胚胎分割技术在肉牛繁育中的研究与应用

本试验采用弗莱维赫肉牛和日本和牛作为供体,本地黄牛为受体,进行同期发情和超数排卵,共得到可用胚胎234枚,分为4个处理:新鲜全胚113枚、新鲜双半胚55枚、冷冻全胚56枚和冷冻双半胚10枚,进行移植。结果表明,新鲜双半胚的妊娠率为65.45%,略高于新鲜全胚移植的妊娠率(63.72%),差异不显著(P＞0.05)。冷冻双半胚产犊率为20.00%,低于冷冻全胚移植的妊娠率(39.29%),差异不显著(P＞0.05),而冷冻组的妊娠率均显著低于新鲜组(P＜0.05)。新鲜双半胚的产犊率为95.56%,双犊率为35.55%,犊牛存活率为72.09%。新鲜全胚组的产犊率为63.16%,显著低于新鲜双半胚组(P＜0.05),但犊牛存活率却显著高于新鲜半胚组(90.00%和72.09%)(P＜0.05)。此外,全胚与双半胚移植后产生的犊牛初生重之间并无显著差异(P＞0.05)。     

Critical differences of clinical-chemical parameters in blood from Red Danish dairy cows.

The purpose of the present study was to calculate the critical differences between two analytical results for 23 routinely used bovine clinical-chemical parameters. The critical difference can be used to judge whether the difference between two consecutive analytical results from the same animal is due to natural variation or not. From 20 cows, blood samples were collected once daily for five days, and the interindividual variance, the intra-individual variance, and the analytical variance were calculated using nested analysis of variance. The critical difference both in absolute values and in percentages was calculated from these variances. The main conclusions were that the critical difference in percentages was in general between 10 and 40 per cent but varied from 10.6 per cent for sodium to 280.2 per cent for lactate and that the analytical variation was generally in accordance with accepted standards, although the analytical variation for alkaline phosphatase, total serum protein, urea and fructosamine should be improved.     

Morphology and size of embryos recovered from superovulated cows.

A total of 691 normal embryos were recovered from 183 superovulated donor cows on the 5th and 6th days after the first insemination, and were examined for their morphology and size in relation to their developmental stage. There was no significant difference in the thickness of the zona pellucida, the diameter of the cell mass, and the overall diameter of the embryos among zygotes, 2-, 4-, 8- and 16-cell embryos, and morulae. In the blastocyst stage, however, the diameter of the cell mass and the overall embryo diameter were significantly greater and the zona pellucida was significantly thinner than in the earlier-stage embryos. The volume of the blastomere significantly decreased from zygote to morula in proportion to the increase in the number of blastomeres. The volume of the cell mass of 2-cell embryos was decreased by about 30% compared with that of zygotes and no increase in the volume of the cell mass was observed during the progression from 2-cell stage to morula. The diameter of the cell mass and the overall diameter of morulae recovered on the 6th day after the first insemination were significantly greater than those of morulae recovered on the 5th day.     

奶牛胚胎分割与半胚裸露冷冻保存试验

本试验研究了①Ａ、Ｂ、Ｃ三个质量等级胚胎的分割效果；②五种不同冷冻方法冷冻裸露半胚的存活情况；③冻前培养３．０、１．５～２．０、１．０小时的裸露半胚存活率。结果①Ａ、Ｂ、Ｃ级胚胎的分割成功率分别为９５．８％（９２／９６）、７且．４％（７０／９８）、３０％（６／２０），三者间差异极显著（Ｐ＜０．０１）；②半胚冷冻前以２０％ＦＣＳ－ＰＢＳ液培养３．０小时后，以五种不同冷冻方法冷冻，均未获得存活半胚；③半胚冷冻前培养３．０、１．５～２．０、１．０小时后，以添加１０％而聚糖的１０％甘油＋０．１Ｍ蔗糖／ＰＢＳ液冷冻，分别获得了０％（０／４５）、６８．８％（１１／１６）、７５．０％（９／１２）的存活率。结果表明，胚胎质量是影响胚胎分割成功率的关键因素，半胚冷冻前的培养时间是影响裸露半胚存活的重要因素。     

Loser cows in Danish dairy herds: definition, prevalence and consequences

During the last few years, many Danish dairy farmers have expressed increasing concerns regarding a group of cows, which we have chosen to term ‘loser cows’. Until now, a loser cow has not been described scientifically. We defined a loser cow on the basis of a clinical examination of the cow. A total of 15,151 clinical examinations were made on 6,451 individual cows from 39 randomly selected, large Danish dairy herds with loose-housing systems using a clinical protocol. Scores for the clinical signs lameness, body condition, hock lesions, other cutaneous lesions, vaginal discharge, condition of hair coat and general condition were converted into a loser cow score. Cows with a loser cow score of 8 or more were classified as loser cows. The overall prevalence of loser cows was 2.15%, 4.50% and 2.98% during the first, second and third round of herd visits, respectively.

