The effects of age, route of exposure, and coinfection with infectious bursal disease virus on the pathogenicity and transmissibility of chicken anemia agent (CAA)

Specific-pathogen-free (SPF) chickens were inoculated with several different concentrations of chicken anemia agent (CAA) by the intra-abdominal, intratracheal, or oral routes. Based on lowered hematocrit values, the birds were most susceptible to CAA introduced by the intra-abdominal route. When SPF chickens were infected with infectious bursal disease virus (IBDV) at 1 day of age, they remained susceptible to CAA up to at least 21 days, whereas birds inoculated with CAA alone were susceptible only at 1 day of age. Infectious bursal disease virus introduced at 1 day of age also increased the susceptibility of birds to contact infection with CAA and resulted in increased mortality rates in CAA inoculates. The response of SPF birds to CAA infection varied following exposure at 1 day of age to two different strains of IBDV (STC and Variant-E). Chicken anemia agent contacts and inoculates infected with the Variant-E strain were affected 1 week earlier by CAA than by STC inoculates, as evidenced by depressed hematocrits. However, the total number of birds affected was similar for both the Variant-E and STC-inoculated chickens. Commercial broiler chickens inoculated at 1, 7, 10, and 14 days of age by non-parenteral routes with CAA or a combination of CAA and IBDV had mean hematocrits that were lower than controls. Several CAA-inoculated birds were considered anemic, with hematocrit values of 25 or less, while uninoculated birds remained within normal ranges.     

禽腺病毒血清4型贵州流行株分离及其致病性分析

为了解贵州地区禽腺病毒血清4型(FAdV-4)分离株的致病性和生物学特性,本研究对经PCR检测为FAdV-4核酸阳性的肝脏样本应用鸡肝癌细胞(LMH)进行FAdV-4分离培养,通过观察FAdV-4感染细胞病变,并应用PCR、间接免疫荧光试验(IFA)、电镜等技术对分离病毒进行鉴定;然后通过动物感染试验分析分离病毒的致病...     

禽呼肠孤病毒鸡源分离株的鉴定与人工感染试验

从临床患病鸡关节病料中分离到一株病毒，在电镜下观察到病毒粒子无囊膜，有双层衣壳，外衣壳直径约75nm，内核约50nm；分离病毒株对酸、热有较强的抵抗力，对乙醚不敏感；病毒感染鸡胚成纤维细胞上能产生以融合细胞为特征的CPE；爪垫接种1日龄雏鸡表现出明显的病毒性关节炎临床症状，并伴有吸收障碍型和坏死型的病理变化；血清中和交叉试验表明，该病毒分离株与国内的2株番鸭源呼肠孤病毒（YH和YB）分离株和国外S1133呈不同程度的交叉反应。通过对分离病毒株进行电镜观察、理化特性试验、病毒感染力测定、致病性试验、血清学试验，可以确定该分离病毒为禽呼肠孤病毒。     

中国鸡群禽呼肠孤病毒抗体的检测与分析

为了解我国鸡群中禽呼肠孤病毒(Avian reovirus,ARV)的抗体水平,本研究利用ELISA方法对2010-2013年来自全国19个省市的鸡血清样品进行了ARV抗体检测。在检测的17 058份鸡血清样品中共检出ARV抗体阳性血清156 80份,阳性率为92.83%,其中免疫鸡群的阳性率为98.50%,非免疫鸡群的阳性率为74.72%。除山东省和广东省外,其他各个省市的样本阳性率均在90%以上。商品肉鸡、肉种鸡、商品蛋鸡和蛋种鸡的ARV抗体阳性率分别为72.66%,98.48%,93.40%和95.53%。部分地方品种鸡群的ARV抗体水平也较高。结果表明,我国鸡群ARV感染情况普遍存在,主要表现为一定比例的免疫鸡群抗体过高、未免疫鸡群存在较高的抗体阳性率。     

鸭源禽副黏病毒1型YH_(99) V株的毒力测定和致病特性分析

从浙江省某麻鸭养殖基地的产蛋下降病鸭生殖器官内分离到1株致产蛋锐减、不致鸭死亡的病毒,经鉴定该病毒属于禽副黏病毒1型,命名为YH99V株。该毒株经动物回归试验能成功诱导鸭发病并回收同样的病毒,通过SPF鸡胚盲传至第9代时致病性突然增强,第11代出现血凝特性,第15代血凝价趋于稳定,鸡胚半数致死量EF15ELD50为10-4.8。MDT、ICPI和IVPI毒力学指标测定以及毒力相关基因序列结构分析表明,分离株的MDT为112h,ICPI为0.225,IVPI为0.41;YH99 V株F0蛋白裂解位点(112～117位)区域氨基酸推导序列为Gly-Arg-Gln-Gly-Arg-Leu,与弱毒株相一致。证明该毒株为新城疫弱毒株。     

