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PubMed2003

Fertilized eggs obtained from transplantation of frozen ovaries and parthenogenesis in combination with artificial insemination of frozen semen of the silkworm, Bombyx mori.

A reliable method is reported for the long-term preservation of ovaries and spermatozoa of the silkworm (Bombyx mori). Three studies are presented. In the first, ovaries were removed from larvae at either 3rd, 4th, or 5th instar, cryopreserved, and stored in liquid nitrogen. Thawed ovaries were transplanted to surgically castrated female larvae at the same or a different developmental stage. The highest percentage of recipient females producing eggs resulted into either 3rd or 4th instar larvae (respectively, 22.1 and 8.7%). Similarly, the highest levels of other measurements of successful cryopreservation and transplanted ovary, and number of eggs laid, occurred with the same combination of donor and recipient developmental stages. Other combinations of ovary/recipient developmental stages yielded lower results. In the second experiment, semen was collected from male moths, cryopreserved, and then thawed semen was diluted with trypsin solution and artificially inseminated into females obtained from the best conditions of first experiment. A small percentage of inseminated moths laid eggs (8-10.3%) compared to that of controls (100%). In addition, the fertility of eggs from experimental moths was lower than that of control females (respectively, 40.3-88% and 97.5%). In the third experiment, eggs were surgically removed from ovarian tubules of moth following transplantation of thawed ovaries and subjected to parthenogenetic activation and artificial hatching. As expected, all resulting moths were female and, following natural mating or artificial insemination with thawed semen, yielded normal offspring at high rates.

Animals↗
PubMed2003

Seminal lead concentrations negatively affect outcomes of artificial insemination.

OBJECTIVE: To determine the relationships among seminal lead levels, acrosome status, and artificial insemination cycle fecundity (AI f) in semen donors. DESIGN: Longitudinal analysis of seminal lead levels, sperm function testing, and fecundity. SETTING: University medical center andrology and research laboratories. PATIENT(S): Semen donors (n = 15) participating in a therapeutic donor insemination program. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Seminal plasma lead levels, acrosome sensitivity to progesterone (P) and voltage-gated potassium channel inhibitors (e.g., charybdotoxin [CBTx]), and AI f. RESULT(S): Seminal plasma lead levels and AI f were strongly negatively correlated. Semen donors were divided into three groups by acrosome response to P: normal (CBTx sensitive [Rs] or CBTx resistant [Rr]: responders) and reduced (nonresponders [NR]) (Rs > Rr >> NR). Seminal lead differed among the three groups (NR > Rr > Rs). Comparison of 330 artificial insemination cycles from four Rs, four Rr, and two NR demonstrated that cycle AI f also differed significantly between groups (Rs >Rr >>NR). CONCLUSION(S): Measurements of seminal plasma lead, P-stimulated acrosome loss, and sensitivity to CBTx may provide prognostic information on the fertility status of potential donors as well as male infertility patients. Such evaluations may assist in donor acceptance, or in the case of patients, in selection of the appropriate treatment regimen.

Acrosome Reaction↗
PubMed1992

Spermatozoal characteristics from fresh and frozen donor semen and their correlation with fertility outcome after intrauterine insemination.

OBJECTIVE: To determine if conventional sperm parameters, specific characteristics of sperm motion determined by computer-aided semen analysis (CASA), sperm penetration assay (SPA), and/or spontaneous acrosome reaction assay could best predict fertility outcome after intrauterine insemination (IUI) from frozen donor sperm. DESIGN: A retrospective analysis of 2,245 cycles of therapeutic donor IUIs were initially studied; 1,147 cycles that met selection criteria were used in this report. SETTING: A university-based assisted reproductive technology center. PATIENTS, PARTICIPANTS: All IUIs were performed on women with documented patency of at least one fallopian tube, ovulatory cycles, and who did not receive human menopausal gonadotropins. Sperm donors had to be used for at least four different recipients (mean of 15) and at least 14 different cycles of insemination (mean of 41). INTERVENTIONS: None. MAIN OUTCOME MEASURE: Pregnancy. RESULTS: Statistical comparisons were made between donors of different relative fertility by using the Mann-Whitney test, Spearman's rank correlation, and multiple regression analysis. These analyses demonstrated that the most significant predictors of the fertility of frozen-thawed donor sperm were curvilinear velocity, straight line velocity, and the total number of motile sperm inseminated. The number of sperm with spontaneous acrosome reactions negatively correlated with fertility outcome, and SPA provided no predictive value. CONCLUSIONS: Our study supports the hypothesis that the study of sperm motion characteristics using CASA after thawing and washing of cryopreserved sperm is a better predictor of fertile outcome after IUI than analysis of fresh semen.

