Improved Prediction of Ovulation Time may Increase Pregnancy Rates to Artificial Insemination in Lactating Dairy Cattle

A prospective observational study was conducted in two Australian dairy herds to assess the potential for improving pregnancy rates (proportions of inseminations that result in pregnancy) to artificial insemination (AI) if the time of ovulation could be predicted with more certainty. Herd 1 calved year‐round and inseminations were performed during two periods each day. Herd 2 calved during autumn–winter and inseminations were performed only after the morning milking each day. In both herds, the AI to ovulation interval of enrolled cows was determined by trans‐rectal ovarian ultrasonography approximately 0, 12, 24 and 36 h after AI, and pregnancy was assessed by palpation per rectum 35–56 days after AI. Also, in Herd 1 vaginal electrical resistance (VER) measurements were taken at approximately 0, 12, 24 and 36 h after AI, and in Herd 2 cows were fitted with neck mounted activity meters that monitored cow activity count in 2‐h periods. There was substantial variation in the intervals from AI to ovulation within and between herds (mean ± SD 21.2 ± 10.7, n = 102; 14.7 ± 10.4, n = 100 in herds 1 and 2, respectively). Pregnancy rates were higher for inseminations close to, but preceding, ovulation. Using combined herd data (n = 202), the highest pregnancy rate (50.8%) was observed for inseminations between 0 and 16 h before ovulation, a period in which only a modest proportion of inseminations (31.2%) occurred. In contrast, pregnancy rate was significantly lower (28.7%; risk ratio 0.6; 95% CI 0.4–1.0; p = 0.039) for inseminations between 16 and 32 h before ovulation, a period where the highest proportion of inseminations (53.2%) occurred. Thus pregnancy rates could potentially be improved if a greater proportion of inseminations were conducted shortly before ovulation. In Herd 1, mean VER during the peri‐ovulatory period varied with time from ovulation. Lowest values (mean ± SEM, VER = 64.8 ± 1.2, n = 55) occurred approximately 18 h before ovulation and were significantly lower than measurements approximately 6 h before ovulation (67.4 ± 1.0; n = 73; p = 0.003). Further work is required to determine if VER can be used to identify ovulation time and hence the optimal time to inseminate in individual animals. In Herd 2 a modest proportion of inseminations (26.9%) occurred between 24 and 40 h after the onset of increased cow activity where the highest pregnancy rate (67.9%) was observed, whereas a significantly lower pregnancy rate (42.4%; risk ratio 0.6; 95% CI 0.4–0.9; p = 0.036) was observed for inseminations between 8 and 24 h after the onset of increased cow activity where the highest proportion of inseminations (56.7%) occurred. Thus cow activity monitoring may be useful to identify the optimal time to inseminate cows. Results from this study indicate that improved methods of ovulation prediction may allow better insemination timing relative to ovulation and consequently increased pregnancy rates.     

Fertility in Dairy Cows After Artificial Insemination Using Sex‐Sorted Sperm or Conventional Semen

The aim of this study was to compare pregnancy per artificial insemination (P/AI) after timed AI with sex‐sorted sperm (SS) or conventional semen (CS) in lactating dairy cows. Cyclic cows (n = 302) were synchronized by Ovsynch and randomly assigned into two groups at the time of AI. Cows with a follicle size between 12 and 18 mm and clear vaginal discharge at the time of AI were inseminated with either frozen‐thawed SS (n = 148) or CS (n = 154) of the same bull. A shallow uterine insemination was performed into the uterine horn ipsilateral to the side of probable impending ovulation. Pregnancy per AI on Day 31 tended (p = 0.09) to be less for SS (31.8%) than CS (40.9%). Similarly, P/AI on Day 62 was less (p = 0.01) for cows inseminated with SS (25.7%) compared with CS (39.0%). The increased difference in fertility between treatments from Days 31 to 62 was caused by the greater (p = 0.02) pregnancy loss for cows receiving SS (19.2%) than CS (4.8%). Cow parity (p = 0.02) and season (p < 0.01) when AI was performed were additional factors affecting fertility. Primiparous cows had greater P/AI than multiparous cows both on Day 31 (41.7% vs 25.0% in SS and 53.0% vs 31.8% in CS groups) and on Day 62 (33.3% vs 20.5% in SS and 48.5% vs 31.8% in CS groups). During the hot season of the year, P/AI on Day 31 was reduced (p = 0.01) in the SS group (19.6%) when compared with the rates during the cool season (38.1%). In conclusion, sex‐sorted sperm produced lower fertility results compared to conventional semen even after using some selection criteria to select most fertile cows.     