和缓艾美耳球虫保定株的分离鉴定与致病性研究

采用单卵囊分离技术,从河北省保定市暴发球虫病的某鸡场鸡粪便中成功分离出1株和缓艾美耳球虫(Eimeria mitis),并对其生物学特性和致病性进行了研究。该虫株主要寄生于小肠后段,潜隐期为97 h,孢子化时间为22 h,卵囊呈球形,平均大小为15.9μm×14.4μm,卵囊指数为1.10,PCR鉴定为纯种的和缓艾美耳球虫,将其命名为和缓艾美耳球虫保定株。该虫株排卵期为感染后第4～10天,其中第5～6天为高峰期。SPF鸡分别感染5×10~4个、1×10~5和2×10~5个和缓艾美耳球虫保定株孢子化卵囊后,均出现精神不振、排水样粪便等球虫病引起的临床症状,且各组相对增重率分别为78.5%、63.5%和59.3%,明显低于空白对照组,该虫株具有一定的致病性,表现为发病和减重。     

禽呼肠孤病毒广西分离株σ3基因的克隆和表达

采用RT_PCR技术扩增禽呼肠孤病毒广西分离株R1σ3基因,将σ3基因克隆至PGEX_4T_1载体上,测序结果表明插入的片段为σ3目的基因。切下目的基因σ3,重组到含有谷胱甘肽(GST)的融合蛋白质粒表达载体PGEX_4T_1,获得重组质粒经PCR酶切以及序列分析鉴定,表达σ3基因插入的位置,大小和阅码框架均正确,表明成功构建了融合表达载体PGEX_R1_σ3。构建好的重组质粒,经1mmol/LIPTG诱导,在大肠杆菌DH5α中得到了表达。融合蛋白GST_R1_σ3的分子量为60ku,以包涵体形式存在。Westernblot分析表明,融合蛋白能够与ARV阳性血清发生特异性反应,表明该重组蛋白具有良好的抗原性和特异性。     

Association of avian reovirus M and S genes with viral behavior in vivo. II. Viral pathogenicity

A group of avian reoviruses comprising serially passaged S1133 strains and their vaccine derivatives was examined biochemically to study the temporal evolution of the viruses and biologically to assess their relative pathogenicities. The strains fell into three groups of differing virulence, the viruses becoming less pathogenic the longer they were passaged. Protein and RNA profiles of the strains showed no distinct patterns of evolution nor any trend that could be correlated with pathogenicity. Nucleic acid hybridization studies of the strains indicated that all the genes were altered to some extent during passage. The S1 and M3 genes appeared to change the most during the first half of passage history, but later, as the virus was cold-adapted or passaged extensively, the M2, S2, and S3 genes also appeared to vary. When viruses were grouped according to virulence, the greatest changes were seen in the S1, M2, and M3 genes, suggesting that these may be associated with the virulence of a given avian reovirus strain.     

Effects of age, sex and housing on the trabecular bone of laying strain domestic fowl.

To determine the effects of age, sex and housing on trabecular bone volume, samples were collected from groups of male and female domestic fowl housed in cages or floor pens from four to 60 weeks old. Between 25 and 60 weeks old, trabecular bone volume decreased by 25 per cent in sections of free thoracic vertebrae (T5) from female birds, the loss occurring at an earlier age in caged birds. Over the sample period, TBV in male caged birds diminished by 35 percent, but male floor birds showed no reduction in trabecular bone volume. At 60 weeks, trabecular bone volume was 30 per cent greater in male caged birds and 40 per cent greater in male floor birds than in the corresponding females. In reproductively active females, no trabecular osteoid was observed, indicating no new trabecular bone formation. However, trabecular osteoid was present in two birds aged 60 weeks which had regressed ovaries. Osteomalacia was not seen in any of the bone samples.     

猪链球菌2型HA0609的分离及致病性研究

从江苏省某屠宰场猪的扁桃体中分离到1株细菌,通过培养特性、菌体形态、菌落形态、染色特性、生化试验以及荚膜多糖(cps)基因的PCR检测,确定为猪链球菌2型,命名为HA0609。本试验针对猪链球菌7种主要毒力因子——谷氨酸脱氢酶(gdh)、溶菌酶释放蛋白(mrp)、胞外因子(epf)、溶血素(sly)、纤连蛋白/血纤蛋白原结合蛋白(fpbs)、次黄嘌呤核苷酸脱氢酶(impdh)及毒力相关序列orf2,进行PCR检测。与已知强毒株比较,该菌株2种主要毒力因子sly和epf均为阴性。动物试验显示HA0609对猪、兔和Balb/c鼠均无致病性。     

Interaction of age,strain, sex and food restriction on plasma creatine kinase activity in turkeys

1. Plasma creatine kinase activity was determined at 4, 8, 12, 16, 20, 24, 28, 38 and 48 weeks of age in male and female traditional turkeys fed ad libitum , in male and female turkeys of a sire-line fed ad libitum or restricted to 0.5 during rearing and subsequently to 0.8 of sex-specific ad libitum -fed body weight, and in sire-line males fed ad libitum to 18 weeks and 0.8 of ad libitum body weight thereafter. 2. Plasma creatine kinase activity was low in traditional turkeys and increased rapidly after 12 weeks of age in males and females of the sire-line of turkeys. 3. Food restriction decreased the activity of plasma creatine kinase. 4. There was no difference in plasma creatine kinase activity between the sexes during rearing. After the onset of lay, the activity in plasma from females decreased at 38 weeks of age and rose dramatically in restricted females at 48 weeks when the birds had ceased laying. 5. The changes in plasma creatine kinase activity in females were associated with concomitant changes in ovarian activity as reflected in altered plasma triglyceride concentrations.     

