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Molecular epidemiology of Campylobacter fetus subsp. fetus on bovine artificial insemination stations using pulsed field gel electrophoresis.

The presence of Campylobacter fetus subspecies fetus (Cff) on bovine artificial insemination (AI)-stations can have major economical consequences. More knowledge on the epidemiology of C. fetus is needed to control Cff infections at AI-stations. We assessed the epidemiology of Cff on AI-stations and the molecular relationship between Cff strains isolated from outbreaks on AI-stations. Thirteen Cff strains (two Cff strains per outbreak and one sporadic case) isolated from bulls housed on different AI-stations were selected and compared with ten unrelated bovine and ovine Cff isolates from different geographical regions. Molecular typing by pulsed field gel electrophoresis (PFGE) with the restriction enzymes SmaI, SalI and KpnI, yielded unique profiles for most unrelated strains but indistinguishable profiles for all isolates from the same outbreak. Computer aided analysis using a composite data set of SmaI, SalI and KpnI restriction profiles revealed separate clusters for outbreak strains. Thus, PFGE profiling of Cff strains is a valuable tool to discriminate between strains derived from separate outbreaks and to identify routes of infection.

Animals↗

Application of timed artificial insemination protocols to grazing Japanese black cattle with long open period.

To investigate the viability of timed artificial insemination (TAI) protocols in grazing Japanese Black cattle with long open period, ovarian status and progesterone and estradiol-17 beta profiles of the animals during the protocol were monitored. In 1998, prosta-glandin F(2a) (PG) was administered to 36 animals seven days after GnRH injection. Three out of the 36 animals were inseminated after detection of estrus and did not receive further treatment. The second GnRH was injected to the remaining 33 animals 48 hr after the PG injection and TAI was performed 24 hr later. In 1999, PG was injected to 25 animals six days after GnRH, the second GnRH was injected to 22 animals 48 hr after PG, and TAI was performed 16 hr later (The other three animals were inseminated before the time of TAI). The percentage of the animals with at least one functional corpus luteum and one follicle equal to or greater than 10 mm in diameter at PG injection was similar between the groups in 1998 and 1999. Likewise, the hormonal profiles were similar between the two groups. Pregnancy rates (PR) after the TAI protocols and natural mating in 1998 and 1999 were 75.0% and 88.0%, respectively. These figures were comparable to the PR obtained by conventional estrus synchronization protocols using PG (in 1995; 69.4%) or CIDR (in 1996; 59.1%). In conclusion, the TAI protocol can be applicable into grazing Japanese Black cattle with long open period.

Animals↗

[Factors influencing conception rate after synchronization of ovulation and timed artificial insemination--a review].

This review describes factors that affect conception rate after synchronization of ovulation and timed artificial insemination. Intervals of 7 days between GnRH and PGF2alpha, 48 hours to the second GnRH treatment and a further 16 to 20 hours to the timed insemination have been proven to be most effective. Conception rates (CR) increase as lactation progresses up to 100 days in milk. Primiparous cows have higher CR than older cows. Anovular cows at the start of the synchronization protocols have poor CR. These are highest for cows started in early dioestrus. While poor body condition and some post partum and post insemination health disorders have negative effects on the CR, a significant effect of postpartum chronic endometritis could not be demonstrated. High milk yield was also not shown to have a negative effect on CR in almost all studies, while the negative effect of heat stress on fertility is also found in Ovsynch cows. However, the negative effects of high milk yields and heat stress on AI submission rates are overcome by the timed insemination protocol.

Animals↗

Influence of short-term relocation and male exposure on sexual receptivity and reproduction in artificially inseminated lactating doe rabbits.