Repeatability and Heritability of Ovulation Number and Embryos in Dam-daughters Pairs in Superovulated Holstein–Friesian Cows

Holstein-Friesian dams (n = 28) and daughters (n = 28) were superovulated (total number of observations was 235) to determine the repeatability and heritability of ovulation number and embryo collection result for FSH treatment. The donor cows were superovulated with FSHp, artificial insemination was performed and embryo collection was carried out 7 days later. For the analysis, the raw corpdata of the number of corpora lutea (CL), the number of collected embryos (EM) and their log-transformed values were used (log CL, log EM). The genetic parameters were calculated by using the VCE4 software. For calculating heritability, the number of embryo collection was used as a random effect; for calculating repeatability, the permanent environment was fitted. The additive genetic variance of CL was 8.91 and that of the EM was 9.23. The additive genetic variance for the log CL and log EM were 0.457 and 0.340, respectively. The estimated heritability for CL and EM were 0.234 and 0.159, and repeatability were 0.386 and 0.301, respectively. Higher heritabilities but lower as the previous repeatabilities were observed for the log-transformed data, 0.266, 0.194 and 0.294, 0.208 for log CL and for log EM, respectively.     

Comparison of Computer-assisted Sperm Motility Analysis Parameters in Semen from Belgian Blue and Holstein–Friesian Bulls

Subjective microscopic sperm motility results have recently been demonstrated to differ between Holstein-Friesian (HF) and Belgian Blue (BB) bulls. However, such assessments are rather imprecise. In the present study, sperm motility was assessed objectively by means of the Hamilton Thorne CEROS version 12.2c computer-assisted sperm motility analyser (CASA), and differences between the BB and HF breed could also be demonstrated. Higher percentages of both totally (p < 0.0001) and progressively (p < 0.0001) motile spermatozoa were encountered in the HF breed compared with the BB breed. Furthermore, a lower kinetic efficiency of the BB spermatozoa, evidenced by a lower beat cross-frequency (p = 0.0007) combined with a higher lateral head displacement (p = 0.0015), was the basis for the lower velocity of BB sperm cells. Additionally, BB spermatozoa move less straight forward, resulting in a lower straightness (p < 0.0001). No sperm motility differences were observed between age groups within the BB breed. The breed differences were observed in the examined bull populations residing at AI centres, in Belgium for the BB bulls and in the Netherlands for the HF bulls. However, these bull populations are selected for fertility. A similar pattern was observed in an unselected bull population of both breeds, although these differences were mostly non-significant for the different CASA parameters. Nevertheless, these data suggest that a genetic component might be responsible for the observed sperm motility breed differences.     

用荷斯坦奶牛性控精液生产体内性控胚胎

采用荷斯坦奶牛X性控冻精和超数排卵技术结合生产奶牛性控胚胎,以建立高效、低成本的体内性控胚胎生产方法。研究发现,使用CIDR和Cue-Mate进行奶牛同期发情处理,发情率分别为90.9%和100%(P<0.05),且Cue-Mate的发情时间更加集中;加拿大的FOLLTROPIN-V和中国科学院动物研究所的FSH对奶牛超排效果影响不显著,分别为头均胚胎数10.08枚±8.08枚、头均可用胚胎数2.38枚±3.07枚和头均胚胎数6.57枚±4.31枚、头均可用胚胎数2.00枚±1.88枚,无显著差异(P>0.05);奶牛发情后12h～14h和16h～19h人工授精,分别获得头均胚胎数10.25枚±5.53枚、头均可用胚胎数5.33枚±4.52枚和头均胚胎数10.13枚±6.93枚、头均可用胚胎数5.88枚±6.26枚,无显著差异(P>0.05)。但是未受精胚胎数差异显著(27对7,P<0.05)。     

Effect of Acute Bull Exposure Around the Time of Artificial Insemination on Serum Oxytocin and Progesterone Concentrations and Pregnancy Rates in Dairy Cows

