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.     

Intravaginal insemination of bitches with fresh and frozen-thawed semen with addition of prostatic fluid: use of an infusion pipette and the Osiris catheter

One hundred fifty-two bitches of seven breeds were vaginally inseminated with fresh or frozen-thawed semen of 10 stud dogs of respective breeds. The semen was supplemented with prostatic fluid before insemination. In experiment 1 bitches of each breed were randomly assigned to three treatment groups, consisting of 29 females (group 1), 33 females (group 2) and 32 females (group 3). In group 1 bitches were inseminated into vagina with fresh semen using a bovine infusion pipette. In group 2 bitches were inseminated into vagina with fresh semen using the Osiris catheter. In group 3 bitches were inseminated with frozen-thawed semen with the Osiris catheter. The number of sperms in each insemination dose was adjusted to 300 x 10(6). In experiment two bitches were randomly assigned to two treatment groups, consisting of 30 females (group A) and 28 females (group B). In group A bitches were inseminated with fresh semen, whereas in group B with frozen-thawed semen. Osiris catheter was used in both groups. The total number of sperms was adjusted to provide 250 x 10(6) of progressively motile spermatozoa in each insemination dose. In experiment 1 the pregnancy rates/whelping rates were 86.2/82.8%, 81.8/81.8% and 59.4/59.4% for groups 1, 2 and 3, respectively. The differences between group 1 and 3 were statistically significant (p < 0.05). The litter sizes at birth/litter sizes at weaning were 5.8+/-2.3/5.4+/-2.0, 6.3+/-1.4/5.7+/-1.0 and 3.9+/-1.2/3.5+/-1.5 in groups 1, 2 and 3, respectively. The litter size at birth and at weaning was reduced (p < 0.05) when frozen-thawed semen was used for insemination (group 3). There were not significant (p > 0.05) differences in the litter size between groups 1 and 2. In experiment 2 pregnancy rates/whelping rates and litter sizes at birth/litter sizes at weaning were 86.7/86.7%, 60.7/57.1% (p < 0.05) and 6.1+/-1.6/5.7+/-1.7, 4.0+/-1.4/3.8+/-1.4 (p < 0.05) in groups A and B, respectively. This study shows that results of AI with a fresh semen using a bovine infusion pipette and the Osiris catheter are equivalent. The results of the use of the Osiris catheter for vaginal insemination of frozen-thawed dog semen extended with prostatic fluid after thawing are not encouraging. The pregnancy rate, whelping rate and litter size are reduced when frozen-thawed, prostatic fluid-supplemented semen is vaginally deposited using the Osiris catheter.     

The effect of sperm number on fertility in blue fox vixens (Alopex lagopus ) artificially inseminated with frozen silver fox (Vulpes vulpes ) semen

During the breeding seasons of 1989 and 1990, a total of 617 blue fox vixens aged 1 to 6 years (mean +/- SEM, 2.6 +/- 0.1) were inseminated with frozen silver fox semen with either 150 million (n = 213, 1989 + 1990), 100 million (n=172, 1990), 75 million (n = 119, 1989) or 37.5 million (n = 113, 1989) spermatozoa per insemination. Two intrauterine inseminations, each with an insemination volume of 1.0 ml, were performed at 24-hour intervals on the first and second days after maximum vaginal electrical resistance was measured. Conception rates were 87% (186 of 213) with 150 million spermatozoa per insemination, 85% (146 of 172) with 100 million, 77% (91 of 119) with 75 million and 68% (77 of 113) with 37.5 million. The mean numbers of cubs per litter +/- SEM for the four groups were 7.6 +/- 0.2 (168 registered litters), 7.5 +/- 0.3 (115 litters), 6.4 +/- 0.4 (86 litters) and 6.4 +/- 0.4 (75 litters). A negative effect on both the conception rate and mean litter size at whelping was observed with decreasing sperm numbers (conception rate percentage: p = 0.0001, Chi-square, litter size: p = 0.02, Kruskal-Wallis Test). Only the two larger numbers of spermatozoa gave litter sizes comparable to those obtained by artificial insemination (AI) with fresh semen.     

