奶牛乳汁孕酮现场快速测定技术的发展

本文描述了近年来奶牛乳汁孕酮现场快速测定技术的发展概况。对测定原理和方法学作了详细论述；讨论了影响测定结果准确性的因素；同时介绍了乳汁孕酮现场快速测定技术研究的新动态及其应用前景。     

奶牛早孕诊断乳汁孕酮临界值的确定

随机选择健康荷斯坦母牛６３头，其中发情周期牛５头，配后１８－２４天牛３０头和通过直接确定的怀孕牛２８头，按程序采集末乳乳样。用放射免疫测定法（ＲＬＡ）测定乳汁孕酮（ＭＰ４）浓度。以发情第３天ＭＰ４浓度加上２倍标准差为未孕临界值的土限，以配后１８－２４天怀孕牛ＭＰ４浓度的平均值减去１倍标准差为怀孕临界值的下限。结果表明，奶牛ＭＰ４早孕诊断临界值为：≥７．１９ｎｇ／ｍｌ为怀孕，〈４．７２ｎｇ／ｍｌ为未     

高产奶牛产后乳汁孕酮含量变化与卵巢机能恢复和子宫复旧的关系

高产奶牛产后乳汁孕酮含量变化与卵巢机能恢复和子宫复旧的关系刘崇立，王运亨，周玉云（北京市西郊农场１０００９４）李强（北京市北郊农场１００２０９）于德洪（北京奶牛中心１０００８５）前言高产奶牛产后卵巢机能恢复延迟引起的“乏情”或异常发情，是造成繁殖力下...     

隐性乳腺炎影响奶牛乳汁中蛋白质含量变化的研究

隐性乳腺炎是致病因子侵入乳房组织感染,引起乳腺实质和间质组织的炎症。乳腺受到侵害直接影响到泌乳机能,进而影响奶的产量和质量。近年来,此病的危害性已引起国内许多国营奶牛场和奶牛专业户的普遍重视,给奶牛业造成较大的经济损失。据国外报道,如在美国此病每年引起的损失达10亿美元以上。     

奶牛乳汁中孕酮的浓度值与受胎率之间关系的研究

调查24头54例奶牛在人工授精后，乳汁中孕酮的浓度值与受胎率的关系，得到4种变化类型。（1）正常型：孕酮的浓度值迅速增加并维持较高水平的占59%。（2）二峰型：孕酮的浓度值在黄体中其突然下降又回升的占24%。（3）低值型：孕酮的浓度在低值俳徊的占11%。（4）迟延型：孕酮的浓度值缓慢性升的占6%。其中正常受胎的15例中14例为正常型。19-26d复发情的29例中，正常型11例，占38%；二峰型11例，占38%，低值型5例，占17%；迟延型2例，占7%。怀疑死胎10例中正常型7例占70%。由此可见正常受胎的奶牛人工授精后乳汁中孕酮的浓度值多为正常型，空怀的奶牛多为异常型。     

清宫液治疗奶牛化脓性子宫内膜炎时乳汁孕酮的变化

中西药合剂“清宫液”治疗奶牛化脓性子宫内膜炎,分析乳汁孕酮含量变化的结果表明,治疗前卵巢有黄体和无黄体的患牛,经治疗恢复正常滤泡—黄体周期的时间分别为13.8±8.4天和15.8±9.1天,均较临床发情表现早25天左右。卵巢恢复正常性周期的时间与子宫内膜炎的严重程度呈正相关。说明清宫液中药成分有明显的调节卵巢机能的作用。     

奶牛乳汁孕酮免疫检测的方法

孕酮(P4)是奶牛和黄牛在发情周期(一般为21天)中由卵巢黄体中产生的激素,可以进入血液、乳汁和唾液中。在母牛配种后21～24天期间采集奶测定孕酮浓度,可以判断牛是否受孕。发情期间奶中孕酮浓度可判断奶牛是否处于发情期。临床上,孕酮     

检测奶牛乳汁孕酮的免疫生物传感器优化

为了完善已建立的孕酮免疫生物传感器的检测性能,本试验应用碳化二亚胺法将11α-羟基孕酮半琥珀酸和OVA制备成孕酮完全抗原,其浓度达0.6 mg/m L。在此基础上建立了以乳汁为基质的孕酮生物传感器的标准曲线(y=-4.9461x+8.7982,R2=0.993 2),进一步确立了检测乳汁孕酮的线性范围为0.31 ng/m L~50ng/m L,检测限为0.31 ng/m L,灵敏度为0.15 ng/m L,而且孕酮与皮质酮、皮质醇、雄烯二酮的免疫交叉反应较低,表明特异性较好。批内、批间变异系数分别为7.7%和7.5%,回收率为95.8%~115.7%,生物传感器稳定性在4℃保存4 d。该优化的奶牛乳汁孕酮免疫生物传感器的性能参数符合规定的标准,为检测妊娠、乏情和繁殖障碍疾病的奶牛乳汁孕酮含量提供了工具。     

A new dried milk sampling technique and its application for progesterone detection in cows

A new method for milk sample collection and storage, based on a dried milk sampling technique, is proposed. The method includes application of a whole milk sample to a porous membrane followed by drying. One hundred whole milk samples (dried and liquid) taken on day 21 post insemination were analysed for progesterone by ELISA and results for both dried and liquid samples were well correlated (r = 0.911). Milk progesterone ELISA accuracy for pregnancy diagnosis in cows was 87%.     

