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Microencapsulation of bovine spermatozoa for use in artificial insemination: a review.

A technique for microencapsulation of bovine spermatozoa has been developed with minimal spermatozoal injury and thus of potential use in artificial insemination. The polymers poly-l-lysine, polyvinylamine and protamine sulfate have proven best for membranes. Encapsulation has been successful with capsules ranging in size from 0.75 to 1.5 mm, and with sperm concentrations from 45 to 180 x 10(6) cells mL-1. Successful extenders include CUE, CAPROGEN, and egg yolk-citrate-glycerol (maximum 10% v/v egg yolk for normal capsular shape). Capsule fragility (ability to rupture under ageing and physical stress) is negatively related to membrane thickness which ranges from 1.92 to 5.32 microns (depending on the concentration of polymer used) and positively related to concentration of sperm encapsulated. Heterospermic studies have shown that encapsulated sperm are capable of fertilization in vivo, but are at a disadvantage to unencapsulated sperm when cows are inseminated at conventional times. Uterine retention of inseminates is favoured by capsules having a 'sticky' membrane. Using current procedures, preliminary homospermic fertility studies indicate that sperm encapsulated with poly-l-lysine or protamine sulfate may achieve normal fertility.

Animals↗

A case of successful use of reproductive technology for a couple with subfertility due to retrograde ejaculation, where artificial insemination had failed.

A case history of a couple where the infertility was due to retrograde ejaculation is presented; when conventional treatment with artificial insemination using sperm isolated from postejaculatory urine failed, IVF and GIFT technology was applied. This resulted in a pregnancy which unfortunately failed at 3 months, unrelated to the method of conception. This case report highlights the possible use of IVF/GIFT for yet another type of male subfertility.

Adult↗

Influence of PMSG and time of artificial insemination on fertility of progestogen-treated sheep in confinement.

The need for pregnant mates' serum gonadotropin (PMSG) in breeding confined sheep by artificial insemination (AI) at progestogen-synchronized estrus was assessed in 152 adult crossbred ewes brought into season by a controlled light regimen. One-half of the ewes received 500 IU PMSG after intravaginal progestogen treatment; all ewes were inseminated either 54, 57 or 60 h after sponge removal or at 54 and again at 60 h. Based on progesterone determinations 18 d after AI, conception rates with single insemination 54, 57, or 60 h and double insemination at 54 and 60 h were 76, 72, 47 and 72%, respectively, among ewes receiving PMSG, compared to 17, 22, 47 and 43%, respectively, among ewes not give PMSG (P less than .01) Lambing rates were higher (P less than .01) with PMSG (67, 67, 37 and 61%) than without PMSG (11, 11, 26 and 33%). While there was only a small increase (.06 less than P less than .05) in litter size with PMSG, fecundity decreased (P less than .01) from 1.4 to .3 when PMSG was not used. These data indicate that, even with controlled lighting to induce estrous activity, additional stimulation of ovulation by PMSG at progestogen-synchronized estrus is necessary for normal fertility when confined sheep are bred by AI.

Animals↗

Treatment with gonadotropin-releasing hormone after first timed artificial insemination improves fertility in noncycling lactating dairy cows.

Lactating Holstein cows were assigned randomly to treatments to improve fertility after first postpartum timed artificial insemination (TAI). In Experiment 1, cows received no treatment (control; n = 9), a controlled internal drug releasing (CIDR) insert from 5 to 12 d after TAI (CIDR; n = 9), or 100 microg of GnRH 5 d after TAI (G5; n = 7). Although treatments did not affect circulating progesterone (P4) concentrations from 5 to 19 d after TAI, there was a tendency for CIDR cows to have greater P4 compared with control or G5 cows within 24 h after treatment. In 2 field trials, cows received either control (n = 223), CIDR (n = 218), or G5 (n = 227) treatments (Experiment 2), or control (n = 160), G5 (n = 159), or treatment with 100 microg of GnRH 7 d after TAI (G7; n = 163; Experiment 3). Treatment did not affect pregnancies per AI (P/AI) in Experiments 2 or 3; however, when data were combined to compare control (n = 383) and G5 (n = 386) treatments, P/AI tended to be greater for G5 (49.1%) than for control (45.8%) cows. This effect resulted from a GnRH treatment x cyclicity status interaction in which P/AI for noncycling cows receiving G5 was greater than for noncycling control cows (45.5 vs. 31.1%). In conclusion, treatment with CIDR inserts after TAI had no effect on P/AI, whereas treatment with GnRH 5 d after TAI improved P/AI for noncycling, but not for cycling cows.