A study was conducted to evaluate the effect of short-term relocation and male exposure on receptivity rate, kindling rate and total born per litter in lactating does under an artificial insemination (AI) programme. Thirty-two, 2-month-old New Zealand White rabbits were randomly allocated to one of four treatments: (1) relocation and male exposure; (2) relocation without male exposure; (3) no relocation with male exposure; (4) no relocation without male exposure (control). Relocation and male exposure were done 8-10 h before the time of service. First insemination was when does reached 3200 g body weight and does were bred 4-13 days after parturition across parities during a 6 month reproduction period. Of all breeding records, 125 inseminations and 91 kindlings were from nursing does. The mean interval from parturition to insemination for nursing does was 10.3 days. Relocation of lactating does resulted in greater (P<0.01) receptivity rate at service (74.8%) as compared with no relocation (55%). Receptivity rate was not influenced by male exposure. However, the interaction of relocationxmale exposure tended to be significant (P=0.07). Receptivity rate in relocated does exposed to males was 62.8 and 86.7% without exposure while in non-relocated does male exposure showed no effect. Kindling rate was not influenced by relocation or male exposure. The mean total born per litter in relocated and non-relocated does was 8.05 +/- 0.33 and 7.39 +/- 0.36, respectively, but no significant difference was observed. There was no effect of male exposure on total born per litter (7.85 +/- 0.34 versus 7.59 +/- 0.34 without male exposure). However, interaction of relocationxmale exposure on this variable was significant (P=0.009). Male exposure in relocated does decreased the size of the litter (7.52 +/- 0.46 versus 8.58 +/- 0.47 without male exposure) whereas mean values in non-relocated does increased when they were exposed to males (8.18 +/- 0.52 versus 6.60 +/- 0.49). Short-term relocation improved receptivity rate and reproduction in lactating does under an artificial insemination programme. Preliminary results indicated that male exposure in non-relocated does improves the total born per litter at a similar level than relocated does without male exposure. Relocation combined with male exposure decreased receptivity rate and total born per litter as compared with relocated does without male exposure, but the reproductive performance in the former was greater as compared with those does where no relocation occurred without male exposure.

Animals↗

Changes in the localization of antigen presenting cells and T cells in the utero-vaginal junction after repeated artificial insemination in laying hens.

The goal of our present study was to observe whether the populations of antigen presenting cells (Ia+ cells) and T cell subsets (CD4+ and CD8+ T cells) change in the utero-vaginal junction (UVJ) of Rhode Island Red laying hens that showed dramatic declines in fertility after repeated artificial insemination (AI). Rhode Island Red laying hens were divided into two groups: a virgin group (R-V) and artificial inseminated group (R-AI), which was exposed to weekly AI for a period of 3 mo. Undiluted fresh semen collected from healthy Tosa-Jidori roosters, a native Japanese breed maintained in Kochi Prefecture, was used for AI. The UVJ tissues were processed for frozen sections, and Ia+ cells and CD4+ and CD8+ T cells were identified by immunohistochemistry. The Ia+ cells and CD4+ and CD8+ T cells were observed in the stroma and mucosal epithelium of UVJ in both the R-AI and R-V birds. The frequencies of them in the stroma were significantly higher in R-AI than R-V. The higher frequency of Ia+ cells in the UVJ of R-AI group indicated a greater potential capability for antigen presentation to CD4+ cells. The significant increase in CD8+ and CD4+ T cells in the UVJ of R-AI birds might be the result of a homing process of lymphocytes, which may affect sperm survivability and fertility.

Animals↗

The separation of X-and Y-spermatozoa with regard to the possible clinical application by means of artificial insemination.

With the sephadex gel-filtration method according to Steeno and Adimoelja (1974), X-spermatozoa have been successfully separated from the Y-spermatozoa in semen samples from 81 donors with the possibility of practical use with the aid of artificial insemination to promote pregnancy. A better quality of spermatozoa is obtained after this method of filtration, which will support these spermatozoa in having the potency of fertilization. There is indeed a decrease in concentration of spermatozoa after filtration, but the samples still fulfil the minimal concentration required in artificial insemination.

Cell Separation↗

Birth of offspring following artificial insemination in the common marmoset, Callithrix jacchus.

