竹丝鸡鸡白痢沙门菌的分离鉴定与药敏试验

为查找广东某鸡场竹丝鸡雏鸡发病原因,对疑似鸡白痢沙门菌病残鸡进行细菌分离培养、聚合酶链式反应(PCR)鉴定及药敏试验。结果显示:该鸡场共分离获得2株鸡白痢沙门菌(S1、S2),鸡白痢沙门菌是鸡场此次疫病的主要病原;头孢他啶、头孢曲松、阿米卡星、卡那霉素、氟苯尼考、盐酸四环素、甲氧苄啶等抗菌药为2株分离株的首选药物,临床治疗建议避免使用阿莫西林、氨苄西林、链霉素。试验表明,对鸡白痢沙门菌的治疗应重视多重耐药性问题,通过药敏试验选择高敏药物指导临床合理用药。     

黄羽肉种鸡场死胚中鸡白痢沙门菌的分离鉴定及耐药性分析

为了解广东某黄羽肉种鸡场鸡白痢沙门菌的感染及耐药情况,随机采集该场孵化至21日胚龄时的117个死亡胚进行鸡白痢沙门菌分离,通过生化鉴定、PCR、血清分型及rfbS基因序列测定等鉴定鸡白痢沙门菌分离株,并采用琼脂稀释法测定分离株对16种抗菌药物的最小抑菌浓度(MIC)。结果显示:共分离鉴定出18株鸡白痢沙门菌,分离率为15.38%;18个鸡白痢沙门菌分离株对头孢噻肟、头孢吡肟、亚胺培南、多黏菌素B、庆大霉素、阿米卡星、氟苯尼考、链霉素均敏感,而对磺胺异吡恶唑(100%)、萘啶酸(72.2%)、氨苄西林(55.6%)、四环素(44.4%)耐药率较高,且分离株均有多重耐药性。研究表明该黄羽肉种鸡场已受到鸡白痢沙门菌感染的困扰,有必要尽快启动鸡白痢沙门菌防控与净化。     

河北省保定地区雏鸡鸡白痢沙门菌的分离鉴定及耐药性检测

为了有效防控雏鸡白痢沙门菌病,试验自保定某蛋种鸡场疑似为鸡沙门菌病雏鸡采取肝脏病料进行细菌的分离培养、生化鉴定、鸡白痢沙门菌多价血清学鉴定及沙门菌inv A基因PCR鉴定,并对分离菌进行致病性检测及对7种抗菌药物的耐药性检测。结果表明,10株分离菌的形态特征、生化特性、血清学鉴定结果符合鸡白痢沙门菌的特征,说明从病死雏鸡组织中分离出的细菌为鸡白痢沙门菌。沙门菌inv A基因PCR扩增出373 bp目的片段,进一步证实分离菌为鸡沙门菌。用分离菌株接种雏鸡,其死亡率高达90%,表明该菌株有很高的致病性。10株分离菌株对阿莫西林和青霉素耐药率达100%,对阿齐霉素的耐药率达90%,而对阿米卡星、头孢唑林和庆大霉素耐药性较弱、对氧氟沙星较为敏感。3耐菌株多重耐药比例为42.86%,4耐菌株多重耐药比例为57.14%,5耐菌株多重耐药比例为71.43%,说明雏鸡沙门菌分离株对所试抗菌药物均表现出不同程度的耐药性,且表现出多重耐药现象。本试验为鸡白痢沙门菌病的防治提供参考。     

2010～2012年苏皖两省鸡白痢沙门菌的分离鉴定及耐药性分析

通过培养特性、生化反应、血清学试验及PCR检测,对2010年10月～2012年6月江苏、安徽两省送检的禽源病料进行鸡白痢沙门菌的分离鉴定,并对分离株进行药敏试验.结果从1 420份样品中分离鉴定出55株鸡白痢沙门菌;药敏试验结果表明,分离株对萘啶酸、羧苄青霉素、链霉素、磺胺异恶唑和氨苄西林等抗菌药物具有较高的耐药性,而对庆大霉素、头孢曲松、氯霉素和卡那霉素具有较强敏感性.上述鸡白痢沙门菌分离株中,有49株(89.09％)为多重耐药菌株,其中以四耐菌株比例最高(25.45％).本研究对于指导临床合理用药及控制鸡白痢沙门菌发生与流行具有重要的现实意义.     

