored (P less than 0.05) after intrauterine insemination (50% of ova from 8/11 ewes).

Animals↗
PubMed1986

Interstrain competition amongst mouse spermatozoa inseminated in various proportions, as affected by the genotype of the Y chromosome.

Nonagouti (KP X C57BL)F1 hybrid females were artificially inseminated with a mixture of spermatozoa from males of the KE (nonagouti) and CBA (agouti) strains and the genotype of young was estimated by fur pigmentation. When KE and CBA spermatozoa mixed in the ratios of 1:1, 2:1 and 4:1 were inseminated after ovulation, 87%, 56% and 29% of progeny, respectively, were sired by CBA males, i.e. proportions of CBA progeny were significantly higher than ratios of CBA spermatozoa in the mixture. The surplus of CBA progeny was significantly less in females inseminated before ovulation, which may suggest that more rapid capacitation of CBA spermatozoa is partly responsible for their competitive advantage. In preparations from oviducal flushings of females killed 2-3 h after insemination, CBA spermatozoa (recognized by their shape) were found in similar proportions as in the inseminated mixture. There was therefore no evidence of their preferential selection at the uterotubal junction. No competitive advantage of CBA spermatozoa occurred when they were inseminated with spermatozoa from males of the KE.CBA strain, congenic with KE but with the Y chromosome derived from the CBA strain. This indicates that genetic factors linked with the Y chromosome may influence competitive ability of spermatozoa.

Animals↗
PubMed1994

Effect of exogenous progesterone on superovulatory response in heifers inseminated with fresh or frozen semen.

Superovulation in cattle normally involves the administration of gonadotrophins at specific times of the oestrous cycle, followed by the induction of luteolysis and insemination with high quality semen. The first aim of this experiment was to examine the effect of supplementary progesterone when used in conjunction with porcine FSH (pFSH) to induce superovulation in heifers. The methods compared were PGF2 alpha given at mid-cycle or a progesterone-releasing intravaginal device (PRID) inserted at different phases of the cycle. The second aim was to determine whether site of insemination or use of fresh or frozen semen affected embryo production. A factorial design was used involving 185 beef heifers. The main factors were (i) synchronization methods (PGF2 alpha or PRID); (ii) semen type (fresh or frozen); (iii) insemination regimens (involving two inseminations and variations in the sites) and number of straws used (one or two) at the second insemination. Eight injections of pFSH were given twice a day for 4 days starting either on days 9, 10 or 11 of the oestrous cycle or on the fourth day after insertion of a PRID. Heifers were checked for oestrus, inseminated twice and embryos were recovered on day 7 of the superovulated cycle. There was no difference between heifers given either PRID or PGF2 alpha in the oestrous response (93% versus 96%), number of ovulations (15.9 +/- 1.11 versus 13.4 +/- 1.06), large follicles (2.5 +/- 0.24 versus 2.3 +/- 0.23) or embryos recovered (9.1 +/- 0.77 versus 9.1 +/- 0.74).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Effects of time of insemination relative to ovulation, as determined by ultrasonography, on fertilization rate and accessory sperm count in sows.

The effects of the timing of insemination relative to ovulation on fertilization rate, accessory sperm count and early embryo development were studied in sows. Oestrus detection was performed at intervals of 8 h. Sows were artificially inseminated once with 3 x 10(9) spermatozoa. Transrectal ultrasonography was performed at intervals of 4 h to determine when ovulation occurred and sows were killed at 120 +/- 6 h after ovulation. For each insemination-ovulation interval of 8 h, fertilization rates were as follows: > 48 h, 35% (n = 1); 48-40 h, 51 +/- 36% (n = 6); 40-32 h, 54 +/- 36% (n = 14); 32-24 h, 79 +/- 32% (n = 19); 24-16 h, 94 +/- 11% (n = 24); 16-8 h, 92 +/- 21% (n = 24); 8-0 h, 95 +/- 22% (n = 21) and for the sows that were inseminated after ovulation: 0 to -8 h, 75 +/- 38% (n = 26); -8 to -16 h, 74 +/- 43% (n = 15) and < -16 h, 0% (n = 1). The median accessory sperm count differed among the groups from 1 (insemination 40-48 h before ovulation) to 126 (insemination 0-8 h after ovulation) (P = 0.0001). Within each 8 h time interval, the normal embryos from sows with less than 90% normal embryos were less developed and had a lower sperm count than did the normal embryos from sows with more than 90% normal embryos (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1997

Influence of insemination-ovulation interval and sperm cell dose on fertilization in sows.

