眼镜蛇腐皮病病原菌分离鉴定与药敏试验

为了研究眼镜蛇腐皮病的致病菌,试验采用细菌分离培养的方法,在无菌条件下,从患腐皮病眼镜蛇皮肤溃烂处挑取少许内容物划线接种于普通琼脂平板、兔血平板、SS琼脂平板及真菌培养基上,继而选取优势菌落进行分离、纯培养。结果表明:得到9种细菌,没有发现真菌。根据细菌形态、菌落生长特征和生化特性得出,这9种细菌分别为:黏质沙雷菌1株、奇异变形杆菌3株、铜绿假单胞菌3株、克氏耶尔森菌1株、肺炎克雷伯菌1株、雷极普鲁威登斯菌1株、嗜麦芽假单胞菌1株、金黄色葡萄球菌1株和腐生葡萄球菌3株。经致病性试验证实,奇异变形杆菌、铜绿假单胞菌、克氏耶尔森菌、雷极普鲁威登斯菌、嗜麦芽假单胞菌和金黄色葡萄球菌具有致病性,黏质沙雷菌、肺炎克雷伯菌和腐生葡萄球菌无致病性。药物敏感性试验证实:在15株致病菌中,除克氏耶尔森菌外,均对妥布霉素和庆大霉素高度敏感;嗜麦芽假单胞菌对妥布霉素不敏感,对庆大霉素高度敏感;除金黄色葡萄球菌外,均对阿米卡星高度敏感;15株致病菌均对头孢类抗生素不同程度敏感;哌拉西林、氨曲南和卡那霉素对大部分菌有抑制作用。说明眼镜蛇腐皮病主要由致病性细菌引起,敏感药物治疗后有良好效果。     

腹泻兔病原菌的分离鉴定及耐药性检测

为了确定娄底地区兔腹泻的病原菌、致病性及耐药性,对采集的203份患腹泻的兔肛拭子、粪便进行病原菌分离鉴定,结果分离得到了30株大肠埃希氏菌、20株铜绿假单胞菌、17株产气荚膜梭菌、15株金黄色葡萄球菌;人工接种小鼠试验、KB药敏纸片法分别检测病原菌的致病性及耐药性,结果表明分离得到20株大肠埃希氏菌、15株铜绿假单胞菌、12株产气荚膜梭菌、10株金黄色葡萄球菌对小鼠具有致病性;分离的15株致病性铜绿假单胞菌对阿莫西林、氨曲南、庆大霉素等7种药物耐药率在46.7%以上,分离的12株致病性产气荚膜梭菌对环丙沙星、磺胺嘧啶、氟苯尼考等8种药物耐药率在41.7%以上,分离的10株致病性金黄色葡萄球菌对磺胺嘧啶、氟苯尼考、庆大霉素等7种药物耐药率在80.0%以上,分离的20株大肠埃希氏菌对磺胺嘧啶、多黏菌素、强力霉素等7种药物耐药率在45.0%以上,4种病原菌对其他药物具有不同的耐药性。     

铜绿假单胞菌flgE基因PCR检测方法的初步建立

为建立一种基于铜绿假单胞菌flgE基因的PCR检测方法,本试验根据GenBank中已发表的铜绿假单胞菌flgE基因序列,设计合成了1对特异性引物,由铜绿假单胞菌基因组DNA中扩增获得了目的基因。从退火温度、循环次数、Mg2+浓度、dNTPs浓度和引物浓度5个方面优化了反应条件,并检测了该方法的特异性和敏感性。结果成功扩增出了铜绿假单胞菌1 400 bp的flgE特异性基因片段,最佳退火温度、循环次数、Mg2+浓度、dNTPs浓度和引物浓度分别为56℃、30个循环、1.6 mmol/L、0.2 mmol/L和0.3μmol/L。特异性试验表明,仅铜绿假单胞菌中可扩增出目的片段,而多杀性巴氏杆菌、鼠伤寒沙门氏菌、志贺氏菌、致病性大肠杆菌和金黄色葡萄球菌中均未扩增出相应片段。敏感性试验结果表明,本方法可检测到最低10 pg/μL的铜绿假单胞菌基因组DNA以及100 CFU/mL的病原菌。     

