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).     

Unilateral intrauterine horn insemination of fresh semen in cats

The sperm count required were investigated to obtain a conception rate of 80% by unilateral intrauterine insemination (UIUI) of fresh semen in cats. The conception rates obtained by insemination before and after ovulation were also examined. Thirty-six female cats aged 1-7 years were used in the experiments, and the number of experimental cases was 44. Seven male cats aged 2-12 years from which semen could be collected by the artificial vagina method were used. In artificial insemination, 100 iu x 2 or 250 iu of hCG was administered on days 2-4 of estrus, and sperm were introduced into the uterine horn with a greater number of ovulations (or mature follicles) 15, 20 and 30 hr after hCG administration by laparotomy. The inseminated sperm counts were 2 x 10(6) (Exp. 1). 4 x 10(6) (Exp. 2), and 8 x 10(6) (Exp. 3). As a result, ovulation was induced in 42 of 44 cases (induction rate: 95.5%) regardless of the dosage of hCG. Conception was obtained by UIUI in two of 16 animals (conception rate: 12.5%) in the Exp. 1, five of 16 animals (31.3%) in Exp. 2, and eight of 10 animals (80.0%) in Exp. 3. Regarding the relationship between the ovulation state at insemination and conception, the conception rate obtained by insemination before ovulation was clearly higher than that obtained by insemination after ovulation (p<0.05). Regarding the number of kits compared to the number of ovulations on the inseminated side, the percentages of cases in which the number of kits exceeded the number of ovulations on the inseminated side were similar in all groups inseminated with a different number of sperm. It is therefore necessary to investigate conception rates obtained by bilateral insemination to increase the fertility rate. Based on the above findings, it was shown that the sperm count required for fertilization by UIUI is 8 x 10(6).     

Bitch fertility after natural mating and after artificial insemination with fresh or frozen semen

The birth rate and fecundity of 92 bitches of 32 different breeds mated naturally or following artificial insemination of fresh or frozen semen were compared. The birth rate after natural service was 92 per cent compared with 84 per cent when fresh semen was deposited in the uterine body. When frozen semen was inseminated intra-uterine a birth rate of 67 per cent was obtained whilst a figure of 25 per cent was obtained following intra-vaginal insemination of fresh semen. There were no differences in litter size.     

Laparoscopic intra-uterine insemination of fallow deer with frozen-thawed or fresh semen after synchronisation with CIDR devices

This study investigated the efficacy of fixed-time laparoscopic intra-uterine insemination of farmed fallow deer (Dama dama) with frozen-thawed or fresh semen. In the trials with frozen-thawed semen, a total of 547 mature non-lactating does across five New Zealand farms were used. For oestrous synchronisation and artificial insemination, a standard control regimen was applied to at least 30% of the does on each farm, involving the insertion of single CIDR type-G devices intravaginally for 14 days, deposition of 50 x 10(6) frozen-thawed spermatozoa at 65 hours after withdrawal of the CIDR device and the continuous presence of vasectomised bucks from the insertion of the CIDR device until 10 days after insemination. Various aspects of this protocol were changed for the remaining does on each farm, including inseminations at 60 or 70 hours, the absence of vasectomised bucks, insemination with 25 x 10(6) or 10 x 10(6) spermatozoa, synchronisation with CIDR type-S devices and synchronisation with prostaglandin. The conception rate, based on rectal ultrasonography at 45 days after insemination, was 67% across all treatments (n=547). Corrected conception rates (+/-s.e.), calculated following between-farm adjustments, were 67+/- 3% for the control regimen, 67+/- 9% and 73 +/- 8% for inseminations at 60 and 70 hours respectively, 61 +/- 9% for absence of bucks, 80 +/- 8% and 74 +/- 9% for inseminations with 25 x 10(6) and 10 x 10(6) spermatozoa respectively, 62 +/- 10% for CIDR type-S device synchronisation, and 49 +/- 10% for prostaglandin synchronisation. Despite apparent differences, none of the treatments resulted in adjusted conception rates that were significantly different from the control regimen (P>0.01). In the trials with fresh semen, 216 does in the USA were inseminated at 69-71 hours after withdrawal of the CIDR device using either cryopreserved semen from New Zealand (n=158; 25 x 10(6) spermatozoa per inseminate) or fresh semen (n=58; 7.5 x10(6) to 20 x 10(6) spermatozoa per inseminate) collected less than 10 hours earlier. The overall conception rates were 77% and 81% respectively, with no significant differences between semen type (frozen v. fresh) or fresh spermatozoa number per inseminate (P>0.01). A further 102 does in New Zealand similarly received fresh semen from 3/4 Mesopotamian buck. Doses of 10 x 10(6) (n=35), 5 x 10(6) (n=32) or 2.5 x 10(6) (n=35) spermatozoa per inseminate were delivered at 69-71 hours after withdrawal of the CIDR device. The conception rates were 77%, 66% and 51% respectively, reflecting a dose effect (P<0.05). However, 1/4 Mesopotamian does in the group (n=19) exhibited higher conception rates (95% overall) irrespective of semen dose, possibly indicating a semen/recipient genotype interaction. It is concluded that laparoscopic intra-uterine insemination of fallow deer with frozen-thawed or fresh semen at fixed intervals after removal of a CIDR device can give acceptable conception rates under a range of on-farm management options and semen doses.     