Animals↗

Prevalence of bovine viral diarrhea virus infection in bulls in artificial insemination centers in Poland.

This study was directed at the evaluation of the prevalence of bovine viral diarrhea virus (BVDV) infection in bulls in artificial insemination centers. Both serological and virological examinations were performed. Blood samples were tested in micro-seroneutralization test for BVDV antibodies. Virus isolation was performed in cell culture and BVDV antigen was detected in an indirect immunofluorescence assay with monoclonal antibodies. One hundred and seventy-five serum samples and 219 whole blood samples for virus isolation were tested. Neutralizing antibodies were found in 86% of the bulls. Persistent infection (PI) was detected in 0.9% of the analyzed blood samples.

Animals↗

Effect of day of the estrous cycle at the initiation of a timed artificial insemination protocol on reproductive responses in dairy heifers.

Our objectives were to identify stages of the estrous cycle at which initiation of a timed artificial insemination (Ovsynch/TAI) protocol may reduce pregnancy rates and to monitor ovarian follicle dynamics and corpus luteum development after initiation of the Ovsynch/TAI protocol at different stages of the cycle. Cycling Holstein heifers (n = 24) were injected twice with prostaglandin F2alpha to induce estrus and were scanned by ovarian ultrasonography to determine the day of ovulation (d 0). Heifers were assigned to initiate the Ovsynch/TAI protocol at d 2 (n = 5), 5 (n = 5), 10 (n = 4), 15 (n = 5), or 18 (n = 5) of the cycle. The Ovsynch/TAI was initiated with an injection of gonadotropin-releasing hormone agonist followed 7 d later with an injection of prostaglandin F2alpha. At 36 h after injection of prostaglandin F2alpha, heifers were injected with gonadotropin-releasing hormone agonist and inseminated 16 h later. Heifers were scanned daily during the Ovsynch/TAI protocol and every other day after insemination until 16 d later. Blood samples were collected daily starting at the 1st day heifers were scanned and continued until 16 d after insemination. Initiation of the Ovsynch/TAI protocol at d 15 of the estrous cycle caused heifers to ovulate prior to insemination. A shortened return to estrus (< 16 d) was caused by ovulation failure to the second gonadotropin-releasing hormone injection, by incomplete regression of the corpus luteum, and by short life-span of the induced corpus luteum. Day of the cycle in which the Ovsynch/TAI protocol is initiated affects dynamics of follicular development, plasma progesterone profiles, and occurrence of premature ovulation. Size of the pre-ovulatory follicle was associated positively with subsequent progesterone concentrations following insemination.

Animals↗

Evaluation of two hormonal protocols for synchronization of ovulation and timed artificial insemination in dairy cows managed in grazing-based dairies.