Acrosome↗
PubMed1994

Correlation of semen variables and pregnancy rates for donor insemination: a 15-year retrospective.

OBJECTIVE: To determine the validity of the criteria currently used to select fertile sperm donors. DESIGN: Retrospective study of the outcome of donor insemination treatment cycles over a 15-year period. SETTING: The Reproductive Medicine Clinic of Prince Henry's Institute of Medical Research (formerly Medical Research Centre, Prince Henry's Hospital, South Melbourne, Victoria, Australia). PATIENTS: Couples (1,299) presented with severe male factor infertility (azoospermia or severe oligozoospermia) and were treated by donor insemination (10,796 treatment cycles) using cryopreserved semen from 292 normal healthy men aged between 18 and 46 years as sperm donors. MAIN OUTCOME MEASURES: Semen characteristics, including volume, count, motility, motility index, normal morphology, live cells, and post-thaw motility, were analyzed by logistic regression analysis to determine which groups of factors were related independently and significantly to pregnancy rates. RESULTS: The pregnancy rate for donor insemination was 8.9% per cycle. By logistic regression count, normal morphology, post-thaw motility (%), and post-thaw motility index together explained 28% of the variance of the pregnancy rates. Other factors, including prethaw motility characteristics (percent motility and motility index), donor age, weight, LH, FSH, and T levels, were not significant. CONCLUSIONS: The standard parameters of sperm count, morphology, and post-thaw motility are the most important factors for the selection of highly fertile donors. As most of the variability in the pregnancy rates resulted from other undetermined factors related both to the donor and the recipient, it is suggested that the pragmatic approach of discarding donors after a reasonable number of treatment failures (20 to 50) should continue.

Cryopreservation↗
PubMed1988

Spermatozoa selection in discontinuous Percoll gradients for use in artificial insemination.

In a preliminary study of the ejaculate of 93 men, changes in motility and hypo-osmotic swelling before and after sperm separation by discontinuous Percoll gradients were evaluated. Both parameters improved significantly (P less than 0.01) in Percoll-separated spermatozoa. In a second stage, 99 couples underwent intrauterine insemination of separated semen by Percoll gradients. The population had infertility for a mean of 5.6 years. In a 1-year period, a total of 344 intrauterine insemination cycles were completed. Failure was considered when conception did not occur within four treatment cycles. The mean number of cycles per successful couple before pregnancy was 1.52. An overall 21% pregnancy rate was achieved (16% for oligoasthenospermia and 24% for asthenospermia). Sperm separation in Percoll gradients is a useful technique for intrauterine insemination in some cases of male subfertility.

Adult↗
PubMed1989

Differences in the fertility of donor insemination recipients--a study to provide prognostic guidelines as to its success and outcome.

Data on 207 couples attending for donor insemination (DI) were analyzed to determine the differences in fertility of insemination recipients and to provide prognostic guidelines as to its success. This study has shown that DI recipients with no infertility factors whose partners were azoospermic had the highest fecundability (0.178), whereas those with no infertility factors whose partners were subfertile had the lowest fecundability (0.036). Donor insemination recipients with treated endometriosis and ovulatory dysfunction have significantly higher fecundability (0.093 and 0.091, respectively) than those with other infertility factors (0.021) such as fibroids, tubal disease, or a combination of infertility factors. This study provides prognostic guidelines for DI recipients as to its success.

