Timed artificial insemination in beef cattle using GnRH agonist, PGF2alpha and estradiol benzoate (EB)

The present work evaluated low-cost protocols for timed artificial insemination (TAI) in beef cattle. In Experiment 1, cycling nonlactating Nelore cows (Bos indicus, n=98) were assigned to the following groups: GnRH-PGF (GP) and GnRH-PGF-GnRH (GPG), whereas cycling (n=328, Experiment 2) or anestrus (n = 225, Experiment 3) lactating (L) cows were divided into 3 groups: GP-L, GPG-L and GnRH-PCF-Estradiol benzoate (GPE-L). In Experiment 4, lactating cows (n=201) were separated into 3 groups: GP-L, GPE-L and G 1/2PE-L. Animals from Experiment 1, 3 and 4 were treated (Day 0), at random stages of the estrous cycle, with 8 mug of buserelin acetate (GnRH agonist) intramuscularly (im), whereas in Experiment 2 half of the cows received 8 and the other half 12 mug of GnRH (im). Seven days later (D 7) all animals were treated with 25 mg of dinoprost trometamine (PGF2 alpha, im) except those cows from the G 1/2PE-L group which received only 1/2 dose of PGF2 alpha (12.5 mg) via intravulvo-submucosa (ivsm). Alter PGF2 alpha injection the animals from the control groups (GP and GP-L) were observed twice daily to detect estrus and Al was performed 12 h afterwards. The cows from the other groups received a second GnRH injection (D 8 in GPG-L and d9 in GPG groups) or one injection of estradiol benzoate (EB, 1.0 mg, D 8 in GPE-L group). All cows from GPG and GPG-L or GPE-L groups were AI 20 to 24 or 30 to 34 h, respectively, after the last hormonal injection. Pregnancy was determined by ultrasonography or rectal palpation 30 to 50 days after AI. In the control groups (GP and GP-L) percentage of animals detected in heat (44.5 to 70.3%) and pregnancy rate (20 to 42%) varied according to the number of animals with corpus luteum (CL) at the beginning of treatment. The administration of a second dose of GnRH either 24 (Experiment 2) or 48 h (Experiment 1) after PGF2 alpha resulted in 47.7 and 44.9% pregnancy rates, respectively, after TAI in cycling animals. However, in anestrus cows the GPG treatment induced a much lower pregnancy rate (14.9%) after TAI. The replacement of the second dose of GnRH by EB (GPE-L) resulted in a pregnancy rate (43.3%) comparable to that obtained after GnRH treatment (GPG-L, 47.7%, Experiment 2). Furthermore, the use of 1/2 dose of PGF2 alpha (12.5 mg ivms, Experiment 4) resulted in pregnancy rate (43.5%) similar to that observed with the full dose (im). Both protocols GPG and GPE were effective in synchronizing ovulation in cycling Nelore cows and allowed approximately a 45% pregnancy rate after TAI. Additionally, the GPE treatment is a promising alternative to the use of GPG in timed Al of beef cattle, due to the low cost of EB when compared to GnRH agonists. (C) 2001 by Elsevier B.V.

Treatments to optimize the use of artificial insemination and reproductive efficiency in beef cattle under tropical environments

Bos indicus cattle, the preferred genetic group in tropical climates, are characterized by having a lower reproductive efficiency than Bos taurus. The reasons for the poorer reproductive efficiency of the Bos indicus cows include longer lengths of gestation and postpartum anestrus, a short length of estrous behavior with a high incidence of estrus occurring during the dark hours, and puberty at older age and at a higher percentage of body weight relative to mature body weight. Moreover, geography, environment, economics, and social traditions are factors contributing for a lower use of reproductive biotechnologies in tropical environments. Hormonal protocols have been developed to resolve some of the reproductive challenges of the Bos indicus cattle and allow artificial insemination, which is the main strategy to hasten genetic improvement in commercial beef ranches. Most of these treatments use exogenous sources of progesterone associated with strategies to improve the final maturation of the dominant follicle, such as temporary weaning and exogenous gonadotropins. These treatments have caused large impacts on reproductive performance of beef cattle reared under tropical areas. Copyright © 2011 O. G. Sá Filho and J. L. M. Vasconcelos.

