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Oestrous synchronization, semen preservation and artificial insemination in the Mohor gazelle (Gazella dama mhorr) for the establishment of a genome resource bank programme.

Gazella dama mhorr is an endangered species with an extant population of about 190 animals distributed between several zoos. Semen was collected by electro-ejaculation from 12 adult males, and cryopreserved in TEST-yolk diluent containing 6% glycerol. The effects of the concentration of egg yolk (5%, 10% and 20%) and the presence or absence of sodium triethanolamine lauryl sulfate (equex) on sperm motion and acrosomal integrity after thawing were examined. Increasing concentrations of egg yolk resulted in more acrosomal damage and poorer motility after thawing. The presence or absence of equex had no effect on either parameter. The frozen spermatozoa were used in an insemination trial, in which 13 females were treated with intravaginal progesterone-releasing devices to synchronize oestrus. Seven females were inseminated with frozen-thawed semen 48 h after removal of the devices, and six were inseminated after 60 h. Three females in the first group and one in the second group became pregnant. However, only one pregnancy (from the 48-h group) was carried to term. The study demonstrated the feasibility of applying artificial insemination in this species, but revealed that a number of outstanding technical problems remain to be solved.

Acrosome↗

Effect of artificial insemination on submission rates of lactating dairy cows synchronised and resynchronised with intravaginal progesterone releasing devices and oestradiol benzoate.

This study investigated the hypothesis that a reduction in submission rates at a resynchronised oestrus is not due to the resynchrony treatment involving intravaginal progesterone releasing devices (IVDs) and oestradiol benzoate (ODB) but is associated with artificial insemination (AI) at the first synchronised oestrus. In Experiment 1, cows were synchronised for first oestrus with IVDs, with ODB administered at the time of device insertion (Day 0, 2 mg IM) and 24 h after removal (Day 9, 1 mg IM) and PGF(2alpha) injected at the time of device removal. Cows were then either inseminated (I) for 4 days or not inseminated (NI) following detection of oestrus (first round of AI). Every animal was resynchronised for a second round of AI by reinsertion of IVDs on Day 23 with administration of ODB (1 mg IM) at the time of insertion as well as 24 h after removal (Day 32). Cows detected in oestrus and inseminated for 4 days at the second round of AI were resynchronised for a third round by repeating the resynchrony treatment starting on Day 46 and inseminating cows on detection of oestrus for 4 days. In Experiment 2 the same oestrous synchronisation and resynchronisation treatments were used, but the timing of treatments differed. The cows had their cycles either presynchronised (treatment start Day -23) without AI and then resynchronised, starting on Day 0, for the first round of AI for AI at detected oestrus for 4 days, or they were synchronised (treatment start Day 0) for the first round of AI. In Experiment 1, 91.4% (64/70) and 92.6% (63/68) (P = 0.79) of cows in the I and NI treatments, respectively, were detected in oestrus after the initial synchronisation. At the second round of AI, submission rates for insemination were lower in the I group compared to the NI cows (74.5%, 35/47 versus 92.6%, 63/68, respectively; P = 0.007). Pregnancy rates (proportion treated that were classified as becoming pregnant) in I and NI cows 4 weeks (61.4%, 43/70 versus 63.2%, 43/68) and 7 weeks (77.1%, 54/70 versus 69.1%, 47/68) after the AI start date (AISD) did not differ significantly between treatments. In Experiment 2, presynchronisation and then resynchronisation of oestrous cycles before the first round of AI did not affect oestrous detection rates at the first round of AI (100%, 44/44 versus 98.0%, 50/51; P = 0.54), or pregnancy rates 1 week (63.6%, 28/44 versus 60.8%, 31/51; P = 0.70), 4 weeks (72.7%, 32/44 versus 76.5%, 39/51; P = 0.76) and 7 weeks (81.8%, 36/44 versus 88.2%, 45/51; P = 0.40) after AISD compared to cows that had their cycles synchronised for the first round of AI. These findings support our hypothesis that a reduction in submission rates at a resynchronised oestrus is associated with AI at the first synchronised oestrus and not due to a resynchrony treatment involving IVDs and ODB. This study supports the concept that early embryonic loss following AI at a synchronised oestrus could cause a reduction in submission rates following resynchronisation of oestrus, although investigation of the effect of passing an AI catheter or semen components were not studied per se.