This study examined the effect of acute bull exposure around the time of artificial insemination (AI) on oxytocin and progesterone concentrations, and pregnancy rates in dairy cows. Ninety six dairy cows, stratified according to parity into primiparous and pluriparous, were divided into three groups; short bull exposure (SBE; 10 min, n = 32), long bull exposure (LBE; 4 h, n = 32) or no bull exposure (NBE; n = 32). On day 45 post‐partum, all cows were treated with PGF2α on three occasions 11–14 days apart to synchronize oestrus. They were submitted to fixed time AI 80 h after the third PGF2α injection. Cows in the SBE and LBE groups were artificially inseminated 5 min after the introduction of the bull. From a subset of cows (n = 6 per group; three primiparous and three pluriparous), blood samples were collected once every 5 min starting 15 min before AI until 15 min after AI and analysed for oxytocin concentrations. Additional blood samples were collected for measurements of progesterone (P4) concentrations once daily for 4 days starting on the day of AI and once every 3 days thereafter until day 22. The effects of bull exposure, time, parity, difficulty of AI, and pregnancy on oxytocin and P4 concentrations were analysed using the mixed linear model procedure. Mean oxytocin concentrations or change in oxytocin concentrations after bull exposure or AI were not different among groups. Pregnancy rates for the NBE, SBE and LBE groups were 55.5%, 33.3% and 44.4%, respectively, and were not different among groups. In conclusion, acute bull exposure around the time of AI did not affect oxytocin and progesterone concentrations and did not improve pregnancy rates in dairy cattle under these farms conditions.     

OvSynch法处理进行黄牛定时人工授精研究

利用国产GnRH、PGF2α和EB等外源激素对本地黄牛进行处理，观察了OvSynch处理进行定时人工授精的繁殖效果，并探讨了卵巢状况对GnRH作用是否存在影响。结果显示：（1）各试验组同期发情率与对照组差异不显著（P＞0．05），但发情同期化程度显著高于对照组，且能够集中定时进行人工授精；（2）试验Ⅰ、Ⅱ组的第一情期受胎率分别为60．0％、58．3％，与对照组（42．9％）之间无显著差异（P＞0．05）；（3）GnRH类似物能够调控卵泡发育波，其作用不受卵巢状况的影响。试验表明，采用国产GnRH对本地黄牛进行OvSynch处理，能够达到定时人工授精且不降低受胎率的目的。     

奶牛性控冻精人工授精技术的推广效果调查研究

通过对性控冻精人工授精技术推广,并将其推广效果作了调查研究,结果利用性控冻精进行人工授精,可大幅度提高奶牛优良母畜的比例,加快牛群品质改良的进程。通过技术推广3年共受配奶牛254头,怀孕161头,受胎率平均63.8%,生产牛犊130头,生产母犊120头,生母率平均89.3%,该项技术在奶牛生产中有很大的应用空间,可提高奶牛养殖业经济效益。     

犬的采精与人工授精技术的研究与效果

目前,饲养肉用犬在我国各省区越来越多,所用公犬大多数为体型较大的圣伯纳、藏獒、高加索等品种.由于公母犬之问体型差异太大,以及犬择偶性较强,影响肉用犬的繁殖.     

First Service Pregnancy Rates Following Post‐AI Use of hCG in Ovsynch and Heatsynch Programmes in Lactating Dairy Cows

Lactating dairy cows (n = 667) at random stages of the oestrous cycle were assigned to either ovsynch (O, n = 228), heatsynch (H, n = 252) or control (C, n = 187) groups. Cows in O and H groups received 100 μg of GnRH agonist, i.m. (day 0) starting at 44 ± 3 days in milk (DIM), and 500 μg of cloprostenol, i.m. (day 7). In O group, cows received 100 μg of GnRH (day 9) and were artificially inseminated without oestrus detection 16–20 h later. In H group, cows received 1 mg oestradiol benzoate (EB) i.m., 24 h after the cloprostenol injection and were artificially inseminated without oestrus detection 48–52 h after the EB injection. Cows in C group were inseminated at natural oestrus. On the day of artificial insemination (AI), cows in all groups were assigned to subgroups as follows: human Chorionic Gonadotrophin (O‐hCG) (n = 112), O‐saline (n = 116), H‐hCG (n = 123), H‐saline (n = 129), C‐hCG (n = 94) and C‐saline (n = 93) subgroups. Cows in hCG and saline subgroups received 3000 IU hCG i.m. and or 10 ml saline at day 5 post‐AI (day 15), respectively. Pregnancy status was assessed by palpation per rectum at days 40 to 45 after AI. The logistic regression model using just main effects of season (summer and winter), parity (primiparous and pluriparous), method₁ (O, H and C) and method₂ (hCG and saline) showed that all factors, except method₁, were significant. Significant effects of season (p < 0.01), hCG and parity (p < 0.01), and a trend of parity and season (p < 0.1) were detected. A clear negative effect of warm period on first service pregnancy rate was noted (p < 0.01). The pregnancy rate was the lowest in the H protocol during warm period (p < 0.05). Treatment with hCG 5 days after AI significantly improved pregnancy rates in those cows that were treated with the H protocol compared with saline treatments (41.5% vs 24.8%; p < 0.01). O and H were more effective in primiparous than in pluriparous cows (46.1% vs 29.9%; p < 0.1 and 43.6% vs 24.6%; p < 0.01). First service pregnancy rates were higher in primiparous hCG‐treated than in pluriparous hCG‐treated cows (57.9% vs 32.3%; p < 0.01). The pregnancy rate was higher for the hCG‐treated cows compared with saline‐treated cows during warm period (37.9% vs 23.6%; p < 0.001).     