Strategies for Artificial Insemination of Cattle at Synchronized Ovulation or Synchronized Estrus

Two synchronization protocols in lactating dairy and beef cows and in dairy heifers were tested for efficacy of breeding by artificial insemination (AI) with or without estrus detection. Controls received three prostaglandin F2a (PGF2a) injections 14 days apart before AI at observed estrus. Pregnancy rates were compared with animals on the Ovsynch protocol that combined gonadotropin releasing hormone (GnRH) and PGF2a treatments with a timed AI 16 to 20 hours after the second GnRH injection. The pregnancy rates were similar at synchronized ovulation to fixed-time AI in lactating cows, but not effective in heifers because of the lack of synchronization.

The artificial insemination of dairy cattle : a handbook and laboratory manual /

Includes bibliographical references.

Comparison of the pregnancy rates and costs per calf born after fixed-time artificial insemination or artificial insemination after estrus detection in Bos indicus heifers

This study compared pregnancy rates (PRs) and costs per calf born after fixed-time artificial insemination (FTAI) or AI after estrus detection (i.e., estrus detection and AI, EDAI), before and after a single PGF2α treatment in Bos indicus (Brahman-cross) heifers. On Day 0, the body weight, body condition score, and presence of a CL (46% of heifers) were determined. The heifers were then alternately allocated to one of two FTAI groups (FTAI-1, n = 139) and (FTAI-2, n = 141) and an EDAI group (n = 273). Heifers in the FTAI groups received an intravaginal progesterone-releasing device (IPRD; 0.78 g of progesterone) and 1 mg of estradiol benzoate intramuscularly (im) on Day 0. Eight days later, the IPRD was removed and heifers received 500 μg of PGF2α and 300 IU of eCG im; 24 hours later, they received 1 mg estradiol benzoate im and were submitted to FTAI 30 to 34 hours later (54 and 58 hours after IPRD removal). Heifers in the FTAI-2 group started treatment 8 days after those in the FTAI-1 group. Heifers in the EDAI group were inseminated approximately 12 hours after the detection of estrus between Days 4 and 9 at which time the heifers that had not been detected in estrus received 500 μg of PGF2α im and EDAI continued until Day 13. Heifers in the FTAI groups had a higher overall PR (proportion pregnant as per the entire group) than the EDAI group (34.6% vs. 23.2%; P = 0.003), however, conception rate (PR of heifers submitted for AI) tended to favor the estrus detection group (34.6% vs. 44.1%; P = 0.059). The cost per AI calf born was estimated to be $267.67 and $291.37 for the FTAI and EDAI groups, respectively. It was concluded that in Brahman heifers typical of those annually mated in northern Australia FTAI compared with EDAI increases the number of heifers pregnant and reduces the cost per calf born.

A field investigation of a modified intravaginal progesterone releasing device and oestradiol benzoate based ovulation synchronisation protocol designed for fixed-time artificial insemination of Brahman heifers

Pregnancy rates (PR) to fixed-time AI (FTAI) in Brahman heifers were compared after treatment with a traditional oestradiol-based protocol (OPO-8) or a modified protocol (OPO-6) where the duration of intravaginal progesterone releasing device (IPRD) was reduced from 8 to 6 days, and the interval from IPRD removal to oestradiol benzoate (ODB) was increased from 24 to 36 h. Rising 2 yo heifers on Farm A: (n = 238 and n = 215; two consecutive days AI); B (n = 271); and C (n = 393) were allocated to OPO-8 or OPO-6. An IPRD was inserted and 1 mg ODB i.m. on Day 0 for OPO-8 heifers and Day 2 for OPO-6 heifers. On Day 8, the IPRD was removed and 500 μg cloprostenol i.m. At 24 h, for OPO-8 heifers, and 36 h, for OPO-6 heifers, post IPRD removal all heifers received 1 mg ODB i.m. FTAI was conducted at 54 and 72 h post IPRD removal for OPO-8 and OPO-6 heifers. At Farm A, OPO-6 heifers, AI on the second day, the PR was 52.4 to FTAI (P = 0.024) compared to 36.8 for OPO-8 heifers. However, no differences were found between OPO-8 and OPO-6 protocols at Farm A (first day of AI) (39.9 vs. 35.7), or Farms B (26.2 vs. 35.4) and C (43.2 vs. 40.3). Presence of a corpus luteum at IPRD insertion affected PR to FTAI (43.9 vs. 28.8; P < 0.001). This study has shown that the modified ovulation synchronisation protocol OPO-6 may be a viable alternative to the OPO-8 protocol for FTAI in B. indicus heifers.