Animals↗

Reestablishment of a transgenic rabbit line by artificial insemination using cryopreserved semen.

The production of recombinant proteins in the milk of transgenic animals is an alternative to traditional cell culture methodology. Transgenic rabbits can serve in the small-scale production of recombinant proteins, underscoring the need to maintain valuable transgenic lines. In this study, the authors used cryopreserved transgenic rabbit semen to artificially inseminate does, demonstrating the utility of this method for the reestablishment of a transgenic rabbit herd.

Animal Husbandry↗

Factors associated with first service conception in artificially inseminated nulliparous Holstein heifers.

Animal and management factors associated with first service conception in nulliparous dairy heifers were determined in 601 Holstein heifers from a dairy farm in north central Florida. Animal data collected included body weight, height at the withers and tail head, body condition score at 6 months of age and just prior to first artificial insemination (AI), and pelvimetry measurements taken just prior to first AI. Management data included season of first AI, inseminator, service sire, method of estrus detection, whether the estrus of first insemination was induced using prostaglandin F(2alpha) (PGF(2alpha)), and whether the heifer received a modified live virus (MLV) vaccine within 21 days of first insemination. Data were analyzed using multivariable logistic regression. Heifers inseminated in the summer were more than four times less likely to become pregnant to first insemination than heifers bred during the rest of the year (odds ratio (OR)=0.24; 95% CI=0.14, 0.41). Using secondary signs for estrus detection instead of standing estrus resulted in significantly reduced odds of conception to first service (OR=0.37; 95% CI 0.13, 1.02). Also, heifers inseminated at estrus induced by PGF(2alpha) were approximately one-third less likely to conceive than those heifers inseminated to a naturally occurring estrus (OR=0.66; 95% CI 0.46, 0.95). An interaction between pelvic size and breeding season was found indicating that large pelvic size had a significant positive effect on fertility in the summer, but was not associated with conception to first service in the winter.

Analysis of Variance↗

Effect of sire fertility and timing of artificial insemination in a Presynch + Ovsynch protocol on first-service pregnancy rates.

Sire fertility may influence pregnancy rate (PR) by differences in sperm survival in the female reproductive system and time required for capacitation and transport of sperm to site of fertilization. A predicted fertility index, Estimated Relative Conception Rate, was used to select 3 high-fertility artificial insemination (AI) sires (+3) and 3 average AI sires (-1). Ovulation can be predicted to occur at approximately 28 h following GnRH administration when used in an Ovsynch protocol. The objective of this study was to determine if AI at 2 times, 0 or 24 h after GnRH administration, in a Presynch + Ovsynch protocol resulted in different first-service PR when average or high-fertility sires were used. Lactating Holstein cows (n = 1,457) from 2 dairy herds located in the Piedmont region of North Carolina were utilized for 12 mo. Timing of AI did not affect first AI PR and no interaction of sire-fertility group and timing of AI was detected. First AI PR did not differ between sire-fertility groups (23.2 vs. 29.4%) for average and high-fertility groups, respectively. First-lactation cows were 53% more likely to conceive than older cows, and cows bred during April through June were 66% less likely to become pregnant compared with cows bred from October through January. No interactions were detected among parity, season, sire-fertility group, or time of AI. Using only 3 sires per group based on Estimated Relative Conception Rate estimates resulted in large variability of sire conception within groups, although group averages differed by 6 points.

Animals↗

State of the art in artificial insemination of pigs in the United States.

Currently, the United States ranks second in pork production worldwide. In 1999 the breeding herd consisted of approximately 6.2 million breeding females with an annual production of 100 million market animals having an average market live weight of 118.6 kg. The objective of this paper is to briefly describe the current US swine artificial insemination (AI) industry and to profile semen production and insemination practices that are currently in use.

Animals↗

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.

Animals↗

Evaluation of a fixed-time artificial insemination protocol for postpartum suckled beef cows.