The objective of the study was to develop a method for artificial insemination (AI) in the common marmoset, a New World primate species. For AI to be successful, sperm must be deposited at an appropriate site and time in the female reproductive tract, details of which are currently not available for Callitrichid species. Epididymal sperm were deposited in the cervix of 18 marmoset monkeys (Callithrix jacchus) around the time of expected ovulation using either 3, 2, or 1 inseminations. Six out of 18 females conceived, resulting in the first reported births following AI in this species. These pregnancies show that the presence of coagulum in the vagina and the stimulus of the female reproductive tract by natural mating are not essential for effective sperm transport in this species. Although 3 different timing regimes for sperm deposition relative to ovulation were employed, no protocol was demonstrably better than the others in terms of number of conceptions. The proportions of motile, live, and morphologically normal sperm in the suspensions used for AI were comparable with published values for ejaculates from fertile male macaques. These preliminary results indicate that births are possible following AI in marmosets: the technique could be used to aid effective genetic management of the species and possibly to facilitate captive breeding of endangered Callitrichids.

Animals↗

Conception rates and serum progesterone concentration in dairy cattle administered gonadotropin releasing hormone 5 days after artificial insemination.

The objectives of this study were to determine the effect of administration of exogenous GnRH 5days after artificial insemination (AI) on ovarian structures, serum progesterone concentration, and conception rates in lactating dairy cows. In experiment 1, 23 Holstein cows were synchronized using the Ovsynch protocol. Five days after AI (day 0) cows were assigned randomly to receive either saline (saline; n=11) or 100microg GnRH (GnRH; n=12). To examine ovarian structures, ultrasonography was performed on day 1 and every other day beginning on day 5 until day 13. On days 5 and 13 blood samples were obtained to measure serum progesterone concentrations. All cows in the GnRH-treated group developed an accessory corpus luteum (CL), whereas cows in the saline group did not. Mean serum progesterone concentrations did not differ between GnRH and saline groups on day 5 (1.64+/-0.46ng/ml versus 2.04+/-0.48ng/ml). On day 13 serum progesterone concentrations were greater (P<0.05) in the GnRH group compared with saline (5.22+/-0.46ng/ml versus 3.36+/-0.48ng/ml). In experiment 2, 542 lactating cows, at two different commercial dairies, were used to test the effect of administering GnRH 5 days after AI on conception rates. Cows were synchronized and detected for estrus according to tail chalk removal. Cows detected in estrus received AI within 1h after detection of estrus. Five days after AI, cows were assigned randomly to receive either GnRH (n=266) or saline (n=276). Pregnancy status was determined by palpation per rectum of uterine contents approximately 40 days after AI. There was no effect of farm on conception rate. There was no effect of treatment as conception rates did not differ between GnRH and saline groups (26.7% GnRH versus 24.3% saline). Regardless of treatment, days in milk, parity, milk yield, and number of services had no effect on the odds ratio of pregnancy. In summary, the results of this study indicated that GnRH administered 5 days after AI increased serum progesterone by developing an accessory CL but did not improve conception rates in dairy cattle.

Animals↗

Uterine leucocyte infiltration after artificial insemination in bitches.

In the present study, polymorphonuclear neutrophils (PMN) were enumerated to evaluate acute uterine inflammation after artificial insemination in the bitch. It was concluded that the canine seminal plasma possessed an immunomodulating action. However, the most commonly used extender for freezing canine semen (Tris glucose with egg yolk and glycerol) was a potential inducer of uterine inflammation.

Animals↗

Anaphylactic reaction after artificial insemination.