Fertility Rates Following Fixed-Time Artificial Insemination in Dairy Heifers in a Practical Progesterone-Based Protocol

Background: In bovines, more efficient management practices are important for maximizing profitability. In order to increase the pregnancy rates in artificial insemination (AI) programs, several hormonal protocols were developed to synchronize the follicular wave and the moment of ovulation in beef and dairy cattle. In dairy cattle, detection of estrus can be difficult due to a number of factors including the incidence of silent estrus. Hormonal treatments designed to control both luteal and follicular function has permitting efficient synchronizations of time of ovulation. Thus, the AI can be performed in a large number of animals on a fixed schedule without the need for detection of estrus. Using these management techniques, the fixed-time artificial insemination (TAI) can overcome the problem of accurate estrus detection and help in reducing the incidence of repeat breeding. In addition, with TAI in cattle operations, it is possible to facilitate management practices and commercialization, and to reduce the time and semen wasting with animals inseminated at incorrect times. The investigation of practical and efficient TAI protocols is important for reducing the labor and animal handling of TAI in dairy cattle, as well as for increasing the profitability of the cattle management system. This study was carried out in order to investigate the effectiveness of TAI in dairy heifers treated with a practical progesterone-based protocol.Materials, Methods & Results: This experiment was conducted at the university farm located in southwestern Brazil, during May 2009. Thirty-nine cycling crossbred dairy heifers were employed in this study. All animals received a single intramuscular injection of estradiol benzoate and intravaginal progesterone releasing device in a random stage of the estrous cycle (Day 0). on day 7 the animals were treated with PGF2a analogue and on day 9 the device was removed. Forty-eight hours after the device removal (day 11) a synthetic analogue of GnRH was administered and the animals were fixed-time artificially inseminated at the time of GnRH injection. The inseminations were performed using four different batches from the same Holstein bull. Among the heifers that were synchronized (87.2%), 30.8% ovulated until 24 h after TAI and 56.4% ovulated between 24 and 32 h after TAI. The conception rate was 61.5%. No effects of ovulation time in conception rates were detected. The conception rate from heifers that ovulated until 24 h after TAI was 58.3% and from heifers that ovulated between 24 and 32 h after TAI was 77.3%. The mean of ovulatory follicle in heifers that ovulated until 24 h was 14.3 mm and in heifers that ovulated between 24 and 32 h was 11.9 mm.Discussion: Taking together, the findings of the present study, along with those of others, emphasize the concept that development of practical methods for TAI offers significant advantages to dairy producers if conception rates are close or greater to those obtained after breeding at detected estrus. Thus, the results of the present study reinforce the possibility of making dairy cattle production more cost-effective using TAI. In conclusion, with the progesterone-based TAI protocol of the present experiment all synchronized animals ovulated up to 32 h after GnRH+TAI and no effects of ovulation time related to conception rate was detected. The exogenous control of luteal and follicular development facilitated the reproductive management and animal handling. Also, inseminating the heifers at the moment of GnRH injection in a progesterone-based TAI protocol is a practical strategy and provided satisfactory results regarding ovulation and conception rates in dairy heifers.

Neither plasma progesterone concentrations nor exogenous eCG affects rates of ovulation or pregnancy in fixed-time artificial insemination (FTAI) protocols for puberal Nellore heifers