家蚕精子顶体酶活性分析及冷冻精液的人工授精

采用改良Kennedy s法测定了家蚕精子顶体酶活性,得到了适合家蚕精子顶体酶活性测定的最佳条件,即:孵育温度为25℃,孵育时间为4 h;精液体外4℃存放随时间延长其顶体酶活性呈下降趋势,酶活性的测定应在15 h内进行。虽然冷冻精液随着冷冻时间的延长其酶活性下降,但是不同冷冻方式之间酶活性变化趋势不同,-80℃保存时酶活性直线下降,120 d后酶活性下降了61%。液氮中保存的精液虽然在30 d后的酶活性为冷冻前的58%,但在其后的几个月中酶活性下降不明显。此外,顶体酶活性的高低与精液人工授精产卵数及受精卵率相关,酶活性越低其产卵数越少,受精卵率也越低,提示顶体酶活性的高低可以在一定程度上反映精液的品质。     

Pregnancy Rate Obtained with Short-term Protocol for Timed Artificial Insemination in Goats

Recent studies demonstrated that the Short-term Protocol of 5 days of progestogen treatment plus one dose of prostaglandin F(2alpha) and equine chorionic gonadotrophin (eCG) resulted in a close synchronized ovulation (60 h after the end of treatment approximately). In addition, oestradiol benzoate (ODB) is effective in synchronizing ovulation in goats and could be an alternative to eCG. This study was performed to determine the pregnancy rate using the Short-term Protocol comparing: (i) two different moments of timed artificial insemination (TAI) after eCG treatment (trial 1) and (ii) ODB as an alternative to eCG treatment (trial 2). In trial 1, 250 IU of eCG was given at the end of progestogen exposure, and cervical TAI with fresh semen was performed 48 h (n = 156) or 54 h (n = 168) later. In trial 2, 250 IU of eCG was given at sponge withdrawal (eCG group, n = 154) or 200 mug of ODB was given 24 h later (ODB group, n = 119). TAI was performed 54 h after the end of progestogen treatment. Pregnancy rate was determined by transrectal ultrasonography. In trial 1, the pregnancy rate for goats with TAI performed at 54 h (107/168, 63.7%) was higher than for those with TAI performed 48 h (77/156, 49.4%; p < 0.05) after sponge withdrawal. In trial 2, pregnancy rate was higher in eCG (94/154, 61.0%) than in ODB (49/119, 40.3%; p < 0.05) treated goats. In conclusion, the highest pregnancy rate was achieved using Short-term Protocol associated with eCG and TAI performed 54 h after treatment.     

哺乳牦牛同期发情/定时授精技术试验研究

本研究旨在对不同繁殖体况的母牦牛在补饲的基础上,分别从营养、隔离断奶、激素选择考察影响母牦牛繁殖率及同期发情处理效果。选用青海省环湖地区480头5～10岁繁殖体况不同的母牦牛,按照试验要求将母牦牛分成不同的试验组别,采用相同的同期发情处理方案,通过不同程度的补饲,补饲组与对照组的排卵率和妊娠率对比差异显著（P〈0.05...     

奶牛“同期发情-定时输精”技术综述

奶牛发情鉴定在规模化奶牛养殖中是一个难题,容易因发情鉴定不及时而错过配种期,延长产犊间隔,降低繁殖效率。使用同期发情、同期排卵和定时输精技术可以有效地克服这一缺点,提高繁殖效率,便于配种员的操作。本文对同期发情-定时输精技术的原理和国内外在奶牛上的研究作一综述,供奶牛场有关技术人员参考。     

Successful Production of Offspring Using Cryopreserved Sperm via Nonsurgical Artificial Insemination in Rats