Adult↗
PubMed1998

A randomized, prospective analysis of five sperm preparation techniques before intrauterine insemination of husband sperm.

OBJECTIVE: To evaluate pregnancy rates (PRs) in women undergoing artificial insemination with sperm alternately prepared by one of five techniques: sperm washing, Percoll gradient centrifugation, swim-up, swim-down, or refrigeration/heparin treatment. DESIGN: Each treatment group alternated in a different order through the five sperm preparations. Pregnancy rates were compared for each sperm preparation. SETTING: Two infertility centers, one located in an academic institution and the other a regional hospital. PATIENT(S): Three hundred sixty-three women undergoing 898 artificial inseminations with husband semen with a progressive motile sperm count of >20 million sperm per mL were randomly placed in the five treatment groups. MAIN OUTCOME MEASURE(S): Pregnancy rates. RESULT(S): The overall ongoing PR per insemination was 9.7% (87/898), including 6.12% for natural cycles (n = 196), 12.8% for clomiphene citrate-stimulated cycles (n = 101), and 10.3% for gonadotropin-stimulated cycles (n = 601). The highest ongoing PRs for sperm preparations followed the swim-up technique (13.2%, 26/197) and the Percoll gradient centrifugation technique (12.7%, 26/204). CONCLUSION(S): These data suggest that the swim-up and Percoll gradient preparations result in higher PRs than the wash, swim-down, and refrigeration/heparin techniques.

Centrifugation, Density Gradient↗
PubMed1997

Effect of motile sperm count after swim-up on outcome of intrauterine insemination.

OBJECTIVE: To analyze the prognostic value of motile sperm count after swim-up in IUI with husband's sperm in a large group of subfertile couples. DESIGN: Retrospective study. SETTING: University hospital. PATIENT(S): Nine hundred two couples undergoing 3,037 treatment cycles. INTERVENTION(S): Intrauterine insemination with husband's sperm after swim-up was performed after mild ovarian stimulation with clomiphene citrate and hCG under hormonal and ultrasonographic control of follicle development. MAIN OUTCOME MEASURE(S): Pregnancy rate (PR) in correlation to motile sperm count after swim-up. RESULT(S): A nonlinear increase in PR per treatment cycle was observed with increasing numbers of motile sperm used for IUI. Insemination with < 0.8 x 10(6) motile sperm after swim-up resulted in a PR of < 1% per treatment cycle. When the motile sperm count was above this level, the PR per cycle reached a plateau of 6.9% to 10.2%, with a minor tendency for increase with higher sperm numbers. CONCLUSION(S): Strict analysis of motile sperm count after swim-up is a useful prognostic factor for PRs after IUI. There is a good chance for conception if > or = 0.8 x 10(6) motile sperm are available after appropriate selection methods. Intrauterine insemination performed with considerably higher numbers of motile sperm does not lead to a significant increase in PRs.

Adolescent↗
PubMed2000

Effect of body condition on reproductive efficiency of lactating dairy cows receiving a timed insemination.

Body condition may influence pregnancy rates to a timed insemination (Ovsynch/TAI) protocol and affect the economical performance of dairy farms. The objectives were to compare pregnancy rates using the Ovsynch/TAI protocol for the first service of lactating dairy cows with body condition scores < 2.5 (scale: 1 to 5, low BCS group) versus > or = 2.5 (control group) and to estimate the economic impact of the effect of body condition on reproductive performance. At 63 +/- 3 d post partum, cows were assigned to 2 experimental groups (low BCS = 81; control = 126), and were treated with GnRH at d 0 and with PGF2alpha 7 d later. At 48 h after PGF2alpha, cows received an injection of GnRH and were inseminated 16 h later. Pregnancy rates to the Ovsynch/TAI protocol were lower for the low BCS group than for the control group at 27 d (18.1 +/- 6.1% < 33.8 +/- 4.5%; P<0.02) and at 45 d (11.1 +/- 5.4% < 25.6 +/- 4.1%; P<0.02) after insemination. Economic analysis indicated that reducing the percentage of the herd in low body condition increases net revenues per cow per year. Body condition influenced pregnancy rates to the Ovsynch/TAI protocol.