Comparison of a vaccine strain and field isolates of avian reovirus by T1-oligonucleotide mapping.

Total RNA of eight avian reovirus isolates and the S1133 strain were compared by RNase T1-oligonucleotide mapping. The viruses were propagated in Vero cell cultures, and viral genomes were extracted from purified virions for comparison. Pairwise comparisons of the oligonucleotide maps showed genetic variation among reovirus isolates ranging from 78% to 99%. The T1 fingerprints of the RNA of isolates 1103, 724, 615, and 684 differed slightly from the standard S1133 strain, suggesting that the vaccine strain might have changed and became part of the circulating reoviruses. In contrast, when compared with the vaccine strain, isolates 902, 644, and 6207 showed greater differences in the fingerprint pattern. This genomic diversity may be due to the differences in immunological status of the affected avian population and/or due to simultaneous coinfection with different reovirus strains.     

鸡源圣雷利气单胞菌(Aeromonas sanarellii)的分离鉴定及致病性研究

对1例鸡鼻炎样病例进行细菌学诊断,从病鸡鼻腔中分离到1株革兰阴性杆菌,编号为ZY17105,对其进行形态学、生理生化、分子生物学鉴定,对其致病性和耐药性进行分析,同时探讨gyrB基因在气单胞菌鉴定中的意义。16S rDNA进化分析显示ZY17105与圣雷利气单胞菌模式菌株LMG24682同源性为99.4%;gyrB基因进化分析显示ZY17105与各圣雷利气单胞菌株形成同一进化支,与圣雷利气单胞菌模式菌株LMG24682和A2-67之间的同源性为分别为97.8%和98.2%。根据形态学、生理生化特性、16S rDNA和gyrB基因进化分析,将分离株ZY17105鉴定为圣雷利气单胞菌,分离株ZY17105对小鼠和鸡有较强致病性;另外,进化分析表明gyrB基因在气单胞菌属各种间的区分能力高于16S rDNA。本研究首次从中国大陆分离到圣雷利气单胞菌,并证实gyrB基因可替代16S rDNA用于气单胞菌属各菌种间的进化分析鉴别。     

A comparison between the effect of an avian reovirus and infectious bursal disease virus on selected aspects of the immune system of the chicken

Reovirus 81-176 was inoculated subcutaneously into day-old specific-pathogen-free leghorns and evaluated for its effects on the immune system over a 3-week period. Structural criteria included organ weights of the bursa of Fabricius (BF) and spleen (SP), scoring of histological lesions in the BF, SP, and thymus, and hematological analyses of the circulating leukocytes. Alterations in the functional capacity of the immune system were measured using the graft-versus-host reaction, the response of peripheral blood lymphocytes (PBLs) to mitogens, the ability of circulating monocytes to phagocytize latex beads, and the serological responses to Newcastle disease virus, sheep red blood cells, and Brucella abortus antigens. For comparison, infectious bursal disease virus (IBDV) was similarly evaluated by most of the same tests. Structurally, reovirus 81-176 altered BF and SP organ weights, the total numbers of white blood cells in circulation, and the degree of follicular atrophy in the BF. Functionally, reovirus inoculation reduced both the response of PBLs to the phytohemagglutinin-P stimulation and monocyte uptake of latex beads. According to the protocols used here, no significant alteration in B-cell function could be detected in reovirus-infected chicks. With the exception of leukocyte hematology, IBDV-infected chicks had significantly altered responses in all tests used. By way of comparison, the effects of IBDV were more persistent and pronounced than were those seen with reovirus. The graft-versus-host reaction indicated an elevated and/or uninhibited response of T-cells in the blood of IBDV-infected chicks.     

Effect of age and sex on the eviscerated yield,muscle and edible offal of a commercial broiler strain

1. Comparisons of the effects of age (up to 364 d) and sex on the rate of growth, eviscerated yield, blood loss and yield of skin, breast, thigh and drumstick muscles, heart, liver and gizzard have been made on a commercial British broiler strain.

2. Male broilers grew linearly up to 76 d whereas the females grew more slowly after 35 d. At 364 d the difference in live body weight was small.

3. Up to 56 d, males had a slightly higher eviscerated yield. There was no significant difference between the sexes beyond this age.

4. The proportional yield of breast muscle was variable with age, whereas that of thigh and drumstick increased with age in the male, particularly at 175 and 364 d.

5. After 35 d the proportion of skin was larger in females, whereas the gizzard as a proportion of live body weight in both sexes decreased rapidly between 21 and 76 d. Blood loss during killing was always higher in male broilers after 21 d.