奶牛乳房炎自动检测技术研究进展

奶牛乳房炎是影响奶牛健康的主要疾病之一,发病率高、发病范围广、经济损失严重。目前奶牛乳房炎的检测大多是采集奶牛乳汁进行理化性质检测,该方法对检测环境有着较高要求,且检测周期长。随着信息技术的迅速发展,奶牛乳房炎的自动检测技术取得了较好的研究成果。该研究根据数据的传感器类型,从视觉传感器、自动挤奶系统与其他传感器3个方面阐述了奶牛乳房炎自动检测的研究进展。基于视觉传感器的奶牛乳房炎自动检测方法包括基于乳房热红外图像和基于乳汁图像的检测方法,该方法较大程度上保障了动物福利,但检测精度有待提升;基于自动挤奶系统（automatic milking systems, AMS）的奶牛乳房炎自动检测方法利用AMS获取乳汁信息,然后构建乳房炎检测模型,该方法检测误差较小,但成本较高;基于其他传感器的奶牛乳房炎检测方法采用独立研发的传感器获取乳汁数据,预测乳房炎发病情况,该方法操作简便,但使用不同传感器构建的检测模型精度差异较大。该文还探讨了目前奶牛乳房炎自动检测领域存在的精度低、实时性差、自动化不足等问题,并展望了该领域未来的发展趋势,以期为开展奶牛乳房炎自动检测技术与方法研究提供参考。     

奶牛隐性乳房炎便携式计算机视觉快速检测系统设计与试验

为解决奶牛隐性乳房炎难以防治的问题,构建了一种基于计算机视觉技术的快速检测系统。通过电脑与USB摄像头采集牛奶p H测试纸图像,提出了一种融合颜色特征与形态学处理的分割方法,分割化学反应区并获取RGB值,使用Foss5000牛乳体细胞分析仪得到牛奶体细胞实测值,采取幂回归法建立RGB值与牛奶体细胞数的预测模型,并基于Android技术开发了便携式移动终端设备。牛场实测20组数据试验结果显示,牛奶体细胞数估测值与实测值相关系数为0.970,估测平均相对误差为3.67%,标准差为1.88%。系统估测牛奶体细胞数较为准确,可用于奶牛隐性乳房炎快速检测。     

基于热红外图像的奶牛乳区温度分布与乳房炎识别方法

乳房炎是影响奶牛健康与牛奶品质的主要疾病之一，是健康养殖的监控重点，该研究提出了一种基于热红外图像的奶牛乳区温度分布测量与乳房炎识别方法。通过现场采集的健康与患病共189头荷斯坦奶牛挤奶前后的后乳区热红外图像样本，提出了左右后乳区自动识别方法，确定了后乳区的特征区域及识别乳房炎的数据最佳采集时间为挤奶前。通过线剖法获取奶牛特征区域内的温度值点，建立乳区温度分布拟合方程，经分析发现91.9%的健康奶牛乳区温度拟合线的斜率小于0，斜率范围为-0.083～-0.001；92.1%的患病奶牛乳区温度拟合线的斜率大于0，斜率范围为0.001～0.093，可根据温度拟合线斜率的正负实现奶牛乳房炎的自动识别。与加州乳房炎检测法（California Mastitis Test，CMT）对比试验，结果表明：健康奶牛左右后乳区的识别准确率均值为76%，患病奶牛的左右后乳区识别准确率均值为75%。该研究提出的方法为牧场奶牛健康管理与乳房炎的快速在线识别提供了参考。     

Conversion of cephapirin to deacetylcephapirin in milk and tissues of treated animals

Cephapirin is one of six beta-lactam antibiotics approved for use in the treatment of food-producing animals in the United States. When used for treatment of mastitis by intramammary infusion, it is partially converted to a microbiologically active metabolite identified as deacetylcephapirin (DACEP). The degradation was followed in four cows with naturally acquired mastitis which were treated with cephapirin. DACEP persisted longer than the parent compound in the milk. When a calf was treated with cephapirin by intramuscular injection, the compound was almost completely converted to DACEP in tissues. The deacetyl form must be considered in the determination of residues in treated animals.     

