乳酸菌对苜蓿草粉发酵品质的影响

通过为期60d的发酵,探讨添加乳酸菌对苜蓿(Medicago sativa)草粉发酵品质的影响。试验设4个处理,分别为添加无菌水(对照)、植物乳杆菌(Lactobacillus plantarum,LP)、屎肠球菌(Enterococcus faecium,EF)和植物乳杆菌+屎肠球菌(LP+EF)发酵苜蓿草粉,设5个时间点采样进行相关项目测定。结果表明,与对照组相比,发酵60d时各试验组发酵苜蓿草粉的评分提高,总酸含量显著升高(P0.05),pH显著降低;LP、LP+EF和对照组的干物质含量明显低于EF组;各试验组酸性洗涤纤维含量明显低于对照组;有氧暴露7d后,各试验组的pH均显著低于对照组、总酸含量均显著高于对照组(P0.05),且只有对照组中检测到酵母菌(Saccharomycetes)和霉菌(Mycete)。综上可知,添加乳酸菌对苜蓿草粉的发酵品质、稳定性及营养组成具有良好的改善作用。     

The effect of temperature on survival of pathogenic bacteria in biogas plants

The paper deals with the hygienic advantages of sanitation to treat dairy manure in full‐scale biogas plants. The slurry samples were collected from two thermophilic biogas plants (55°C) and two mesophilic biogas plants (38°C) in Hokkaido Japan. A detectable number of Coli‐aerogenes group and Enterococcus in the slurries after anaerobic digestion (AD) could not be found in either thermophilic biogas plants. However, in both mesophilic biogas plants the viable numbers of Coli‐aerogenes group and Enterococcus were detected in the slurries even after anaerobic digestion. The mean decimation reduction time (T₉₀) values of the Coli‐aerogenes group and Enterococcus in the slurries during mesophilic digestion were 13.3 days and 16.7 days, respectively.     

北京市伴侣动物源肠球菌的抗菌药物耐受情况调查

为研究北京市伴侣动物源肠球菌对常见抗菌药物的耐受情况,于2015年5月至2016年1月以北京市某宠物医院门诊就诊犬、猫作为采样对象,采集犬、猫自然排出的粪便作为样品,共得到样本320份。采用选择性培养基进行菌株分离和VITEK-2型全自动微生物分析仪进行肠球菌的鉴定;对分离鉴定的菌株进行8类11种常见抗菌药物敏感性试验;同时进行高水平庆大霉素和高水平链霉素耐受的检测试验,对受试肠球菌的耐药性和多重耐药性进行分析。共得到菌落形态及生化特性不同的非重复的肠球菌318株,其中粪肠球菌（49.06%）和屎肠球菌（29.87%）分离率较高。受试肠球菌对高水平庆大霉素和高水平链霉素的耐药率分别为39.94%和43.40%,对四环素（78.62%）、红霉素（67.30%）和奎诺普丁-达福普汀耐药率（43.71%）较高,对替加环素全部敏感,对万古霉素和呋喃妥因非耐药率也很高（均为98.74%）。受试肠球菌对1类到7类抗菌药物有不同程度的耐药性,多重耐药现象普遍,多重耐药率高达57.23%。耐受4类抗菌药物的数量最多（n=65株）。经分析共得到44种不同的耐药谱,以耐受4类药物的红霉素-奎诺普丁-达福普汀-四环素-高水平氨基糖苷类耐药谱占比最高（45/318,14.15%）。本研究证实受试的北京市伴侣动物源肠球菌对常见抗菌药物的耐药程度较高,多重耐药情况普遍存在,应加强对其耐药性的监测工作。     

Multi-drug-resistant Enterococcus spp. as a cause of non-responsive septic synovitis in three horses

Abstract

CASE HISTORY: Three Thoroughbred horses, a 6-week-old filly (Case 1), a 15-year-old broodmare (Case 2) and a yearling filly (Case 3), sustained synovial sepsis secondary to trauma.