奶牛子宫内膜炎与隐性乳房炎之间的相关性研究

为探讨奶牛子宫内膜炎和隐性乳房炎之间的相关性,在宁夏地区3个大型奶牛场(根据临床型子宫内膜炎的诊断标准)选择子宫内膜炎患牛111头,用兰州隐性乳房炎检测法检测(LMT)乳样,其中82.5%的病牛同时患有隐性乳房炎,然后进行了子宫样和乳样的采集,并用常规细菌分离、培养结合高智能全自动细菌鉴定及药敏分析系统(VITEK 2 Campact),确定引起宁夏奶牛子宫内膜炎的病原菌有18种110株,引起乳房炎的病原菌有23种107株,其中大肠杆菌、停乳链球菌、克氏库克菌、金黄色葡萄球菌、假单胞杆菌、乳房链球菌、产色葡萄球菌、蜡样芽孢杆菌、荧光假单胞杆菌、产酸克雷伯菌、副溶血弧菌、克雷伯氏菌可以同时引起奶牛子宫内膜炎和隐性乳房炎。     

长春市某奶牛场乳腺炎病原菌的分离鉴定及耐药性分析

奶牛乳腺炎是制约奶牛养殖效益和奶品质的主要疾病之一，为了解当前长春市某大型集约化奶牛场奶牛乳腺炎病原菌流行和耐药情况，本试验对该场奶牛乳腺炎发病情况进行了调查，对采集的119份临床型乳腺炎乳样进行细菌分离鉴定并对分离鉴定的病原菌进行小鼠致病性试验和药敏试验。结果显示：该场奶牛乳腺炎发病率在4%左右，初产牛乳腺炎发病率明显高于经产牛，乳腺炎发病率还与胎次、年龄等因素相关。本试验共分离鉴定出木糖葡萄球菌49株(42.2%)、松鼠葡萄球菌39株(33.6%)、大肠杆菌15株(12.9%)、金黄色葡萄球菌5株(4.3%)、无乳链球菌3株(2.6%)、菠萝泛菌3株(2.6%)、产酸克雷伯菌1株(0.9%)和铜绿假单胞菌1株(0.9%);其中菠萝泛菌、无乳链球菌和铜绿假单胞菌对小鼠具有较强致病性。药敏试验结果显示，分离菌株的耐药性普遍存在，大肠杆菌和铜绿假单胞菌耐药种类最多。除无乳链球菌外，其他菌株均对庆大霉素或阿米卡星敏感，可作为该场奶牛乳腺炎治疗首选药物。本试验通过对该场乳腺炎调查、分离菌鉴定和耐药性分析，为该场乳腺炎的防治提供了参考依据。     

浅谈奶牛乳房炎的临床防制

奶牛乳房炎是乳房受到机械的、物理的、化学的和生物学的因素作用而引起的炎症.近年来笔者对治疗临床乳房炎总结了一些实践经验,供大家参考. 1 病因 1.1病原微生物 感染乳房炎的致病菌有150多种,各种革兰氏阴性菌和阳性菌以及支原体都可致病.如大肠杆菌、环境链球菌、金黄色葡萄球菌、停乳链球菌、绿脓杆菌、沙雷伯氏菌、变形杆菌、化脓性棒状杆菌、诺卡氏菌、假单胞苗、支原体、结核杆菌、放线菌等等.     

枯草芽孢杆菌和沼泽红假单胞菌对鲫鱼免疫机能的影响及其代谢产物的抑菌活性研究

为了检测枯草芽孢杆菌和沼泽红假单胞菌对鲫鱼免疫机能的影响及其代谢产物对小肠耶尔森氏菌、金黄色葡萄球菌和大肠杆菌的抑菌活性,试验将鲫鱼分成3组,分别为沼泽红假单胞菌组、等比例沼泽红假单胞菌和枯草芽孢杆菌的复合菌组和未加菌处理的对照组,在养殖水体中分别加入不同的微生物,在第30天静脉采血,测定血浆中碱性磷酸酶(ALP)、总超氧化物歧化酶(SOD)、溶菌酶(LZM)的活性以及丙二醛(MDA)的浓度;利用超声波破碎细胞,旋蒸浓缩得到枯草芽孢杆菌和沼泽红假单胞菌胞内外代谢产物,采用牛津杯法以小肠耶尔森氏菌、金黄色葡萄球菌和大肠杆菌为指示菌株,测定代谢产物的抑菌圈直径。结果表明:沼泽红假单胞菌组与复合菌组的鲫鱼血浆中ALP的活性显著高于对照组(P0.05),LZM活性高于对照组,MDA的浓度低于对照组,SOD的活性与对照组的差异不显著(P0.05);两种菌的胞外代谢产物对金黄色葡萄球菌的抑菌活性较强,沼泽红假单胞菌胞内代谢产物对金黄色葡萄球菌的抑菌活性较强,枯草芽孢杆菌胞内代谢产物对小肠耶尔森氏菌抑菌活性较强。说明沼泽红假单胞菌和复合菌对鲫鱼的免疫机能有一定的增强作用,两种益生菌胞内外代谢产物对小肠耶尔森氏菌、金黄色葡萄球菌、大肠杆菌均有抑制作用。     

