宿主防御肽在奶牛乳腺中的表达及应用

宿主防御肽是生物体内产生的一类具有抗菌作用的阳离子多肽,是生物体非特异性免疫功能的重要组成部分。宿主防御肽具有广谱、抗菌、耐热、无毒、不易产生耐药性等诸多特性,在畜牧生产中具有广阔的应用前景。本文主要对宿主防御肽在先天免疫系统中的作用、奶牛乳腺中的表达及乳房炎诊断与治疗应用的研究进展进行综述,为宿主防御肽作为抗生素替代品在畜牧生产中的研究与应用提供理论支持。     

新疆、内蒙古部分地区乳房炎奶样中金黄色葡萄球菌的流行性与耐药性分析

为确定患乳房炎奶牛乳中金黄色葡萄球菌的流行性和耐药性状况,试验自2018年4月至2018年5月采集了新疆地区70例和内蒙古地区50例患乳房炎奶牛生鲜乳样。通过对奶样中金黄色葡萄球菌进行增菌培养、分离纯化,KB纸片扩散法药敏试验及PCR耐药基因检测发现,新疆和内蒙古地区奶样中金黄色葡萄球菌分离率分别为28.6%和54.0%;新疆地区分离的金黄色葡萄球菌菌株对青霉素G的耐药性最高(75.0%),其次是磺胺异噁唑(60.0%)、林可霉素(55.0%);内蒙古地区的金黄色葡萄球菌菌株同样对青霉素G高度耐受,耐药率为70.4%,其次为林可霉素(63.0%)、克林霉素(51.9%);两地区所分离菌株中,88.9%以上携带多种耐药基因,呈现多重耐药性;两地区所检出的高频耐药基因分别是新疆地区的ermA(30.0%)、dfrS1(30.0%)基因和内蒙古地区的lnuA(46.2%)基因。     

凉州区奶牛乳房炎主要病原分离鉴定及药敏试验

乳房炎是奶牛最常见的疾病,随着奶牛产业的蓬勃发展,乳房炎已经成为制约该产业的瓶颈之一。从甘肃省武威市凉州区5个奶牛养殖场采集了1 000头荷斯坦奶牛共计3 991份奶样,对临床型和非临床型(隐性)乳房炎分别通过临床症状和兰州隐性乳房炎诊断液(LMT)进行诊断。结果显示:该地区奶牛临床型乳房炎发病率18.20%,亚临床型乳房炎发病率36.50%,总阳性率54.70%,总阳性乳区率54.72%。从阳性奶样中分离纯化得到的主要病原菌为链球菌属、金黄色葡萄球菌和大肠杆菌,且在不同类型乳房炎中的分布情况不同,临床型中链球菌属最多(54.37%),亚临床型中金黄色葡萄球菌最多(48.56%)。对主要病原菌进行药敏试验,结果表明治疗凉州区奶牛乳房炎的最佳药物应为喹诺酮类和庆大霉素类。     

应用生化黄腐酸对奶牛乳房炎和生产性能的影响

通过32头泌乳奶牛饲喂生化黄腐酸的对比试验。结果表明,生化黄腐酸对奶牛乳房炎、隐性乳房炎有效率达100％,治愈率达80％;试验组比对照组产奶量提高13．84％（P〈0．05）;校正体重后试验组比对照组产奶量提高14．94％（P〈0．05）;提高饲料转化率13．79％（P〈0．05）;每头奶牛每年增加的经济效益=产奶量提高效益＋牛奶级别提高效益＋防治乳房炎、隐性乳房炎减少的牛奶损失=881．45元＋400元＋377．86元=1659．31元,增收效益显著。     

Exfoliation rate of mammary epithelial cells in milk on bovine mastitis caused by Staphylococcus aureus is associated with bacterial load

The exfoliation rate of mammary epithelial cells (MECs) in milk is affected by physiological, breeding and environmental factors. Little is known about the relationship between the MEC exfoliation into milk and mammary‐infected Staphylococcus aureus (S. aureus) load on bovine mastitis caused by S. aureus. The aim of this study was to investigate the relationship between S. aureus load and the proportion of MEC exfoliation in milk using five substantial bovine mastitis models. In 64 randomly extracted milk samples from udders at 3–21 days after S. aureus infusion, there were various samples with different numbers of S. aureus counts and somatic cell counts. No significant correlations were found between the S. aureus counts and somatic cell count (r = 0.338). In contrast, a significant correlation was noted between S. aureus counts and the proportion of cytokeratin‐positive cells in the milk from the infused udders (r = 0.734, P < 0.01). In conclusion, the increasing MEC exfoliation rate in milk from mastitis udders caused by S. aureus may contribute to reduced milk yield.     