The associations between the loser cow state and milk production, mortality, morbidity, culling and workload for the farmer were evaluated using data from herd visits and from the Danish Cattle Database and a number of different statistical techniques. It was concluded that the loser cow state has significant negative consequences for both the farmer and the cow. On average, loser cows yielded 0.61 to 2.24 kg energy corrected milk less per day than non-loser cows depending on parity. Hazard ratio for death or euthanasia was 5.69 for loser cows compared to non-loser cows. Incidence rate ratio for disease treatments was 0.69 for non-loser cows compared to loser cows. Loser cows were often culled in an ‘unfavourable’ way and generally caused extra workload for the farmer.

A simplified version of the loser cow score was evaluated and is recommended for future research and use in practice.     

奶牛牛病毒性腹泻病毒与冠状病毒混合感染的PCR诊断

为了对河北省石家庄市某奶牛场发病及死亡奶牛进行确诊,试验对腹泻奶牛的粪样进行了牛病毒性腹泻病毒(BVDV)和牛冠状病毒(BCoV)PCR检测,同时对病死奶牛脏器样品进行BVDV、BCoV、传染性鼻气管炎病毒(IBRV)、牛轮状病毒(BRV)及牛细小病毒(BPV)的PCR检测。试验结果显示,6份粪样BVDV阳性率为50%,BCoV阳性率为33.3%,同一份粪样中可同时检测出BVDV和BCoV两种病毒。小肠、肺脏和肝脏中均检出BVDV和BCoV,未检测出IBRV、BRV和BPV。结果表明,BVDV和BCoV为阳性,IBRV、BRV和BPV结果为阴性。最后确诊为BVDV和BCoV的单一感染和混合感染。     

Effect of feeding on reproductive performance in Finnish dairy cows

The effect of 2 types of feeding on the involution of the genital tract and on the fertility was studied in 79 Finnish dairy cows on a research farm. The cows were fed twice a day with home-produced feed in accordance with the Finnish feeding standard. The cows were divided according to the type of feed into a hay-urea group and a silage group. The cows were examined clinically by rectal palpation 3 times a week during 8 weeks postpartum. The time required for the complete involution of uterus and cervix was recorded. The cows in the silage group had a significantly longer time in uterine involution, a lower fertility rate at first insemination and a longer interval from calving to conception than those in the hay-urea group. The pregnancy rate in the first insemination was in the hay-urea group 91% and in the silage group 57%. The interval between calving and the first insemination was shorter in the hay-urea group than in the silage group: 71.4 +/- 10.6 and 74.0 +/- 10.3 days, respectively. The interval between calving and conception was in the hay-urea group significantly (p less than 0.01) shorter than in the silage group; 74.8 +/- 15.1 and 89.5 +/- 24.4 days, respectively.     

Track way distance and cover as risk factors for lameness in Danish dairy cows

This study investigates the effect of length and cover of track ways between barn and pasture on lameness in Danish dairy cows. We hypothesised that short track distances would be associated with a lower lameness probability of dairy cows compared to longer distances and that track ways with prepared cover (asphalt, gravel, slag, concrete, rubber) compared to no prepared cover (sand, soil and/or grass) would be associated with a lower lameness probability of dairy cows in grazing herds.     

高产奶牛繁殖性能低的原因分析

在过去的几十年里,随着奶牛产奶量增加,奶牛的繁殖能力出现下降现象。人们从包括遗传学、生理学、营养学和管理学等方面去努力解释这一现象,并从动物、器官和细胞的角度进行研究。本文综述了影响奶牛繁殖性能的原因和后果。     

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis.

METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present.

RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected.

CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis.

CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.     

Direct enzyme immunoassay of fecal estrone derivatives in dairy cows

The present study was undertaken to develop a novel, practical and simple procedure for enzyme‐linked immunosorbent assay of fecal estrone derivatives (estrone, estrone sulfate and estrone glucuronide) in dairy cows. Fecal solution, prepared by mixing feces with borate buffer, was applied directly to wells without extraction, and incubated with anti‐estrone antibody and horseradish peroxidase‐labeled estrone. Estrone sulfate was used as the standard. The sensitivity of the assay was estimated as 0.15 ng/mL (0.6 ng/g). The intra‐assay and inter‐assay coefficients of variation were 5.3–8.1% and 13.4–15.7%, respectively. The recovery rate was 78–102%. Only 4 h were needed to complete an assay to measure fecal estrone derivative concentrations. A significant positive correlation was established between plasma estrone sulfate concentrations and fecal estrone sulfate equivalent concentrations. When fecal estrone sulfate equivalent concentrations were measured in pregnant dairy cows, a gradual increase from day 150 of pregnancy, and subsequent drastic increases from day 240 to calving date were observed. These results suggest that the direct enzyme immunoassay procedure developed in the present study is a practical and reliable method for measuring fecal estrone derivative concentrations.     

