一例肉种鸡腹泻引起种蛋沙门氏菌污染的诊治

沙门氏菌可引起鸡白痢、禽伤寒和禽副伤寒等沙门氏菌病,对我国养鸡业产生了巨大威胁.动物养殖中抗生素的广泛使用导致了沙门氏菌耐药性的出现,这给沙门氏菌病的防治增加了难度.本研究从一家肉种鸡养殖场的孵化场采集孵化早期死胚、出壳前2天活胚以及腹泻肉种鸡泄殖腔拭子样本进行沙门氏菌株的分离鉴定,通过药敏试验了解细菌耐药状况.结果表...     

不同来源沙门氏菌的毒力基因检测与耐药性分析

[目的]了解不同畜禽中沙门氏菌的携带状况及其毒力和耐药性,为动物源沙门氏菌的流行病学分析和防控提供数据和依据。[方法]从山东、吉林、江西、新疆和云南等地区采集样本,用国标法分离鉴定沙门氏菌,用沙门氏菌血清型试剂盒检测血清型,用PCR方法检测毒力基因,最后用微量肉汤稀释法(MIC)对分离菌株进行耐药性分析。[结果]健康畜禽和发病畜禽样本中的沙门氏菌分离率分别为20.67%(141/682)、6.13%(45/734)。发现4种沙门氏菌优势血清型,分别为德尔卑(42.6%)、印第安纳(26.2%)、肠炎(17.7%)和汤普森(17%)。对分离菌株进行毒力基因分析,检测SPI-1~SPI-5上的5个核心蛋白基因和spv A、B、C、D和R毒力质粒基因,其中健康畜禽样本中沙门氏菌毒力岛上的基因携带率与发病畜禽样本中的携带率基本一致,但发病畜禽中的菌株毒力质粒基因携带率明显高于健康畜禽。耐药性检测结果表明,除多西环素和庆大霉素外,发病动物中的沙门氏菌对抗生素的耐药率均比健康动物中的高。猪源与鸡源沙门氏菌对庆大霉素、头孢噻呋、氧氟沙星、粘杆菌素的耐药率差异显著,对其他抗菌药物略有差异,但不显著。健康畜禽中的多数沙门氏菌株耐药1~2种,占分离菌株总数的55.32%;发病畜禽中耐药10种以上的沙门氏菌占分离株总数的44.44%。[结论]发病畜禽中的沙门氏菌分离率比健康畜禽中的低,但其毒力质粒基因的携带率较健康畜禽中的高,且耐药情况较严重,多重耐药率高。该分析结果可为沙门氏菌的危害评估和防控措施制定提供依据。     

仔猪源沙门氏菌对抗菌药物及消毒剂的耐药性调查

为了解仔猪源沙门氏菌对抗菌药物和消毒剂的耐药情况,从广西崇左市某猪场采集仔猪肛拭子样品150份,采用常规的细菌分离方法分离沙门氏菌,运用营养肉汤稀释法测定了分离菌对常用的13种抗菌药物和4种常用消毒剂的敏感性。结果表明:150份样品中共分离出13株沙门氏菌,分离率为8.7%;13株分离菌对链霉素、硫酸新霉素、乙酰甲喹、土霉素、庆大霉素有较高的耐药性,耐药率分别为92.3%、69.2%、61.5%、53.8%、53.8%,对头孢曲松、头孢他啶和磺胺间甲嘧啶钠的耐药率为0;多重耐药类数最多为耐9类抗菌药物,多重耐药率为76.9%;13株分离菌对消毒剂百毒杀的耐药率为23.1%,而对新洁尔灭消毒液、聚维酮碘溶液及病毒灭敏感。     

河南省猪源沙门氏菌分离鉴定、血清分型及药物敏感性分析?