Treatment with melengestrol acetate (MGA), an oral progestin, prior to administration of gonadotropin-releasing hormone (GnRH) and prostaglandin F2alpha (PG) effectively synchronizes estrus and maintains high fertility in postpartum beef cows. The objective of this experiment was to determine whether treatment with MGA prior to a GnRH-PG-GnRH protocol would improve pregnancy rates resulting from fixed-time artificial insemination (AI). Multiparous crossbred beef cows at two University of Missouri-Columbia farms (n = 90 and n = 137) were assigned by age and days postpartum to one of two treatments. Cows were fed carrier (1.8 kg x animal(-1) x d(-1)) with or without MGA (0.278 mg x kg(-1)) for 14 d. All cows were administered GnRH (100 microg; intramuscularly) on d 12 after MGA or carrier withdrawal and 7 d before PG (25 mg; intramuscularly). All cows received a second injection of GnRH and AI 72 h after PG. Mean days postpartum for MGA and control cows at the initiation of treatment were 39.6 and 38.9 d for herd 1; and 51.9 and 50.9 d for herd 2, respectively (P > 0.70 within herds). Blood samples were collected from all cows at 10 and 1 d before the feeding of MGA or carrier began and at the times GnRH and PG were administered. Concentrations of progesterone in serum at the initiation of treatment were elevated (>1 ng/mL) in 0% of MGA and 7% of control cows in herd 1, and 54% of MGA and 49% of control cows in herd 2 (P > 0.05 within herds). Pregnancy rates to fixed-time AI were determined by transrectal ultrasonography 50 d after AI. Pregnancy rates in herd 1 were 58% (26/45) and 51% (23/45) for MGA-treated and control cows, respectively (P = 0.52), and 63% (44/70) and 45% (30/67) for MGA-treated and control cows in herd 2, respectively (P = 0.03). Differences in pregnancy rates to fixed-time AI were significant (P = 0.04) when data from the two herds were combined (with MGA = 70/115 [61%]; control = 53/112 [47%]). There was no difference (P > 0.20) in final pregnancy rates (timed AI plus 45 d exposure to bulls) between treatments, within herds, or when herds were combined. In summary, pregnancy rates resulting from fixed-time AI may be improved with treatment of MGA prior to a GnRH-PG-GnRH protocol.

Animals↗

Improved effectiveness of artificial insemination of turkey hens associated with ahemeral light-dark cycles and age at photostimulation.

In lighting studies with turkey hens, long ahemeral (AH; non-24 h) light-dark cycles have consistently resulted in greater, although not statistically significant, fertility than control 24 h light-dark cycles. The present study was designed to further evaluate AH lighting effects on fertility by an evaluation of the effectiveness of artificial insemination (AI) under less than optimal conditions for normal fertility, that is, single AI and early age at lighting. Turkey hens had greater percentage fertility of eggs when photostimulated at 30 wk (95.0) than 26 wk (76.3) of age following single, but not double, AI. Ahemeral lighting dramatically improved the effectiveness of a single AI of hens photostimulated at 26 wk of age as compared to controls (89.3 vs 76.3% fertility, respectively). However, with multiple AI, benefits of AH lighting on fertility exceeding that of control hens was not significant. It may be concluded that AH lighting can dramatically improve the effectiveness of AI under certain adverse conditions, such as early age at lighting and reduced exposure of the hen to sperm.

Animal Husbandry↗

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.

Animals↗

[Some aspects of fertility following artificial insemination and natural service in dairy cattle (author's transl)].

The difference between the non-return percentage (60-90 days) following 17,086 first inseminations and that after 9,776 first matings in Dutch-Friesian cattle was studied in a random sample of the Netherlands Herd Book records in 1969. The effects of the following factors on this difference in the cases studied are discussed: -- age of bulls and cows, -- frequency of natural service and artificial insemination during each oestrus, -- use of deep-frozen semem --housing and grazing periods. The difference having been corrected for these factors, it was estimated at not more than 9 per cent of non-return 60-90 days in favour of natural service.

Age Factors↗

Evaluation of two different oestrus-synchronisation methods with timed artificial insemination and resynchronisation of returns to oestrus in lactating Holstein cows.