BACKGROUND: Bovine seroalbumin is known as an allergen for human beings, but reactions to it in an artificial insemination procedure are much rarer. We report a case of anaphylaxis after intrauterine insemination (IUI) in which sensitization to bovine serum albumin (BSA) is demonstrated. OBJECTIVE: Report the allergy evaluation performed in a patient who suffered a severe reaction immediately after an IUI procedure. METHODS: A 33-year-old woman was referred because of an anaphylactic reaction after a second trial of IUI. She developed pruritus, abdominal pain, nausea and vomiting, bronchospasm, and generalized urticaria. She had an atopic medical history of pollen allergy and sensitization to cat epithelium. She had never had trouble with minor surgery and she usually uses latex material. She had never received heterologous sera before. Her husband's semen for the IUI was processed in a standard fluid medium called upgraded INRA B 2 (Laboratoires CCD, Paris, France), which contains amino acids, lipids, vitamins, BSA, penicillin, and streptomycin in addition to inorganic salts. RESULTS: Skin prick tests with the medium and BSA 10 mg/mL were positive. In vitro studies demonstrated an immunoglobulin E binding protein of 60 to 65 kDa and mast cells and basophil activation (CD63 expression) against BSA contained in the medium. Cutaneous and challenge tests with penicillin and streptomycin were negative. CONCLUSIONS: We consider the BSA in the semen culture medium to be the factor which triggered the anaphylactic reaction. This case supports the authors who state that media free from heterologous proteins should be used for human application, especially on atopic patients, to avoid sensitization.

Adult↗

Factors affecting pregnancy rate in artificial insemination with frozen semen during non-breeding season in Murciano-Granadina goats: a field assay.

An artificial insemination programme was carried out to study the effect of factors such as depth of semen deposition, inseminator skill, farm, sire and expression of oestrus on pregnancy rate in Murciano-Granadina (MG) goats during non-breeding season and using frozen semen. Frozen-thawed semen from six males was applied by three technicians to inseminate a total of 551 goats in 17 farms distributed throughout the Mediterranean area of Spain. Pregnancy rate was determined at 6 weeks after insemination by transabdominal ecography. Overall pregnancy rate was 57%. Farm and depth of semen deposition affected pregnancy rate, whereas the sire and the technician had no effect. The deeper the semen was deposited in the genital tract, the higher was the rate of pregnancy obtained, being greater when the catheter reached the uterus. In spite of the relevant difference observed (48.2% vs 59.0%), pregnancy rate of females not coming into oestrus until 30 h after sponge removal was not significantly different, compared with those showing oestrus during the OD procedure. In conclusion, our field assay data on AI in MG goat with frozen-thawed semen showed that post-cervical insemination presented significantly greater pregnancy rate in comparison to when semen is deposited in the vagina or in the caudal part of the cervix.

Animals↗

Artificial insemination for oligospermia. A critical review.

The study concerns the review of the literature concerning the effectiveness of artificial insemination in cases of oligospermia. Differences in AIH techniques, the lack of standardized methodology explain the wide range of discrepancies in the resulting pregnancy rates. Still, in the course of time, a few certainties have been acquires: A detailed and thorough investigation of the female partner is mandatory. The use of fresh ejaculates is today totally obsolete, even with the help of cups, intra-cervical or eventual intra-uterine insemination. The standardization of seminal parameters still remains an unsolved problem. At the present time, intrauterine insemination with capacitated sperm is the method of choice. Still a basic condition is mandatory: a minimum of 1 M motile capacitated spermatozoa is required. Timing of insemination determined by the LH peak seems the most valuable procedure. The overall benefit is low, +/- 5% conception rate per cycle, and short lived because of the strain involved in an optimalized procedure. The policy to plan 3 to 6 AIH cycles before going over to an IVF procedure is the more debatable.

Adult↗

Campylobacter fetus in artificial insemination unit and slaughterhouse bulls in Ontario.

Preputial fluid samples were collected from 90 bulls in two Ontario artificial insemination units using a penial glove swab technique previously developed by one of us for use in donor bulls. No Campylobacter fetus organisms were identified from the prepuce or from samples of semen collected at the same time from these bulls. The distal genitalia of 200 bulls were collected at a slaughter house. One isolation of a Campylobacter fetus subspecies venerealis was obtained on a culture from the fornix area of the prepuce of one of these bulls.