In rats, artificial insemination (AI) is surgically performed as a general tool to obtain offspring using cryopreserved spermatozoa. Nonsurgical AI is a more desirable technology because it does not require any surgical procedures. However, there has never been a successful nonsurgical AI since frozen-thawed rat spermatozoa show low motility. We show here for the first time successful nonsurgical AI in rats using oxytocin treatment. Intraperitoneal injection of oxytocin (1/800 IU) immediately before nonsurgical AI significantly increased the number of sperm collected from the oviducts compared with that without oxytocin treatment. Therefore, to obtain pups, oxytocin was intraperitoneally injected into females mated with vasectomized males, and the rats were then used for nonsurgical AI. Seven of the 12 oxytocin-treated rats became pregnant after nonsurgical AI, and 37 pups were obtained. Only one rat (1/13) without oxytocin treatment was pregnant after nonsurgical AI, and only 1 pup was delivered. These results show success for the first time in obtaining offspring using frozen-thawed rat spermatozoa via nonsurgical AI. Our results also suggest the possibility that oxytocin treatment is effective for improvement of nonsurgical AI even in other species.     

张掖市甘州区能繁母牛零费用配种情况实地调查报告

[目的]为调查落实张掖市基础母牛养殖户补贴政策过程中存在的问题。[方法]张掖市畜牧管理人员随机入乡镇配种站,母牛养殖户,对甘州区基础母牛养殖户补贴政策落实情况进行走访调查,并对调查结果进行了统计、分析。[结果]表明：＂零费用配种＂政策对养殖户起到了积极作用,但同时存在政策资金不及时到位,会影响养牛生产的矛盾。[结论]＂...     

Milk Progesterone Profile at and after Artificial Insemination in Repeat-Breeding Cows: Effects on Conception Rate and Embryonic Death

The aim of this study was to investigate whether the skim milk progesterone concentrations at artificial insemination (AI) and day of rise of post-ovulatory progesterone concentration thereafter affect the conception and embryonic death rates in repeat-breeding cows. Milk samples were obtained from 96 repeat-breeding cows that failed to conceive to three or more AIs. The samples were taken from the cows at the day of AI and three times/week until day 45 post-AI. Skim milk was obtained after centrifugation and used for progesterone assay. The cows with a progesterone concentration more than 0.5 ng/ml at AI showed a significantly higher incidence of late embryonic death than those having a progesterone concentration<0.5 ng/ml at AI (p<0.01). As the progesterone level at insemination rose, conception rate declined. A negative correlation was shown between conception rate and skim milk progesterone level at AI. Of 56 cows showing a rise of progesterone to 1 ng/ml or more within 6 days after AI, 28 cows (50%) conceived. On the contrary, only eight of 39 cows (20.5%) conceived when the progesterone rose up to 1 ng/ml after day 6 post-AI. We concluded that increased progesterone concentration at the time of AI and delayed rise of progesterone post-AI might lead to decrease in fertility in repeat-breeding cows.     

Pharmacokinetic Profile of Erythromycin after Intramammary Administration in Lactating Dairy Cows with Specific Mastitis

The pharmacokinetics of erythromycin was studied in five lactating dairy cows following single intramammary infusion of 300 mg erythromycin in each of two quarters per cow with specific mastitis. Levels of erythromycin in plasma and quarter milk samples were measured by agar plate diffusion assay using Micrococcus luteus (ATCC 9341) as the test organism. Erythromycin level in plasma reached a peak concentration value (C _max) of 0.07 ± 0.01 μg/ml at 30 min; thereafter, levels declined gradually to reach 0.05 ± 0.00 μg/ml 12 h post drug administration. The pharmacokinetic profile of the drug revealed mean absorption half life (t _1/2ka) as 0.26 ± 0.05 h. The drug was eliminated slowly with elimination half-life (t _1/2β) of 13.75 ± 0.35 h and elimination rate constant (k _el) of 0.04 ± 0.00 h⁻¹. The volume of distribution based on the zero-time plasma concentration intercept of the least-squares regression line of the elimination phase (V _d(B)) was 0.032 L/kg. The drug crossed to untreated quarters also; mean drug levels of 0.20 ± 0.07, 0.23 ± 0.07, 0.17 ± 0.04, and 0.17 ± 0.04 μg/ml were found at 3, 6, 8 and 12 h, respectively. The mean drug concentration for treated quarters was measured as 22.97 ± 2.31 μg/ml milk at first milking (12 h) following drug infusion. No apparent adverse reaction was seen in cows administered erythromycin. It is concluded that following intramammary infusion erythromycin diffuses readily and extensively in various body fluids and tissues and adequate concentration is maintained in udder tissues for at least 12 h post intramammary administration. Thus, erythromycin may be recommended for local therapy of acute mastitis caused by Gram-positive bacteria in lactating dairy cows.