The objective was to evaluate the effects of plasma progesterone (P4) concentrations and exogenous eCG on ovulation and pregnancy rates of pubertal Nellore heifers in fixed-time artificial insemination (FTAI) protocols. In Experiment 1 (Exp. 1), on Day 0 (7 d after ovulation), heifers (n = 15) were given 2 mg of estradiol benzoate (EB) im and randomly allocated to receive: an intravaginal progesterone-releasing device containing 0.558 g of P4 (group 0.5G, n = 4); an intravaginal device containing 1 g of P4 (group 1G, n = 4); 0.558 g of P4 and PGF2α (PGF; 150 μg d-cloprostenol, group 0.5G/PGF, n = 4); or 1 g of P4 and PGF (group 1G/PGF, n = 3). On Day 8, PGF was given to all heifers and intravaginal devices removed; 24 h later (Day 9), all heifers were given 1 mg EB im. In Exp. 2, pubertal Nellore heifers (n = 292) were treated as in Exp. 1, with FTAI on Day 10 (30 to 36 h after EB). In Exp. 3, pubertal heifers (n = 459) received the treatments described for groups 0.5G/PGF and 1G/PGF and were also given 300 IU of eCG im (groups 0.5G/PGF/eCG and 1G/PGF/eCG) at device removal (Day 8). In Exp. 1, plasma P4 concentrations were significantly higher in heifers that received 1.0 vs 0.588 g P4, and were significantly lower in heifers that received PGF on Day 0. In Exp. 2 and 3, there were no significant differences among groups in rates of ovulation (65-77%) or pregnancy (Exp. 2: 26-33%; Exp. 3: 39-43%). In Exp. 3, diameter of the dominant ovarian follicle on Day 9 was larger in heifers given 0.558 g vs 1.0 g P4 (10.3 ± 0.2 vs 9.3 ± 0.2 mm; P < 0.01). In conclusion, lesser amounts of P4 in the intravaginal device or PGF on Day 0 decreased plasma P4 from Days 1 to 8 and increased diameter of the dominant follicle on Day 9. However, neither of these nor 300 IU of eCG on Day 8 significantly increased rates of ovulation or pregnancy. © 2011.

Impact of the temperament of Nellore cows on the quality of handling and pregnancy rates in fixed-time artificial insemination

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effects of two estradiol esters (benzoate and cypionate) on the induction of synchronized ovulations in Bos indicus cows submitted to a timed artificial insemination protocol

The effects of estradiol benzoate (EB) and estradiol cypionate (EC) on induction of ovulation after a synchronized LH surge and on fertility of Bos indicus females submitted to timed AI (TAI) were evaluated. In Experiment 1, ovariectomized Nelore heifers were used to evaluate the effect of EB (n = 5) and EC (n = 5) on the circulating LH profile. The LH surge timing (19.6 and 50.5 h; P = 0.001), magnitude (20.5 and 9.4 ng/mL; P = 0.005), duration (8.6 and 16.5 h; P = 0.001), and area under the LH curve (158.6 and 339.4 ng/mL; P = 0.01) differed between the EB and EC treatments, respectively. In Experiment 2 (follicular responses; n = 60) and 3 (pregnancy per AI; P/AI; n = 953) suckled Bos indicus beef cows submitted to an estradiol/progesterone-based synchronization protocol were assigned to receive one of two treatments to induce synchronized ovulation: 1 mg of EB im 24 h after progesterone (P4) device removal or 1 mg of EC im at P4 device removal. There was no difference (P > 0.05) between EB and EC treatments on follicular responses (maximum diameter of the ovulatory follicle, 13.1 vs. 13.9 mm; interval from progesterone device removal to ovulation, 70.2 vs. 68.5 h; and ovulation rate, 77.8 vs. 82.8%, respectively). In addition, P/AI was similar (P < 0.22) between the cows treated with EB (57.5%; 277/482) and EC (61.8%; 291/471). In conclusion, despite pharmacologic differences, both esters of estradiol administered either at P4 device removal (EC) or 24 h later (EB) were effective in inducing an LH surge which resulted in synchronized ovulations and similar P/AI in suckled Bos indicus beef cows submitted to TAI. (C) 2012 Elsevier Inc. All rights reserved.

Special report on artificial breeding of dairy cattle in California.

Includes bibliography.

Artificial insemination in penaeid shrimps

Artificial Insemination (AI) is a tool for genetic manipulation in the shrimp stocks. It is seen as one of the means for propagating shrimp culture to new areas by controlled reproduction. Attempts at artificial insemination in the dominant closed-thelycum penaeid shrimps species of the area viz. Metapenaeus affinis and Metapenaeus brevicornis were induced in wild adult stocks collected off Mumbai coast. Female specimens were subjected to unilateral eyestalk ablation by pinching so as to induce moulting and maturation. AI was performed two days after moulting on these females when the cuticle was still soft and flexible. Moulting also ensured rejection of initial spermatophores, if present. Response of males to electrical stimulation for spermatophore expulsion was spontaneous. Use of tissue glue for spermatophore retention was found to be unnecessary. Latency period ranged between10-16 days, while spawning occurred within 10-12 days of spermatophore transfer. Three partial spawning were recorded viz., two in Metapenaeus affinis and one in Metapenaeus brevicornis with an average spawning and hatching rates of 30% and 72.3% respectively. Average survival from first nauplius (N1) to one-day old post-larva (PLI) was a meager 3.43%. Use of AI in genetic manipulation of shrimp stocks for aquacultural purposes is indicated.