鸡白痢沙门菌等位基因特异性PCR检测方法的建立

根据鸡白痢沙门菌与鸡伤寒沙门菌的rfbS基因在第237和第598位碱基的不同，设计和合成了等位基因特异性PCR引物，建立了快速检测鸡白痢沙门菌的PCR方法，并应用该方法对鸡白痢沙门菌临床分离样品进行了PCR鉴定。结果显示，该PCR方法能够特异性地鉴定鸡白痢沙门菌，检测灵敏度达100PgDNA。对35个经常规方法鉴定的鸡白痢沙门菌分离株应用等位基因特异性PCR方法进行鉴定，鉴定出33株鸡白痢沙门菌，符合率为94．3％。表明，建立的等位基因特异性PCR方法能够准确而快速地鉴定鸡白痢沙门菌。     

华东地区禽源沙门菌的分布及血清型分析

本文旨在研究中国华东地区沙门菌分布情况及其优势菌的血清型变异情况.收集2010年10月-2012年8月来源于江苏、安徽、上海等地区的1 730份禽源样品,进行增菌、纯化、PCR鉴定、血清学试验及生化试验,并对7株血清型有变异的疑似鸡白痢沙门菌进行进一步序列测定,对血清型进行分析.结果表明,本次调查中共分离沙门菌87株,包括鼠伤寒沙门菌、德尔卑沙门菌、圣保罗沙门菌、鸡白痢沙门菌及阿巴特图巴沙门菌,并且在鸡白痢沙门菌血清型H相存在Hd、Hg阳性的现象.结果提示,中国禽类沙门菌感染以鸡白痢沙门菌感染为主,且存在血清型变异情况,变异概率呈上升趋势.     

禽沙门菌分离鉴定方法的建立与优化

本研究旨在建立和优化禽沙门菌分离鉴定的方法。选择山东省某地方品种鸡疑似发生沙门菌病的未出壳死胚样品24份,通过4组不同的培养基组合比较沙门菌的分离情况,选择沙门菌通用引物两对和鸡白痢特异性引物一对进行分离株的PCR检测,比较其特异性,然后进行血清分型和MLST分型,确定沙门菌类型并研究其相关性。结果表明,该批样品中TTB比SC增菌效果好,XLD和XLT4选择培养基对沙门菌分离效果相同,最终通过PCR方法鉴定出沙门菌阳性率为58.3%;发现两对通用引物特异性一致,表明可任选一对引物进行PCR鉴定;用鸡白痢沙门菌特异性引物完成的PCR鉴定结果显示,14株沙门菌中有7株为鸡白痢沙门菌;血清型鉴定结果表明,14株沙门菌中有7株鸡白痢沙门菌、7株肠炎沙门菌,共两种血清型;MLST分子分型结果表明,14株沙门菌分为3个ST型,分别是7株ST11、3株ST92和4株ST2151,其中ST11为肠炎沙门菌,ST92和ST2151为鸡白痢沙门菌;本研究表明,疑似沙门菌感染样品经BPW和TTB增菌、XLD或XLT4选择培养基分离培养,最后通过PCR方法可准确完成沙门菌的分离鉴定;同时,应用该方法可完成对鸡白痢沙门菌的鉴定,并与血清分型和MLST分子分型结果一致。     

鸡白痢与鸡伤寒沙门菌双重PCR方法的建立和初步应用

分析不同血清型/生物型沙门菌全基因组序列筛选出鸡白痢和鸡伤寒沙门菌特异性基因组序列,设计两对引物,建立双重PCR方法鉴别检测鸡白痢和鸡伤寒沙门菌不同生物型,并进行初步的临床应用。双重PCR方法结果显示,鸡白痢沙门菌显示417 bp条带,鸡伤寒沙门菌显示417 bp和636 bp两个条带,而阴性对照未出现条带,与预期设计相符。双重PCR体系对56株不同血清型沙门菌鉴定结果与细菌学分离的血清型鉴定结果完全一致,说明本试验建立的双重PCR体系特异性良好,应用上述方法检测鸡场疑似20份临床样本,结果发现8株鸡白痢沙门菌阳性,1株鸡伤寒沙门菌阳性。上述结果表明,已建立双重PCR方法特异性检测鸡白痢和鸡伤寒沙门菌。本试验为鸡白痢和鸡伤寒不同生物型沙门菌的检测提供了一种简洁、敏感、特异的新方法。     