Single, transcervical insemination using frozen-thawed semen in the Greyhound: a case series study

Transcervical insemination (TCI) has generated recent interest as an assisted reproductive technique in the dog. A case series study was performed to determine if TCI using frozen-thawed semen was a viable technique to offer in a general veterinary practice setting. Over a period exceeding 28 months, 137 Greyhound bitches were presented for assisted breeding. A single, timed insemination using a rigid cystoscope to aid in transcervical deposition of a frozen-thawed semen dose was given within 72 h after the behaviorally estrual bitch had a > 4 ng/mL serum progesterone concentration and estrus-categorized vaginal cytology. Litter size, pregnancy and whelping rate were collected; their association to semen center and stud dog were quantified. Of the 137 bitches, 117 were bred for one cycle and 20 were bred for two or more cycles, giving a total of 161 single, timed inseminations. Pregnancy rate was 89.4%, with 141 (87.5%) whelping. Litter size was 6.9+/-2.7 (mean+/-S.D.) pups. Semen center (P=0.84) and stud (P=0.79) had no effect on pregnancy. These results were quite favorable when compared to prior TCI studies, and are possibly due to the use of a single breed (i.e., Greyhound) with good fertility. This study supported the application of TCI, in Greyhounds, as a successful and viable service to offer in private practice. Additionally, these results have value in their use for benchmarking future breed-specific and TCI research. Serendipitously, the apparent fecundity results obtained in this observational study suggests a possible greater appreciation be given to breed composition and choice in assisted reproductive technique studies.     

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).     

Laparoscopic intrauterine insemination in the bitch

A technique for laparoscopic intrauterine insemination in bitches is described. During natural estrus, 5 beagle bitches were inseminated and S others were naturally mated (control group) twice at a 48-h interval on Days 3 and S (n = 4) or Days 4 and 6 (n = 6) after the increase in plasma progesterone considered to be indicative of the day of the preovulatory LH peak. All the inseminations were with fresh semen and under general anesthesia. The technique involved the introductions of 1) a Verres needle to insufflate the abdominal cavity by direct punction on the middle line 1 cm over the umbilicus, 2) a laparoscope to visualize the abdominal cavity by a 1 cm puncture on the middle line 1 cm under the umbilicus, 3) a forceps used to manipulate the uterus by a 0.5 cm puncture at 2 to 3 cm lateral to the mammary glands, and 4) an 18-g catheter used to puncture the uterus on the middle line between the 3rd and 5th mammary gland. The uterine body was grasped by the forceps and elevated against the ventral abdominal wall. The 18-g catheter was then inserted through the abdominal wall directly into the uterine lumen, and 1.0 ml of fresh semen containing 250 to 480 x 10(6) spermatozoa/ml was injected. The inseminations resulted in pregnancies in all animals. Litter size was similar in the artificially inseminated and naturally mated bitches (5 +/- 1.8 and 4.8 +/- 1.6 pups per litter, respectively). Bitches in the artificially inseminated group delivered at 65.2 +/- 0.8 d and in the natural mated group at 65.4 +/- 0.5 d after the LH peak. In conclusion, this paper gives the first results of intrauterine laparoscopic insemination in bitches, indicating interesting perspectives for this technique in dog's reproduction.     

