禽流感HA7基因DNA疫苗的构建及免疫保护效果研究

为了研究禽流感HA7亚型DNA疫苗,将HA7基因插入高效真核表达载体pCAGGS,构建了DNA疫苗质粒pCAGGHA7,然后将pCAGGHA7以100μg剂量一次免疫3周龄SPF鸡,免疫SPF鸡后两周抗体水平达到3Log2,4周后用100 LD50的H7N1鼻腔途径进行攻击毒,发现100μg pCAGGHA7可形成8/8的免疫保护。表明禽流感DNA疫苗pCAGGH7诱导的保护性抗体免疫反应水平较高,能达到免疫保护效果。     

H5亚型禽流感灭活疫苗的研制

以H5N1亚型禽流感病毒接种鸡胚，收取鸡胚液为抗原研制了禽流感灭活疫苗，并对制苗抗原以及疫苗的物理性状、安全性、免疫效力、免疫剂量和保护期等进行了试验。结果表明，获得了较高效价的制苗抗原．灭活前后HA分别为2^10.0-11.0和2^9.0-10.0。试制疫苗物理性状符合要求，安全性良好。试验鸡在免疫14d后血清抗体达到2^7.5，210d血清HI抗体仍维持在2^5.0左右；免疫后18d使用高致病性H5亚型禽流感病毒攻毒，获得100％的保护。分别使用0．1，0．3和0．5mL免疫鸡，发现0．3～0．5mL剂量组免疫后血清抗体达到2^7.4-8.0。疫苗在2～8℃保存540d，疫苗物理性状没有改变，免疫后血清HI抗体可以达到2^7.5。这提示研制的疫苗可以有效控制禽流感H5亚型的流行。     

三黄鸡临床多发性肿瘤相关的J亚群禽白血病病毒的分离鉴定及其病理学观察

为研究蛋鸡多发性肿瘤的病因,本实验对病鸡肿瘤组织进行病毒分离、培养及PCR检测,均扩增出针对J亚群禽白血病病毒(ALV-J)gp85基因序列的阳性条带;组织病理学观察显示发病鸡呈现髓细胞瘤、血管瘤以及纤维肉瘤等多发性肿瘤的病理变化;肿瘤细胞通过血液转移、浸润,在肝和脾组织形成局灶性或弥漫性肿瘤病灶。免疫组化染色显示肿瘤组织内,部分肿瘤细胞呈现阳性反应,表明只有部分肿瘤细胞存在ALV-J感染,而大部分肿瘤细胞检测结果呈阴性。这些病毒检测为阴性的肿瘤细胞可能是正常细胞转化为肿瘤细胞大量克隆化增殖的结果。     

禽传染性喉气管炎病毒体内外感染模型中鸡Toll样受体21mRNA转录水平的研究

为研究病毒与机体的相互作用,本研究参考GenBank中鸡Toll样受体21 (ChTLR21)的基因序列设计实时定量PCR特异性引物,以鸡核糖体蛋白L4 (RPL4)为内参基因,建立检测ChTLR21 mRNA相对转录水平的实时定量PCR方法,分析ChTLR21在禽传染性喉气管炎病毒(ILTV)感染的鸡胚成纤维细胞(CEF)中和感染SPF雏鸡免疫器官脾脏、法氏囊和胸腺组织中的转录水平.结果显示:ILTV感染CEF后2h、4h、8h和18h时间点ChTLR21 mRNA转录水平分别为未感染对照细胞的1 540.53、0.98、1.19和3.70倍.但仅在ILTV感染2h时引起ChTLR21转录水平升高,与对照组相比差异显著(P＜0.05).ILTV感染SPF雏鸡后6h、24 h和30 h脾脏中ChTLR21 mRNA转录水平分别为未感染对照的56.34、59.85和3.61倍;法氏囊中分别为0.03、25.98和3.08倍;胸腺中分别为2.52、50和7.32倍.在感染初期,脾脏中ChTLR21转录量显著升高,随后有所降低,但均高于未感染对照(p＜0.05);法氏囊中仅在感染6h时呈显著抑制(p＜0.01);胸腺中呈波动性转录水平升高,但与对照组无统计学差异(p＞0.05).本研究证明ChTLR21参与了鸡体对ILTV感染的应答,并在体内外感染模型中呈现不同的表达规律.     