鸡白痢沙门菌和鸡伤寒沙门菌PCR-RFLP鉴别

根据鸡白痢沙门菌和鸡伤寒沙门菌flic基因可变区两端的保守序列设计1对引物,PCR扩增出约866 bp的产物,用HinplI对PCR产物进行酶切,经RFLP分析区分鸡白痢沙门菌和鸡伤寒沙门菌.利用该技术对1株鸡白痢沙门菌标准株及2株鸡伤寒沙门菌标准株进行分子鉴别,结果与预计的RFLP模式相符,证明该方法可行.在此基础上...     

黄羽肉鸡鸡白痢沙门菌的分离鉴定

文章通过增菌、选择性培养、形态学染色、生化鉴定、glg C和spe C二重PCR鉴别、ERIC和Xba-IPFGE谱型分析、代表株体内致病性试验等方法,对一家规模化黄羽肉鸡养殖企业开展5年(从2013～2018年)鸡白痢沙门菌的分离和鉴定。从2013～2018年共分离鸡白痢沙门菌82株。对乌骨鸡源的临床代表菌株LHSP001以1×106～1011cfu口服感染8日龄的健康慢速型草鸡,各个感染组在感染后持续观察8 d,鸡群表观正常。病理剖检出现肝脏坏死点、心脏肉芽肿等。上述所分离的细菌将为后续黄羽肉鸡鸡白痢的净化提供有意义的支撑。     

Horizontal transmission of a pancreas-passaged avian encephalomyelitis virus in chicks

Pancreas-passaged avian encephalomyelitis (AE) virus was transmitted horizontally in a group of 40 (1-day-old) chicks within 3 weeks after they were intermingled with two orally infected 1-day-old chicks. Viral antigen was detected in the pancreas of these contact-exposed chicks. After 5 weeks, contact-exposed chicks developed high titers against AE virus, but the chicks did not develop clinical signs of AE. The passaged virus could not be recovered from feces of six immunized chicks.     

应用PCR从AA肉种鸡鸡胚和1日龄雏鸡体内检测出CIAV和ALV-J

为了了解鸡传染性贫血病病毒（CIAV）和J亚群禽白血病病毒（ALV-J）对AA肉种鸡鸡胚和1日龄雏鸡的感染情况,试验直接从山东省3个不同AA肉种鸡场的鸡胚和1日龄雏鸡的肝、脾、肾、胸腺、骨髓、法氏囊等组织（器官）提取DNA,进行了PCR扩增及PCR产物的克隆和序列测定。结果显示,被检的3个肉种鸡场的鸡胚和1日龄雏鸡体内均有这2种病毒核酸的检出,其中CIAV的阳性率是20.42%,ALV-J的阳性率是15.83%,二者共感染的阳性率是6.25%;在阳性检出率中弱雏〉死胚〉健康雏〉正常胚。病毒核酸在各感染组织（器官）的含量也有所差异,CIAV以脾的含量最多,ALV-J以肾的含量最多。对肝进行细菌分离鉴定时发现,3个鸡场还存在大肠杆菌、沙门氏菌和霉形体的混合感染。结果提示,在AA肉种鸡鸡胚和雏鸡体内存在CIAV、ALV-J的感染和二者的共感染以及继发性大肠杆菌、沙门氏菌和霉形体的混合感染。     

Experimental infection in specific-pathogen-free chicks with avian reovirus and avian nephritis virus isolated from broiler chicks showing runting syndrome

Avian reovirus (ARV) and avian nephritis virus (ANV) were individually isolated from runty 10-day-old broiler chicks. The ARV isolate, IR-R, the ANV isolate, IR-N, and the reference strain of ANV, G-4260, were inoculated orally into 1-day-old chicks of two specific-pathogen-free (SPF) chicken lines, 151 and PDL-1. Growth retardation without the presence of gross lesions was clearly observed at 7 and 14 days postinoculation (PI) in chicks of both lines inoculated with the IR-R strain. On the other hand, in chicks inoculated with IR-N strain, growth retardation was observed only in the chicks of line 151 at 7 and 14 days PI. Microscopically, nephritis was observed in both chicken lines at 7 and 14 days PI. When chicks that were inoculated with the IR-N strain at 1 day of age were inoculated with the IR-R strain at 3 days of age, growth retardation was observed in the chicks of line PDL-1 at 10 and 17 days PI. However, the growth retardation was less severe than in the group receiving a single inoculation of the IR-R strain.     