To evaluate the efficacy of two hormonal protocols for synchronization of ovulation and timed artificial insemination (TAI) in dairy cows managed in grazing-based dairies, lactating dairy cows (n = 142) from two grazing-based dairies were randomly assigned to one of three treatment groups. Cows in the first group (Ovsynch) received 50 microg of GnRH (d -10); 25 mg of PGF2alpha (d -3), and 50 microg of GnRH (d -1) followed by timed AI on d 0. Cows in the second group (PGF + Ovsynch) received a modified Ovsynch and timed AI similar to Ovsynch but with the addition of 25 mg of PGF2alpha 12 d (d -22) before initiation of Ovsynch. Cows in the third group (control) received standard reproductive management in place on each farm. Luteolysis occurred in 90.5% of cows exhibiting luteal function on d -22 in the PGF + Ovsynch treatment group, whereas none of the cows in the Ovsynch group underwent luteolysis on d -22. Synchronization rate (i.e., ovulatory response at 48 h after the second GnRH injection), conception rates at TAI and pregnancy rates after 35 d of breeding were similar for cows in the Ovsynch and PGF + Ovsynch groups. The proportion of anovular cows at the first GnRH injection of the synchronization protocols (d -10) was similar for cows receiving Ovsynch (28.0%) and PGF + Ovsynch (30.7%), and conception rate at TAI was similar for cycling (45.8%) and anovular (30.0%) cows. The cumulative pregnancy rate was greater for cows receiving TAI compared with control cows after 7 d of breeding (41.2 vs. 20.0%) but did not differ at 35 d of breeding (54.9 vs. 60.0%). Administration of PGF2alpha 12 d before initiation of Ovsynch did not improve synchronization, conception, or pregnancy rate compared with the standard Ovsynch protocol. Synchronization of ovulation to initiate timed AI at the onset of the breeding season resulted in earlier establishment of pregnancy compared with standard reproductive management.

Animals↗

Parameters affecting the results in a program of artificial insemination with donor sperm. A 12-year retrospective review of more than 1800 cycles.

PURPOSE: We aimed to establish the influence of the parameters affecting artificial insemination (AI) results in order to describe the ideal situations to achieve the best results as well as to adequately counsel the patients undergoing these treatments about their pregnancy chances. METHODS: We performed a controlled retrospective clinical study over more than one decade in a total of 1858 cycles in 710 patients. Clinical histories and computer registers were systematically reviewed between January 1990 and June 2002. We analyzed the influence of diverse factors affecting AI results such as patient's age, ovarian stimulation, and seminal characteristics to offer a detailed description of the technique. RESULTS: Less than 35-years-old, smooth ovarian stimulation and 5 million of progressive motile sperm inseminated two consecutive days are the optimum conditions for achieving good results. Also, period of time that sperm remained frozen do not affect the result. Furthermore, we present the likely or expected outcomes of these treatments depending on the male and female etiologies. CONCLUSIONS: We discourage AI in aged patients, and strongly recommend undergoing ovarian stimulation. Nonetheless, we must reach an adequate amount of sperm with good motility in order to inseminate with maximum guaranties of success.

Age Factors↗

Use of estradiol cypionate in a presynchronized timed artificial insemination program for lactating dairy cattle.

Experiment 1 evaluated pregnancy rates when estradiol cypionate (ECP) was used to induce ovulation as part of a timed artificial insemination (TAI) protocol in comparison to Ovsynch for lactating dairy cows in Florida (n = 371) and Texas (n = 321). Cows were presynchronized with two injections of PGF2, (25 mg, im) given 14 d apart with TAI protocols beginning 14 d after the second injection of PGF20. The TAI protocols consisted of an injection of GnRH (100 microg, im) followed by PGF2alpha 7 d later. Then, cows either received an injection of GnRH (Treatment I, Ovsynch) at 48 h after PGF2alpha and inseminated 16 to 24 h later or received an injection of ECP (1 mg, i.m.) at 24 h after PGF2alpha, (Treatment II; Heatsynch) and inseminated 48 h later. In Florida, pregnancy rates after TAI were 37.1 +/- 5.8% for Ovsynch compared with 35.1 +/- 5.0% for Heatsynch. In Texas, pregnancy rates were 28.2 +/- 3.6% for Ovsynch and 29.0 +/- 3.5% for Heatsynch. Overall pregnancy rates did not differ between Ovsynch and Heatsynch treatments. In Experiment 2, estrus and ovulation times were determined in lactating dairy cows submitted to the Heatsynch protocol. Frequencies of detected estrus and ovulation after ECP were 75.7% (28/37) and 86.5% (32/37), respectively. Mean intervals to ovulation were 55.4 +/- 2.7 h (n = 32) after ECP and 27.5 +/- 1.1 h (n = 27) after onset of estrus. Estrus occurred at 29.0 +/- 1.8 h (n = 28) after ECP. It is recommended that any cow detected in estrus by 24 h after ECP injection be inseminated at 24 h and all remaining cows be inseminated at 48 h because 75% (n = 24/32) of the ovulations occurred between > or = 48 h to < or = 72 h after ECP. Synchronization of ovulation and subsequent fertility indicated that estradiol cypionate could be used to induce ovulation for successful timed insemination.