Bayesian inferences for receiver operating characteristic curves in the absence of a gold standard

Sensitivity and specificity are used to characterize the accuracy of a diagnostic test. Receiver operating characteristic (ROC) analysis can be used more generally to plot the sensitivity versus (1-specificity) over all possible cutoff points. We develop an ROC analysis that can be applied to diagnostic tests with and without a gold standard. Moreover, the method can be applied to multiple correlated diagnostic tests that are used on the same individual. Simulation studies were performed to assess the discrimination ability of the no-gold-standard method compared with the situation where a gold standard exists. We used the area under the ROC curve (AUC) to quantify the diagnostic accuracy of tests and the difference between AUCs to compare their accuracies. In particular, we can estimate the prevalence of disease/infection under the no-gold-standard method. The method we proposed works well in the absence of a gold standard for correlated test data. Correlation affected the width of posterior probability intervals for these differences. The proposed method was used to analyze ELISA test scores for Johne’s disease in dairy cattle.     

泰州市奶牛乳房炎调查及其病原菌的分离

奶牛乳房炎是奶牛场的常发病和难以防治的疾病,为进一步了解引起奶牛乳房炎的病原微生物,本研究通过对泰州市3个奶牛场572头泌乳牛的乳房炎发病率进行了调查并对乳房炎主要病原菌进行分离和鉴定。结果表明,奶牛临床型乳房炎的发病率为7.69%,隐性型乳房炎发病率为56.64%,乳区阳性率为33.8%,其中临床型：隐性=1：7.36。从65份确定患有乳房炎的奶样中共分离获得8种细菌、156株分离株,且引起乳房炎的主要病原菌是金黄色葡萄球菌和表皮葡萄球菌,其次是停乳链球菌、大肠杆菌、乳房链球菌和无乳链球菌,其检出率分别为42.31%、23.08%、8.97%、6.41%、5.13%和1.28%。该项调查结果初步明确了泰州市乳房炎发病情况,同时为进一步综合防治奶牛乳房炎和研制乳房炎治疗药物提供了科学依据。     

Effect of feeding intensity and milking system on nutritionally relevant milk components in dairy farming systems in the North East of England

There is increasing concern that the intensification of dairy production reduces the concentrations of nutritionally desirable compounds in milk. This study therefore compared important quality parameters (protein and fatty acid profiles; α-tocopherol and carotenoid concentrations) in milk from four dairy systems with contrasting production intensities (in terms of feeding regimens and milking systems). The concentrations of several nutritionally desirable compounds (β-lactoglobulin, omega-3 fatty acids, omega-3/omega-6 ratio, conjugated linoleic acid c9t11, and/or carotenoids) decreased with increasing feeding intensity (organic outdoor ≥ conventional outdoor ≥ conventional indoors). Milking system intensification (use of robotic milking parlors) had a more limited effect on milk composition, but increased mastitis incidence. Multivariate analyses indicated that differences in milk quality were mainly linked to contrasting feeding regimens and that milking system and breed choice also contributed to differences in milk composition between production systems.     

Estimation of sensitivity and specificity of three conditionally dependent diagnostic tests in the absence of a gold standard

This article presents a model to evaluate the accuracy of diagnostic tests. Data from three tests for the detection of EF-positive Streptococcus suis serotype 2 strains in sows were analyzed. The data were collected in a field study in the absence of a gold standard, that is, the true disease status (noninfected or infected) of the tested animals was unknown. Two tests were based on a polymerase chain reaction (PCR); one test was applied to a tonsil swab (taken from the live animal), and the other test was applied to the whole tonsil (collected at slaughter). The third test was based on a bacterial examination (BE) of the whole tonsil. To reduce experimental cost BE was performed only for a subset of the animals in the sample. The model allows for dependence between tests, conditional upon the unknown true disease status of the animals. Accuracy was expressed in terms of sensitivity and specificity of the tests. A Bayesian analysis was performed that incorporated prior information about the accuracy of the tests. The model parameters have a simple interpretation and specification of priors is straightforward. Posterior inference was carried out with Markov chain Monte Carlo (MCMC) methods, employing the Gibbs sampler, as implemented in the WinBUGS program. Different parameterizations to allow for selection and missing values, use of different priors, practical problems in the analysis, and some interesting issues in a joint analysis of the binary (positive or negative) results of PCR and BE and two additional continuous enzyme-linked immunosorbent assays (ELISA) are discussed.     