进口奶牛感染铜绿假单胞菌的微生物学诊断

从病死奶牛脏器中分离出1株革兰氏阴性短杆菌,通过染色镜检、培养特性分析及生化鉴定,证明为铜绿假单胞菌。动物试验证明,分离株对小鼠有较强的致病性,并从死亡鼠体内分离到相应细菌。药敏试验结果显示,铜绿假单胞菌对多种抗菌药物有耐药性。     

致水貂出血性肺炎铜绿假单胞菌胞外产物生物活性及免疫保护力分析

胞外产物在细菌致病过程中发挥重要作用,为探索胞外产物在铜绿假单胞菌致病过程中的作用,首先从铜绿假单胞菌培养液提取其出其胞外产物,利用小鼠模型分析其致病性,并通过细胞证实了该产物对巨噬细胞功能的抑制作用,进一步测定其胞外产物的酶活性并利用LC-MSMS方法对铜绿假单胞菌胞外产物蛋白成分和功能进行分析。结果表明,铜绿假单胞菌胞外产物对小鼠有致病性,能够显著抑制巨噬细胞吞噬细菌活性,其具有淀粉酶、脂肪酶、蛋白酶、卵磷脂酶活性和溶血活性可能在这一过程中发挥关键作用。LC-MSMS分析鉴定所提纯的胞外产物中有153种蛋白成分,涉及多种酶类、细胞结构蛋白和功能性蛋白。用低剂量胞外产物免疫动物对于铜绿假单胞菌感染具有较明显的保护率,可达90%。本试验为揭示铜绿假单胞菌致病机理,研制新型疫苗奠定基础。     

鸡绿脓杆菌病的诊断与治疗

<正>绿脓杆菌病是由假单胞菌科假单胞菌属的铜绿假单胞菌引起的一种人畜共患病,铜绿假单胞菌又称绿脓杆菌,广泛存在于土壤、水、正常人和动物的肠道、皮肤上,可以感染多种动物和人,人主要是外伤、烧伤尿道感染,多种动物都容易感染,鸡鸭引起心包炎肝周炎,牛羊引起乳房炎,水貂引起出血性肺炎,狐狸引起子宫内膜炎,通常情况只感染受伤动物,由于机体抵抗力低     

A retrospective study of the aetiology and temporal distribution of bovine clinical mastitis in smallholder dairy herds in the Dar es Salaam region of Tanzania

A 31-year record-based retrospective study was carried out to determine the aetiology and temporal distribution of bovine clinical mastitis in smallholder dairy herds in the Dar es Salaam region of Tanzania over the period November 1971-December 2002. Laboratory information on 1964 quarter samples from 1365 cows in 281 smallholder dairy herds were retrieved, compiled and studied. Eighty-eight percent of the quarter samples were culture-positive and the predominant mastitis pathogens isolated were Staphylococcus aureus (25.7%), Streptococcus agalactiae (15.4%), Klebsiella pneumoniae (14.3%) and Escherichia coli (14.1%). Other isolates included Pseudomonas aeruginosa (7.5%), Streptococcus dysgalactiae (5.2%) and Streptococcus uberis (4.2%). Contagious mastitis pathogens were isolated from 45.6% of the culture-positive samples, whereas environmental and miscellaneous pathogens were isolated from 48.2% and 5.7%, respectively. Thirty percent of the miscellaneous mastitis pathogens were Candida species. The results demonstrate a steady increase in clinical Candida albicans mastitis. The prevalence of Candida albicans has increased from 1% in 1971 to 17.0% in November 2002. Conversely, despite some fluctuations, the prevalence of Staphylococcus aureus, Streptococcus agalactiae, E. coli and K. pneumoniae remain above 10%. The possible risk factors for these observations are discussed.     

