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PubMed2000

Artificial intravaginal insemination using fresh semen in cats.

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

Animals↗
PubMed2003

Application of timed artificial insemination protocols to grazing Japanese black cattle with long open period.

To investigate the viability of timed artificial insemination (TAI) protocols in grazing Japanese Black cattle with long open period, ovarian status and progesterone and estradiol-17 beta profiles of the animals during the protocol were monitored. In 1998, prosta-glandin F(2a) (PG) was administered to 36 animals seven days after GnRH injection. Three out of the 36 animals were inseminated after detection of estrus and did not receive further treatment. The second GnRH was injected to the remaining 33 animals 48 hr after the PG injection and TAI was performed 24 hr later. In 1999, PG was injected to 25 animals six days after GnRH, the second GnRH was injected to 22 animals 48 hr after PG, and TAI was performed 16 hr later (The other three animals were inseminated before the time of TAI). The percentage of the animals with at least one functional corpus luteum and one follicle equal to or greater than 10 mm in diameter at PG injection was similar between the groups in 1998 and 1999. Likewise, the hormonal profiles were similar between the two groups. Pregnancy rates (PR) after the TAI protocols and natural mating in 1998 and 1999 were 75.0% and 88.0%, respectively. These figures were comparable to the PR obtained by conventional estrus synchronization protocols using PG (in 1995; 69.4%) or CIDR (in 1996; 59.1%). In conclusion, the TAI protocol can be applicable into grazing Japanese Black cattle with long open period.

Animals↗
PubMed2003

Intratubal insemination with fresh semen in dogs.

The number of spermatozoa required to obtain conception by intratubal insemination in dogs was examined. Three groups consisting of 5, 8 and 8 dogs received 0.5 x 10 (6), 2.0 x 10(6) and 4.0 x 10(6) spermatozoa, respectively, into each uterine tube. No conception occurred in the 5 animals inseminated with 0.5 x 10(6) spermatozoa, but conception occurred in 6/8 (75.0%) and 3/8 (37.5%) dogs inseminated with 2.0 x 10(6) and 4.0 x 10 (6) spermatozoa, respectively. Among the pregnant animals, three aborted (33.3%) and the mean number of newborns was small, 2.5 +/- 0.5 (SE). One acardiacus anceps was observed with normal fetus in one animal with a Caesarean delivery.

Abortion, Veterinary↗
PubMed2004

Artificial insemination of frozen epididymal sperm in beagle dogs.

Freeze-storage of epididymal sperm is an important technique for the preservation of gametes in animals, including those becoming extinct. We froze canine sperm recovered from the cauda epididymis and investigated the fertility. The qualities of sperm from the cauda epididymis before freezing were: mean sperm motility, 89.4 +/- 1.6 (SE) %; sperm viability, 89.1 +/- 1.1%; and these were significantly higher than those of sperm from the caput-corpus epididymis (P<0.01, P<0.05). The number of sperm recovered from both cauda epididymides varied among animals: 6.3-122.3 x 10(7), mean 61.5 +/- 10.0 x 10(7). Freezing was used only for sperm recovered from the cauda epididymis. The sperm motility and viability after thawing were 19.5 +/- 2.5% and 53.1 +/- 3.3%, respectively. These were slightly lower than those of frozen-thawed ejaculated sperm, but the differences were not significant. When 2 x 10(8), 3 x 10(8), or 4 x 10 (8) sperm were inseminated in the unilateral uterus, only one animal inseminated with 3 x 10(8) sperm was fertilized (1/16, 6.3%). When 1 x 10(8) sperm were inseminated in the bilateral uterine tubes, one of six animals (16.7%) was fertilized. Therefore, although the qualities of epididymal sperm after thawing were similar to those of ejaculated sperm, the conception rate obtained with frozen-thawed epididymal sperm was low in beagle dogs. It is necessary to investigate the differences in damage between epididymal sperm after thawing and ejaculated sperm and to develop a method for improving the conception rate.

Animals↗
PubMed2005

Unilateral intrauterine insemination with cryopreserved caudal epididymal sperm recovered from refrigerated canine epididymides.