This experiment was conducted to determine the effects of sperm dose at insemination on fertilization rates and accessory sperm cells attached to day 5 embryos. Multiparous sows (n = 115) were artificially inseminated once with 1 x 10(9), 3 x 10(9) or 6 x 10(9) sperm cells between 3 h and 48 h before ovulation. Transrectal ultrasonography was performed at intervals of 4 h to determine the time of ovulation and sows were killed at 120 +/- 5 h after ovulation to assess the results of fertilization. The insemination-ovulation interval had a major influence on the fertilization rate and accessory sperm count. A nonsignificant but consistent increase in fertilization rate and in number of accessory sperm cells due to the sperm dose was observed. During the insemination-ovulation interval of 12-24 h, the median fertilization rates were 95%, 100% and 100%, and the median accessory sperm counts were 11, 17 and 31 for the 1 x 10(9), 3 x 10(9) and 6 x 10(9) doses, respectively. During the insemination-ovulation interval of 24-36 h, the median fertilization rates were 88%, 95% and 97%, and the median accessory sperm counts were 6, 8 and 11 for the 1 x 10(9), 3 x 10(9) and 6 x 10(9) doses, respectively. No direct relationship was detected between embryo quality and the accessory sperm count but there was a relationship between insemination-ovulation interval and accessory sperm count. The fertilization rate was positively correlated with the breeding value for litter size of the sows. In conclusion, the effects of sperm dose on fertilization rate and on accessory sperm count in sows were small and nonsignificant, indicating only small effects of sperm dose on the functioning of the sperm reservoir in the sow.

Animals↗
PubMed1998

Intraperitoneal insemination of the guinea pig with synchronized estrus induced by progesterone implant.

Female guinea pigs with synchronized ovulation by means of implantation of progesterone-filled tubing (P-tube) followed by a progesterone injection, were inseminated by intraperitoneal injection with sperm suspension. First, to obtain the optimum conditions for insemination, the females were inseminated singly over the range of 1-10 x 10(7) spermatozoa before and after the synchronized ovulation. The incidence of conception and implantation was 100% in the females given more than 5 x 10(7)/animal at 9:00 h on the 5th day after removal of the P-tube. Second, the reproductive ability of the inseminated females under this optimal condition was observed throughout the pregnancy to delivery. Inseminated females had a mean +/- S.D. gestation period of 68.7 +/- 0.5 days, a litter size of 2.8 +/- 0.6 pups and body weight of 110 +/- 14 g. These data were comparable to those of naturally-mated females. Our findings suggest that the artificial insemination by intraperitoneal injection in combination with the synchronized estrus technique is very useful for production control in a small colony of guinea pigs.

Animal Husbandry↗
PubMed2002

Producing progeny from endangered birds of prey: treatment of urine-contaminated semen and a novel intramagnal insemination approach.

Wild raptors brought into an ex situ environment often have poor semen quality that is further compromised by urine contamination. Generally, it is believed that in birds, artificial insemination into the cloaca or caudal vagina of females requires large doses of high-quality spermatozoa to maximize fertility. In an effort to define and overcome some of the challenges associated with reproduction in wild raptors, the objectives of this study were to 1) evaluate the frequency, impact, and remediation of urine contamination in fresh ejaculates for the purpose of maintaining sperm motility and viability in vitro, and 2) develop a deep insemination method that allows low numbers of washed sperm to be placed directly into the magnum to increase the probability of producing fertilized eggs. The species evaluated include golden eagle (Aquila chrysoetos), imperial eagle (A. adalberti), Bonelli's eagle (Hiernaetus fasciatus), and peregrine falcon (Falco peregrinus). Semen samples were collected and pooled by species, and a minimum of 25 pooled ejaculates per species were evaluated for urine contamination, pH, sperm viability, and sperm motility; the samples were either unwashed or washed in neutral (pH 7.0) or alkaline (pH 8.0) modified Lake's diluent. Female golden eagles and peregrine falcons were inseminated via transjunctional, intramagnal insemination with washed spermatozoa from urine-contaminated samples. Urine contamination occurred in 36.8 +/- 12.8% (mean +/- SEM) golden eagle, 43.1 +/- 9.1% imperial eagle. 28.7 +/- 16.1% Bonelli's eagle, and 48.2 +/- 17.3% peregrine falcon ejaculates. The pH in urine-contaminated semen samples ranged from 6.48 +/- 0.3 to 6.86 +/- 0.2, and in noncontaminated samples it ranged from from 7.17 +/- 0.1 to 7.56 +/- 0.1. Sperm viability and motility were reduced (P < 0.05) in all species for unwashed vs. washed sperm after 30 min incubation at room temperature. Two peregrine falcon chicks and one golden eagle chick hatched after intramagnal insemination. This study demonstrates that urine contamination, a common and lethal acidifier in manually collected raptor ejaculates, can be circumvented by immediate, gentle seminal washing. Furthermore, these processed sperm, when deposited by transjunctional intramagnal insemination, can produce live young.

Animals↗