新疆垦区奶牛乳房炎致病菌的调查

本试验选取新疆垦区七个试验牛场,临床调查和试剂诊断,检测奶牛712头,采集奶牛乳房炎阳性乳样314份。其中,奶牛隐性乳房炎阳性检出率平均为36.3%,临床性奶牛乳房炎为16.7%;采集阳性奶样经试验分离鉴定,革兰氏阳性菌仍是引起奶牛乳房炎的主要致病菌,但金色葡萄球菌已基本取代链球菌成为主要的致病菌,其检出率为78.98%。另外,常见环境性致病菌如沙门氏菌、大肠杆菌已明显减少,检出率仅为21.1%和9.24%。相反,绿脓杆菌和蜡样芽孢杆菌的检出率明显增加,其检出率为5.41%和11.15%,而且绿脓杆菌和隐球菌已经成为引起临床性乳房炎爆发的两种新的病原,其中隐球菌已经成为造成顽固性乳房炎的又一种新型病原。     

Resistance of mastitis pathogens in northern Germany

The data of 1692 susceptibility tests acquired from April 2003 through March 2004 in the mastitis laboratory of the Institute for Food Quality and Safety were summarized in order to help veterinarians confronted with acute mastitis in choosing the appropriate antibiotic. Two thirds of the milk samples were infected with gram-positive cocci. One third of these were identified as Streptococcus (S.) uberis, one fourth as Staphylococcus (S.) aureus. All isolates (100%) of S. uberis, S. dysgalactiae, S. agalactiae and Arcanobacterium pyogenes were susceptible to Penicillin and Ampicillin. Concering S. aureus, nearly 100% of the isolates were susceptible to Oxacillin, Cephalothin, Cefacetril, Cefquinom and Neomycin, but only 88% of the isolates were sensitive to Penicillin, Ampicillin and Cefoperazon. The gram-negative rods (Escherichia (E.) coli, Klebsiella spp. and Pseudomonas spp.) displayed an irregular resistance pattern. More than 93% of all examined isolates including Pseudomonas spp. were susceptible to Colistin. The sensitivity of E. coli and Klebsiella spp.to Marbofloxacin, Enrofloxacin and Cefquinom exceeded 96%. Thus, the susceptibility of gram-positive mastitis pathogens to common antibiotics is favourable. Because the highly effective Colistin is no longer approved for local therapy, the situation for gram-negative bacteria is more difficult.     

犬脓皮病病原菌的分离鉴定与药敏试验

本试验对犬脓皮病致病菌进行分离鉴定,发现主要致病菌为金黄色葡萄球菌和表皮葡萄球菌,采用18种抗菌药物及7味中药进行体外药敏试验.结果显示:两种致病菌对18种抗菌药物中的头孢拉定、阿米卡星、环丙沙星等6种药物表现极敏,对复方新诺明不敏感;7味中药中黄芩、黄连对两种致病菌均有较强的抑菌效果,平均抑菌圈达到了18.5 mm.本试验结果可为临床筛选高效低毒的治疗药物提供理论依据.     

Antibacterial effect of bovine lactoferrin against udder pathogens

The antibacterial effect of lactoferrin (Lf) was tested on isolates of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and coagulase-negative staphylococci (CNS) as well as on Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae), originally isolated from bovine mastitis. Concentrations of Lf used were 0.67 mg/ml, 1.67 mg/ml, and 2.67 mg/ml. Growth of udder pathogens was monitored by turbidometry either in broth culture or in whey prepared from normal milk. We focused on 3 different growth variables: lag time, slope, and maximum absorbance of bacterial growth curves. Growth inhibition was seen in the broth but hardly at all in whey. The isolates of E. coli and CNS did not grow sufficiently well in whey to draw any conclusions. The most effective inhibitory activity of Lf was seen against E. coli and P. aeruginosa. All 5 E. coil isolates had similar growth patterns. Inhibition of growth by Lf was concentration-dependent. The concentration of 0.67 mg/ml in broth and whey was generally too low for a significant inhibitory effect.     

Investigation and control of mastitis outbreaks caused by Pseudomonas aeruginosa in a sheep flock and a goat herd

Outbreaks of mastitis in a sheep flock and a goat herd caused by Pseudomonas aeruginosa, and their control are described.The morbidity rates of mastitis in a sheep flock and in a goat herd were 18.7 and 18.2 per cent, respectively. The lethality and involuntary culling rates of the animals with sub-clinical mastitis correspond to the morbidity rate.The highest morbidity rates were noted in yearlings--22.5 and 30 per cent, as well as in animals pre- and post partum (7 and 20 per cent, in sheep and goats, respectively). The morbidity periods in the flock and in the herd were 20 and 27 months, respectively. A strict-hygiene milking programme and maintenance of the milking devices was introduced. In addition, prevention of crowding, provision of dry bedding and thorough cleaning of water troughs have also been instituted. Ruthless culling of all infected and carrier animals was an essential part of the control program against Pseudomonas aeruginosa mastitis.     