为了解河南省猪源沙门氏菌的血清分型和耐药性,从郑州、开封、焦作等5市生猪屠宰场抽取猪盲肠内容物样品840份进行沙门氏菌分离。采用PCR、BD PhoenixTM-100全自动微生物鉴定系统和血清凝集反应对分离菌株进行鉴定,并通过微量肉汤稀释法对分离菌株进行药物敏感性分析。结果显示:从840份样品中共分离沙门氏菌45株,分离率为5.36%(45/840);分离的沙门氏菌共分为6个血清型,其中德尔卑(Derby)沙门氏菌为优势血清型;分离的沙门氏菌对四环素、磺胺异恶唑、大观霉素、氨苄西林耐药较严重,耐药率分别为82.22%、75.56%、73.33%、73.33%。结果表明:河南省存在一定的猪源沙门氏菌污染,尤其是德尔卑沙门氏菌,需要重点加强控制;沙门氏菌耐药情况较为严重,应进一步规范养殖环节抗菌药物的使用。     

成都某种禽场鸡胚沙门氏菌的检测

对四川省成都市某种禽场的156个死胚进行沙门氏菌的分离鉴定及药物敏感性检测.本试验采用沙门氏菌显色培养基、肠杆菌科生化鉴定管、三糖铁试验、沙门氏菌多价血清和16S rRNA PCR鉴定5种方法对疑似菌株进行鉴定,并用6种毒力岛基因将分离的沙门氏菌进行PCR鉴定.结果显示,沙门氏菌的分离率为15.4%(24/156),其中伤寒沙门氏菌占58.3%(14/24);fimY、invA和mgtC毒力基因的检测率均为100%;本试验分离菌对大部分沙门氏菌临床药物表现出明显的耐药性.     

上海市活禽交易市场鸡群沙门氏菌的分离鉴定与耐药性分析

为了解上海市活禽交易市场鸡群中沙门氏菌的携带情况,我们于2016年对市区的8个活禽交易场所进行了调查。共采集肛门拭子500份,分离培养沙门氏菌,运用诊断血清、VITEK 2Compact和VITEK MS分别对分离菌株进行血清型、生化和质谱鉴定,使用PCR和药敏板分别对分离菌株的耐药基因和耐药性进行检测。结果显示:从500份肛门拭子中分离鉴定出28株沙门氏菌,分离率为5.6%,以肠炎沙门氏菌、印第安纳沙门氏菌和鼠伤寒沙门氏菌为主,说明在市场交易环节应当加强对沙门氏菌的防控;耐药基因检出率较低,仅有喹诺酮类和磺胺类抗生素耐药基因有检出,分别为21.4%和7.1%,I类整合酶基因检出率达到28.6%;从耐药性检测结果看,分离菌株对磺胺类抗生素、头孢噻肟和庆大霉素的耐药率达到50%以上,对环丙沙星、美罗培南、萘啶酸、头孢他啶和氨苄西林的耐药率在40%以下,这些药可用于鸡群沙门氏菌的防控;耐药基因和耐药性之间无明显相关性。     

贵州省猪源沙门氏菌对β-内酰胺类药耐药性及耐药基因分析

为了解贵州省猪源沙门氏菌对β-内酰胺类抗菌药物耐药性及其耐药基因的流行情况,本试验从贵州省9个地区规模养猪场中分离鉴定130株沙门氏菌,采用微量肉汤稀释法测定其对常用的8种β-内酰胺类抗菌药物的敏感性,并用PCR法对β-内酰胺酶耐药基因进行检测。结果显示,沙门氏菌对常用的β-内酰胺类抗菌药物耐药性十分严重,其中对头孢他啶的耐药率为100%,其次是氨苄西林和阿莫西林,耐药率分别为80.77%和76.15%,耐药率最低的是头孢噻呋和头孢氨苄,均为46.15%。所有菌株均为多重耐药,其中最少为二重,占总数的2.31%,最多为八重,占总数的4.62%,多重耐药主要集中在四至七重,占总数的88.46%。PCR结果显示,SHV耐药基因未检出,TEM、OXA、CTX-M 3种基因检出率分别是85%、75%和46%,细菌的耐药性与相关耐药基因的检出率基本呈正相关。结果表明,猪源沙门氏菌对β-内酰胺类药物具有普遍耐药性,其中头孢他啶尤为严重。TEM、OXA、CTX-M基因是贵州省猪源沙门氏菌主要耐药基因,临床日益严重的耐药现象与耐药基因的普遍存在有很大的关系。     