Effect of green buffer storage on the fertility of fresh camel semen after artificial insemination

Artificial insemination (AI) is one of the most widely used reproductive technologies, and there is considerably interest in commercializing this technology in camels. Storage of semen extender frozen (at -20 °C) is of considerable interest to scientists working with camels, as transportation of diluents at refrigeration temperature is not always possible given the hot, arid and remote conditions that dromedary camels exist in. Therefore, this study was conducted to compare the fertility of fresh camel semen, after dilution in fresh or frozen-thawed green buffer (GB), after AI into single and multiple ovulating female camels. No differences were observed in any sperm characteristics (motility, membrane integrity, acrosome integrity or morphology) when semen was diluted in fresh or frozen-thawed GB (p>0.05). Sperm motility was increased by dilution (fresh: 70.7 ± 4.9% and frozen: 68.8 ± 3.1%) compared with the motility of sperm in neat semen (35 ± 2.85%; p<0.05), and sperm motility changed from oscillatory to forward progressive after dilution. Pregnancy rates were higher (p<0.05) for single ovulating camels inseminated with semen diluted in fresh (72.7%) compared with frozen-thawed GB (27.3%), and fertilization rates were also higher (p<0.05) for multiple ovulating camels inseminated with semen diluted in fresh (83.3%) compared with frozen-thawed GB (11.1%). These results clearly demonstrate the detrimental effect of freezing and thawing semen diluent on the fertility of fresh camel semen. However, further studies are required to elucidate the mechanism responsible for this reduction in fertility. Moreover, these results demonstrate that the fertility of fresh camel semen diluted in fresh GB is high enough to be considered commercially viable.     