Animals↗

Time and incidence of ovulation and conception rates after incorporating estradiol cypionate into a timed artificial insemination protocol.

Two experiments were conducted to determine the effect of estradiol cypionate (ECP), when incorporated into a conventional GnRH-PGF(2alpha)-GnRH timed artificial insemination protocol (Ovsynch), on systemic estradiol (E(2)), time and incidence of ovulation, luteal development, and conception rate in Holstein cows. Our objective was to determine if administration of 0.25 mg of ECP at the time of the second GnRH injection would effectively synchronize ovulation and increase conception rate. In Experiment 1, lactating Holstein cows (n = 23; 58.7 +/- 1.2 d in milk) were synchronized with PGF(2alpha) (at d -10). Ten days later, Ovsynch was initiated with the administration of 100 mug of GnRH (d 0) followed by PGF(2alpha) on d 7. On d 9, cows were assigned randomly to be treated with either GnRH + 0.25 mg of ECP (OVS-ECP; n = 11) or GnRH and 1 mL of cottonseed oil (OVS-C; n = 12). Ovarian activity was monitored by ultrasonography on d 0, 7, and 9. To determine the time of ovulation, ultrasound examinations were conducted at 12 and 20 h posttreatment and then at least every 3 h until either 36 h posttreatment or ovulation was observed. Blood samples were collected on d 0, 7, 9, and 16 for progesterone analysis. Blood samples also were collected at the time of treatment (d 9, 0 h) and at 6, 12, 20, and 28 h for E(2) analysis. Incidence of ovulation did not differ between treatments. Mean ovulation time relative to the second GnRH administration was similar between treatments. Serum progesterone concentration did not differ between treatments at any time. Serum E(2) concentration was not different at the time of treatment (0 h); however, mean E(2) concentration was greater for the OVS-ECP group at 6 and 12 h after treatment compared with OVS-C. In Experiment 2, lactating dairy cows (n = 333) in 3 commercial herds were randomly assigned to OVS-ECP (n = 169) or OVS-C (n = 164). Cows were inseminated 22 to 24 h posttreatment. Conception rates did not differ between treatments. Estradiol cypionate treatment was successful in increasing serum E(2) when administered at the time of the second dose of GnRH in the Ovsynch protocol. Conception rates, however, were not affected by treatment.

Animals↗

Successful artificial insemination in a case of retrograde ejaculation.

A case is described of retrograde ejaculation due to bladder neck resection followed by ileal conduit with a non-functioning bladder, where, after unsuccessful attempts at artificial insemination with spermatozoa retrieved from the bladder, ejaculate was obtained at defecation followed by artificial autoinsemination by the male, leading to two consecutive pregnancies.

Adult↗

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

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

Animals↗

Pregnancy outcome after artificial insemination or IVF with frozen semen donor: a collaborative study of the French CECOS Federation on 21,597 pregnancies.