禽波氏杆菌病原性研究及防治初探

本文介绍了禽波氏杆菌的培养特性、药物敏感性及生化特性，重点进行了禽波氏杆菌对雏鸡的致病性研究，结果证明本菌主要引起雏鸡的急性死亡，主要病变为肺脏出血，肝脏出血、边缘坏死，肾脏出血，腺胃、肌胃浆膜出血，肠道出血。不同日龄的雏鸡对禽波氏杆菌的抵抗力不同，与日龄成正相关。同时还用不同来源的8株禽波氏杆菌制成蜂胶灭活苗，并分别免疫1日龄和11日龄雏鸡，结果发现本疫苗对雏鸡的保护率低。     

不同断喙方式对蛋雏鸡生长性能和免疫功能的影响

为研究不同断喙方式对雏鸡生长性能和免疫功能的影响,将720只1日龄蛋雏鸡随机分为不断喙、红外线断喙和热刀断喙组。测量不同时间雏鸡喙长度、体重、采食量和饲料浪费量,计数血液中异嗜性粒细胞和淋巴细胞数,通过生化分析和ELISA试剂盒检测血浆中蛋白和细胞因子含量。致死雏鸡后,采集脾和法氏囊并称重,对脾进行组织学和Real-time PCR检测。结果显示,断喙显著降低雏鸡喙长度,且红外线断喙效果更明显;断喙显著减少饲料浪费量,但不影响采食量;红外线断喙组4日龄雏鸡体重显著低于不断喙组;红外线断喙组29日龄雏鸡传染性法氏囊病毒抗体滴度极显著低于其他组;断喙对鸡血液中异嗜性粒细胞和淋巴细胞数、血浆中的蛋白、T细胞分化簇和细胞炎性因子的含量、脾和法氏囊器官指数、脾显微结构及其炎性因子和凋亡因子表达无显著影响。结果表明,断喙可显著减少饲料浪费,且红外线断喙比热刀断喙可更有效地保证断喙效果;红外线断喙可暂时性地引起雏鸡体重降低,但对后期体重无显著影响;1日龄时雏鸡在红外线断喙后又进行疫苗接种,可能会缩短疫苗对雏鸡的保护时间。     

Genetic variations in maternal transfer and immune responsiveness to infectious bursal disease virus

The immune responsiveness to infectious bursal disease virus (IBDV) in four native and crossbred chicken lines was compared. ELISA IBDV antibody titers in hen serum samples, yolk from matched eggs and sera from matched 1-day-old chicks from each chicken line with an identical vaccination program were measured, and plotted. There was considerable variation between lines in the measured IBDV specific antibodies, in vaccinated parent hens and in the amounts of inherited maternally derived antibodies in both yolk and progeny chicks. Differences in ratios of the inherited antibody level from hen to 1-day-old chicks were also found among different chicken lines. Breed differences in regressions of IBDV antibody levels in yolk to that of hen or progeny chicks' sera were also found, so prediction of serum titer of hen and/or progeny chicks from yolk are varied among chicken lines.     

Peripheral leptin effect on food intake in young chickens is influenced by age and strain

The acute effect of leptin on the regulation of food intake was investigated in layer and broiler chickens. In an initial study, we observed that a single intraperitoneal injection of recombinant chicken leptin (1 mg/kg BW) dramatically reduced (38%) food intake in 56-day-old layer chickens, more moderately reduced (15%) food intake in 9-day-old layer chicks, and had no significant effect in 9-day-old broiler chicks. In a subsequent study, body weight and plasma concentrations of leptin were measured weekly in layer and broiler chicks from day 1 to 35 of age and brain leptin receptor and neuropeptide Y (NPY) mRNA expression were analyzed at 1, 9, and 35 days of age. At day 1 of age, peripheral concentrations of leptin were significantly greater in layer than broiler chicks. Subsequently, despite increases in body weight and differences in growth rates between layer and broiler chicks from day 8 to day 35 of age, peripheral concentrations of leptin were constant and similar in both genotypes. Leptin receptor and NPY mRNA were expressed in brain from day 1 in chicks of both genotypes and increased significantly to day 35 of age. These observations provide evidence that the inhibitory effect of leptin on the regulation of food intake in growing chicks is an age dependent process. Furthermore, acquisition of the anorectic effect of leptin is likely to be associated with greater expression of the leptin receptor and NPY mRNAs than to changes in blood levels of leptin. Finally, this study provides evidence that chickens selected for high growth rates may be less sensitive or responsive to peripheral concentrations of leptin than chickens with low growth rates (layers), suggesting that the faster growth of broiler chicks may be related to a lessened responsiveness to anorexigenic factors.     