Fertility in the ewe following cervical insemination with fresh or frozen-thawed semen at a natural or synchronised oestrus

Artificial insemination (AI) in sheep is currently limited by the poor fertility obtained following non-surgical intracervical insemination of frozen-thawed semen. An exception to this general finding is the non-return rate of around 58% reported for large scale on-farm AI in Norway. The objective of the present study was to determine if similar results could be obtained under Irish conditions. Comparisons were made between semen collected, and frozen, from rams in Norway (NOR) and Ireland (IRL). The effects of synchronisation and inseminator were also examined. Parous ewes (n=297) of various breed types were inseminated to a natural (N) or synchronised (S) oestrus with either fresh (from Irish rams) or frozen-thawed (IRL and NOR) semen. Ewes were randomly assigned, within breed, to the following treatment groups: (i) Fresh-N: n=28, (ii) Fresh-S: n=30, (iii) IRL-N: n=62, (iv) IRL-S: n=50, (v) NOR-N: n=68, (vi) NOR-S: n=59. Within each group, ewes were inseminated by an experienced Norwegian or by an Irish inseminator. Pregnancy rate did not differ significantly between ewes inseminated to a natural or synchronised oestrus nor between Norwegian and Irish frozen semen. The proportion of ewes pregnant after insemination with fresh semen was 0.82 and 0.70 (treatments i and ii) compared with 0.40, 0.52, 0.34 and 0.37 (treatments (iii)-(vi)) for frozen semen (P<0.001). Corresponding litter sizes (+/-S.E.), adjusted for ovulation rate, were 2.9+/-0.22, 3.3+/-0.23, 2.2+/-0.21, 1.7+/-0.21, 2.2+/-0.21 and 2.1+/-0.21 (fresh versus frozen; P<0.001). There was an interaction between semen type (fresh or frozen) and oestrus type (N or S) for litter size due to an increased adverse effect of frozen semen on litter size in synchronised ewes (P<0.05). Pregnancy rate was significantly influenced by breed of ewe (P<0.01) and inseminator (P<0.05). These results suggest that ewe breed may be a critical determinant of the potential for the exploitation of cervical insemination of frozen-thawed semen in sheep breeding programmes.     

The optimum time for single artificial insemination of blue fox vixens (Alopex lagopus) with frozen-thawed semen from silver foxes (Vulpes vulpes)

During the breeding season of 1991 a total of 608 blue fox vixens aged 1 to 6 years (2.3 +/- 0.1 years, mean +/- SEM) from 2 farms were artificially inseminated intrauterine once with frozen-thawed silver fox semen (1 ml dose containing a total of 150 million spermatozoa). The vixens were allocated to 3 different groups according to the time of insemination. Vixens in Group 1 (n = 203), Group 2 (n = 198), and Group 3 (n = 207) were inseminated on the first, second or third day after the peak value of vaginal electrical resistance, respectively. An overall conception rate of 75% (456 of 608) and 6.0 +/- 0.1 (mean +/- SEM) cubs per litter was obtained. Conception rates and mean litter sizes were significantly different between groups of vixens with respect to day of insemination (P = 0.02, Chisquare, Kruskall-Wallis Test). Vixens inseminated on the second day (Group 2) had the highest conception rate (81%) and the largest mean litter size (7.0 +/- 0.2 cubs) of the three groups, while those inseminated on the third day (Group 3) had the lowest conception rate and mean litter size (70%, 5.4 +/- 0.3 cubs).     

Lambing rates and litter sizes following intrauterine or cervical insemination of frozen/thawed semen with or without oxytocin administration

Intrauterine insemination by laparoscopy is required to achieve acceptable lambing rates in ewes when using frozen semen but the procedure has evoked welfare concerns. Oxytocin has been used to dilate the cervix as a means of accessing the uterus during conventional cervical insemination, but its effect on fertility is not well documented. Three hundred crossbred ewes were synchronised in estrus and randomly allocated to one of three insemination procedures using frozen/thawed semen containing 400 x 10(6)/ml progressively motile sperm: single cervical (0.2 ml), multiple cervical (4 x 0.05 ml) or laparoscopic (0.05 ml per uterine horn). The effects of each insemination procedure on lambing rate (percentage of treated ewes lambing) and litter size (lambs per ewe lambing) were tested with and without oxytocin (10 IU given i.m.) prior to fixed-time insemination. Oxytocin did not permit complete cervical penetration in any ewes and neither lambing rate nor litter size was influenced by the number of inseminations. Lambing percentages were 69 and 42 (P < 0.01) for the laparoscopic and cervical insemination methods, respectively, and oxytocin reduced these to 58 (NS) and 10 (P < 0.001) percent, respectively. Corresponding litter sizes for ewes not receiving oxytocin were 1.91 and 1.51 and for those receiving oxytocin, 1.83 and 1.41 (laparoscopic versus cervical, P < 0.02). Thus, in the absence of complete cervical penetration at insemination, 10 IU oxytocin decreased the number of ewes lambing but had no effect on their litter size.     