OBJECTIVE: To assess pregnancies and conceptus after artificial insemination (AID) or IVF with frozen semen donor (IVF-D) on sufficiently large study population in order to distinguished minor variations. STUDY DESIGN: From 1987 to 1994, all pregnancies obtained after AID or IVF-D were registered prospectively in the French CECOS Federation data base. Different factors were recorded for this study: first menarche age of the recipient women, cycle length, insemination date in the conception cycle, maternal age at delivery, hormonal treatments, donor age, sperm conservation length and follow up of the pregnancy: miscarriage, tubal pregnancy, time at delivery, sex of the foetus, weight, malformation. RESULTS: 21,597 pregnancies obtained after AID and 3381 after IVF-D were registered. 2% were lost to follow up. Foetal loss rate is 18% after AID and 21.5% after IVF-D (p < 0.001). The tubal pregnancy rate is 0.9% after AID and 1.7% after IVF-D (p < 0.0001). 18,128 children were born after AID and 3313 after IVF-D. After AID, the twin pregnancy rate is 6.9% and the multiple pregnancy (> or = 3 foetus) rate is 0.7%. After IVF-D, these rates are 24.8% and 4.2% respectively (p < 0.0001). After AID the mean weight at delivery, sex ratio, premature rate, intra uterine growth retardation rate are not different from national rates published in 1995. The foetus malformation rate (including medical abortions) is 1.9% after AID and 2.7% after IVF-D (p < 0.009). After AID the trisomy 21 rate increases with the mother age but also with the donors age if the maternal age is equal. The birth defects rate is not different from those registered in Paris, Strasbourg and Marseille. The birth defects rate observed after IVF-D is not different from the rate observed after IVF with husband semen. (2.74% versus 2.99%; p = 0.16). CONCLUSION: After AID the miscarriage and tubal pregnancy rate, the children's weight, the premature rate is not different from that of the general French population. Sex ratio is normal as is the global malformation rate. The multiple pregnancy rate (x 7 for twin and by 10 for multiple pregnancies more than 3 foetus) is high, showing the influence of ovulation induction treatment. The birth chromosomal abnormalities rate is normal and correlated not only to the mother's age but also to the donor's age. This result without clear biological explanation will require further verification in a greater population. Practically speaking, these observations encourages lowering the age limit for semen donors less than 45 years. IVF-D practice instead of AID doubles the tubal pregnancy rate (0.9% versus 1.7% and increases the twin pregnancy rate by 2.5% and the multiple pregnancy (> or = 3 fetus) rate by 3. It is necessary to promote good practice for AID for which the pregnancy rate is very different from one centre to another within the centres with AID low results a too high rate of IVF-D. Finally we can say that pregnancies from IVF-D or IVF with husband semen are not significantly different. In other words pregnancy outcome is not changed after sperm cryopreservation.

Adult↗

A reciprocal translocation between autosomes 8 and 10 in a boar used for artificial insemination service and its effects on litter size.

The low number of piglets per litter was the reason why a Finnish Yorkshire breed boar used for artificial insemination (AI) was taken for cytogenetic study. The boar had a reciprocal translocation between autosomes 8 and 10, resulting in a reduction in its litter size of about 19%. In addition the litter sizes were counted from 16 female offspring of this 2n = 38,XY; rcp(8;10)(p1.1;q1.3) boar.

Animals↗

Fertilization and embryonic mortality rates in beef heifers after artificial insemination.

A total of 256 beef heifers, in 2 experiments, was used to establish fertilization rate and subsequent embryo survival rates. Fertilization rate following a single artificial insemination was 90%. Pooled estimates of embryo survival showed high survival rates up to Day 8 (93%) but markedly reduced (P < 0.001) survival at Days 12 (56%), 16 (66%) and 42 (58%). It is suggested that most embryonic mortality occurs between Days 8 and 16.

Animals↗

Artificial inseminations with frozen semen and application of DNA fingerprint for parents/offspring conformation on Great Pyrenees.

Frozen semen of Great Pyrenees, air-shipped from France, was artificially inseminated (AI) to two Great Pyrenees bitches. Blood LH and P concentrations and vulval width principally determined optimal breeding time of bitches. AI were performed on 17 to 21 days from the onset of estrous bleeding, and 25 days after final Al, pregnancy of one bitch, with two fetuses, was identified by ultrasound image diagnostic equipment. On 62 days, the bitch whelped one male pup and one female pup. Blood samples were obtained from four parents and offspring, and their DNA was extracted. DNA samples digested with four kinds of restriction endonucleases, and resulting fragmented DNA were used as test samples for DNA fingerprinting with Myo prove. As a result bands from pups, displayed by any enzyme, were inherited from either parent, and clear parent-offspring relationships were identified.