2014-2016年广州市屠宰场分离沙门氏菌血清学鉴定和耐药性分析

2014-2016年从广州市屠宰场分离沙门氏菌151株,鉴定出12种血清型,前3位优势血清型分别为德尔卑沙门氏菌、罗森氏沙门氏菌、鼠伤寒沙门氏菌。采用微量肉汤稀释法测定分离菌株对13种抗菌药物的敏感性。结果显示,沙门氏菌对四环素、多西环素、磺胺异恶唑、氨苄西林、大观霉素的耐药率都在60%以上,对奥格门丁、恩诺沙星、氧氟沙星、头孢噻呋、多粘菌素E敏感（耐药率≤16%）。多重耐药率为79.20%。研究表明,沙门氏菌对部分药物耐药性较严重,亟需加强养殖业抗菌药物的合理应用以控制耐药性的发展。     

禽源大肠杆菌和沙门氏菌耐药性研究

本试验主要对山东、河南及安徽共5个养禽场分离的大肠杆菌和沙门氏菌进行耐药性研究．研究结果表明：所有分离菌株对大多数抗菌药物的耐药率都非常高,并且多重耐药情况严重．大肠杆菌和沙门氏菌对磺胺类药的耐药性几乎达到100％,对氨苄西林、四环素、氟喹诺酮类的耐药性也非常严重。     

内蒙古地区奶牛源致病性沙门氏菌耐药性分析及ESBLs基因检测

为研究内蒙古地区奶牛源致病性沙门氏菌的耐药特性及ESBLs基因流行特征,试验采用微量肉汤稀释法测定了22种兽医临床常用抗菌药物对临床分离沙门氏菌的最低抑菌浓度(MICs),并对其多重耐药特性进行了分析;采用PCR法对具有β-内酰胺类药物耐药表型的菌株进行了8种动物源沙门氏菌常见ESBLs基因的检测。结果显示,内蒙古地区奶牛源致病性沙门氏菌对甲氧苄啶(97.4%)及磺胺甲基噁唑(94.7%)的耐药率最高,对多数β-内酰胺类、氨基糖甙类、氯霉素类及四环素类药物的耐药性也较为严重(耐药率为40%~80%),所有菌株对氟喹诺酮类药物均敏感;菌株的多重耐药率为94.7%,有29株菌(76.3%)同时对6类抗菌药物具有耐药性;35株对β-内酰胺类药物耐药的菌株中,有30株(85.7%)为ESBLs基因阳性,共检出6种ESBLs基因,其中CTX-M型基因检出率最高(40.0%),未检出SHV和PSE型基因;共发现15种ESBLs基因型,9种ESBLs基因型组合,有16株菌(45.7%)同时携带两种或两种以上ESBLs基因。结果表明,内蒙古地区奶牛源致病性沙门氏菌对兽医临床常用抗菌药物的耐药性较为严重,且ESBLs基因的流行模式较为复杂,提示兽医临床抗菌药物不合理使用可能是造成该地区奶牛源沙门氏菌耐药性产生的原因。     

Incidence and tracking of Clostridium perfringens through an integrated broiler chicken operation

Clostridium perfringens has been shown to be widespread in the broiler chicken hatchery, grow-out, and processing operations. In a previous study, ribotypes of certain strains of C. perfringens isolated from processed chicken carcasses were shown to match ribotypes isolated from paper pad lining trays used to transport commercial chicks from the hatchery to the grow-out facility on the farm. These results suggest that C. perfringens contaminating the processed product could originate from facilities in the integrated poultry operation prior to grow out. In this study, samples were collected from the breeder farm, hatchery, previous grow-out flock, during grow out and after processing. In the first trial, C. perfringens was recovered from the breeder farms, the hatchery, previous grow-out flock, grow-out flock at 3 weeks of age, grow-out flock at 5 weeks of age, from processed carcasses, and from the breeder farm after processing in 4%, 30%, 4%, 0%, 2% and 16%, and 4% of the samples, respectively. In the second trial, the incidence of C. perfringens in samples collected from breeder farms, the hatchery, previous grow-out flock, grow-out flock at 3 weeks of age, grow-out flock at 5 weeks of age, and fromprocessed carcasses was 38%, 30%, 32%, 8%, 4%, and 8%, respectively. The genetic relatedness of the isolated strains as determined by ribotyping suggests that C. perfringens may be transmitted between facilities within the integrated broiler chicken operation.     