Assessing reproductive patterns and disorders in free-ranging dogs in Jodhpur, India to optimize a population control program

The objectives were to test the hypothesis that estrus and pregnancy are seasonal in free-ranging female dogs (>3 mo old) in Jodhpur, India, and to determine litter size, and the prevalence of fetal resorption in this population. The prevalence of estrus and pregnancy was determined in 5400 free-ranging bitches (trapped and released) at the time of ovariohysterectomy. In a separate study, the uteri and ovaries of 246 free-ranging bitches were examined to determine litter size and fetal resorption. The bitches exhibited seasonal estrus and pregnancy (P < 0.00001), with a higher percentage of bitches in estrus or pregnant during the late monsoon season (September to November) compared to the other three seasons. The mean litter size based on embryo/fetal counts was 4.6 (95% CI = 4.0-5.3; n = 40) and based upon placental site counts was 4.4 (95% CI = 3.9-4.8; n = 105). Prevalence of fetal resorption was 32.6% (95% CI = 20.5-47.5; n = 43) with a mean of 2.8 resorptions per litter in those with at least one resorption (95% CI = 1.8-3.8; n = 14). This was the first study to estimate previous litter size of non-pregnant, free-ranging dogs based upon placental sites. Litter size data from this study will be used in a population demographic model to predict the long-term impact of animal birth control (ABC) on the free-ranging dog population in Jodhpur. Increasing the efforts to surgically sterilize bitches prior to the time of year of peak pregnancy or whelping will help maximize the impact of an ABC program on the Jodhpur free-ranging dog population.     

Comparison of endoscopic-assisted transcervical and laparotomy insemination with frozen-thawed dog semen: A retrospective clinical study

The objective of this retrospective clinical study was to compare pregnancy rates obtained after the use of endoscopic-assisted transcervical catheterization (EIU) or laparotomy (SIU) for insemination of frozen-thawed dog semen. Healthy bitches from various breeds were inseminated with semen from multiple donors processed by different freezing centers. Data from 118 inseminations (78 EIU and 40 SIU) performed between 2009 and 2011 were analyzed. Insemination timing was based on vaginal cytology, serum progesterone concentrations, and vaginoscopy. A ureterorenoscope and a CH-5 Transcervical insemination catheter were used for EIU; 28 of the bitches in this group were inseminated twice with the second insemination less than 12 hours after the first. The numbers of live morphologically normal sperm (LMNS) were determined to characterize insemination doses. Overall, pregnancy rate was greater (P < 0.05) in the EIU group (65%) than in the SIU group (45%). Pregnancy rates were greater (P ≤ 0.06) when more than 100 × 10⁶ LMNS were inseminated regardless of insemination method; the greatest pregnancy rate was observed in the EIU group when this insemination dose was used (38/49; 78%). There was no significant difference in pregnancy rate whether one (69%) or two inseminations (64%) were performed in the EIU group. Complications in the SIU group included anesthetic-induced bradycardia during surgery, significant postsurgery pain, seroma formation over the abdominal incision, and delayed wound healing. No complications were noted during or after insemination in the EIU group. In conclusion, these results support the use of EIU as a noninvasive alternative to laparotomy for insemination of frozen-thawed dog semen. In addition, use of more than 100 × 10⁶ LMNS is also recommended for insemination.     