Effects of time of progesterone supplementation on embryo development and interferon-tau production in the cow

We have investigated the effects of the timing of progesterone supplementation on early embryo development in mature, non-lactating Holstein-Friesian cows. Animals were inseminated 72 h (day 1) and 96 h following prostaglandin injection and were either left as untreated controls (n=6) or received progesterone supplementation from either days 5 to 9 (early; n=6) or from days 12 to 16 (late; n=6). Daily plasma samples were collected until day 16, when cows were slaughtered and reproductive tracts recovered and flushed to collect embryos and to measure interferon-tau activity. Both early and later progesterone supplementation resulted in marked increases in plasma progesterone (P<0.01). Early, but not late, progesterone supplementation resulted in a fourfold increase in trophoblast length (P<0.01) and a sixfold increase in uterine concentration of interferon-tau (P<0.05). The results demonstrate that progesterone supplementation during the postovulatory rise, but not later in the luteal phase, increases embryo development and interferon-tau production.     

Culling of dairy cows. Part III. Effects of diseases, pregnancy status and milk yield on culling in Finnish Ayrshire cows

The effects of 15 diseases, pregnancy status and milk yield on culling were studied in 39 727 Finnish Ayrshire cows that calved in 1993 and were followed until culling or next calving. Survival analysis, using the Cox proportional hazards model, was performed with diseases, pregnancy status and milk yield as time-dependent covariates. Effects of parity, calving season and herd were also accounted for.

Pregnancy status was the single most influential factor affecting culling decisions, followed by milk yield. Several diseases also had a significant effect on culling, the most influential ones being mastitis, lameness, teat injuries, and milk fever. The effects of all of these factors varied according to the stage of lactation.

Milk yield had a significant effect on culling decisions, depending on the stage of lactation. At the beginning of lactation, milk production did not have any effect on culling decisions, but later on, the highest producers were at the lowest risk of being culled and the lowest producers had the highest risk. Adjusting for milk yield modified the effects of parity, most diseases and also pregnancy status on culling. Effects of parity increased after including milk yield in the model, indicating that milk yield and parity are interrelated in their effects on culling. The effects of pregnancy status also increased towards the end of lactation when milk yield was accounted for in the model. The effects of mastitis, teat injuries and lameness decreased after adjusting for milk production. These diseases lower milk yield and thus, part of their effect on culling was mediated through milk production. The effects of anestrus and ovarian cysts were mainly modified by pregnancy status, but not by milk yield. The effects of milk fever on culling increased at the beginning of lactation after including milk yield in the model. This suggests that even though cows with milk fever tend to be higher producers, it is the disease as such that triggers the culling decision early in the lactation. The changes in the effects of other diseases after adjusting for milk yield varied, depending on the disease and the stage of lactation.     

traT and CNF2 genes of Escherichia coli isolated from milk of healthy cows and sheep

The objectives of the present study were to isolate Escherichia coli from milk of apparently healthy cows and sheep and to investigate the presence of traT and cytotoxic necrotising factor-2 (CNF2) virulence genes by multiplex polymerase chain reaction (PCR). Milk samples collected from a total of 1028 apparently healthy ruminants (737 cows and 291 sheep) in eastern Turkey were streaked onto 5% sheep-blood agar. E. coli was isolated and identified by biochemical tests in 5.9% (61/1028) of milk samples. Correct amplification with the molecular length of 232 bp was obtained from all E. coli isolates by the species-specific PCR. The isolation rates of the agent were calculated to be 7.6% (56/737) in cows and 1.7% (5/291) in sheep. The difference between these proportions was statistically significant (p < 0.001). Multiplex PCR showed that traT and CNF2 genes were present in 62.3% and 6.6% of all isolates, respectively. Both genes were present in 16.4% of the isolates, with only 14.7% having neither gene.     