不同水平纤维素酶对肉鸡屠宰性能、免疫器官指数及血清生化指标的影响

试验选用1dAA＋肉鸡1050羽作为试验动物,随机分为7个组,每组5个重复,每个重复30羽,分别饲喂不同水平的纤维素酶,试验期42d,分别于21d、42d测定肉鸡的屠宰率、半净膛率和腹脂率;同时测定胸腺、脾脏和法氏囊的器官指数及血清中血糖和尿素氮含量。试验结果表明,添加不同水平的纤维素酶对21d肉鸡的屠宰性能并未产生显著影响;对42d肉鸡的屠宰率和半净膛率效果显著,试验组G屠宰率提高了3.94%（P〈0.05）,试验组E半净膛率提高了2.46%（P〈0.05）。肉鸡胸腺、脾脏、法氏囊器官指数有不同程度的提高,差异不显著。添加不同水平的纤维素酶能够显著提高1～21d和22～42d肉鸡血液内葡萄糖含量,试验组G提高了4.25%（P〈0.05）。添加不同水平的纤维素酶能够降低1～21d肉鸡血清内尿素氮水平,试验组G降低了6.14%（P〈0.05）。     

Comparison of phytohemagglutinin-induced cutaneous hypersensitivity reactions in the interdigital skin of broiler and layer chicks

Cutaneous basophil hypersensitivity (CBH) responses elicited by intradermal injection of phytohemagglutinin-P (PHA-P) were evaluated in the interdigital skin of broiler and layer chicks. Five- and 10-day-old broiler and layer chicks were injected intradermally in the interdigital skin between the third and fourth digits of the right foot with 25, 50, or 100 micrograms of PHA-P. The PHA-P-elicited CBH responses were evaluated by determining interdigital skin thickness before injection and 12 and 24 hr after injection. The effect of treatment with the T-lymphocyte-suppressing drug cyclosporin on the CBH responses was determined in chicks treated daily for 4 days before skin-testing at 10 days of age. Injection of PHA-P caused significant increases (P less than 0.005) in skin thickness at all dose amounts, with the maximum CBH response elicited by 100 micrograms PHA-P. The CBH response was significantly higher (P less than 0.05) in broilers than in layers. Treatment with cyclosporin significantly suppressed (P less than 0.01) the CBH response in 10-day-old chicks of both types. Results indicate that the interdigital skin-test is a rapid, simple means of assessing normal and suppressed T-lymphocyte-mediated immune response in broiler and layer chicks.     

Glycosylation of the envelope protein of West Nile Virus affects its replication in chicks

Birds are important for the transmission of West Nile virus (WNV) in nature, but the significance of the potential N-linked glycosylation at position 154 in the WNV envelope (E) protein with regard to viral replication in young chickens has not been assessed. In this study, the effect of glycosylation of the WNV E protein on viral pathogenicity in birds was investigated using young domestic chicks. A higher viral load was detected in the blood and the peripheral organs, particularly the hearts, of 2-day-old chicks inoculated with a glycosylated WNV variant compared to those inoculated with the nonglycosylated variant. There was no significant difference in the neutralizing antibody titers and cytokine expression profiles in chickens inoculated with the glycosylated and the nonglycosylated WNV variants. In contrast, no virus w as detected in the blood and the tissues of 3-wk-old chicks, although the host immune response was induced to similar levels as in the 2-day-old chicks. These data indicate the utility of young domestic chicks as an animal model of WNV infection; they also indicate that glycosylation of the E protein of WNV enhances multiplication in the blood and peripheral organs, which is associated with the strong pathogenicity of WNV in birds.