CLINICAL FINDINGS: Case 1 presented with a heel bulb laceration communicating with the distal interphalangeal joint. Arthroscopic lavage was performed and treatment commenced using systemic and local broad spectrum antimicrobial drugs. A pure growth of multi-drug-resistant (MDR) Enterococcus gallinarum was cultured from samples of synovium and joint fluid. Antimicrobial treatment was changed according to the susceptibility results. Response to treatment was poor and despite repeat arthroscopic lavage and intra-osseous regional perfusion of antimicrobials the filly was subject to euthanasia 24 days after the initial injury. Post-mortem examination confirmed septic synovitis, cartilage degeneration and osteomyelitis.

Case 2 sustained a full thickness wound to the carpus which was sharply debrided and closed. The wound dehisced with effusion within the tendon sheath. Drainage was established and treatment included systemic broad spectrum antimicrobials, topical lavage with povodine-iodine and manuka honey infusion. A mixed infection including MDR Enterococcus faecalis was cultured from the synovial fluid. Antebrachiocarpal joint effusion developed 21 days after initial injury and joint sepsis was confirmed. Arthroscopic lavage and tendon sheath debridement were performed, followed by treatment with systemic and local antimicrobials. The mare improved and was discharged. Three months later lameness recurred and corticosteroids were administered intra-articularly. The mare became non-weight bearing lame and was subject to euthanasia. Post-mortem examination confirmed joint sepsis of the antebrachiocarpal and intercarpal joint.

Case 3 presented with a complete articular open fracture of the tibial crest. Under general anaesthesia the fracture was stabilised and the wounds debrided and closed. Systemic broad-spectrum antimicrobials were administered. Six days later the wound dehisced and a bone fragment was removed. Three weeks post-surgery the wound deteriorated with a purulent discharge. Culture of the discharge revealed a mixed bacterial infection, including a MDR Enterococcus faecalis. Femoropatellar joint involvement was confirmed, and treatment included joint lavage, local and systemic antibiosis, and manuka honey instilled into the wound. The filly initially improved, and then deteriorated such that euthanasia was performed.

DIAGNOSIS: All three cases had synovial sepsis with MDR Enterococcus spp.

CLINICAL RELEVANCE: Increased awareness of MDR pathogens in equine wound infections is essential. Prompt diagnostic testing, appropriate therapy, infection control strategies and on-going monitoring and management are vital to limit the clinical impact of these organisms.     

鸡源粪肠球菌对雏鸡致病力试验和毒力基因gelE及efaA的检测

为了解凉山地区鸡源粪肠球菌毒力基因gelE和efaA的基因表达情况,对毒力基因进行检测和致病力试验。通过实验室提供的粪肠球菌10株,进行致病力试验,结果显示,雏鸡死亡率为0～100%,存在较大差异;进行毒力基因gelE(432 bp)、efaA(591 bp)的检测,10株粪肠球菌阳性率均为100%。根据NCBI的基因序列进行粪肠球菌毒力基因gelE、efaA的同源性比对结果均为100%。     

屎肠球菌及其微生态制剂的研究进展

屎肠球菌作为一种潜在的新型饲用抗生素替代品,具有耐热、耐胃酸、耐胆盐、粘附力强、抑菌性和抗生素耐受性等优良的生物学特性,还能提高饲料的转化效率,增强动物机体的免疫力,降低腹泻率及死亡率,从而改善动物产品品质,屎肠球菌微生态制剂在畜禽养殖业中应用已十分广泛。其中微囊化屎肠球菌活菌制剂更是近几年的研究热点,其抗逆性强、稳定性高,可以作为饲用高活性微生态制剂应用于实际生产。本文主要阐述屎肠球菌的生物学特性及其在饲用微生态制剂中的研究进展。     