To examine the outcomes of adding medroxyprogesterone acetate (MAP) to the ovsynch protocol with the traditional ovsynch protocol in both cycling and anoestrus cows, and to evaluate a resynchronisation protocol, 742 cows averaging more than 40 days postpartum were assigned to the following four treatments: (1) ovsynch (OVS): day 0: GnRH; day 7: PGF2alpha; day 9: a similar dose of GnRH; day 10: timed artificial insemination (TAI), approximately 16-20h later; (2) ovsynch+MAP (MAP): the same ovsynch protocol plus an intravaginal insert made of polyurethane sponge impregnated with 300mg of MAP immediately after the first GnRH treatment and on day 7, at the time of the PG treatment, the sponge was removed; (3) resynchronisation (MAP+ODB): 1mg of oestradiol benzoate (ODB) on day 13 after TAI and a new sponge impregnated with MAP was inserted and; on day 20, 1mg of ODB was given and the sponge removed; and (4) no resynchronisation (No MAP): only oestrus detection and AI at any repeat oestrus detected after TAI. Progesterone was measured in milk samples collected on days -17, -10, -3, 13 and 20 (TAI=day 0). Based on milk P4 at days -17 and -10, 27.4% of the cows were still anoestrus. At PG injection, 67.7% of the cycling and 21.3% of the anoestrus cows had elevated P4. Farm, days postpartum and parity variations were detected in both cases. On day 20 after TAI 42.6% of cycling and 8.3% of the anoestrous cows had elevated P4. Pregnancy rates were similar in both pre-breeding treatments (20%), but interactions (P<0.001) were detected between treatment and cycling activity (for anoestrous cows: MAP=34.9%; OVS=11.1%. Average interval from TAI to subsequent AI was 37+/-3 days. Resynchronisation resulted in more (P<0.001) cows in oestrus between days 18 and 25 after TAI. Conception rate in the MAP+ODB treatment was lower (P<0.05) than the No MAP group (22.8% versus 47.4%). It was concluded that the addition of a progestin to the ovsynch protocol resulted in increased pregnancy rates of cows treated during anoestrus. The benefit of including MAP with the ovsynch protocol for cycling cows is equivocal.

Administration, Intravaginal↗

Use of boar spermatozoa for artificial insemination. III. Fecundity of boar spermatozoa stored in Beltsville liquid and Kiev extenders for three days at 18 C.

A field trial was conducted on several hundred farms in The Netherlands to compare the fertilizing capacity of boar spermatozoa stored for 1, 2, or 3 d at 18 C in either Kiev or Beltsville liquid extender (BL-1). Two storage volumes (25 and 100 ml) were used. Twelve Dutch Landrace boars, all in routine commercial artificial insemination service, provided the semen in twice-a-week collections. Over an 8-wk period in March and April, 1979, 2,849 sows and gilts were inseminated. Farrowing rates were higher (P less than .0001) with Kiev extender than with BL-1 extender (69.3 vs 60.5%) when semen was used over a 3 d period. The volume at which the semen was extended and stored had no effect on any parameter. Farrowing rate declined significantly when semen stored at 18 C for more than 2 d was used (74.5% vs 64.7% with semen stored for 1 and 3 in Kiev extender; 65.9% vs 52.7% with semen stored for 1 and 3 d in BL-1 extender). The farrowing rate, total number of pigs/litter and live pigs/litter were significantly higher for sows than for gilts inseminated with similar extended semen. On the basis of the results of this trial, Kiev extender appears to be more suitable as a diluent for liquid semen than does BL-1. AI with semen extended and stored for 3 d at 18 C can be expected to result in a lower farrowing rate than AI with semen stored for 1 or 2 d.

Animals↗

How useful is Leuven personalism in the world of bioethics? The test case of artificial insemination.

Personalism in ethics denotes any system based upon the value of the person. Several versions of personalist morals have been developed over the past 50 years. Some have had particular interest in the field of medical ethics. Here the question is being studied about one such system, the so-called Leuven personalist morals and its usefulness in today's world of bioethics. In order to test the usefulness of this system the case of artificial insemination is examined both in the early 1970s in the context of the Leuven clinics and, subsequently, in the 1990s in a US policy document. The investigation reveals strengths and weaknesses of this personalism. Regarding AID it reveals unresolved oppositions. The conclusion seems to be that this personalism had, no doubt, a profound impact upon medical ethics within its own circle but, as regards the universal usefulness of the system, serious doubts remain.

Bioethics↗

[Superfoetation following artificial insemination in a ewe (author's transl)].

A case of superfoetation following artifical insemination in a ewe is reported. Estimation of the age of the lambs was based on the maturity of the liver, lung, kidney, brown adipose tissue and skin, showing a difference in gestational age of approximately thirty-six days. The possibility of spontaneous superfoetation as well as that of a second fertilization after artificial insemination is discussed.

Animals↗

[Sperm collection, evaluation, preservation and artificial insemination in the dog].