Intrauterine and deep cervical insemination with frozen semen in sheep

Inhalt (Intrauterine und tiefcervicale Insemination mit Gefriersperma beim Schaf). In diesem Versuch wurden 241 Schafe unter Feldbedingungen mit tiefgefrorenem Widdersperma besamt. Nach der Samenentnahme mit Hilfe einer künstlichen Scheide wurde das Sperma ca. 1:6 verdünnt und zwar mit einem Tris-Fruktose-Citronensäure Verdünner, der 8 vol. % Glyzerin and 20 vol. % Eidotter enthielt. Während der ungefähr 2stündigen Equilibrierungszeit wurde der Samen auf 5°C heruntergekühlt, anschlieβend wurde der Samen in P. V. C. Röhrchen abgefüllt, in Stickstoffdampf eingefroren und während ca. 3 Wochen in flüssigen N₂ gelagert. Die Inseminationsdosis betrug 0,3 ml und enthielt etwa 150 × 10⁶ Spermien. Aufgetaut wurde in Wasser von + 75°C während 8,5 Sekunden unmittelbar vor der Sameneinführung. In einer Gruppe von 77 Tieren, die mit Gestagen imprägnierten Vaginaltampons synchronisiert worden waren, wurde nur einmal besamt und zwar 12–20 Stunden nach Brunstbeginn. Bei 53 Schafen dieser Gruppe wurde der Samen mittels eines durch den Cervicalkanal eingeführten Metallkatheters intrauterin deponiert. Bei den verbleibenden 24 Tieren wurde der Samen 2,5–4 cm tief in den Cervicalkanal hinein verbracht. Von den intrauterin besamten Tieren konzipierten 54%, von denjenigen mit intracervicaler Deponierung nur 29%. Von 164 nicht synchronisierten Schafen wurden 83 intrauterin und 60 tiefcervical, auch einmal und im gleichen Zeitpunkt der Brunst wie die synchronisierten Tiere, besamt. Es konzipierten 89% bzw. 45%. Die restlichen 21 Tiere wurden wie üblich mittels der Röhrchenmethode im orificium externum, normalerweise zwei mal mit etwa 12stündigen Intervallen, besamt. Von diesen konzipierten 57%. Contents Artificial insemination with frozen ram semen was performed in a field trial including 241 ewes. The semen was diluted about 1:6 with Tris-fructose-citric acid extender containing 8% (v/v) glycerol 20% (v/v) eggyolk, frozen in P.V.C. straws by use of liquid N₂ after an equilibration time of 2 hrs at 5°C and thawed at 75°C for 8,5 sec. The insemination doses contained approximatly 150 × 10⁶ spermatozoa. In one group the semen was deposited via the cervical canal into the uterus in 53 ewes and deep cervically in the remaining 24 ewes once, in the first oestrus after heat synchronization with progestagen impregnated vaginal sponges. The conception rate was 54% and 29% respectively. In another group consisting of 164 normal oestrous ewes, 83 animals were inseminated according to the intrauterine procedure of semen deposition and 60 animals deep cervically once in the heat, resulting in a conception rate of 89% and 45% respectively. In the remaining 21 ewes the semen was deposited just inside the cervical os usually twice in the heat, giving a conception rate of 57%.     

水牛性控冷冻精液授精效果的研究

为优化推广水牛性控冷冻精液人工授精技术,本研究分析了不同类型、胎次、年龄的受体母牛及不输精员对水牛性控冷冻精液人工授精产犊率的影响。收集武鸣县8个村级牛人工授精点3 029头受体母水牛X性控冷冻精液人工授精产犊数据,利用卡方检验方法对所收集数据进行统计分析。结果表明:本研究3 029头受体母牛经人工授精后产犊率为44.2%(1340/3029),产母犊率为82.3%(1103/1340)。以本地水牛及杂交水牛作为受体母牛,杂交水牛人工授精产犊率极显著高于本地沼泽型水牛(P0.01)。不同年龄受体母牛人工授精产犊率差异不显著(P0.05),但存在组间差异。不同胎次受体牛人工授精产犊率差异不显著(P0.05),但存在组间差异。不同输精员人工授精产犊率差异极显著(P0.01)。本研究结果表明不同类型、年龄、胎次的受体母牛及输精员皆会影响到水牛性控冷冻精液人工授精效果。进一步加强受体母牛选择、加强输精员培训是今后水牛性控冷冻精液人工授精大范围推广应用不可忽视的环节。     

Effects of coitus and the artificial insemination of different volumes of fresh semen on uterine contractions in mares

Uterine contractions may play an important role in the transportation of spermatozoa towards the site of fertilisation in the oviduct of mares. M-mode ultrasound was used to measure the number, amplitude and duration of uterine contractions in each uterine horn and the uterine body of oestrous mares for four minutes before and four minutes after either coitus, or the artificial insemination of either 80.0 ml of fresh semen or 10.0 ml of fresh semen. The direction of the uterine contractions in each uterine horn and the uterine body was measured before and after coitus. Coitus and the insemination of 80.0 ml semen significantly increased the total number, mean amplitude and mean duration of contractions in all parts of the uterus. The insemination of 10.0 ml of semen did not affect the total number or the mean duration of contractions in the uterine horns. Their mean amplitude was increased, but largely owing to the results from one mare; it also did not affect the contractions in the uterine body. There was no significant difference between the percentage of contractions moving in a cervicotubal or tubocervical direction after coitus in any part of the uterus examined.     

Intrauterine insemination in ewes with frozen semen (author's transl)]

110 ewes were allocated for intrauterine insemination. One succeeded to deposite semen intra uterine in 43 of the ewes and 37 (86%) of these lambed. In the remaining 67 the semen was deposited in cervix, and of these 15 lambed (22,4%). Totally 52 (47,3%) of the inseminated ewes lambed. Similar results were obtained by one or two inseminations in the heat. Dilution 1:3 gave a significantly (p less than 0.05) higher lambing rate than dilution 1:9 when the semen was deposited in the cervix, while the results were similar for dilution 1:3 and 1:9 by intra interine deposition. A group of 49 ewes was inseminated according to the conventional method on the first day of heat and the intra uterine method on the second day. Intra uterine deposition of semen succeeded in 31 ewes and 27 (87,1%) lambed. In the 18 remaining ewes the semen was depposited in the cervix and 9 (50%) lambed. Totally 73,5% of the ewes in this group lambed.     