乳房炎源性金黄色葡萄球菌生物被膜检测及药敏试验

金黄色葡萄球菌是奶牛乳房炎的最主要病原之一,由其引起的奶牛乳房炎发病率高、防治困难,常常会给奶业造成巨大经济损失,对食品安全造成巨大威胁.以新疆阿克苏地区奶牛乳房炎性金黄色葡萄球菌分离株为研究对象,详细观察了菌株在刚果红培养基上的形态,并通过K-B扩散法分析了5株金黄色葡萄球菌乳房炎分离株对19种抗生素的耐药情况.结果表明,5株金黄色葡萄球菌均具有生物被膜形成能力,且对青霉素、链霉素、氨苄西林和四环素等常用抗生素完全耐药,对万古霉素、环丙沙星、诺氟沙星、阿米卡星等抗生素中度敏感,而对恩诺沙星、头孢西丁和头孢噻肟3种抗生素高度敏感.研究结果提示,乳房炎性金黄色葡萄球菌具有较强的耐药性,而这种耐药性可能与其生物被膜形成有关.     

Antibiotic sensitivity of bovine staphylococcal and coliform mastitis isolates over four years

Eight hundred and forty-eight strains of Staphylococcus aureus and coliforms isolated from milk samples taken from cows with clinical or subclinical mastitis were tested for their sensitivity to a range of antibiotics, comparing strains isolated in 1984, 1985, 1986 and 1987. The only increase in the proportion of resistant strains occurred with coliforms resistant to ampicillin and neomycin.     

Antimicrobial resistance patterns to beta-lactams of gram-positive cocci isolated from bovine mastitis in Lithuania

The aim of the study was to isolate gram-positive cocci from cows with mastitis and to determine their resistance to beta-lactamic antibiotics. Eight hundred and nine strains were isolated and identified as staphylococci (n=516), streptococci (n=199) and enterococci (n=94) from sub-clinical and clinical cases of bovine mastitis in Lithuania. The most common causative agents of udder disease included: S. epidermidis (n=176), S. aureus (n=176), S. agalactiae (n=134), S. hyicus (136) and E. hirae (n=68). Isolates were analysed for antimicrobial resistance to penicillin, ampicillin, amoxicillin, cephalothin, cephalexin, amoxicillin + clavulanate. The susceptibility patterns were analysed using the agar disk diffusion method. S. aureus showed the highest level of resistance to amoxicillin (81.3%), penicillin (76.7%) and ampicillin (78.4%). The corresponding values for CNS strains were 59.7%, 59.7% and 50.6% against penicillin, ampicillin and amoxicillin respectively. Streptococci were the most frequently resistant to amoxicillin (29.3%), and enterococci to penicillin (27%), amoxicillin (27.5%) and amoxicillin/clavulanic acid (23.8%). The resistance of all tested mastitis pathogens to aminopenicillins and penicillin highly correlated (r=0.83). Compared with other antibiotics, amoxicillin and clavulanic acid combination tended to be more effective (p<0.05) against all tested bacteria in vitro. However, S. aureus, in 38.1% of cases, was resistant to this combination of antimicrobials. This study demonstrates that S. epidermidis, S. aureus, S. hyicus, S. agalactiae and E. hirae remain the most frequent mastitis causative agents on Lithuanian cattle farms. The highest resistance in vitro to penicillins was demonstrated by S. aureus, S. hyicus and S. intermedius. Resistance to cephalosporins remains low, irrespective of bacterial species of gram-positive cocci.     

奶牛乳腺炎的细菌学研究

在对山东7个地区14个奶牛场临床型和隐性乳腺炎调查的基础上采集234头临床乳腺炎病牛乳样、241个隐性乳腺炎乳样并分别做了细菌学检查,结果表明:泌乳期临床型乳腺炎病原菌以凝固酶阴性葡萄球菌、金黄色葡萄球菌、链球菌、酵母菌和棒状杆菌为主;干奶期临床型乳腺炎病原菌以大肠杆菌、链球菌、金黄色葡萄球菌、凝固酶阴性葡萄球菌和酵母菌为主;隐性乳腺炎病原菌以凝固酶阴性葡萄球菌、金黄色葡萄球菌链球菌、酵母菌、假单胞菌和棒状杆菌为主;厌氧菌在隐性乳腺炎、干奶期乳腺炎和干奶期乳腺炎乳样的捡出率分别为 5．82％,4．17%,10．16％;隐性乳腺炎、干奶期乳腺炎细菌的共感染率较高,与泌乳期乳腺炎病原菌的差异极显著(P<0．01),隐性乳腺炎与干奶期乳腺炎病原菌共感染率差异不显著(P >0．05)。