Litter size and piglet traits of gilts with different prolactin receptor genotypes

Seventy-seven Large White x Meishan F2 crossbred gilts with prolactin receptor (PRLR) genotype AA (n = 26), AB (n = 36) and BB (n = 15) were compared for teat number (FTm), age at first estrus, gestation length (GL), litter size, and litter means of functional teat number (FTp), birthweight (BW), and pre-weaning growth rate (GR). Own placental information was available for 88% of 620 live-born piglets (62 gilts), since placentae were labeled during farrowing. The effect of PRLR genotype of the mother on average placenta weight (PLW) and placenta efficiency (EFF = BW/PLW), was therefore, also analyzed, PRLR genotype significantly (P < 0.05) affected age at first estrus and, as a result (since the gilts were inseminated at a fixed estrus number), age and bodyweight at insemination. Furthermore, PRLR genotype affected total number of piglets born (TNB, P = 0.056) and number of piglets born alive (NBA, P = 0.072), but it did not affect (P > 0.3) GL, BW or GR, neither before nor after correction for litter size. BB gilts were significantly younger at first estrus and younger and lighter at insemination than AA gilts (P < 0.05). AA gilts had larger TNB (P = 0.047) and tended to have a larger NBA (P = 0.062) than BB gilts. TNB was 11.4 +/- 0.7, 10.8 +/- 0.6, and 8.8 +/- 0.9; NBA was 11.1 +/- 0.6, 10.5 +/- 0.6, and 8.7 +/- 0.9; BW was 1309 +/- 40, 1277 +/- 34, and 1290 +/- 53 g; and GL was 113.6 +/- 0.3, 113.8 +/- 0.3, and 113.5 +/- 0.4 days for AA, AB and BB gilts, respectively. The effects on litter size and age at first estrus are independent effects. PRLR affected PLW (P = 0.050) and EFF (P = 0.066), resulting in a difference between AA and BB gilts. PLW was 160 +/- 9, 181 +/- 7 and 196 +/- 11 g and EFF was 7.6 +/- 0.2, 7.3 +/- 0.2 and 6.7 +/- 0.3 for AA (n = 19), AB (n = 29) and BB (n = 14) gilts, respectively. After correction for TNB, the differences disappeared. Functional teat number of the AA. AB and BB gilts was 15.35 +/- 0.22, 15.53 +/- 0.18, and 15.60 +/- 0.29, respectively, and was not affected by PRLR genotype (P = 0.7). Functional teat number of piglets from AA, AB and BB mothers was 14.20 +/- 0.10, 14.37 +/- 0.08, and 14.63 +/- 0.13, respectively. Piglets from BB mothers had on average larger numbers of functional teats compared to piglets from AA mothers (P = 0.028). In conclusion, PRLR gene is a major gene or marker for age at first estrus, litter size, and litter average of number of functional teats in the Large White x Meishan F2 crossbred gilts studied. The favorable allele for litter size (A allele) is the unfavorable allele for age at first estrus and for litter mean of functional teat number.     

Pregnancy and conception rate after two intravaginal inseminations with dog semen frozen either with 5% glycerol or 5% ethylene glycol