Adipose Tissue↗
PubMed2000

T-cell distribution in two different segments of the equine endometrium 6 and 48 hours after insemination.

The T-cell response after the introduction of semen into the uterine cavity in the mare was studied by examining, immunohistochemically, the distribution of helper T-cells (CD4+) and cytotoxic T-cells (CD8+) in endometrial biopsy specimens. Endometrial tissue samples were obtained from twenty-five gynecologically healthy mares during estrus before and 6 or 48 h after deposition of a single dose of stallion semen. An increase (P=0.04) in the number of helper T-cells (CD4+) compared to pre-insemination values was observed in the uterine body in both groups, 6 and 48 h, after insemination. No significant variations in numbers of CD8+ cells were recorded either 6 or 48 h after insemination. There seems to be an early (6 h) recruitment of helper T-cells to the equine endometrium after semen deposition, which might be related to the activation of the endometritis-like reaction seen as part of the equine uterine immune defense during estrus.

Animals↗
PubMed2000

Comparison of culture and insemination techniques for equine oocyte transfer.

This study was designed to test 3 approaches for insemination and transfer of oocytes to recipient mares. Oocytes were recovered transvaginally from naturally cycling donor mares 24 to 26 h after an intravenous injection of 2500 IU of hCG when follicles reached 35 mm in diameter. Multiple oocytes (1 to 4) were transferred surgically into the oviducts of 4 or 5 recipient mares per group. Three groups of transfers were compared: 1) transfer of oocytes cultured in vitro for 12 to 14 h postcollection with insemination of the recipient 2 h postsurgery; 2) transfer of oocytes into the oviduct within 1 h of collection, with completion of oocyte maturation occurring within the oviduct, and insemination of the recipient 14 to 16 h postsurgery; and 3) transfer of spermatozoa and oocytes (cultured 12 to 14 h in vitro) into the oviduct. Numbers of embryos detected by Day 16 of gestation were not different (P>0. 1) for groups 1, 2, and 3 (57%, 43% and 27%). Therefore, equine oocytes successfully completed the final stages of maturation within the oviduct, and sperm deposited within the oviduct were capable of fertilizing oocytes.

Animals↗
PubMed2001

Post-thaw survival of ram spermatozoa and fertility after insemination as affected by prefreezing sperm concentration and extender composition.

A study was conducted to investigate the effects of prefreezing sperm concentration using two extenders on post-thaw survival and acrosomal status of ram spermatozoa (Experiment 1) and fertility after intrauterine insemination with differing doses of semen (Experiment 2). In autumn (Northern hemisphere), semen was collected by artificial vagina from 8 adult Leccese rams and ejaculates of good quality semen were pooled. Two extender systems for cryopreservation were considered, one based on milk-lactose egg yolk (Milk-LY) and the other based on tris-fructose egg yolk (Tris-FY). Experiment 1 (2 x 6 factorial scheme) examined the in vitro characteristics of spermatozoa in relation to the Milk-LY and Tris-FY extenders and six prefreezing sperm concentrations (50, 100, 200, 400, 500 and 800 x 10(6) spermatozoa/mL). Experiment 2 (2 x 4 factorial) evaluated the influence of the Milk-LY vs Tris-FY extenders and four doses (20, 40, 80 and 160 x 10(6) spermatozoa/0.25 mL) corresponding to prefreezing spermatozoa concentrations of 100, 200, 400 and 800 x 10(6) spermatozoa/mL, on fertility of ewes inseminated in uterus by laparoscope. Prefreezing sperm concentration influenced (P < 0.01) freezability of spermatozoa and affected negatively all the in vitro parameters at 800 x 10(6) spermatozoa/mL. Overall, Milk-LY tended to ensure higher viability and acrosomal integrity of spermatozoa after thawing at the intermediate sperm densities (range 100 to 500 x 10(6) spermatozoa/mL). At 500 x 10(6) spermatozoa/mL concentration corresponded the best condition for survival of spermatozoa (71.2%), acrosome integrity (71.5%) and acrosomal loss (6.0%). At the lowest sperm concentration (50 x 10(6) spermatozoa/mL), Tris-FY resulted in a higher survival rate than Milk-LY (61.3%, P < 0.05) and lower acrosomal loss (9.7%, P < 0.05). Milk-LY supported spermatozoa motility better than Tris-FY after incubation at sperm concentration between 50 and 400 x 10(6) spermatozoa/mL (0.05 > P < 0.01). Semen doses of 20 to 40 x 10(6) spermatozoa/ewe provided satisfactory fertility rates (64 to 81%). The increase of inseminate doses to 160 x 10(6) spermatozoa/ewe failed to improve fertility, actually tending to decrease lambing rates.