源于奶牛蹄部感染的牛源污蝇解壳杆菌的分离鉴定与药敏试验

为确定导致江苏某奶牛场蹄部感染的病原,采集发病奶牛蹄部感染病灶的病料进行细菌分离培养,使用MAL-DI Biotyper微生物快速鉴定系统进行细菌鉴定,并对分离菌株进行药物敏感性试验.结果显示,病灶中的分离株为污蝇解壳杆菌(Wohlfahrtiimonas chitiniclastica),血平板上呈β型溶血.药敏试验...     

Antioxidant status and biomarkers of oxidative stress in bovine leukemia virus-infected dairy cows

Bovine leukemia virus (BLV) is among the most widespread livestock pathogens in many countries. Despite advances in understanding the pathogenesis of this disease, little is known about the involvement of oxidative stress. Therefore, this study examined the antioxidant status and the markers of oxidative stress in BLV-infected dairy cows. BLV infection was associated with an increase in triacylglycerol levels, a decrease in glutathione peroxidase (GSH-Px) activity and a tendency toward lower superoxide dismutase activity in the infected animals. No significant difference was observed in other markers of oxidative stress (i.e., conjugated dienes, hydroperoxides and malondialdehyde) in the infected animals compared to controls. A novel method for the analysis of oxidative stress, Z-scan based on the measurement of the mean-value of θ in low density lipoprotein indicated that the infected animals had low-density lipoprotein particles that were slightly less modified than those from the healthy group. Thus, we conclude that BLV infection is associated with a selective decrease in GSH-Px activity without any alteration in the common plasma markers of oxidative stress.     

A method for preparation of chromosomes from bovine zygotes and blastocysts

A simple technique for making chromosome preparations from zygotes and early blastocysts is described. The morphological features of blastocysts and total number of cells greatly influence the quality of the preparation.     

Prediction of enteric methane production from dairy cows

Abstract

The main objective of this study was to elucidate the potential for prediction of enteric methane (CH₄) emissions from dairy cows by using predicted rumen plus hindgut digested (fermented nutrients) and total tract digested nutrients (by using NorFor) as input variables. Twenty-one experiments (78 dietary treatments) were collected. The data-set was used to develop prediction models and to test their and extant models ability to predict enteric CH₄ emissions. Models were compared based on mean squared prediction error and concordance correlation coefficient (CCC) analysis. Fermented nutrients did not predict enteric CH₄ emissions adequately (CCC < 0.420). Including total digested (td) nutrients in the model [CH₄ (MJ/d) = ?2.13 + 1.64 tdOM (kg/d) ?9.74 tdFat (kg/d) + 1.64 tdNDF (kg/d)] predicted enteric CH₄ emissions more precisely (CCC = 0.733), and showed an improvement in the prediction of enteric CH₄ emissions over the extant models tested.     

Bactericidal activity of standard bovine serum against coliform bacteria isolated from udders and the environment of dairy cows.

Coliform organisms were isolated from bedding, udders of cows exhibiting acute or chronic mastitis, and milk of cows tested for udder pathogens. The bactericidal activity of standard bovine serum was tested on 362 isolates, of which 2.4% were killed at a serum dilution of less than 1:5; 2.7% were killed at a dilution of 1.5; 3.0% were killed at a dilution of 1:12.5; and 4.4% were sensitive to a dilution of 1:25. Organisms isolated from mastitic udders were not killed by serum. Sensitivity to agglutination in 1:500 solution of acriflavine was tested on 157 isolates, and only 54 had any degree of agglutination. Of these 54, only 4 organisms had any susceptibility to killing by serum. Results suggest that most organisms in the environment of the cow are resistant to serum, few serum-sensitive coliforms cause mastitis, and agglutination in acriflavine (indicating organisms in the rough phase) does not correlate with sensitibity to serum.     

LRH-A3 and HCG increase pregnancy rate during timed artificial insemination in dairy cows

The objective of this study was to use luteinizing hormone-releasing hormone A3 (LRH-A3) and human chorionic gonadotrophin (HCG) to improve pregnancy rate of dairy cows during timed artificial insemination (TAI). In experiment 1, the TAI process (0 d, GnRH, 100 μg; 7 d, PGF2α, 0.4 mg; 56 hr, GnRH, 100 μg; 16 hr, AI) was applied to 160 dairy cows on 50th and 60th days after parturition respectively. In experiment 2, 320 postpartum dairy cows were treated with TAI (Group A), TAI + 25 μg LRH-A3 (Group B), TAI + 1,500 IU HCG 5 days after AI (Group C), and TAI + 25 μg LRH-A3 + 1,500 IU HCG 5 days after AI (Group D). In experiment 3, endometrial cells were treated with HCG. The results showed that TAI did not affect the pregnancy rate, while LRH-A3 and HCG increased the pregnancy rate of the cow. HCG of 5 IU/ml and 10 IU/ml increased the expressions of leukemia inhibitory factor but decreased those of interleukin-6, epidermal growth factor and vascular endothelial growth factor in endometrial cells. This study provided a plan for the use of LRH-A3 and HCG to increase pregnancy rate during TAI in dairy cows.