CN102576008A - 用于检测乳腺炎和奶质量的方法以及乳腺炎传感器 - Google Patents
用于检测乳腺炎和奶质量的方法以及乳腺炎传感器 Download PDFInfo
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Abstract
本发明公开了乳腺炎传感器和用于实时检测乳腺炎和测定奶质量(线上)的方法。本发明包括快速非侵袭测定挤出的奶中溶解分子氧的浓度。乳腺炎传感器包括用于测定氧浓度的光导纤维的、电流测量的或电势测量的设备,用于数据获取和处理的设备,乳腺炎指示器和生成自动线上去除受感染动物不合格奶的信号的设备以防止更大量奶的污染。
Description
技术领域
本发明属于兽医和奶生产的领域,特别为原料奶质量的测定。本发明意欲实时快速检测乳腺炎(mastitis)和其它炎性过程,还意欲快速测定原料奶的质量和不合格奶的线上分离以避免乳品厂生产的污染。
背景技术
乳腺炎是奶牛最普遍的感染而且它造成经济损失,成为全世界的重大问题。
现今,乳腺炎的诊断通常基于不同的非侵袭方法:
●通过光谱测定(专利申请US2008000426;Grabek等,2008)或粘度测定检测奶中的体细胞计数(SCC),其中将阴离子表面活性剂加入奶中,致使在体细胞中形成蛋白凝胶,测定其粘度并针对体细胞浓度校准(专利US2,935,384;Schalm,O.,Noorlander,D.,1960);
●检测奶中的乳酸根(lactate)(专利EP1192460,Agresearch,2003)-细菌致使乳房中乳酸根的浓度升高(厌氧环境);
●测定奶的传导性,该方法相对缺乏特异性,这是由于奶的传导性受细菌之外其他因素的影响而且正常生物学上传导性的变化与乳腺炎无关
(http://www.iah.bbsrc.ac.uk/BMC/2004/Papers%202004/Word/FI NAL%20Whyte%20BMC%202004.doc);
●监测奶中各种离子的比率。随着乳腺炎感染程度的发展,钠离子浓度升高而钾离子浓度降低(国际专利申请WO/2006/127921,Westfall,G.,2006);
●检测MAA。作为对感染的应答,哺乳动物免疫系统产生急性期蛋白,例如奶淀粉样A(MAA)蛋白在奶牛的乳房中产生。http://www.trideltaltd.com/maakit.html;
●用于检测导致乳腺炎的细菌的微生物学测试,例如用于检测金黄色葡萄球菌(S.aureus)的RAPIDEC Staph测试(分析时间24h),(Boerlin,P.等,J Clin Microbiol.,2003,41(2):767-771);
●基于化学试剂应用的分光光度计测量方法(专利CN100460866,Ox.Biolog.Tech.Co Ltd,2009和US6979550 Rivas等,2005),其产生含可检测到的化合物的有色产物;
●基于红外温度记录的检测方法(国际专利申请WO0057164,Emerge Interactive Inc.,2000);
●在专门的移液管中将鲜奶离心并且在不同沉降层中通过细胞数目检测病原体(Biomedical Diagnostics Institute;http://www.bdi.ie/research/mastitis chip.html)。
●化学发光测定法用于测定细菌侵袭后吞噬细胞发射光线的能力(Takahashi,H.:Cytokine Therapy for Staphylococcus Mastitis inDairy Cows.Science & Technonews Tsukuba,1999,50:55-56)。
与本发明最接近的是乳腺炎检测的方法,其基于奶中乳酸根的测定并且将该乳酸根水平与健康动物的奶的乳酸根水平相比(US7033836,Pastoral Agric.Res.Inst.Nz Ltd,2006)。
然而,上述方法具有若干缺点:
●传感器的寿命相对较短。例如,乳酸根传感器需要经常更新,这是由于其识别系统是基于酶的。应用于本发明的(光学)氧传感器,可运行很多年;
●奶中的乳酸根浓度取决于许多不同因素-饲养、挤奶频率、泌乳期等;
●大多数上述方法不适用于实时线上过程并且不可能在挤奶的过程中除去不合格奶;
●一些方法需要将不同化合物加入到奶中;
●一些方法需要昂贵的设备和高度合资格人员。