2株鸡源沙门菌主要表观性状及16S rRNA基因系统发育研究

对分离于发病死亡鸡的12株沙门菌（Salmonella）,进行了形态特征、理化特性、血清型、致病作用、药物敏感性等主要表观性状及系统发育的研究。其中包括O4（B）群无动力沙门菌（Salmonellagroup B O form）2株（EU073022）,血清型1,4,5,12：-：-;鸡沙门菌（S.gallinarum）2株（EU073018）,血清型1,9,12：-：-;病牛沙门菌（S.bovismorbificans）5株（EU073019）,血清型6,8：r：1,5;肠炎沙门菌（S.enteritidis）3株（EU073020）,血清型1,9,12：g,m：-。人工感染试验表明,均具有较强的致病作用;药物敏感性试验结果显示,在不同菌株间的敏感、耐药等差异不明显。     

Characterization of Salmonella spp. isolated from an integrated broiler chicken operation in Korea

The purpose of this study was to investigate the biological and genetic characterization of persistent Salmonella isolates in an integrated broiler chicken operation, in an attempt to elucidate the source of contamination. From the breeder farm, the hatchery, the broiler farm and the chicken slaughter house of an integrated broiler chicken operation, a total of 6 serotypes were observed. Although S. Heidelberg was not detected in the broiler farm, it was consistently found in the breeder farm, the hatchery and the chicken slaughter house. Also, S. Enteritidis and S. Senftenberg were found in the hatchery and the chicken slaughter house, and the hatchery and the broiler farm, respectively. S. Gallinarum and S. Blockley were found only in the broiler farm, and S. Virchow was only recovered in the chicken slaughter house. Isolated S. Heidelberg, S. Enteritidis and S. Senftenberg strains were divided into 3, 5 and 7 types, respectively, on the basis of all properties. Especially, S. Senftenberg isolates, divided into four types by their antimicrobial resistance patterns, were all obviously the XbaI PFGE pattern. Also, four S. Enteritidis isolates resistant to nalidixic acid showed a difference in phage type and PFGE pattern. Such a different pattern was shown despite Salmonella isolates originating from an integrated broiler operation, suggesting that further epidemiological studies on many integrated chicken companies in Korea are needed.     

川西北牦牛沙门氏菌的健康带菌调查及药敏实验

本文对来自四川省甘孜藏族自治州和阿坝藏族羌族自治州的临床健康成年牦牛进行了沙门氏菌带菌情况调查,并对其中一些沙门氏菌作了耐药情况检测。用世界卫生组织（WHO）推荐的Kirby-Bauer氏法对9株分离株进行药敏实验。结果从1 262例临床健康成年牦牛中最终分离鉴定出103株沙门氏菌,分离率约为8.17%;且9株分离株对14种抗菌素均表现为敏感,没有发现耐药菌株。     

Studies on feasibility of producing Salmonella-free turkeys

The feasibility of producing salmonella-free turkeys was investigated over a 5-year period. In Phase 1, a hatchery-breeder flock operation was monitored extensively for 4 years. Hatching eggs from a primary breeder over this period (1978-81) resulted in salmonella-free day-old poults from which 7500 hens and 600 tons were selected for breeders each of the 4 years. Approximately 2.5 million poults were produced over the 4 years. Salmonella arizonae was isolated from the hatchery debris over a 2-week period in 1980. The pelleted feed contained no animal protein products except fish solubles. A sample of feed from each delivery was cultured with no salmonella isolations. Environmental samples of dust and litter remained negative for salmonella. Phase 2 involved monitoring seven grow-out flocks initiated with salmonella-free poults with extra precautions directed at the feed and environment. The intestinal tracts of five of seven flocks at the time of marketing were negative for salmonella. Phase 3 involved a primary breeder-hatchery that had a 10-year history of S. sandiego infection in its breeder flocks and poults. A vaccination program using an autogenous oil-adjuvant bacterin supplementing other sanitation and management efforts resulted in elimination of S. sandiego. Because the breeder went out of business, it was not possible to determine if the freedom from salmonella could be sustained over a period of years.     