奶水牛体型线性性状与其第一胎305 d产奶量的相关与通径分析

对广西水牛研究所共84头奶水牛的体型线性性状与其第一胎305 d产奶量进行相关与通径分析。结果显示:奶水牛体型线性性状中,尻长与摩拉水牛、尼里-拉菲水牛、三品杂水牛等三个品种奶水牛的第一胎305 d产奶量均有较强的相关,分别是0.3141、0.3343和-0.2184;此外,摩拉水牛的体高(-0.3410),尼里-拉菲水牛的后房高(0.3847)、后房宽(0.4750),三品杂水牛的后房高(-0.2852)、乳房深(0.2662)与其对应的第一胎305 d产奶量有较强的相关。结果表明:奶水牛体型线性性状与其第一胎305 d产奶量存在一定的相关,但其影响在不同品种间有很大的差异;结果也可能表明对奶水牛体型线性性状的选择标准和要求不同于普通奶牛。     

Evaluation of the Accuracy of Diagnostic Tests From Repeated Measurements Without a Gold Standard

A model is presented to evaluate the accuracy of diagnostic tests from data from individuals that are repeatedly tested in time. Repeated measurements from three diagnostic tests for foot-and-mouth disease, applied to vaccinated and experimentally infected cattle, were analyzed. At any time the true disease status of the individuals was unknown, i.e., no gold standard was available. The model allows for correlation between repeated test results, in consequence of the underlying structure for the unknown true disease status, but also by the distribution of the test results conditional upon true disease status. The model also allows for dependence between the different diagnostic tests conditional upon true disease status. Prior information about the structure of the prevalence and the specificity of the tests was incorporated in a Bayesian analysis. Posterior inference was carried out with Markov chain Monte Carlo. Simulated data were analyzed to gain insight into the performance of the posterior Bayesian inference. The simulated data are typical for the expensive and, therefore, modestly sized infection experiments that are conducted under controlled conditions.     

中国荷斯坦牛κ-酪蛋白基因两SNP位点基因型组合对产奶性状的影响

研究κ-酪蛋白基因单核苷酸多态性及其组合与中国荷斯坦牛泌乳性状的关联性,为进一步加快高乳蛋白及高乳脂率奶牛育种进程提供参考依据。采用PCR-RFLP、CRS-PCR和DNA测序技术,对435头中国荷斯坦牛的κ-酪蛋白基因的外显子4和5上两个SNP位点进行了研究,计算其基因频率和基因型频率,确定其基因型组合,并对其基因型和基因型组合与泌乳性状进行了关联分析。结果发现,在435头试验牛群中,共有8种基因型组合。单个SNP位点关联分析表明：T10996A突变位点3种基因型间TA基因型个体乳脂率极显著高于AA基因型个体(P＜0.01), TA基因型个体乳蛋白率显著高于AA基因型个体(P＜0.05)。T12980C突变位点的3种基因型间TC基因型个体乳脂率极显著高于TT基因型个体(P＜0.01),TC和CC基因型个体乳蛋白率显著高于TT基因型个体(P＜0.05)。T10996A/T12980C两种基因型组合分析中,在乳脂率上,TTCC基因型组合个体最高,TATC基因型组合个体位居第三位;在乳蛋白率上,TATC基因型组合个体显著高于AATT基因型组合个体(P＜0.05)。【结论】κ-酪蛋白基因不同基因型和基因型组合与中国荷斯坦牛乳脂率和乳蛋白率存在相关性,但与产奶量和脂蛋白比不相关。     