Canine epididymides were excised and immediately stored at 4 degrees C for 48 hr, and the qualities of caudal epididymal sperm after recovery and cryopreservation were evaluated. To confirm the fertility of the cryopreserved caudal epididymal sperm, artificial intrauterine insemination was performed. The sperm motility (61.0%) immediately after recovery from caudal epididymis stored at 4 degrees C for 48 hr was significantly lower than those of sperm stored for 0 and 24 hr (88.6 and 80.7%, respectively), but there was no significant difference after freeze-thawing (0-, 24-, and 48-hr storage groups: 27.9, 24.3, and 28.3%, respectively). The incidence of abnormal sperm immediately after recovery was significantly higher in the 24-hr and 48-hr storage groups (19.3 and 27.7%, respectively) than in the 0-hr storage group (5.6%), and a significant difference was also observed after freeze-thawing. The incidence of immature sperm with cytoplasmic droplets was significantly higher in the 48-hr storage group (18.4%) than in the 0-hr storage group (4.7%), but there was no difference after freeze-thawing. By unilateral intrauterine insemination (2x10(8) sperm), 4 of 5 bitches (80%) conceived. The above findings demonstrated that sperm motility was good even enough the incidence of abnormal sperm was high in canine epididymal sperm that were recovered from the epididymis stored at 4 degrees C for 48 hr and cryopreseved, and that artificial intrauterine insemination resulted in a high conception rate.

Analysis of Variance↗
PubMed1979

Comparison of the embryo survival rate in heifers following artificial insemination, non-surgical blastocyst transfer or both.

In Exp. 1, embryo survival rates of 45 and 47% were recorded after artificial insemination and ipsilateral transfer respectively. In Exp. 2, pregnancy rates of 62 and 60% were recorded after artificial insemination and contralateral transfer to inseminated recipients respectively. In this experiment the contralateral transferred embryo survival rate was 44%. Transferred embryo survival was lower overall when donors and recipients were out of phase by 1 day than when exactly synchronous.

Animals↗
PubMed1980

Distribution of small- and medium-sized molecules within the genital tract of artificially inseminated gilts.

Radiolabelled compounds of different molecular size were used as tracers mixed with seminal plasma to investigate whether seminal plasma enters the oviducts of inseminated gilts. The substances were 125I-labelled human serum albumin for 3 gilts, 131I-labelled polyvinylpyrrolidone plus 59Fe citrate in 2 gilts and 131I-labelled human serum albumin plus 59Fe citrate in 2 gilts. The gilts were slaughtered 1 h after insemination and the radioactivity was measured in 4 parts of the oviduct, 9 parts of the uterine horns, the body of the uterus and the cervix. Different quantities of the compounds used were found in all oviducts, but there was close agreement between the distributions of the two compounds in the genital tract after simultaneous insemination. There was a difference (60-80%) in total amount of recovered radioactivity between left and right uterine horns. It is concluded that all compounds used entered the oviducts regardless of their molecular sizes.

Animals↗
PubMed1981

Detection of the effects of ingested caffeine on fertility of cocks by homospermic and heterospermic insemination.

Cocks were fed diets containing 0, 0.025, 0.05, 0.075 or 0.1% caffeine during a 14-day treatment period. The number of spermatozoa produced by cocks fed 0.075 or 0.1% caffeine declined sharply at 12 days after onset of treatment. Hens were inseminated with a constant number of spermatozoa from individual cocks. The fertility of cocks fed 0.05, 0.075 or 0.1% caffeine declined during the 17-day post-treatment period and then returned to pretreatment levels. Cocks whose offspring were distinguishable were paired and relative fertility was assessed in a heterospermic test. One cock in each pair was fed 0.05% caffeine during the treatment period. Hens were inseminated with semen mixed within pairs. The proportion of chicks sired by cocks fed caffeine decreased during treatment and remined at that level until 17 days after treatment, when it increased to pretreatment levels. The percentage of total eggs hatched declined concomitantly with the reduction in the proportions of chicks sired by treated cocks. These results indicate that the effect of low levels of a toxin could be detected by reduced numbers of eggs hatched after heterospermic insemination with semen of normal appearance.

Animals↗
PubMed1984

Cell-mediated immunity to male-strain histocompatibility alloantigens detected after natural insemination and systemic immunization in the female mouse using the cell-mediated microcytotoxicity test.