我国部分地区蛋鸡ALV-J分离株gp85基因遗传进化分析

为了解我国蛋鸡J亚型白血病病毒(ALV-J)株来源及其遗传进化关系,本研究对2009年从我国6个省区蛋鸡场分离到的19株ALV-J的gp85基因进行克隆和测序,并与11个ALV-J参考株gp85基因作了比较分析.结果表明:19个ALV-J分离株的gp85基因长度为894bp～924 bp不等,分别编码298～308个氨基酸;各病毒株间gp85推导氨基酸的同源性为71.3%～100%.遗传进化分析表明,目前我国蛋鸡ALV-J分离株来源复杂,其中13个分离株与英国原型株HPRS-103、国内麻黄肉鸡株SCAU-0901亲缘关系较近;3个分离株与美国株ADOL-7501及国内白羽肉鸡株HN0001处在同一大的分支;而另外3个分离株则各自形成独立的分支,表明其gp85基因发生了较大变异.本研究表明,19个分离株与国内早期肉鸡分离株亲缘关系较远,提示我国当前蛋鸡ALV-J株可能并非源自国内早期肉鸡ALV-J株,其来源有待进一步研究.     

J亚群禽白血病病毒四川株SCMS1的分离及其囊膜基因序列分析

为研究四川首例分离株SCMS1株J亚群禽白血病病毒(ALV-J)囊膜(env)基因是否产生新的变异,本实验通过将病毒接种鸡胚成纤维细胞、RT-PCR及PCR检测技术,对包括env基因和部分3'UTR在内的片段进行PCR扩增、克隆和序列测定.结果发现该病毒gp85基因出现6个碱基的缺失,与HPRS-103原型株6995位～7242位对应的核酸序列也发生248个碱基的缺失,包括整个rTM区域.这些碱基的缺失在病毒致病性变化中可能具有潜在的意义.     

STATIC AND DYNAMIC 18FDG‐PET IN NORMAL HISPANIOLAN AMAZON PARROTS (AMAZONA VENTRALIS)

Positron emission tomography (PET) is often used to stage and monitor human cancer and has recently been used in a similar fashion in veterinary medicine. The most commonly used radiopharmaceutical is 2‐Deoxy‐2‐[¹⁸F]‐Fluoro‐d ‐glucose (¹⁸F‐FDG), which is concentrated and trapped within cells that use glucose as their energy substrate. We characterized the normal distribution of ¹⁸F‐FDG in 10 healthy Hispaniolan Amazon parrots (Amazona ventralis) by performing whole body PET scans at steady state, 60 min after injection. Significant variability was found in the intestinal activity. Avian species are known to reflux fluid and electrolytes from their cloaca into their colon. To evaluate reflux as the cause of variability in intestinal distribution of ¹⁸F‐FDG, dynamic PET scans were performed on the coelomic cavity of six Hispaniolan Amazon parrots from time 0 to 60 min postinjection of radiotracer. Reflux of radioactive material from the cloaca into the colon occurred in all birds to varying degrees and occurred before 60 min. To evaluate the intestinal tract of clinical avian patients, dynamic scans must be performed starting immediately after injection so that increased radioactivity due to metabolism or hypermetabolic lesions such as cancer can be differentiated from increased radioactivity due to reflux of fluid from the cloaca.     