Animals↗

A comparative study on the oestrous response to PGF2 alpha analogue treatment, and conception rates according to time of artificial insemination, in Zebu (Bos indicus) and Baoulé (Bos taurus) cattle.

The oestrous response, interval and conception rates were studied after synchronization with a prostaglandin analogue (cloprostenol) and artificial insemination (AI) performed at different times in 50 Zebu (Bos indicus) and 83 Baoulé (Bos taurus) cattle indigenous to Burkina Faso. The overall proportion of cows responding to synchronization was 70% (93/133). Although the response was higher for the Baoulé cattle, at 73.5% (61/83), than for the Zebu, at 64% (32/50), the difference was not statistically significant (p > 0.05). The mean oestrous interval from treatment to the onset of oestrus (TOI) was shorter in the Zebu (54.1 h, SD 6.7) than in Baoulé (65.2 h, SD 12.9) cattle (p < 0.001). Of the Zebu (n = 32) that responded, 65.7% presented oestrus over a period of 12 h ranging from 48 h to 60 h after treatment. For the Baoulé cows, the highest proportion of animals in oestrus over a period of 12 h was 41% between 60 h and 72 h after treatment. The frequency distribution of onset of the oestrus indicated that up to 64.5% of the Zebu and 79.5% of the Baoulé cattle showed onset of oestrus during the daytime. For Zebu and Baoulé cows inseminated 13 h or 18 h after the onset of oestrus, conception rates were 56% and 57% (p > 0.05) and 33% and 64% (p < 0.05), respectively. Based on these findings, it appears that the oestrous response to synchronization was adequate for both Zebu and Baoulé cattle and that the time to onset of oestrus varied according to genotype. It was also concluded that conception rates were satisfactory for both genotypes but that, for Baoulé cattle, AI performed 18 h after oestrus significantly increased conception rates compared to AI at 13 h after oestrus.

Animals↗

Sensitivity to exogenous gonadotropins for ovulation induction and laparoscopic artificial insemination in the cheetah and clouded leopard.

Ovarian sensitivity to exogenous gonadotropins was assessed in the cheetah (Acinonyx jubatus) and clouded leopard (Neofelis nebulosa) to help optimize artificial insemination (AI). Eighteen female cheetahs were used on 29 occasions and were given i.m. injections of 100, 200, or 400 IU eCG and 100 or 250 IU hCG 80 h later. Twenty-three female clouded leopards were treated i.m. on 27 occasions with 25, 50, 75, 100, 200, or 400 IU eCG followed 80 h later with 75, 140, or 280 IU hCG. Ovaries were examined laparoscopically at 43-48 h after hCG in cheetahs and 39-50 h in clouded leopards. All gonadotropin dosages stimulated ovarian activity in both species, but ovulation success and corpus luteum (CL) morphology varied (p < 0.05) with treatment. For both species, the highest and intermediate eCG dosages resulted in ovulation in a high proportion (72-100%) of females. The lowest eCG dosage, although capable of stimulating follicular development, compromised ovulation and resulted in few (< 26%) postovulatory females. For each species, small CL (2-4-mm diameter) were observed with the highest and lowest eCG dosage, and large CL (5-8-mm diameter) were associated with intermediate eCG dosages. Aged CL (10-12 mm diameter) were observed in 4 of 23 (17.4%) clouded leopards with no prior male exposure, indicating occasional spontaneous ovulation. Nineteen laparoscopic intrauterine AI procedures were performed in eCG/hCG-treated postovulatory cheetahs. Eighteen AI procedures were conducted in eCG/hCG-treated postovulatory clouded leopards. Six of the 13 cheetahs (46%), all in the 200-IU eCG/100-IU hCG group, became pregnant, in contrast to none of the clouded leopards. This study has revealed differences in ovarian activity in two wild felid species as a result of changes in exogenous gonadotropin dosage. Because of this dose-effect response, this comparative approach is necessary to identify a gonadotropin regimen that can mimic "normalcy." Even then, the relatively high AI success in the cheetah compared to the clouded leopard suggests that factors other than ovarian response can dictate the efficiency of assisted reproduction in this taxon.

Acinonyx↗