Female cell-mediated immunity to allogeneic spermatozoa after repeated natural insemination, in the absence of pregnancy, was compared with that after systemic challenge using the cell-mediated microcytotoxicity test to measure cytotoxic cell alloreactivity. After multiple (3-6) inseminations the majority of females (11 out of 13) showed a significant degree of lymphocytotoxicity to male-strain histocompatibility alloantigens in the para-aortic lymph nodes, and to a lesser extent in the spleens, while a single insemination was usually not sufficient to evoke a specific cytotoxic cell response. This differed from the low and highly variable degree of female sensitization after multiple systemic challenge with allogeneic spermatozoa via the intraperitoneal route. By contrast, a single systemic challenge via the footpad proved to be the most highly consistent and effective route for eliciting cell-mediated immunity to male-strain histocompatibility alloantigens in all 9 female mice. This alloreactivity appeared to be directed at alloantigens other than the male-specific H-Y antigen. These findings show that the precise route of immunization is a major factor in the development of female cell-mediated immune responsiveness to allogeneic spermatozoa.

Animals↗
PubMed1988

Turkey hen fertility and egg production after artificial insemination and multiple oviduct eversion during the pre-laying period.

The onset of egg production (mean 18.3 days after the onset of photostimulation) and the rate of egg production (flock averaged 4.9 eggs per bird per week for the first 8 weeks of egg production) were not affected by 5 days of twice daily oviduct eversion ('venting') in the pre-laying period when compared to unvented controls. After the onset of photostimulation, pre-laying hens were inseminated twice daily on Days 12 to 16 with 3 microliter semen containing 15 x 10(6) spermatozoa, and compared with groups of hens inseminated once daily on Days 15 and 16 with 15 microliters semen containing 75 x 10(6) spermatozoa or 41 microliter semen containing 200 x 10(6) spermatozoa. Fertility remained high for the first 5 weeks of egg production. However, by Week 6 the fertility of the hens receiving frequent low doses of semen dropped significantly below that of the others, which suggests that multiple inseminations with a low semen volume containing relatively low numbers of spermatozoa does not lead to an increase in the efficacy of sperm transport and storage in the oviduct.

Animals↗
PubMed1988

Duration of fertility of turkeys inseminated at different times after the onset of photostimulation.

To investigate the relationship between photostimulation, ovary and oviduct weights, and oviductal sperm-storage tubule (SST) development and functional capacity, 33-week-old turkey hens were placed in four groups of 15 hens each. Each group was inseminated three times in 30 min with 50 x 10(6) spermatozoa (a total of 150 x 10(6) spermatozoa) on the day of photostimulation, or on Days 7, 14, or 21 after the onset of photostimulation (Groups A, B, C and D, respectively). From weeks 4 to 8 after photostimulation, 5 additional hens were inseminated each week and then killed 24 h later to determine ovary and oviduct weights and SST status. Regardless of ovary and oviduct weights and insemination times in relation to photostimulation, 76 of 78 hens possessed morphologically differentiated SST, and 73 of the 76 hens had some SST which contained spermatozoa. For Groups A, B, C and D, the duration of fertility was approximately 7-8 weeks. It is concluded that morphologically differentiated SST are functional in 33-week-old hens regardless of the status of the ovary and oviduct.

Animals↗
PubMed1996

Responsiveness of ovaries to exogenous gonadotrophins and laparoscopic artificial insemination with frozen-thawed spermatozoa in ocelots (Felis pardalis).

Adult female ocelots (Felis pardalis) were treated with one of four dosages of equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG) (100 iu eCG/75 iu hCG, n = 3; 200 iu eCG/150 iu hCG, n = 4; 400 iu eCG/150 iu hCG, n = 5; 500 iu eCG/225 iu hCG, n = 5); hCG was administered 80 h after eCG. Ovaries of each animal were evaluated by laparoscopy 39-43 h after hCG, and blood was collected for progesterone and oestradiol analysis. With progressive increases in gonadotrophin dosage, female ocelots produced more (P < 0.05) unovulated follicles (> or = 2 mm in diameter), ranging from 1.3 +/- 0.7 (mean +/- SEM) follicles per female at the lowest dosage to 8.8 +/- 2.8 follicles per female at the highest dosage. Similarly, ocelots produced more (P < 0.05) corpora lutea with increasing gonadotrophin dosages, with mean values ranging from 0-5.0 +/- 1.2 corpora lutea. However, across treatment groups, a similar proportion (P > 0.05) of females ovulated in response to each dosage. At laparoscopy, serum concentrations of oestradiol (overall mean, 330.2 +/- 62.2 pg ml-1) and serum concentrations of progesterone (overall mean, 18.5 +/- 6.4 ng ml-1) in ovulating females did not differ (P > 0.05) across treatment groups. Ten ovulating ocelots were laparoscopically inseminated with fresh (4.7 +/- 0.2 x 10(6); n = 2 females) or frozen-thawed (10.7 +/- 1.8 x 10(6); n = 8 females), motile spermatozoa. One female treated with 500 iu eCG/225 iu hCG and inseminated with 7.5 x 10(6) motile, frozen-thawed spermatozoa conceived and gave birth to a healthy male kitten after a gestation of 78 days. We conclude that ocelots are relatively insensitive to exogenous gonadotrophins, requiring much higher dosages (on a per body mass basis) to elicit an appropriate ovarian response than do any other felid species studied to date. Nonetheless, the gonadotrophin-treated female can become pregnant and carry offspring to term after laparoscopic intrauterine insemination with frozen-thawed spermatozoa.

