Fertility after different artificial insemination methods using a synthetic semen extender in sheep

The present study aimed to investigate the fertility of ewes artificially inseminated with three different methods using a synthetic semen extender, AndroMed. The three methods of artificial insemination (AI) were cervical AI with fresh-diluted or frozen-diluted semen at observed estrus, and an intrauterine AI with frozen-thawed semen. A total of 80 ewes were treated with a controlled internal drug release (CIDR) containing 0.3 g progesterone per device for 12 days. In Experiment 1 (26 Suffolk ewes), superovulation was induced with 20 mg follicle-stimulating hormone and 250 IU equine chorionic gonadotropin (eCG) two days and one day before CIDR removal, respectively, during the non-breeding season. In Experiment 2 (54 Suffolk and Suffolk crossbred ewes), an intramuscular injection of 500 IU eCG was administered one day before CIDR removal to synchronize estrus and ovulation during the breeding season. In Experiment 1, fresh-diluted or frozen-thawed semen was deposited into the cervical orifice after estrus detection, and an intrauterine AI with frozen-thawed semen was performed by laparoscopy at a fixed-time basis without estrus detection. Embryos were recovered by uterine flushing 6 days after AI, and the rates of recovered, fertilized (cleaved) ova and embryos at the morula or blastocyst stage were compared among the three AI methods. In Experiment 2, the pregnancy rates after the three AI methods were compared. In Experiment 1, the rates of recovered ova were not significantly different among the three AI methods (52.5-56.7%). The rate of fertilized ova (81.0%) by laparoscopic AI with frozen-thawed semen was significantly higher compared with cervical AI of fresh-diluted (25.5%) or frozen-thawed (3.5%) semen, but the rate of embryos at the morula or blastocyst stage (17.6%) was significantly lower than that of the cervical AI with fresh-diluted semen (69.2%). The rates of ewes yielding fertilized ova were not significantly different among the three groups (44.4, 11.1 and 62.5% for cervical AI with fresh-diluted and frozen-thawed semen and intrauterine AI with frozen-thawed semen). In Experiment 2, the pregnancy rate of ewes intrauterinally inseminated with frozen-thawed semen (72.2%) was significantly higher than those of ewes inseminated cervically with fresh-diluted (5.5%) or frozen-thawed (0.0%) semen. The present results showed that acceptable fertilization and pregnancy rates could be obtained by an intrauterine AI with frozen-thawed semen using a synthetic semen extender (AndroMed), but not sufficient by the cervical AI with either fresh or frozen semen.     

Laparoscopic artificial insemination in sheep

The goal of any AI program is to create improved offspring, and the achievement of this objective will depend on the breeding value of the ram and ewe selected. Laparoscopic AI is being utilized in the sheep industry to extend the use of superior rams, and it offers the producer the opportunity to maximize the reproductive potential of superior sheep. Rapid genetic trait infusion of known superior stud rams into the flock is the primary economic benefit of laparoscopic AI. The success of laparoscopic AI depends on events and factors that interrelate in a complex way. Once the selection and preparation of the ewe have been accomplished, one of the more important steps in the program is the successful synchronization of the ewe to deliver the necessary ova to the site of fertilization at a specific time. One of the best methods of synchronization for laparoscopic AI is the use of a progesterone product for a controlled time period and the administration of PMSG upon its removal. Detecting the onset of estrus is critical, and the addition of sterile (e.g., vasectomized) males is helpful, even essential, to accurately determine when each ewe begins her estrus. The ram effect has been shown to stimulate ovulation and estrus. Ewes must be inseminated within a narrow window of time after the synchronization product is removed. Ewes should be inseminated in the order in which they begin to exhibit signs of behavioral estrus, but age, stage of lactation, duration of behavioral estrus, and breed must be taken into account when this order is established. Fresh-extended semen works well throughout this preferred time frame established for laparoscopic AI, but frozen semen gives best results when used near the end. Advancement in manufacturing technology today removes equipment as a variable factor. It is important, therefore, that the inseminator develop a level of expertise in laparoscopy to ensure maximum fertilization rates. If available, fresh-extended semen is preferred over frozen semen, using at least the minimal number of spermatozoa necessary for fertilization. Evaluation of the post-thaw frozen or fresh semen is necessary to determine motility, morphology, and concentration, all of which help determine the volume of the insemination dose. The minimum necessary for laparoscopic AI in fine-wooled breeds is 20 X 10(6) normal motile spermatozoa; however, the more seasonal and less fertile American sheep need approximately 40 to 50 X 10(6) normal motile sperm to achieve acceptable fertility rates.(ABSTRACT TRUNCATED AT 400 WORDS)     

Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin

The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal. Ejaculates from a suffolk ram were diluted with a Tris-based extender containing either 15% (v/v) egg yolk or 10% (w/v) BSA, and the diluted semen was frozen in 0.25 ml straws. A fixed-time intrauterine artificial insemination (AI) was performed 43-47 h after CIDR removal, regardless of incidence of estrus. There was no significant difference in pregnancy rates at 60 days after AI between the extenders containing egg yolk (66.7%, 20/30 animals) or BSA (65.5%, 19/29 animals). Furthermore, there were no significant difference in the lambing rates (66.7% and 62.1%) and prolificacy (1.25 and 1.56) between the two semen extenders. The present study indicates that a semi-defined semen extender containing 10% BSA produces fertility after intrauterine AI that is similar to that achieved with semen extender containing egg yolk.     

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.     

Inactivation of bovine herpesvirus 1 in semen using a hyperimmune egg yolk semen extender

Hyperimmune egg yolk semen extender was used for the inactivation of bovine herpesvirus (BHV-1) in experimentally infected bovine semen. As much as 5 x 10(4) TCID50/ml of virus was inactivated in semen as assayed by tissue culture. Moreover the hyperimmune egg yolk semen extender did not produce any adverse effect on the quality of the semen after being frozen/thawed in comparison with normal egg yolk semen extender (P > 0.05). The hyperimmune egg yolk semen extender is considered an important tool for containing the spread of BHV-1 from infected semen.     

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。     

Rabbit reproductive performance after insemination with buserelin acetate extender

Rabbit farmers and insemination centres have established new requirements relating to males (genetic, nutritional and environmental factors) and rabbit insemination techniques. The aim of this study was to evaluate buserelin acetate on the reproductive performance of nulliparous, multiparous lactating and multiparous non-lactating does inseminated on three commercial farms. Two thousand two hundred and three commercial crossbred does belonging to three commercial farms were inseminated with pooled semen from males of two selected meat lines; R line (UPV rabbit selection centres, Spain) in farms 1 and 2, and PS Hyplus 59 line (Grimaud Frère, France) on farm 3. Ejaculates from 12–20 males from each line were pooled and diluted to twelve million sperm per millilitre by adding TRIS–citrate-extender. Diluted semen from each male line was split into two fractions, the first without buserelin acetate added to semen (Control group) and the second fraction was supplemented with 10 μg of buserelin acetate per millilitre (Buserelin group). Receptive females (red colour of vulvar lips) were inseminated with 0.5 mL using a standard plastic curved pipette. Artificial insemination with semen extended with buserelin resulted in lower pregnancy and kindling rates irrespective of the physiological status of females or the farm (78.7% and 76.0% vs 85.8% and 83.4%, buserelin extender group and control respectively, P < 0.01), although does from both treatment groups had a similar litter sizes (10.4 and 9.6 total and alive born, respectively). This study opened up new prospects for changing rabbit insemination procedures.     

Fertility in zebu cattle (Bos indicus) after prostaglandin administration and artificial insemination.

A total of 137 cycling zebu cows, each receiving a single dose of prostaglandin PGF(2alpha)were used in an oestrus synchronization programme on three different farms. Of the cows on the three farms, 60.6 and 90.5% showed overt oestrus and luteolysis, respectively. Pregnancy rate to fixed time inseminations following single injection of PGF(2alpha)was 61.4% for farm 1, significantly higher than the values of 45.7 and 46.9% for farms 2 and 3, respectively. The pregnancy rates to second service of rebred cows were 53.3, 50.0 and 50.0% for the three farms, respectively, with no significant differences between each.Fertility classification of the cows based on progesterone (P(4)) concentration showed that 6.6% of cows on the three farms were incorrectly diagnosed as having corpora lutea; 2.9% of them had incomplete luteolysis and 5.1% may have lost their embryos between days 21 and 45 post-insemination. The pregnancy rate was 10% higher in the rainy season than in the dry season. Cows with body condition scores of 3 and 4 had a higher overall pregnancy rates than those with a body condition score of 2. The findings of this study further confirm the luteolytic efficacy of prostaglandin in inducing oestrus in zebu cattle and indicate that the nutritional status of the cows must be satisfactory before embarking on oestrus synchronization programmes.     

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).     

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.