Aroma compounds of fresh milk from New Zealand cows fed different diets

Volatile compounds were extracted from fresh milk produced by New Zealand cows using the newly developed solvent-assisted flavor evaporation (SAFE) technique. The two samples that were used came from cows that had been fed on different diets and represented the considerably different flavors of Northern hemisphere and New Zealand milk. Using gas chromatography-olfactometry (GC-O), 71 aroma compounds were found from the milk extracts, 66 of which were identified. Nearly all of the aroma compounds were common to both extracts, despite the two milk samples having quite different flavors. Only one compound, gamma-12:2 lactone, was significantly odor-active for the extract of milk from cows fed a supplement diet, but was not found for the extract of milk from cows fed a pasture diet. Thus, differences in milk flavor are primarily caused by concentration differences of a common set of flavor compounds, rather than by the occurrence of compounds uniquely associated with a particular feed.     

香蕉茎叶饲喂泌乳牛试验

4头一胎泌乳 5个月的荷斯坦试验牛 ,在其日粮中用经青贮处理的香蕉茎叶 2 0 kg替代 60 %象草。经 60 d的试验结果表明 :试验组和对照组的泌乳量分别为 1 6.0 6kg和 1 5.36kg,乳脂率、乳蛋白质和乳的密度为 3.34、 3.0 9、 1 .0 2 51和 3.4 1、 3.0 7、 1 .0 2 65,两组牛的泌乳量和营养成分差异不显著 ( P>0 .0 5)。试验牛每产 1 kg鲜乳比对照组降低饲料费 0 .0 6元 ,试验期间两组牛体重变化和健康状况相似     

基于融合坐标信息的改进YOLO V4模型识别奶牛面部

为实现奶牛个体的准确识别,基于YOLO V4目标检测网络,提出了一种融合坐标信息的奶牛面部识别模型。首先,采集71头奶牛面部图像数据集,并通过数据增强扩充提高模型的泛化性能。其次,在YOLO V4网络的特征提取层和检测头部分分别引入坐标注意力机制和包含坐标通道的坐标卷积模块,以增强模型对目标位置的敏感性,提高识别精度。试验结果表明,改进的YOLO V4模型能够有效提取奶牛个体面部特征,平均精度均值（mAP）为93.68%,平均帧率为18帧/s,虽然检测速度低于无锚框的CenterNet,但mAP提高了10.92%;与Faster R-CNN和SSD模型相比,在检测速度提高的同时,精度分别提高了1.51%和16.32%;与原始YOLO V4相比,mAP提高0.89%,同时检测速度基本不变。该研究为奶牛精准养殖中的牛脸图像识别提供了一种有效的技术支持。     

Nonparametric estimation of ROC curves based on Bayesian models when the true disease state is unknown

We develop a Bayesian methodology for nonparametric estimation of ROC curves used for evaluation of the accuracy of a diagnostic procedure. We consider the situation where there is no perfect reference test, that is, no “gold standard”. The method is based on a multinomial model for the joint distribution of test-positive and test-negative observations. We use a Bayesian approach which assures the natural monotonicity property of the resulting ROC curve estimate. MCMC methods are used to compute the posterior estimates of the sensitivities and specificities that provide the basis for inference concerning the accuracy of the diagnostic procedure. Because there is no gold standard, identifiability requires that the data come from at least two populations with different prevalences. No assumption is needed concerning the shape of the distributions of test values of the diseased and non diseased in these populations. We discuss an application to an analysis of ELISA scores in the diagnostic testing of paratuberculosis (Johne’s Disease) for several herds of dairy cows and compare the results to those obtained from some previously proposed methods.     

Methodology for quantifying residues of chlorhexidine in raw dairy milk

A residue method was developed as part of a pharmacokinetics study to determine the elimination of chlorhexidine in raw milk after intramammary infusion into dairy cows affected with bovine mastitis. The developed liquid/liquid and solid-phase extraction procedures effectively reduced sources of milk product interferences in the final extract. By optimizing mobile-phase pH buffer/acetonitrile gradient conditions and employing an end-capped reverse-phase polar embedded-phase chromatographic column, excellent peak resolution was achieved without the additional need of mobile-phase amine modifiers or ion-pairing reagents. The combined cleanup and chromatographic method steps reported herein were sensitive and reliable for determining the pharmacokinetic elimination of chlorhexidine following intramammary infusion. The residue method was found to be rugged with a lower detection limit of 0.1 ppm.     