Animals↗
PubMed1996

Timing of ovulation after gonadotrophin induction and its importance to successful intrauterine insemination in the tiger (Panthera tigris).

The ovarian response to equine chorionic gonadotrophin (eCG) and human chorionic gonadotrophin (hCG), the effect of timing of ovulation relative to hCG injection and the use of laparoscopic intrauterine artificial insemination (AI) were examined in two subspecies of tiger (Panthera tigris). Adult female tigers were subjected to the same eCG/hCG treatment followed by laparoscopy under xylazine/diazapam/ketamine HCl anaesthesia at 39-42 h (Group I, n = 9), 46-49 h (Group II, n = 5) or 51-55 h (Group III, n = 5) after hCG. Six of these females, observed to be postovulatory at the time of laparoscopy (Group II, n = 3; Group III, n = 3), were subjected to intrauterine AI. The number of preovulatory follicles observed on the ovaries of Group I females was twofold greater (P < 0.05) than the number observed on ovaries of females in Group II and III. Fewer (P < 0.05) corpora lutea were observed on ovaries of Group I females (1.3 +/- 0.6) compared with the number of corpora lutea in Group II and III (combined average, 7.8 +/- 0.8 corpora lutea per female). Only one of ten females in Groups II and III failed to ovulate by the time of laparoscopy. Four Group I females never ovulated, based on a laparoscopic re-evaluation 4 weeks later. One female inseminated 46 h after hCG (Group II) became pregnant and delivered a healthy cub after a normal gestation. There were no apparent differences between subspecies in response to the same ovulation induction protocol. Results demonstrate the importance of the relationship between exogenous gonadotrophin treatment and onset of anaesthesia for laparoscopic examination and AI in tigers. Data clearly indicate that anaesthesia/laparoscopy conducted too early (39-42 h after hCG) compromises the number of females and proportion of follicles ovulating. In contrast, ovulation success is high if anaesthesia/laparoscopy is performed after this time, and intrauterine insemination can result in healthy young.

Animals↗
PubMed2005

Estrous cycle characterisation and artificial insemination using frozen-thawed spermatozoa in the bottlenose dolphin (Tursiops truncatus).

The reproductive endocrinology of the bottlenose dolphin, Tursiops truncatus, was characterized to facilitate the development of artificial insemination using cryopreserved spermatozoa. Specific objectives were: (i) to determine the excretory dynamics of urinary luteinizing hormone (LH) and ovarian steroid metabolites during the estrous cycle; (ii) to evaluate the effect of an exogenously administered synthetic progesterone analog (altrenogest) on reproductive hormone excretion; (iii) to correlate follicular growth and ovulation (as determined by transabdominal ultrasound) to urinary LH and ovarian steroid metabolites; (iv) examine the in vivo fertilisation capacity of cryopreserved semen, and (v) to develop an intrauterine insemination technique. Based on urinary endocrine monitoring of natural estrous cycles (2 consecutive cycles) and nine post altrenogest cycles in ten females, estrous cycles were found to be 36 days long and comprised of an 8 day and 19 day follicular and luteal phase, respectively. Peak estrogen conjugates (EC; 5.4+/-3.8 ng/mg creatinine (Cr)) occurred 8 h prior to the LH surge (70.9+/-115.7 ng/mg Cr). The time of ovulation, as determined by ultrasonography, occurred 32.1+/-8.9 h and 24.3+/-7.0 h after the onset of the LH surge and LH peak, respectively. Mean preovulatory follicular diameter and circumference were 2.1+/-0.5 cm and 6.5+/-1.5 cm, respectively. Of the 27 estrous synchronisation attempts, 13 resulted in an ovulatory cycle, with ovulation occurring 21 days post-altrenogest treatment. Intrauterine (4 of 5) and intracornual (1 of 3) inseminations conducted across eight estrous cycles resulted in five pregnancies (63%), one pregnancy resulted from the use of liquid stored semen, whereas four were achieved using cryopreserved semen. These data provide new information on female bottlenose dolphin reproductive physiology, and demonstrate that the combination of endocrine monitoring and serial ultrasonography contributed to successful AI using liquid-stored and cryopreserved semen.