The present paper explains the conditions for semen collection in the dog in principal and the technique of semen collection by manual stimulation of the penis in detail. The evaluation of the most important ejaculate parameters is pointed out concerning their dependence of the partially striking variation of the body size of dogs from different breeds. Furthermore some methods of semen preservation and the results of the laboratory examinations are presented. The notes upon artificial insemination contain the time, frequency, interval and technique of semen application. Better conception rates can be obtained by a double than by a single insemination. Using fresh semen an insemination interval of 48 hours has proved favorable whereas with preserved semen the insemination interval should not exceed 24 hours. While freshly ejaculated semen can be applied with good success intravaginally, the intrauterine insemination offers better conditions for conception using liquid or frozen stored semen.

Animals↗

Transmission of bovine virus diarrhoea virus (BVDV) by artificial insemination (AI) with semen from a persistently-infected bull.

Twelve heifers that did not have antibodies to bovine virus diarrhoea virus (BVDV) were inseminated with semen from a bull that was persistently infected with the virus and contained 10(4.0)-10(6.5) TCID50 0.1 ml-1. All 12 became infected, as indicated by seroconversion within 2 weeks of insemination. Four control heifers were inseminated with virus-free semen. The virus was not transmitted to these animals in spite of close contact with the heifers inseminated with the infected semen. All the heifers became pregnant and gave birth to clinically normal calves at term. However, one calf was born persistently infected with BVDV. After the birth of this persistently-infected calf the control heifers and their calves seroconverted. The study demonstrates that BVDV may be transmitted in cattle by artificial insemination (AI). Therefore entry of persistently-infected animals into AI centres should be prevented.

Animals↗

Artificial insemination, hybridization and pregnancy detection in sika deer (Cervus nippon ).

Artificial insemination (AI) was performed on sika hinds (Cervus nippon ) receiving various dosages of pregnant mare serum gonadotropin (PMSG; Year 1: 0, 50 and 100 IU; Year 2: 100 and 150 IU) and using semen collected from elk and 1 2 elk x 1 2 sika stags. The time from synchronization device removal (CIDR vs norgestomet) to estrus was determined through observations of mounting activity. Methods for pregnancy detection, serum progesterone (P4), estrone sulfate (E1S), pregnancy-specific protein B (PSPB) and ultrasonography, following AI (Year 1: AI, Days 28 and 48 after AI; Year 2: AI, Days 42, 53 and 100 after AI) and a 90-d natural breeding season were investigated. From available production data, body weights were compared among sika and 1 4 elk x 3 4 sika hybrids relative to age. Pregnancy rates tended (P < 0.10) to differ relative to PMSG treatment and sire; administration of 0 IU PMSG resulted in fewer hinds becoming pregnant to AI than 50 or 100 IU of PMSG. Hinds receiving 100 IU of PMSG had higher (P < 0.05) pregnancy rates than hinds receiving 150 IU PMSG. Time to standing estrus did not differ (P > 0.10) between the CIDR and norgestomet groups. Pregnancy rates 50 d after a 90-d breeding season were similar (P > 0.10) between ultrasound (70.9%) and PSPB (61.6%). Serum P4 after 90 d in breeding groups and 50 d after stag removal were higher (P < 0.05) for pregnant than open hinds. Pregnancy rates (Year 1) 48 d after AI were similar (P > 0.10) between ultrasound (49.0%) and PSPB (37.3%). Serum P4 28 and 48 d after AI were higher (P < 0.05) for pregnant than open hinds. Serum E1S was higher (P < 0.01) for pregnant than open hinds 48 d after AI. Pregnancy rates (Year 2) 100 d after AI did not differ (P > 0.10) between ultrasound and PSPB (66.7%). Serum P4 was higher (P < 0.03) in pregnant than open hinds at 42, 53 and 100 d after AI. At 100 d after AI, pregnant hinds had higher (P < 0.002) serum E1S than open hinds. At 6 to 8 and 11 to 13 mo of age, 1 4 elk x 3 4 sika males tended (P < 0.08) to be heavier than sika males, while 1 4 elk x 3 4 sika females were heavier (P < 0.05) than sika females at all ages. In summary, this study documents the use of AI and methods for pregnancy detection in sika hinds as well as preliminary information regarding the production of elk-x-sika hybrids.

Journal Article↗