The primary goal of this study was to compare the effects of 5% ethylene glycol (EG) and 5% glycerol (G) on fertility of frozen–thawed dog semen following intravaginal insemination. The sperm-rich fraction of the ejaculate of three male dogs was collected, pooled and divided into two aliquots, and then frozen with a Tris–glucose–egg yolk–citric acid extender containing either 5% G or 5% EG. A total of 10 bitches were inseminated twice, five with G-frozen–thawed semen and five with EG-frozen–thawed semen; intravaginal inseminations were performed the 4th and the 5th day after the estimated LH peak; four straws, thawed in a 37 °C water bath for 1 min and diluted in a Tris buffer, were used for insemination (200 × 10⁶ spermatozoa); the insemination dose was introduced in the cranial vagina of the bitch using a sterile plastic catheter. Ovariohysterectomy was performed in all bitches between days 29 and 31 after the calculated LH surge, and pregnancy status, and the number of conceptuses and corpora lutea were recorded. All bitches were pregnant. Neither the number of conceptuses, nor the ratio of conceptuses to corpora lutea (conception rate) was significantly different between groups. In this first screening, with a limited number of bitches, EG-frozen semen did not show a higher fertility than G-frozen semen when used for two intravaginal inseminations. Irrespective of the semen used, conception rate was 0.50.     

Field fertility with exported boar semen frozen in the new flatpack container

The present study tested the field fertility of frozen-thawed (FT) Swedish boar semen packaged in flat plastic containers (FlatPacks) and exported for artificial insemination (AI) to overseas nucleus herds. Semen from 47 Swedish boars of Landrace (L), Yorkshire (Y), and Hampshire (H) breeds was frozen using a lactose-egg yolk-based extender with 3% glycerol and 10(9) spermatozoa/ml in 5 ml FlatPacks. For all breeds, FT sperm membrane intactness averaged 60%, while mean FT sperm motility ranged from 49 to 53%. A total of 308 litters resulted from 421 overseas inseminations with FT semen, with a mean farrowing rate (FR) of 73% and 10.7 mean number total piglets born. In a within-sow analysis for the purebred L and Y breedings, the FR and litter size of FT semen were compared with natural matings (NM) and on-farm AI with liquid semen (NW/AI breedings) at the same farms. Farrowing rate was 72.3 and 78.8% (P = 0.23), total piglets 11.3 and 11.6 (P = 0.44), and live piglets 10.1 and 10.2 (P = 0.77), for the FT semen and NM/AI breedings, respectively. The present results suggest that this freezing protocol and FlatPack container maintains high sperm viability post-thaw. Further the fertility levels when inseminated at overseas nucleus herds seem to be similar to those achieved with (NM/AI breedings) at the same farms. This freezing method may be a reliable alternative for the freezing/thawing of boar semen under commercial AI conditions.     

Fertility in bitches artificially inseminated with extended, chilled semen

Sixteen bitches were artificially inseminated with either fresh, 24 h-chilled or 48 h-chilled extended semen over 38 estrous cycles. A commercial system for extending, chilling and transporting semen commonly used in the equine industry was used Pregnancy rates and litter sizes of the bitches inseminated with extended, chilled semen (19/20, 95%; litter size = 7.1) were not significantly different from those observed in bitches inseminated with fresh semen (17/18, 94%; litter size = 7.2; P > or = 0.89). These results show that a commercial system for extending, chilling and transporting equine semen is an attractive and efficient method of shipping canine extended chilled semen.     

Fertility of weaned sows after deep intrauterine insemination with a reduced number of frozen-thawed spermatozoa