A practical approach on boar sperm cryodamage. Morphofunctional and immunocytochemical study of cryopreserved boar sperm intended for use in artificial insemination

L'ús d'esperma criopreservada en la inseminació artificial (IA) d'espècies d'interès productiu permet un major control sanitari i la creació de bancs de germoplasma d'alt valor genètic, entre d'altres avantatges. En el mercat porcí la major part de les inseminacions són encara realitzades amb semen refrigerat degut a l'èxit de l'aplicació de diluents de llarga durada i també a causa de la sensibilitat de l'esperma porcina a la criopreservació. Malgrat que aquesta sensibilitat ve donada per característiques particulars de la fisiologia espermàtica en l'espècie, algunes ejaculacions mantenen els paràmetres de qualitat espermàtica després de la criopreservació (ejaculacions amb bona "congelabilitat", GFEs) enfront d'altres que no sobreviuen al procés (ejaculacions amb mala "congelabilitat", PFEs). El primer objectiu de l'estudi va ser comparar ambdós grups en termes de fertilitat in vivo. El segon objectiu va ser testar l'eficiència de la inseminació postcervical (post-CAI) amb l'esperma criopreservada. El tercer objectiu va ser buscar predictors de la congelabilitat de les ejaculacions, tant en les GFEs com en les PFEs i en tres passos del procés de criopreservació (a 17ºC, a 5ºC i a 240 min postdescongelació). Aquest objectiu es va dur a terme mitjançant l'avaluació de paràmetres convencionals de qualitat espermàtica i a través de l'estudi de la localització i la reactivitat sota el microscopi de tres proteïnes (GLUT3, HSP90AA1 i Cu/ZnSOD) relacionades amb la fisiologia espermàtica i amb possibles rols en la congelabilitat. El quart objectiu va ser quantificar l'expressió de les tres proteïnes per transferència western, tant en espermatozoides d'ejaculacions GFEs com en els d'ejaculacions PFEs i en els tres passos abans esmentats, per tal de determinar el seu potencial com a predictores de la congelabilitat. Pel primer i el segon objectiu, 86 truges van ser inseminades per post-CAI amb 26 ejaculacions de mascles Piétrain dividides en una porció refrigerada a 17ºC (tractament control) i una porció criopreservada, ambdues porcions classificades alhora com a GFEs o PFEs. Els resultats més rellevants van demostrar que les probabilitats d'embaràs eren dues vegades menors en inseminacions amb esperma criopreservada d'ejaculacions PFEs (P < 0.05) que en inseminacions amb esperma criopreservada d'ejaculacions GFEs, fet que indica que les ejaculacions amb percentatges elevats d'espermatozoides mòbils progressius i d'integritat de membrana (per sobre del 40% en les GFEs) són més favorables a provocar embarassos que no pas aquelles ejaculacions amb una pobra funció espermàtica in vitro (PFEs). Ni el nombre de truges que van donar a llum, ni la quantitat de garrins, ni el risc de reflux espermàtic van ser significativament diferents entre les inseminacions amb esperma criopreservada d'ejaculacions GFEs i les inseminacions control amb semen refrigerat, la qual cosa demostra la bona aplicabilitat de la inseminació post-CAI amb l'esperma criopreservada. Finalment, pel tercer i quart objectius van ser criopreservades 29 i 11 ejaculacions de mascles Piétrain, respectivament. Dos paràmetres cinètics espermàtics, la linealitat (LIN) i la rectitud (STR), van mostrar una hiperactivació de la mobilitat superior en les ejaculacions PFEs que en les GFEs després de 30 min a 5ºC durant la criopreservació. A més, la combinació d'ambdós paràmetres va donar una fiabilitat propera al 72% en la predicció de la congelabilitat de les ejaculacions porcines. Tot i que no va ser possible predir la congelabilitat mitjançant l'avaluació de les tres proteïnes al microscopi, els resultats de transferència western van revelar diferències en l'expressió de la HSP90AA1 en l'esperma a 17ºC, molt possiblement relacionades amb la millor supervivència a la criopreservació dels espermatozoides d'ejaculacions GFEs. Aquests resultats suggereixen que la promoció de la criopreservació d'esperma porcina per la seva aplicació en IA passa pel desenvolupament de tests per la predicció de la congelabilitat en semen refrigerat.