驴乳房炎大肠杆菌的分离鉴定及生物学特性研究

为了查明导致新疆某规模化奶驴场乳房炎的主要病原菌及其生物学特性,试验采用常规微生物学方法对病原菌进行分离,采用生化鉴定管和16S rRNA的PCR扩增方法对病原菌进行鉴定,采用K-B药敏纸片法测定其耐药性,D_{600 nm}值绘制细菌生长曲线,最后分析同源性并构建系统进化树。结果显示,分离株为革兰氏阴性短杆菌,能在麦康凯培养基上长出粉红色菌落,在伊红－美兰培养基上长出有金属光泽的菌落;生化鉴定分离株的葡磷胨水、赖氨酸、鸟氨酸、棉子糖、山梨醇、侧金盏花醇、木胶糖试验均呈阳性,而苯丙氨酸和硫化氢试验均为阴性;PCR扩增后测序得到205 bp的1号菌株和203 bp的2号菌株,其特性符合大肠杆菌的特性;药敏结果表明,分离株对链霉素和氯霉素高度敏感,对青霉素、阿莫西林和庆大霉素耐药;生长曲线表明,分离株在2~16 h时为高度繁殖期,16~32 h处于稳定期,32 h以后开始进入衰退期;1号菌株与2号菌株同源性高达99%,1号菌株与大肠杆菌11J和RCB800的同源性最高（100%）;2号菌株与大肠杆菌675SK2和DTU-1的同源性最高（100%）;由系统进化树分析可知,1号菌株与大肠杆菌11J和RCB800处于同一个小分支,2号菌株与大肠杆菌675SK2和DTU-1处于同一个小分支。因此,导致本次驴乳房炎的病原菌为大肠杆菌,本研究结果为进一步系统研究导致驴乳房炎大肠杆菌的致病机制提供参考。     

金黄色葡萄球菌诱导型乳腺炎对中国荷斯坦奶牛乳中脂肪酸组成的影响

本文旨在研究金黄色葡萄球菌诱导型乳腺炎对中国荷斯坦奶牛乳中脂肪酸组成及其绝对含量的影响。根据奶牛乳房结构,试验组利用通乳针将金黄色葡萄球菌细菌悬液经乳头管注射入奶牛乳房内,建立诱导型金黄色葡萄球菌型乳腺炎模型,对照组灌注磷酸盐缓冲液,采用气相色谱法测定乳中的脂肪酸绝对含量。结果表明:乳中共检出34种脂肪酸,金黄色葡萄球菌诱导对感染乳区脂肪酸合成产生直接影响。按不饱和度和碳链长度划分的6大类脂肪酸中,试验组在感染后11 h与感染前24 h相比,短链脂肪酸、中链脂肪酸和饱和脂肪酸的绝对含量均显著降低(P0.05);对照组有相同变化趋势,但除短链脂肪酸外均未达到显著水平(P0.05)。感染后23 h试验组6大类脂肪酸的绝对含量均低于对照组。结果提示:金黄色葡萄球菌诱导型乳腺炎影响了奶牛脂肪酸的合成过程,降低了脂肪酸各组分的绝对含量。     

Prototheca zopfii genotype 2: the causative agent of bovine protothecal mastitis?

In order to clarify the epidemiology of bovine protothecal mastitis, 30 Prototheca zopfii mastitis isolates were genetically investigated. Based on the 18S rDNA, which allows a differentiation of the former species P. zopfii in two distinct P. zopfii genotypes and Prototheca blaschkeae sp. nov., newly developed genotype-specific PCR-assays as well as RFLP-assays were applied.

All mastitis isolates investigated could be assigned to P. zopfii genotype 2 suggesting that this genotype is the aetiological agent of bovine Prototheca mastitis.     

Assessment of Lactate Dehydrogenase,Alkaline Phosphatase and Aspartate Aminotransferase Activities in Cow’s Milk as an Indicator of Subclinical Mastitis

This study examined the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in the milk of lactating Holstein cows in association with subclinical mastitis (SCM). A total of 94 milk samples were collected from 58 lactating dairy cows representing stages of lactation from the second to the tenth week after calving. Those which were classified as positive by California mastitis test (CMT) were deemed to have subclinical mastitis. All the milk samples were skimmed by centrifugation at 10 000g at 0 degrees C and were used for enzyme activities estimations. The mean activities of LDH and ALP were higher in the milk from udders with SCM than in the milk from healthy udders (p < 0.05). There were no significant differences in AST values. The maximum agreement rates between the CMT results and LDH and ALP values were seen at thresholds of > 180 IU/L and > 40 IU/L respectively (kappa values 0.65 and 0.79, respectively). However, the sensitivity of the tests for identifying SCM at these thresholds was higher for ALP (96.4%) than for LDH (68.5%). In this study, LDH and ALP tests were standardized for cow's milk and results showed that only the ALP test was reliable in the early diagnosis of subclinical mastitis.