Animals↗
PubMed2002

The effect of interval from day of administration of bovine somatotropin (bST) to synchronization of ovulation and timed-insemination on conception rate of dairy cows with and without ovarian cysts.

The objective of this study is to evaluate the effect of the interval from the day of administration of bovine somatotropin (bST) to the day of initiation of synchronization of ovulation (Day 0 and 7) and timed-insemination (TAI) on conception rate (CR) of dairy cows with and without ovarian cysts, respectively. Lactating dairy cows (n = 359) were divided into two groups. Cows in Group 1 (n = 238, without ovarian cysts) were treated with 100 microg, i.m. GnRH on Day 0; 25 mg, i.m. PGF2a on Day 8. 100 mirog. i.m. GnRH on Day 10; and inseminated 16 h later without detection of estrus. Cows in Group 2 (n = 121, with ovarian cysts) were treated with 100 microg, i.m. GnRH on Day 0; 100 microg, i.m. GnRH on Day 7; 25 mg, i.m. PGF2alpha on Day 14, 100 ug, i.m. GnRH on Day 16; and inseminated 16 h later without detection of estrus. Between 60 and 63 days postpartum, all cows in the herd were given bST every 14 days for the duration of the study. Logistic regression was used to assess the risk of nonpregnancy associated with interval from bST treatment to Day 0 for cows without ovarian cysts. and both Day 0 and 7 for cows with ovarian cysts adjusting for parity and days in milk. The CR for cows in Group 1 was significantly higher when the interval from last treatment with bST to Day 0 was between 1 and 3 days (28%) compared to 4-6 days (14%). In addition, the risk of nonpregnancy was 2.19 times greater in cows 4-6 days after bST treatment compared to 1-3 days after adjusting for parity and days in milk. The CR for cows in Group 2 was not significantly different when the interval from last treatment with bST to both Day 0 and 7 was between 1 and 3, 4 and 6, and 7 and 14 days. In conclusion, the results of this study suggested bST treatment closer to Day 0 had a positive effect on CR of cows without ovarian cysts, but bST treatment closer to both Day 0 and 7 had no effect on CR of cows with ovarian cysts. This was interpreted to mean that bST had a beneficial effect on either, or both, the preovulatory follicle and the oocyte in dairy cows without ovarian cysts, but not in dairy cows with ovarian cysts.

Animals↗
PubMed2003

Fertility of ram semen frozen in Bioexcell and used for cervical artificial insemination.