Abattoirs↗

Endometrial IL-1beta, IL-6 and TNF-alpha, mRNA expression in mares resistant or susceptible to post-breeding endometritis. Effects of estrous cycle, artificial insemination and immunomodulation.

Endometrial mRNA expression of the pro-inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) was assessed in mares resistant (RM) or susceptible (SM) to persistent post-breeding endometritis (PPBE). Eight RM and eight SM, were selected based on reproductive records and functional tests out of a herd of 2,000 light cross-type mares. Three experiments were done to study transcription patterns in (i) basal conditions; (ii) after artificial insemination (AI); and (iii) after administration of an immunomodulator at time of artificial insemination. Endometrial biopsies were taken during consecutive cycles: (i) at estrus, when follicles reached 35 mm and at diestrus (7 +/- 1 days after ovulation); (ii) at 24 h post-AI, with dead semen (estrus) and in diestrus; (iii) at 24 h after treatment with a Mycobacterium phlei cell-wall extract (MCWE) preparation and AI (with dead semen), and at diestrus. mRNA expression was quantitated by real time PCR. Under basal conditions, SM had significantly higher mRNA expression of all cytokines in estrus and of IL-1beta and TNF-alpha in diestrus, compared to RM. After AI, there were no differences between RM and SM in estrus; however, mRNA expression for all three pro-inflammatory cytokines was higher than under basal conditions. In diestrus, RM showed significantly lower IL-1beta and TNF-alpha mRNA expression than SM. When MCWE was administered at time of AI, no differences between cytokine induction from RM and SM were found. Globally, mRNA expression for all three cytokines correlated well among themselves when expression was high. The present study showed that (i) in basal conditions RM had lower mRNA expression of pro-inflammatory cytokines than SM with no effect of estrous cycle; (ii) AI upregulated mRNA expression for all three cytokines in both RM and SM, with persistance in diestrus in the latter; (iii) treatment with MCWE at time of AI down-regulated mRNA expression of IL-1 with significant effects in SM which behaved like RM. Immunomodulation with MCWE could be of help in restoring homeostatic local inflammatory mechanisms, thus assisting in the prophylaxis of post-breeding endometritis in mares.

Adjuvants, Immunologic↗

Synchronization of estrus in goats: the relationship between time of occurrence of estrus and fertility following artificial insemination.

The fertility rate for goats following artificial insemination (AI) is usually analyzed according to herd or treatment groups. However, these general information are insufficient to allow identification of specific factors which affect this individual reproductive performance. In the present experiment 640 dairy goats were used to analyze to what extent the interval from sponge removal to estrus affects the results of AI, performed at a predetermined time following sponge removal. Estrus occurred in 98.1% of experimental animals between 24 and 72 hours after sponge removal. The fertility rate was lower for goats that came into estrus later than 30 hours after sponge removal (33.3%, n = 108 than for goats that exhibited estrus earlier (65.0%, n = 520; P<0.001). The occurrence of late estrus is not age dependent, but it increases with the number of treatments that an individual animal has previously received. These results show that the low fertility rate observed in some herds after synchronization of estrus and AI may be related to the high proportion of goats with a late occurrence of estrus, and this phenomenon increases in animals that are treated repeatedly.

Journal Article↗

Pregnancy rates per artificial insemination for cows and heifers inseminated at a synchronized ovulation or synchronized estrus.