Artificial insemination with canine semen stored at a low temperature

Cooled canine semen solutions for storage were investigated with three stock solutions: egg yolk-citrate-glycine-glucose solution, egg yolk Tris-fructose citrate solution (EYT-FC), and egg yolk sodium citrate dihydrate solution (EYCD). For the control group, the second fraction of semen was examined. Nine male beagles and 37 female (47 experimental cases) beagles for artificial insemination (AI) were used. The qualities of semen stored at 4 degrees C deteriorated earlier in the control and EYCD groups. In the other two groups, sperm motility was 60% or higher after storage for 6 days and 20% or higher after storage for 12 days. On a comparison of these two groups, the sperm motility and viability were slightly higher in the EYT-FC group. A high conception rate was obtained by AI using semen stored at a low temperature for a maximum of two days in the control group and four days in the EYT-FC group.     

Artificial insemination of gilts with bovine viral diarrhea virus-contaminated semen

Bovine viral diarrhea virus (BVDV) is a pestivirus that infects swine and other species and has genetic and antigenic similarity to classical swine fever virus. The objective of this study was to mimic the infection of swine by contaminated semen and evaluate the effects on their reproductive tracts and litters. Six gilts were artificially inseminated with semen containing BVDV-2 ncp (LVB 16557/15) and 2 were inseminated with BVDV-free semen. Blood samples from all gilts were collected for polymerase chain reaction and virus neutralization tests. No viremia or neutralizing antibodies were detected, and all the litters were born healthy.     

Retention of liquid within insemination equipment using various equine frozen semen insemination methods and two semen freezing extenders

The objective of this study was to examine the effect of different insemination techniques and extenders on the volume of liquid dispensed from insemination equipment. The method of insemination has a significant effect on the volume of semen deposited into the mare's uterus when low volumes are used. Insemination pipettes that allow for direct deposit of straw contents into the uterus are preferred. Aspiration of semen into a pipette is preferred over aspiration into a syringe with deposition through a pipette when direct deposit is not possible. Use of a pipette with a smaller lumen and less length of contact with liquid provides better results. Contact of semen with equipment may allow for residual liquid accumulation on the luminal surfaces and a decrease in overall semen dose. Extenders with differing amounts of egg yolk did not influence volume of liquid dispensed.     

Detection and isolation of Toxoplasma gondii from fresh semen of naturally infected dogs in Southern Brazil

The aim of this study was to isolate Toxoplasma gondii and determine the viability of the parasite in fresh semen samples of clinically healthy adult dogs naturally infected. Eleven seropositive dogs with T. gondii IgG antibodies from southern Brazil were selected to confirm the presence and viability of T. gondii in fresh semen samples using in vitro isolation in Vero cell culture, polymerase chain reaction (PCR) and sequencing analysis. The presence of viable T. gondii was confirmed by in vitro isolation and PCR in five semen samples. The ITS1 region of the isolated protozoa (TG S4) was amplified and sequenced. The nucleotide sequence obtained was 99% compatible with the T. gondii DNA sequences stored in the GenBank. It has been shown that T. gondii tachyzoites may be isolated in vitro from fresh semen samples of clinically healthy dogs seropositive for T. gondii.     

Artificial insemination and preservation of semen.

Artificial insemination is an effective technique for improving utilization of stallions in breeding programs. When proper semen handling and insemination procedures are used, optimal pregnancy rates are attainable. When AI techniques are employed for mares and stallions with marginal fertility, pregnancy rates may be improved in comparison with natural mating. Preservation of stallion semen in the liquid or frozen state reduces the costs and potential health hazards incurred by transporting mares and provides easier access to genetic material that may otherwise be unavailable. Acceptable pregnancy rates are consistently obtained with cooled semen. Conversely, techniques for cryopreservation of stallion semen will require more refinement before the procedure can be considered commercially viable on a wide scale.