The current use of ingredients of animal origin, such as egg yolk, in semen extenders presents a risk of microbial contamination, and has led to the search for alternatives. Such an extender is commercially available for bull semen (Bioexcell), IMV, L'Aigle, France), and it has previously been tested in vitro for freezing ram semen, with satisfactory results. The aim of the present study was to compare the fertility results of ewes in Uruguay, after cervical insemination with ram semen that was frozen in Bioexcell versus semen frozen in a conventional milk-egg yolk extender (control). Semen from five Corriedale rams was frozen, using a split sample design, in either milk-egg yolk or Bioexcell extender, using a two-step extension method. The sperm parameters assessed after thawing were subjective motility, membrane integrity (SYBR-14/PI), and capacitation status (CTC). Thawed semen was inseminated intracervically once during spontaneous estrus in 970 Corriedale ewes that grazed in natural pastures, under extensive management conditions. Fertility was recorded as nonreturn rates at 21 days (NRR-21) and 36 days (NRR-36) after artificial insemination (AI), as well as pregnancy rate (PR-US, diagnosed ultrasonographically 50 days after AI of the last ewe). Subjective motility was slightly higher in Bioexcell than in the milk extender (47 vs. 46.5%; NS), as was membrane integrity (38 vs. 37.7%; NS) and the percentage of uncapacitated spermatozoa (28.5 vs. 26.3%; NS). There were no statistically significant differences in fertility rates found between Bioexcell and the control extender: NRR-21 (35.9 vs. 33.2%), NRR-36 (34.8 vs. 32.6%), and PR-US (28.4 vs. 27.2%). In conclusion, Bioexcell appears to be an alternative to the conventional milk-egg yolk extender for freezing ram semen, and provides similar fertility results after cervical AI under extensive management conditions. Thus, Bioexcell, containing no additives of animal origin, can offer a safer alternative when frozen semen is used for introducing new genetic material into a flock or a country.

Animals↗
PubMed2003

Efficacy of two types of vaginal sponges to control onset of oestrus, time of preovulatory LH peak and kidding rate in goats inseminated with variable numbers of spermatozoa.

In small ruminants, progestagen-impregnated vaginal devices (sponges) are useful tools to manage reproduction irrespective of season and to the application of timed artificial insemination (AI). A novel progestagen releasing vaginal-controlled release device (Chronogest CR), loaded with less (20mg) cronolone using proprietary procedures, was developed and its efficacy (synchronising ability, fertility and prolificacy following sponge removal) evaluated versus the existing Chronogest sponge containing 45 mg of cronolone in goats. Females (n=199) were maintained in field conditions and inseminated with graded amounts of spermatozoa at two stages of the year (breeding and non-breeding seasons). The use of the new Chronogest CR sponge was associated with an earlier initiation of the LH surge (28.7h versus 30.8h following sponge removal, P<0.01). A similar degree of synchronisation of the LH surge was obtained with both types of sponges. In both treatment groups, a longer time interval between sponge removal and the LH surge was noted in females with high milk production. Fertility and prolificacy were high and unaffected by the type of sponge used or the amount of spermatozoa inseminated. It is concluded that the new Chronogest CR sponge allows a reduction of the progestagen load from 45 to 20mg without detrimental effects on synchronisation, fertility and prolificacy.

Animals↗
PubMed1998

Delayed insemination is successful with a new extender for storing fresh equine semen at 15 degrees C under aerobic conditions.

Milk-based semen diluents are known to be practical and effective in protecting equine spermatozoa during storage before artificial insemination. Milk is a biological fluid with a complex composition and contains components which are beneficial or harmful to spermatozoa. The aim of this study was to test the fertility of stallion semen after long-term storage using different milk diluents (INRA 82 or Kenney's diluent) vs one diluent chemically defined (INRA 96), which is composed of efficient milk components and optimized for sperm survival and storage temperature. The milk fraction used was that which best maintained spermatozoal survival based on motility measured in previous studies. Four breeding trials were conducted to determine the influence of combination of new diluent and storage conditions on fertility of the stallion. We compared the standard protocol of storing semen in a skim milk diluent (INRA 82 or Kenney's diluent) at 4 degrees C under anaerobic conditions with the experimental protocol which consisted of storing in a chemically defined, milk-free diluent (INRA 96), at 15 degrees C, under aerobic conditions. After 4 breeding trials, in which the semen was stored for 24 h under the 2 protocols, we obtained 57% (n = 178) and 40% (n = 173) of fertility per cycle using the experimental and the standard protocol respectively (p < 0.001). Another breeding trial was conducted to determine the influence of storage time on the fertility of spermatozoa. We have compared the fertility of semen inseminated immediately (68% of fertility per cycle, n = 50) vs the fertility of semen stored under the experimental protocol for 72 h before insemination (48% of fertility per cycle, n = 52). The experimental protocol improved sperm fertility compared to the standard protocol and seems to be a particular alternative for stallions with cold shock sensitive spermatozoa. Storing semen for 72 h under the experimental protocol seems to be useful in the field.