Two synchronization protocols were tested for lactating dairy cows and heifers. Nulliparous dairy heifers (13 to 23 mo; n = 155) and primiparous and multiparous dairy cows (60 to 289 d postpartum; n = 310) were assigned randomly to two treatments. Controls received 25 mg of PGF2 alpha and were artificially inseminated according to the a.m.-p.m. rule following detected estrus. All controls that were not detected in estrus were injected with 25 mg of PGF2 alpha at 14-d intervals until artificial insemination (AI) at a detected estrus or until timed AI at 72 to 80 h after a third sequential injection of PGF2 alpha. Treated cows and heifers received a protocol that used GnRH and PGF2 alpha to synchronize ovulation (Ovsynch). Cows and heifers that were treated with Ovsynch were injected i.m. with 100 micrograms of GnRH at a random stage of the estrous cycle. Seven days later, cows and heifers in this group received 25 mg of PGF2 alpha followed by a second injection of 100 micrograms of GnRH 30 to 36 h later. Subsequently, the treated cows and heifers received AI 16 to 20 h after the second injection of GnRH. Pregnancy rates per AI were similar (38.9% vs. 37.8%) for control cows and cows treated with the Ovsynch protocol, respectively. However, pregnancy rate per AI was greater for control heifers (74.4%) than for heifers treated with Ovsynch (35.1%). Evaluation of serum progesterone concentrations at each hormonal injection indicated that the first injection of GnRH synchronized luteal function of lactating dairy cows but not of heifers. In summary, one fixed-time AI at a synchronized ovulation provided similar pregnancy rates per AI as did AI following the a.m-p.m. rule after estrus had been induced by PGF2 alpha in lactating cows, but the fixed-time AI was not effective for heifers because of the lack of synchronization.

Animals↗

Genetic parameters of reproductive traits in Brown Tsaiya ducks artificially inseminated with semen from Muscovy drakes.

A selection experiment on maximum duration of fertility of Brown Tsaiya ducks after artificial insemination (AI) with pooled Muscovy semen has been conducted since 1992. The Brown Tsaiya ducks were divided into two lines: a control line (T) with no selection and a selected line (S). The traits measured were the number of eggs set that were laid from Days 2 to 15 after one AI (NES), the number of fertile eggs at candling (NEF), the total number of dead embryos (NED), the maximum duration of fertility (MD), and the number of hatched mule ducklings (NEH). The selected trait was NEF. Six generations with a total of 2,127 females were measured. The variance components were estimated for each line in a multiple-trait animal model, using the restricted maximum likelihood (REML) methodology, which yields estimates free of bias caused by selection and inbreeding. Estimates of the heritability and genetic correlation from the two lines were very similar. Heritabilities in the S and T lines, respectively, were 0.14 and 0.10 for NES, 0.30 and 0.26 for NEF, 0.06 and 0.09 for NED, 0.28 and 0.21 for MD, and 0.18 and 0.19 for NEH. High and favorable genetic correlations existed between NEF and MD (0.96 and 0.92), between NEF and NEH (0.86 and 0.91), and between MD and NEH (0.90 and 0.82). The results suggested that selection for NEF could improve the maximum duration of fertility, but it could be useful to check the estimates of genetic parameters in a meat-type female duck.

Animals↗

Field experiences on post-cervical artificial insemination in the sow.

The present study was performed in order to evaluate the effects of post-cervical artificial insemination (post-CAI) in eastern European continental climate with multiparous sows. The sows were randomly allocated into two groups, and were AI by using CAI with 3 x 10(9) spermatozoa per dose (group 1, n = 859) or by post-CAI, using pooled semen with 1 x 10(9) spermatozoa per dose (group 2, n = 924). Wean-to-oestrus intervals, duration of oestrus, day 24 pregnancy rates, farrowing rates, and total pigs born were evaluated. Wean-to-oestrus intervals (CAI 114.3 +/- 4.1 h; post-CAI 115.2 +/- 5.2 h), duration of oestrus (CAI 64.1 +/- 4.1 h; post-CAI 65.0 +/- 5.2 h), day 24 pregnancy rates (CAI 90.2 +/- 1.7%; post-CAI 89.3 +/- 1.8%) and farrowing rates (CAI 88.1 +/- 2.3%; post-CAI 87.8 +/- 2.9%) did not differ significantly between CAI and post-CAI inseminated sows. The total number of pigs born differed significantly (p < 0.01) between the groups (CAI 12.3 +/- 1.1; post-CAI 10.2 +/- 0.9).

Animals↗