Plant‐frugivore interactions in an intact tropical forest in north‐east Thailand

Fleshy‐fruited plants in tropical forests largely rely on vertebrate frugivores to disperse their seeds. Although this plant–animal interaction is typically considered a diffuse mutualism, it is fundamental as it provides the template on which tropical forest communities are structured. We applied a mutualistic network approach to investigate the relationship between small‐fruited fleshy plant species and the fruit‐eating bird community in an intact evergreen forest in northeast Thailand. A minimum of 53 bird species consumed fruits of 136 plant species. Plant‐avian frugivore networks were highly asymmetrical, with observed networks filling 30% of all potential links. Whereas some of the missing links in the present study might be due to undersampling, forbidden links can be attributed to size constraints, accessibility and phenological uncoupling, and although the majority of missing links were unknown (58.2%), many were probably due to a given bird species being either rare or only a very occasional fruit eater. The most common frugivores were bulbuls, barbets and fairy‐bluebirds, which were responsible for the majority of fruit removal from small fleshy fruited species in our system. Migratory birds seemed to be a minor component of the plant‐frugivore networks, accounting for only 3% of feeding visits to fruiting trees; they filled 2% of the overall potential networks. The majority of interactions were generalized unspecific; however, Saurauia roxburghii Wall. appeared to be dependent on flowerpeckers for dispersal, while Thick‐billed Pigeons were only seen to eat figs.     

Vaccination of Poultry Workers: Delivery and Uptake of Seasonal Influenza Immunization

Avian influenza is a highly infectious disease in poultry and although the risk of human infection is low, concerns exist that it could evolve into a new human strain of pandemic potential if reassortment with a human influenza virus occurs. In January 2007, the UK government introduced a programme to vaccinate poultry workers to reduce the potential of such an event. This study evaluates the delivery, uptake and costs of the programme in three counties of England. A questionnaire survey was completed by consultants in public health in all the Primary Care Trusts in Norfolk, Suffolk and Cambridgeshire in May 2007. The delivery of the programme varied between Primary Care Trusts, including being delivered in some cases by clinics in primary care, by general practitioners and occupational health services in others. The uptake of vaccination was low ranging from 7% to 29% at a cost of £29 to £132 per person vaccinated. Vaccination of poultry workers as a public health measure to prevent an influenza pandemic is likely to be ineffective with the level of coverage found in this evaluation in our region.     

鸡大肠杆菌菌苗免疫原性的研究

用不同的时间和温度培养鸡致病性Ｅ．ｃｌｏｉ，４１℃，４８ｈ和３７℃，７２ｈ均可见众多的菌毛，而在４１℃，４８ｈ的条件下，活菌数及菌毛产量均高于３７℃，７２ｈ的。用提取的菌毛、超声波灭活的Ｅ．ｃｏｌｉ制成油乳苗免疫鸡，用同型Ｏ５０菌株经后胸气囊攻毒后，免疫鸡无死亡，而氢氧化铝胶菌苗免疫组的死亡率为２０％，未免疫攻毒组死亡率为３３．３％。各免疫组幸存鸡平均病变级数均显著低于未免疫攻毒组（Ｐ＜０．０１）。Ｅ．ｃｏｌｉ的菌毛剂量为１２０μｇ／只和８０μｇ／只的油乳苗免疫组，在免疫后用同型Ｏ５０菌株攻击时，保护指数最高；当剂量为６０μｇ／只时，保护指数较高；而用３０μｇ／只的剂量免疫时，免疫无效。雏鸡经含菌毛、不含菌毛的Ｅ．ｃｏｌｉ超声波灭活油乳苗和福尔马林灭活油乳苗免疫后用同型Ｏ５０菌株攻毒，含菌毛菌苗的保护指数均高于相应的不含菌毛菌苗的；超声波灭活菌体的油乳苗免疫组幸存鸡平均病变级数低于福尔马林灭活菌体的油乳苗免疫组的；各免疫组与未免疫攻毒组幸存鸡平均病变级数差异极显著（Ｐ＜０．０１）。