苹果酸对泌乳早期奶牛泌乳性能和代谢产物的影响

选用28头经产奶牛,根据泌乳期、上一泌乳期305d的产奶量和预产期,采用随机区组设计分为4组,研究苹果酸(07、0、140和210g/d)对泌乳早期奶牛采食量、泌乳性能、血液代谢产物和尿酮浓度的影响。结果表明:日粮添加苹果酸对奶牛的采食量、乳脂率、乳蛋白率、乳糖率和乳干物质率无显著影响,140g/d组和210g/d组产奶量和饲料转化效率显著高于对照组(P<0.05)。添加苹果酸后,140g/d组和210g/d组血浆葡萄糖浓度显著高于对照组(P<0.05),而血浆游离脂肪酸和β-羟丁酸浓度显著低于对照组(P<0.05);140g/d组和210g/d组尿酮浓度显著低于对照组和70g/d组(P<0.05)。根据试验结果,苹果酸适宜添加量为140g/d。     

奶牛乳腺炎金黄色葡萄球菌临床分离株生化特性与菌体蛋白免疫原性分析

摘要：病原学调查发现金黄色葡萄球菌是奶牛乳腺炎的最主要致病菌之一。从53株金黄色葡萄球菌乳样分离株中挑选了培养和形态典型的六株进行生化试验和传代稳定性试验，进一步研究这六株菌的凝固酶、溶血素等致病性因子和耐药性。对菌体裂解产物进行蛋白电泳图谱分析发现菌体蛋白条带具有很高的相似性,TQ-1株电泳蛋白条带密度较其它几株高。用其中四株生化典型、传代稳定、致病因子较多的菌株免疫兔，制备高免血清，分析全菌抗原的蛋白转印图谱，发现XG-2株在52KD处有特殊保护性抗原条带。综合本文各试验结果，确定以TQ-1和XG-2株为疫苗候选株。     

红外光谱在牛奶生产和检测方面的研究进展

红外光谱不仅可以反应物质的结构组成,还能够随着成分含量变化形成不同响应值,红外技术也被广泛应用于畜产品的化学成分含量检测和质量评估。牛奶是人类膳食结构中的重要组成部分,对其营养成分和质量的准确评估有着非常重要的生产意义。该文介绍了红外光谱技术牛奶生产中各个环节中的应用,通过测定牛奶成分的含量,红外技术被用于产品定价和品质评价;掺假物质在牛奶中会引起光谱的变化,定性和定量模型的建立为牛奶质量快速鉴别诊断提供了便捷途径;牧场生产中,光谱被用于诊断奶牛酮病、机体能量状态。该文对近年来国外利用红外光谱技术在牛奶成分和凝集性能预测、掺假和质量检测、奶牛健康养殖等方面文献进行综述,重点介绍乳蛋白成分、脂肪酸、凝集性能等牛奶性状红外光谱模型以及牛奶光谱特征,对不同研究中模型性能进行比较,以较为全面的评估光谱技术在牛奶性状、质量和奶牛养殖等方面的应用,并为今后的检测和研究发展提供参考。     

通过乳中孕酮、雌二醇含量诊断奶牛早期妊娠的研究

用放射免疫法测定了奶牛配种前后和配种当天妊娠及未妊娠牛乳中孕酮(P)、雌二醇(E2)的含量,并计算其比值。结果表明:配种前妊娠牛的P水平与未妊娠牛差异不显著,但若统计配种前1-18d的结果时,则差异极显著(P<0.01);而E2差异不显著。配种后妊娠牛P含量从第7-9天开始极显著高于未妊娠牛(P<0.01),并随时间增加,差异继续增大。妊娠牛与未妊娠牛配种后19-21d的E2水平差异极显著(P<0.01),未妊娠牛的E2水平比妊娠。妊娠与未妊娠牛,配种当天P含量差异显著(P<0.05),E2含量差异不显著,而P和E2的比值差异极显著(P<0.01)。