Aerobiosis↗
PubMed1998

The effect of reducing hindquarter elevation time after artificial insemination in bitches.

Hindquarter elevation time after artificial insemination in dogs was reduced from the common and arbitrarily used 10 min to only 1 min after insemination. Artificial insemination with fresh undiluted semen was conducted in 32 breedings using 15 hound bitches. The overall pregnancy rate was 91% (29/32), with an average litter size of 7.35 puppies per pregnancy. The pregnancy rate was not altered by reducing the 10-min (n = 14) hindquarter elevation time to 1 min (n = 18; P = 0.30). Similarly, the litter size was not different between groups (P = 0.40).

Animals↗
PubMed1999

Comparison of fertility data from vaginal vs intrauterine insemination of frozen-thawed dog semen: a retrospective study.

Fertility data from 327 artificial inseminations (AIs) using frozen-thawed dog semen are presented here. The AIs were performed in 274 bitches using semen from 185 males of 76 breeds. The data cover all AIs conducted during 1983 through 1995 at Cryogenetic Laboratories (CLONE) in the United States with AKC-registered and research bitches, and all AIs carried out at the Department of Obstetrics and Gynecology at the Swedish University of Agricultural Sciences in Uppsala, Sweden, using semen frozen by CLONE, in 0.5-mL straws. Semen was frozen using a standardized, three-step liquid nitrogen vapor freezing method. Whelping rates > 70% were obtained when post-thaw motility was 40% or higher. The inseminations were made either directly into the uterus using transcervical catheterization with the Norwegian catheter (NIU; 167 AIs) or a fiberoptic endoscope (EIU; 19 AIs), or in the cranial vagina (VAG; 141 AIs). Resulting whelping rates were 84.4% (NIU), 58.9% (VAG; P < 0.001), and 57.9% (EIU). Increasing the number of VAG AIs per cycle from 1 to 2 enhanced the whelping rate (P < 0.05). The mean interval from the first AI to whelping was 61.8 +/- 2.4 d, and was longer for VAG AIs (62.7 +/- 2.7 d) than for NIU AIs (61.2 +/- 2.1 d; P < 0.001). The mean interval from the last AI was 60.1 +/- 1.9 d, and did not differ between VAG AIs (60.2 +/- 2.2 d) and NIU AIs (60.0 +/- 1.6 d). Gestation length was not influenced by breed or litter size. A total of 1158 pups resulted from the 327 AIs. Litter size was 5.4 +/- 3.0 (NIU), 4.0 +/- 2.7 (VAG; P < 0.001), and 6.0 +/- 2.1 (EIU). Litter size was also influenced by breed (P = 0.006) and, for VAG AIs, by the number of inseminations performed per cycle (P = 0.009). This study is the largest that has been carried out on frozen-thawed dog semen AI. It shows that using a good method for cryopreservation, together with nonsurgical intrauterine AI employing the Norwegian catheter, can yield whelping rates and litter sizes similar to those reported from well-controlled natural matings. Furthermore, this is the first study to show that intrauterine deposition of frozen-thawed dog semen results in a significantly higher whelping rate and larger litter size than vaginal deposition.

Animals↗