Animals↗
PubMed1985

[Three successful cases of artificial insemination in chimpanzees].

Artificial inseminations were performed on two female chimpanzees from July 1981 to April 1983 and three conceptions were obtained. Semen samples collected by rectal probe method of electroejaculation were incubated at 37 degrees C for about 20 minutes for liquefaction. Liquefied portion of the semen was sucked up into polyethylene tube about 30 cm in length attached to a syringe and was inseminated into cervix of each female of which pelvic region was raised in prone position under anesthesia. Ovulation time was speculated by swelling of sex skin in earlier two cases and by urinary LH level in last case. Three offsprings were obtained 234, 235 and 235 days after last insemination, respectively. All of three show normal developments.

Animals↗
PubMed1993

Usefulness of a simplified artificial insemination technique in the rabbit for teratology studies.

We describe usefulness of our simplified artificial insemination technique in rabbits for teratology studies. Our technique includes 1) semen collection from 2 or 3 bucks with a simple artificial vagina, 2) deposition of 0.5 microliters of the diluted semen mixture in the vagina with a glass pipette, and 3) injection of 25 U. of hCG through the auricular vein. Applying these techniques and procedures and keeping about 10 proven bucks would enable 2 operators to inseminate 10-15 does daily by only 1 hour work. This artificial insemination technique should be useful for rabbit teratology studies, especially for safety evaluation studies of toxicants, where a relatively large number of pregnant are needed to be prepared in a short period.

Animals↗
PubMed2002

Effect of diet composition on weight gain, sperm transfer, and insemination in the cat flea (Siphonaptera: Pulicidae).

Weight gain by adult cat fleas, Ctenocephalidesfelis (Bouché), was influenced primarily by the concentrations of protein and sodium chloride in the feeding solution. After 48 h of feeding, fleas fed whole blood weighed almost twice as much as fleas fed plasma or hemolyzed blood and 1.25 times as much as fleas fed 0.15 M sodium chloride. When fleas were fed sodium chloride solutions ranging from 0 to 0.5 M, weight gain was greatest on the 0.15- or 0.2-M solutions. Weight gain decreased significantly when fleas were fed plasma, hemolyzed blood or 0.3 or 0.5 M sodium chloride in place of whole blood, but improved when plasma was diluted 100% and when hemolyzed blood was diluted 10% with distilled water. Adenosine-5'-triphosphate did not appear to stimulate weight gain in cat fleas; weight gain was unchanged in fleas fed hemolyzed blood or 0.15 M sodium chloride to which 0.005 M ATP was added. Insemination did not occur in starved fleas or those fed protein-free diets. When fleas were starved or fed distilled water, sodium chloride, or other salt solutions, sperm was transferred from the testes to the vas deferens in 91-94% of males, but no females were inseminated. In contrast, when fleas were fed whole blood, hemolyzed blood, plasma, or bovine serum albumin (3.5 or 7.0 g/deciliter) dissolved in 0.15 M saline, 80, 80, 10, and 10% of the females were inseminated, respectively.

Animals↗
PubMed2000

Birth of Siberian tiger (Panthera tigris altaica) cubs after transvaginal artificial insemination.

A captive 9-yr-old female Siberian tiger (Panthera tigris altaica) at the Lisbon Zoo was treated with equine chorionic gonadotropin followed 80 hr later by human chorionic gonadotropin to stimulate follicular development and ovulation, respectively. Forty-one hours after the second hormone administration, the tiger was subjected to transvaginal artificial insemination with diluted fresh semen containing 500 x 10(6) motile spermatozoa obtained from an 8-yr-old male Siberian tiger via electroejaculation. After anesthesia, the female tiger was placed in an inclined position with the hind limbs elevated during and after the insemination and was maintained in this position for 15 min to minimize semen reflux. An anesthetic reversal agent was then administered. After a 103-day gestation, the female gave birth to three apparently healthy cubs that survived 24-48 hr. These results demonstrate that transvaginal artificial insemination can be successfully performed in the Siberian tiger to produce full-term offspring.

Animals↗