The present study evaluates the effectiveness of the transcervical deep intrauterine insemination (DUI) with a reduced number of frozen-thawed boar spermatozoa in weaned sows. DUI was performed using a specially designed flexible device (length 180 cm, outer diameter 4mm, working channel 1.8mm, working channel's volume 1.5 ml) that was inserted through an artificial insemination spirette to cross the cervix lumen and moved into one uterine horn as far as possible. Spermatozoa diluted in 7.5 ml of BTS were flushed into the uterine horn by a syringe attached to the working channel. In Experiment 1, 111 hormonally treated (eCG/hCG) weaned sows were inseminated once using one of the following three regimens: (1) DUI with frozen-thawed spermatozoa (1000 x 10(6) cells per dose; n=49); (2) DUI with fresh semen (150 x 10(6) cells per dose; n=29, as control of DUI procedure); and (3) cervical insemination with frozen-thawed spermatozoa (6000 x 10(6) cells diluted in 100ml; n=33). No differences (P>0.05) were found for farrowing rates (77.55, 82.76, and 75.76, respectively) or litter sizes (9.31+/-0.41, 9.96+/-0.32, and 9.60+/-0.53 piglets born per litter, respectively) among the groups. In Experiment 2, DUI was performed on the spontaneous estrus in weaned sows (2-6 parity) with 1000 x 10(6) frozen-thawed (40 sows) or 150 x 10(6) fresh spermatozoa (38 sows). The farrowing rate of sows inseminated twice with frozen-thawed spermatozoa (70%) was significantly (P<0.05) lower than with fresh semen (84.21%). No significant difference (P>0.05) was found in litter size between frozen-thawed spermatozoa (9.25+/-0.23 piglets born per litter) and fresh semen (9.88+/-0.21 piglets born per litter). These preliminary results indicate that application of DUI provides acceptable fertility in weaned sows using a relatively low number of frozen-thawed spermatozoa.     

Canine perinatal mortality: a cohort study of 224 breeds

Canine perinatal mortality is known to be relatively high. However, the literature on perinatal mortality in dogs is still sparse and often refers to a single or only a few breeds. The aim of this large-scale observational study was to describe the perinatal mortality in purebred dogs of various breeds at both puppy and litter level. In addition, the influence of breed, breed size, litter size, age of the bitch, litter number and season for whelping on the risk of perinatal mortality at litter level was studied and the mean litter size at eight days and eight wks after birth was calculated. A retrospective cohort study was performed by studying 10,810 litters of 224 breeds registered in the Norwegian Kennel Club in 2006 and 2007. Perinatal mortality was defined as the sum of stillborn puppies and puppies that died during the first wk after birth (early neonatal mortality) and was present in 24.6% of the litters. Eight percent of the puppies died before eight days after birth, with 4.3% as stillbirth and 3.7% as early neonatal mortality. For most breeds the perinatal mortality was low, but for some breeds a higher perinatal mortality was found. The mean litter size at eight days and eight wks after birth was 4.97 (±0.02) and 4.92 (±0.02) puppies, respectively. Of all puppies born, only 1% died during the period from eight days to eight wks after birth. Random effects logistic regression analysis indicated that increasing litter size and age of the bitch were associated with an increased risk of stillbirth, early neonatal mortality and total perinatal mortality at the litter level (P < 0.001). The random breed effect was significant for all outcomes. Litter number also had a significant effect on stillbirth, early neonatal mortality and total perinatal mortality at the litter level, with the highest risk of perinatal mortality found in the first litter (P < 0.001). Further, the risk of early neonatal mortality was doubled in litters with stillborn puppies. No significant effect of whelping season on perinatal mortality at litter level was found. An interaction existed between the age of the bitch and litter number and the risk of stillbirth was three times as high (odds ratio = 3.00) in litters from bitches having their first litter after the age of six y. Breed was a more important determinant of perinatal mortality in litters than breed size. However, more than 90% of the variation in perinatal mortality was found at the individual litter level and efforts to minimize puppy mortality should be targeted at the management of the individual litter rather than at the breed level.     