在科学研究中,已研究乳房(产奶前)中溶解氧浓度,目的在于研究正常奶牛和乳腺炎奶牛的乳房中溶解氧含量是否足够支持正常嗜中性粒细胞的功能以去除金黄色葡萄球菌。嗜中性粒细胞通过两种方法杀死细菌:氧化和非氧化方法。当嗜中性粒细胞受吞噬的刺激时,氧消耗和氧自由基(例如超氧化物)的产生将会增加,这是由于NADPH氧化酶的活化而造成的,其形成电子传递链,将分子O2转化成超氧化物。已发现,乳腺炎导致O2浓度显著下降并且乳房中嗜中性粒细胞的抗微生物活性降低。正常奶牛具有与静脉血中相似的奶溶解O2水平;乳腺炎奶牛中溶解O2水平小于对照值的10%(Mayer SJ,Waterman AE,Keen PM,Craven N,Bourne J.Oxygenconcentration in milk of healthy and mastitic cows and implications oflow oxygen tension for the killing of Staphylococcus aureus by bovineneutrophil.Journal of Dairy Research 1988;55(4):513-9)。
不存在将奶中溶解O2的测定用于检测乳腺炎的公知方法。
发明公开
本发明提出用于实时检测乳腺炎和测定奶质量的快速且可靠的方法以及乳腺炎传感器。
用于检测乳腺炎和测定奶质量的方法基于非侵袭测定奶中溶解分子氧。在氧浓度基本上大于或小于奶中溶解氧的正常浓度(差异为与典型浓度的平均值或其它给定阈值的标准差σ值大于3)的情况下,线上检测动物生物体中乳腺炎或其它炎性过程以及不合格奶。在挤奶系统中,基于真空设备的应用,氧的测定尽可能快的进行,但不晚于挤奶过程开始后60秒,以避免氧从空气中大量转移。测定来自一个或若干乳房的奶中的氧浓度。
乳腺炎传感器包含用于测定氧浓度的光导纤维的、电流测量的或电势测量的设备;用于数据获取和处理的设备;乳腺炎指示器和生成自动线上去除不合格奶的信号的设备。乳腺炎传感器用于应用该方法,即在本发明中描述的用于检测乳腺炎和测定奶质量的方法。
本发明提出的方法和乳腺炎传感器与先前方案相比具有若干优点,这是由于它们实现
-快速检测产奶动物诸如奶牛、山羊、绵羊等在亚临床阶段中潜在乳腺炎来源;
-线上检测乳腺炎和其它炎性过程并且测定奶质量;
-在奶收集槽之前去除受感染动物的不合格奶;
-乳腺炎传感器非常易于应用并且不需要特殊技能。
所提出方法和设备的维护成本低,这是由于传感器的寿命长和以下事实:没有对废物材料或系统组件更新的定期需要。受感染动物的实时检测使得能够降低生产成本,这是由于不合格奶可在挤奶过程的早期去除从而防止更大量奶的污染。不需要耗时和昂贵的分析程序。所提出设备的应用不需要人员的专门训练。在感染早期对患有亚临床乳腺炎动物的检测允许启动动物的早期治疗,效果更有效。因此本方法和设备的应用不仅在降低生产固定成本和运行成本两方面产生经济效应,而且还在改进动物福利方面产生经济效应。
以下实施例说明本发明的应用,尽管本发明不限于以下实施例,但可依据权利要求应用。
附图
图1.新鲜挤出的奶中氧浓度数据的直方图(385项测量结果)以及这些数据与正态分布的近似度(连续线)。
○-与平均值接近的结果(377项测量结果);
●-异常值(8项测量结果)
实施方案描述
用于检测乳腺炎和测定奶质量的方法在农场中应用,在此我们用克拉克型(Clark-type)传感器测定了385头奶牛中奶的溶解氧浓度。从挤奶设备中取出12ml奶探测物并且尽可能快地测定探测物中的氧,但不晚于从设备中取出探测物后60秒以防止氧从周围空气大量转移到奶。
我们分析所得到的结果并计算氧浓度的标准化平均值和标准差σ。结果示于图1和表1。
表1.不同奶牛中奶的标准化平均氧浓度。
依据图1,在377个案例(97.9%)中,所得到的结果在范围内(within span),这在正态分布的情况下应包括99%的结果(图上两箭头之间的面积)。8个案例(12.1%)中的结果在该范围之外。
将奶的氧水平不同于确定的阈值(平均氧浓度值±3σ)的奶牛取出接受特殊观察。在该组中,在观察期期间50%的奶牛诊断为临床乳腺炎。