粪肠球菌R-WL发酵培养基的优化

本试验旨在优化培养基提高粪肠球菌(Enterococcus faecalis)R-WL发酵液的菌体密度,为高效、优质的微生态制剂产品研制奠定基础。首先通过生长曲线的测定、培养条件和培养基成分的单因素试验,确定菌种RWL的最适培养条件为:时间14 h、pH 8.5、温度37℃、接种量3%,最适碳源为乳糖,浓度为10 g/L,最适氮源为蛋白胨+酵母膏,浓度为30 g/L。然后通过Plackett-Burman试验分析培养基中对粪肠球菌发酵影响最重要的主要因素为蛋白胨、乙酸钠和柠檬酸二铵,再通过最陡爬坡试验获得主要因素的最适范围,最后通过Box-Behnken试验和响应面分析得到主要因素的最适添加量。结果表明:最优发酵培养基配方为:乳糖10 g/L、酵母膏15 g/L、蛋白胨12.18 g/L、乙酸钠4.5 g/L、柠檬酸二铵4.5 g/L、硫酸镁0.58 g/L、硫酸锰0.25 g/L、磷酸氢二钾2 g/L、轻质碳酸钙3 g/L,初始pH 8.5。通过试验验证,优化后发酵培养基的发酵菌液活菌数可达(6.44±0.10)×109cfu/mL,比原MRS培养基活菌数提高了71.28%。     

1株鹑鸡肠球菌产ESBLs和AmpC酶的基因型鉴定

为检测1株临床分离的七彩鸟鹑鸡肠球菌的ESBLs和AmpC酶的基因型,探讨其耐药基因的进化机制;采用两倍稀释法测定此菌株对18种常用药物的敏感性,并用9对特异性引物进行PCR扩增、基因克隆及测序分析,确定鹑鸡肠球菌ESBLs和AmpC酶的基因型及基因亚型.结果表明,鹑鸡肠球菌为产ESBLs和AmpC酶菌,除对3代、4代头孢,碳青霉烯类和磷霉素体外敏感外,对其他常用药物均呈现耐药,具有多重耐药特性;该菌所产 ES-BLs的TEM序列与AJ847364(TEM-116)序列相比发生了2处碱基突变,即512T→A和695A→C,引起了相应氨基酸的突变171Ile→Lys、232Lys→Thr,是一种新的TEM亚型,GenBank注册号DQ849329;AmpC酶与序列EF078894(ACT-like型)有97%的同源性,GenBank登录号是DQ849330.这表明该株鹑鸡肠球菌ESBLs为一种新TEM亚型,来源于同时分离的鹑鸡克雷伯菌TEM型的ESBLs;ACT型AmpC酶的存在可能与七彩鸟鹑鸡接触过β-内酰胺类药物有关.     

屎肠球菌T013复合制剂在断奶仔猪生产中的应用研究

[目的]为屎肠球菌T013复合制剂在猪生产中的应用奠定基础。[方法]将90头杜×长×大杂交断奶仔猪随机分为3组:对照组(不添加)、试验Ⅰ组(添加3%屎肠球菌T013复合制剂)、试验Ⅱ组(添加6%屎肠球菌T013复合制剂)。研究不同添加量的屎肠球菌T013复合制剂对断奶仔猪生产性能、表观消化率、粪便微生物、血清指标的影响。[结果]添加3%和6%屎肠球菌T013复合制剂均能提高断奶仔猪的日采食量,降低料重比,但差异均不显著(P0.05)。添加6%屎肠球菌T013复合制剂能显著提高仔猪日增重、减少腹泻率(P0.05)。添加6%屎肠球菌T013复合制剂后断奶仔猪粗蛋白质和钙的表观消化率显著提高(P0.05),2个试验组粗脂肪、粗纤维等饲料养分的表观消化率均有所提高,但差异均不显著(P0.05)。与对照组相比,试验Ⅱ组仔猪粪样的pH极显著降低,大肠杆菌数量极显著减少和乳酸菌的数量极显著增加(P0.01)。试验Ⅱ组仔猪血清尿素氮的含量极显著低于对照组(P0.01),谷草转氨酶和谷丙转氨酶显著高于对照组(P0.05),添加屎肠球菌T013复合制剂对仔猪血清IgG、总蛋白、白蛋白和葡萄糖的含量无显著影响(P0.05)。[结论]屎肠球菌T013复合制剂能改善断奶仔猪的生长性能,提高粗蛋白质的表观消化率,调节肠道微生态环境,促进断奶仔猪健康生长。