由禽波氏杆菌和奇异变形杆菌共感染为主所致鸡胚发病死亡的生物学鉴定

2010年,辽宁铁岭某种鸡孵化场出现孵化率低、死淘率高的现象。通过对送检病死鸡胚进行病原检测,最终检测出5株革兰阴性杆菌和3种病毒,其中禽波氏杆菌和奇异变形杆菌的检出率分别为76．44％、61.36％,两者混合感染率达51．83％,而其他病原菌（大肠杆菌、沙门菌、绿脓杆菌）和病毒（AI,V、REV、CIAV）的检出率均较低。采用1对根据病原菌23SrRNA基因的共同保守序列设计的引物,以及根据临床常见致病菌16s～23SrRNA基因间隔序列（ISR）两端的16S及23SrRNA保守序列而设计的通用引物分别对分离菌进行PCR扩增。并分别测定所得片段的DNA序列。结果显示,所得DNA片段分别与GenBank中收录的登录号为HM545299的禽波氏杆菌和登录号为AY993943的奇异变形杆菌的相应核苷酸序列同源性达99．5％和98．7％。本研究最终确定禽波氏杆菌和奇异变形杆菌的混合感染是引起该场疫情发生的主要原因。     

山东省小规模养鸡场沙门氏菌流行病学调查分析

对近年来山东省内鸡沙门氏菌病进行了调查,并对典型的沙门氏菌做了生化鉴定和药物敏感试验。调查表明沙门氏菌病在小规模养殖场的发生呈上升趋势,常与大肠杆菌病、新城疫和鸡球虫病混合感染。对所分离的沙门氏菌株鉴定表明,70%为副伤寒沙门氏菌,也有鸡白痢沙门氏菌的存在。药敏试验结果表明沙门氏菌对传统的防治细菌性药物产生了多重耐药性,这说明细菌耐药性的产生和发展与抗生素长期反复使用和盲目使用有密切的关系。本文为更有效地防控细菌性疫病提供科学依据。     

NaCl 胁迫对燕麦苗期形态指标的影响

在7个 NaCl 浓度（0%、0.075%、0.175%、0.275%、0.375%、0.475%、0.675%）胁迫下,对10个燕麦（Avena sativa ）品种（VAO-22、VAO-1、VAO-14、MF9715、定莜84、99AS307、QO245-7、紫燕麦、Sylva、AC1038）幼苗的存活率、株高、单株干重、根长、根数、生长胁迫指数（以盐处理下植株干重与对照植株干重的比值）6项指标进行分析测定,并采用灰色关联度对其苗期耐盐性进行了综合评价,结果表明,裸燕麦 MF9715的耐盐性最高,耐盐性依次为：MF9715、99AS307、QO245-7、VAO-14、紫燕麦、Sylva、AC1038、VAO-14、定莜84、VAO-22。     

Titration of Marek's disease cell-associated vaccine virus (CVI 988) of reconstituted vaccine and vaccine ampoules from Dutch hatcheries

Thirty-one outbreaks of Marek's disease (MD) were reported in the Netherlands and retrospectively analyzed. The outbreaks occurred mostly in vaccinated commercial layer and a few breeder flocks of several breeds; however, the cause of the outbreaks could not be established. Therefore, in a prospective study, the occurrence of true vaccine failures was assessed onfive hatcheries. The plaque-forming units (PFU) of MD vaccine per chicken dose were determined through in vitro assays on vacine ampoules (2 to 5 per hatchery) and samples of reconstituted vaccine (approximately 22 per hatchery). All forty reconstituted vaccine samples of hatcheries 1 and 4 showed PFU doses <10(3). In hatchery 4, 14 samples showed extreme low PFU (< or = 10 PFU). In hatcheries 2, 3, and 5, the numbers of MD vaccine suspensions with a titer > or = 10(3) PFU, which is the standard required, were 1 (5%), 17 (77%), and 3 (14%), respectively. Some vaccine ampoules showed < 10(3) PFU per chicken dose. This study shows the usefulness to assess the PFU per chicken dose of reconstituted MD vaccine and vaccine ampoules to unravel true vaccine failures, which could result in disease outbreaks in the field.