Factors affecting gestation duration in the bitch

A retrospective analysis was performed to determine the effects of age, breed, parity, and litter size on the duration of gestation in the bitch. Bitches at two locations were monitored from breeding to whelping. A total of 764 litters whelped from 308 bitches (36 large hounds, 34 Golden Retrievers, 23 German Shepherd Dogs (GSD), and 215 Labrador Retrievers). By breed, the number of whelpings was 152, 72, 58, and 482 for the hounds, Golden Retrievers, German Shepherd Dogs, and Labrador Retrievers, respectively. Whelping was predicted to be 57 d from the first day of cytologic diestrus in the hounds or 65 d from the initial progesterone rise in the other breeds. The average gestation duration (calculated as 8 d prior to Day 1 of cytologic diestrus in hounds or measured from the initial progesterone rise in other breeds) by breed (days +/- S.D.) was 66.0 +/- 2.8, 64.7 +/- 1.5, 63.6 +/- 2.1, and 62.9 +/- 1.3 for the hounds, Golden Retrievers, German Shepherd Dogs, and Labrador Retrievers, respectively. The relationship of age, breed, parity, and litter size with the difference in gestation duration was evaluated using log linear modeling. Age or parity had no effect on gestation duration. Compared to Labrador Retrievers, the German Shepherd Dogs, Golden Retrievers and hounds were more likely to have a longer gestation duration; three, four and nearly eight times as likely, respectively. Bitches whelping four or fewer pups were significantly more likely to have a longer gestation duration than those whelping five or more pups; the prolongation averaging 1 d.     

Favoring the birth of female puppies after artificial insemination using chilled semen diluted with powdered coconut water (ACP-106c)

The objective was to determine the effect of powdered coconut water extender (ACP-106c) on the proportion of female puppies born. Twenty French Bulldog bitches were subjected to natural mating (NM) and, during the subsequent two estrus periods, were bred by intravaginal artificial insemination (AI), using chilled semen (from the same males) diluted in Tris-egg yolk (AI-Tris) or ACP-106c (AI-ACP-106c). Fresh semen was cooled to 5 °C and maintained at that temperature for 6 h, rewarmed (37 °C for 30 s), and used for AI. Pregnancy and whelping rates following NM were both 100% and were both 90.0% following AI with either extender. Litter size (mean ± SD) was 5.4 ±1.1, 4.7 ± 2.0, and 5.1 ± 2.0 (P > 0.05) for NM, AI-Tris, and AI-ACP-106c, respectively. Furthermore, for these groups, the number of female vs. male puppies born were 2.6 ± 0.6 vs. 2.8 ± 1.0, 2.2 ± 1.0 vs. 2.5 ± 1.1, and 3.4 ± 1.6 vs. 1.8 ± 1.2 (P < 0.05 for AI-ACP-106c only). In conclusion, our hypothesis was supported; AI of semen in ACP-106c extender resulted in a significantly higher proportion of female puppies. Furthermore, this extender yielded acceptable litter size and rates of pregnancy and whelping.     

Side of gestation in dairy heifers affects subsequent sperm transport and pregnancy rates after deep insemination into one uterine horn

Effects of side of previous gestation on sperm transport and pregnancy rates after deep cornual insemination were evaluated in 1686 Friesian cows in their first lactational period. Only single ovulating animals were used. At insemination, semen was deposited deep into the uterine horn ipsilateral or contralateral to the preovulatory follicle. A total of 876 cows (52%) ovulated in the ovary ipsilateral to the postgravid horn, and 810 cows ovulated in the contralateral ovary. Semen was deposited into the previously nongravid uterine horn of 832 cows, and into the gravid horn of 854 cows. The pregnancy rate was higher (P < 0.00001) for semen deposition into the previously nongravid horn (46.6%) than for semen deposition into the gravid horn (35.7%). For inseminations ipsilateral to the side of impending ovulation, pregnancy rates were higher (P = 0.0004) when ovulations occurred on the opposite side to the postgravid horn than on the same side. Pregnancy rates were higher (P = 0.002) for contralateral inseminations when ovulations occurred on the same side to the postgravid horn than on the opposite side; they were higher (P = 0.0001) for total ipsilateral than for total contralateral inseminations. There was no difference between ipsilateral and contralateral inseminations (P = 0.64) when ovulation occurred ipsilateral to the postgravid horn, but pregnancy rates were higher (P < 0.00001) when ipsilateral insemination was carried out into the nonpostgravid horn. Results indicate that the side of gestation in dairy heifers affects subsequent pregnancy rates after deep insemination into one uterine horn, possibly by affecting sperm transport.