用克拉克型氧传感器还测定来自感染乳腺炎的奶牛的不同乳房经人工挤出的奶中的溶解氧浓度。恰在挤奶后60秒内测定12ml的奶探测物中的氧浓度。来自乳腺炎动物的受感染乳区(udder quarter)的奶中氧的测定结果与来自健康动物的奶中的结果之间存在显著差异。在相似条件下挤奶,在来自乳腺炎动物的受感染乳区的奶的大多数探测物中,与健康动物的奶相比,氧浓度显著较低(大约低至1/2)。在相似条件下取奶,还有来自受感染乳区的探测物,其中氧浓度显著高于来自健康动物的奶。结论是,来自乳腺炎动物的受感染乳区的所有奶探测物中所测定的氧浓度与来自健康动物的奶中的氧浓度平均值之间存在显著差异。
上述测定奶中氧浓度的方法还用于测定奶质量。在下述情况下:即来自受感染乳区的奶探测物中所测定的氧浓度与来自健康动物的奶中的氧浓度平均值之间存在显著差异(差异大于3σ值),则鉴定为不合格奶并在到达奶管前线上分离该奶并引向废料。
乳腺炎传感器包括氧传感器、用于数字化传感器模拟输出信号的设备、自动数据获取和处理系统和乳腺炎指示器,其中结果与标准化的氧浓度平均值相比较,并且在确定结果有显著差异(超过3σ)的情况下,在指示器面板上信号灯发亮。若必要,乳腺炎传感器使得能够生成信号,其启动线上分离不合格奶与优质奶的系统。将乳腺炎传感器置于挤奶系统的挤奶管或小收集槽中,依据测试地点的温度将传感器校准,并实时测定奶中溶解氧的浓度。
在所测定奶中氧浓度与氧浓度平均值存在显著差异(通常奶中氧浓度是38.6℃时氧饱和浓度的65至75%或相应为4.30至4.95mg/l;38.6℃时氧饱和浓度为6.60mg/l)并且与平均值的差异大于3σ值的情况下,该动物可能患有亚临床或临床乳腺炎;在该差异为2-3σ值的情况下,建议对动物进行另外的检查;在差异更小的情况下,则动物是健康的。
来自患有乳腺炎的动物受感染乳区的奶中的氧浓度低至正常(38.6℃时氧饱和浓度的23-49%)的1/3-1/2或在另一极端等于氧饱和浓度(100%)。
用乳腺炎传感器得到的结果以连续或离散的颜色标度形式显示于设备的屏幕上(例如差异超过3σ产生红色,差异介于2至3之间产生黄色以及差异低于2σ值产生绿色指示色)或作为数字输出。
Claims (9)
1.用于检测动物中乳腺炎的方法,所述方法包括以下步骤:
a)挤出动物的奶并且非侵袭地测定奶中溶解分子氧的浓度;
b)将奶中溶解分子氧的浓度与存在于未受感染的奶中溶解分子氧的典型浓度相比较;
c)在奶中氧浓度与未受感染的奶中典型氧浓度之间存在显著差异的情况下(差异为与典型浓度平均值或其它给定阈值的标准差σ值大于3),在实时过程中检测所述动物中的乳腺炎。
2.权利要求1的方法,其中奶中溶解氧浓度用光导纤维的、电流测量的或电势测量的设备测定。
3.权利要求1-2中任一项的方法,其中用真空挤奶系统挤出动物的奶并且在没有外部空气进入情况下在所述挤奶系统中测定奶中溶解分子氧的浓度。
4.权利要求1-3中任一项的方法,其中人工挤出动物的奶并且在不晚于挤奶开始后60秒测定奶中溶解分子氧的浓度。
5.权利要求1-4中任一项的方法,其中在一个或多个乳区中测定奶中溶解分子氧的浓度。
6.权利要求1-5中任一项的方法,其中将奶中溶解分子氧的典型浓度确定为农场或动物群的平均溶解氧浓度,或确定为所测定动物的溶解氧浓度的平均值,并且基于农场或动物群的溶解分子氧的平均值建立溶解分子氧浓度的典型值的允许偏差。
7.用于实时测定奶质量的方法,其中依据权利要求1-6中任一项在动物的生物体中已检测到乳腺炎,则将所述动物的奶确定为不合格。
8.权利要求1-7中任一项的方法,其中所述动物是奶牛、山羊或绵羊。
9.用于权利要求1-8中任一项方法的乳腺炎传感器,其包括用于测定氧浓度的光导纤维的、电流测量的或电势测量的设备,用于数据获取和处理的设备,乳腺炎指示器和生成自动线上去除不合格奶的信号的设备。
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Also Published As
Publication number | Publication date |
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US20120115184A1 (en) | 2012-05-10 |
EP2440916A1 (en) | 2012-04-18 |
WO2010142301A1 (en) | 2010-12-16 |
AU2009347606A1 (en) | 2011-12-08 |
RU2011152761A (ru) | 2013-07-20 |
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