Radioimmunoassay of milk progesterone as a tool for fertility control in smallholder dairy farms

This study focused on the use of radioimmunoassay of progesterone in milk for the diagnosis of post-partum ovarian cyclicity and accurate detection of oestrus and non-pregnancy in cows in the artificial insemination (AI) programme in Bangladesh. In Investigation 1, milk samples were collected on day 0 (day of AI), day 9–13 and day 21–24 from 444 milking cows of various breeds presented for the first post-partum insemination by 413 farmers living at 182 villages/regions in Mymensingh District from 6 AI centres and sub-centres. Each cow was then examined three times after each AI until it stopped returning to oestrus. Sixty to 90 days after the last AI, the cows were examined per rectum to confirm the pregnancy. Milk progesterone data on day 21–24 contributed to a clear diagnosis with respect to non-pregnancy in 100% cows, indicating a possible use of this progesterone assay for identifying non-pregnant cows in AI programmes. In Investigation 2, milk progesterone was monitored two times in a month with a 10-day interval in 88 cows. The samples were taken between 10 days after calving and the first detected oestrus, followed by two more samples 10 days apart. The proportion of cows accurately detected in oestrus was 30%. Another 30% were stated to be in oestrus when they were not (false positive) and 40% were not detected when they were in oestrus (false negative). The mean intervals between calving and oestrus and between calving luteal activity were 40 to 362 days (median = 120, n = 82) and 34 to 398 (median = 111, n = 64) days, respectively. The body condition scores at calving and at the initiation of luteal activity influenced the interval between calving and luteal activity (p < 0.05). Cows suckled twice daily initiated luteal activity earlier than their counterparts suckled several times daily (p < 0.05). Determination of progesterone in milk on day 21–24 is a good means for detecting non-pregnant cows.     

Milk progesterone enzyme-linked immunosorbent assay as a tool to investigate ovarian cyclicity of water buffaloes in relation to body condition score and milk production

Background

Application of assisted reproductive technologies in buffaloes is limited to some extent by farmers’ inability to detect oestrus because of its poor expression. The present study aimed at investigating reliability of a milk progesterone enzyme-linked immunosorbent assay (ELISA) to assess the ovarian cyclicity during post partum, oestrus and post-breeding periods in water buffaloes.

Methods

Progesterone concentrations were measured by an ELISA in milk of 23 postpartum buffaloes in relation to oestrus, pregnancy, body condition score (BCS) and milk production. Two milk samples were taken at 10 days intervals, every month starting from day 30 and continued to day 150 post partum. BCS and milk production were recorded during sample collection. Milk samples from bred buffaloes were collected at Day 0 (day of breeding), Days 10–12 and Days 22–24. Defatted milk was preserved at −80°C until analysis. Pregnancy was confirmed by palpation per rectum on Days 70–90.

Results

Seventeen buffaloes had 47 ovulatory cycles, one to four in each, 13 were detected in oestrus once (28 % oestrus detection rate). Progesterone concentration ≥1 ng/ml in one of the two 10-day-interval milk samples reflected ovulation and corpus luteum formation. The intervals between calving to first luteal activity and to first detected oestrus varied from 41 to 123 days (n = 17) and 83 to 135 (n = 13) days, respectively. Eight buffaloes were bred in the course of the study and seven were found pregnant. These buffaloes had a progesterone profile of low (<1 ng/ml), high (≥ 1 ng/ml) and high (≥ 1 ng/ml) on Day 0, Days 10–12 and Days 22–24, respectively. Buffaloes cycling later in the postpartum period had fewer missed oestruses (P < 0.05). Buffaloes with a superior BCS had a shorter calving to oestrus interval and produced more milk (P < 0.05).

Conclusions

Milk progesterone ELISA is a reliable tool for monitoring ovarian cyclicity and good BCS may be an indicator of resuming cyclicity in water buffalo.     

Luteal Blood Flow and progesterone concentration during first and second postpartum estrous cycle in lactating dairy cows

The aim of the present study was to determine the differences in corpus luteum (CL) functionality between the first postpartum estrous cycle and the following cycle in lactating dairy cows. Luteal blood flow (LBF), luteal size and blood progesterone (P4) concentration were monitored during the first and second postpartum estrous cycle. During the first and second postpartum estrous cycle, the mean LBF value increased (p < .05) from early to late dioestrus, while it decreased rapidly in proestrus, resulting statistically lower (p < .05) than those registered in all previous phases. Statistically significant differences were not observed between overall LBF during first and second postpartum estrous cycle (p > .05). During the first postpartum estrous cycle, P4 blood concentrations showed a significant reduction (p < .05) from dioestrus to proestrus. A different trend of P4 concentrations was observed during the second postpartum estrous cycle, where mean P4 value registered in proestrus resulted statistically lower than those registered in the previous cycle phases (p < .05). The mean P4 concentration registered over the first postpartum estrous cycle resulted statistically lower (p < .05) than that registered during the second one. A significant correlation between P4 concentrations and LBF was registered only during the second postpartum estrous cycle. Results indicate that during the first postpartum estrous cycle, P4 concentration was independent of luteal blood flow and luteal size.