禽流感病毒分子生物学的研究进展

禽流行性感冒(av ian in fluenza,A I,简称禽流感)是由A型禽流感病毒(av ian in fluenza v irus,A IV)引起的禽类烈性传染病。作为被世界动物卫生组织(O IE)定为A类的传染病,A I不仅给世界养禽业造成了巨大的经济损失,而且对人类健康和生命安全构成了严重威胁。因此,A I已经成为人们关注的焦点,国内外学者也对其进行了大量研究。作者从病原基因组及其编码的蛋白质、致病力、变异性以及对人类感染A IV的分子机制等角度就A IV的分子生物学研究作一综述,为防制A I提供理论基础,并在此基础上探讨了人类禽流感的防治措施,加深人们对A I的认识。     

J亚群禽白血病SYBR Green Ⅰ荧光定量PCR检测方法的建立

为建立一种能够快速、灵敏和特异地检测J亚群禽白血病病毒(ALV-J)的SYBR Green Ⅰ荧光定量PCR方法,本研究以ALV-J原型病毒株HPRS-103的pol基因3′端与gp85编码基因之间的保守区域(5 258 bp～5 802 bp)为检测目的片段,构建重组质粒并作为靶基因,通过对其浓度、引物浓度和退火温度的优化,建立了ALV-J SYBR Green Ⅰ荧光定量PCR方法.结果显示该方法的最低检测限度为2.36×102拷贝/μL,比普通PCR高100倍;与其他禽源病毒无交叉反应;批内和批间变异系数均小于5％.表明本研究建立的方法具有良好的特异性、稳定性和灵敏性.采用该方法对100只临床病鸡进行检测,ALV的检出率为44％.随机选择10只感染阳性鸡,检测病毒基因在心脏、肝脏、脾脏、肺脏和肾脏中的分布与表达水平,结果表明ALV-J在各主要脏器中均有分布,但以肾脏中的病毒表达量最高.     

鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析

H19N2病毒可以感染多种禽类和哺乳动物，包括人类。流感病毒血凝素受体结合位点的氨基酸可以影响受体结合特性。为了解鸡源株与人源株H19N2病毒受体结合部氨基酸的差异，作者对二者进行了比较与分析，结果表明，鸡源株与人源株血凝素受体结合部位氨基酸的第137、183、190、226位点存在差异，鸡源株在这些位点分别为K、N、AorVorT、LorQ而人源株则分别为R、H、E、L。虽然尚不清楚这些位点的改变对病毒与细胞亲和力及宿主范围的影响，但由于H9N2在我国养鸡业的普遍存在，加强H9N2病毒的分子流行病学监测有重要意义。     

A novel herpesvirus associated with chronic superficial keratitis and proliferative conjunctivitis in a great horned owl (Bubo virginianus)

An adult great‐horned owl (Bubo virginianus; GHOW) presented with a history of recurrent corneal ulceration of the right eye (OD). Findings included ulcerative superficial keratitis, proliferative conjunctivitis, and iris pigmentary changes. The ulcer was initially nonresponsive to medical therapy, but showed rapid and appropriate healing following diamond burr debridement. Proliferative conjunctivitis markedly improved following topical antiviral therapy with cidofovir 1%, interferon alpha 2B ophthalmic solutions, and oral l ‐lysine. Histopathologic evaluation of a conjunctival biopsy revealed epithelial features suspicious for viral cytopathic changes and intranuclear structures suspicious for viral inclusions, suggestive of a possible viral‐induced papillomatous conjunctivitis. A novel alphaherpesvirus, referred to as Strigid Herpesvirus 1 (StrHV1), was identified using PCR and gene sequencing. This case represents a new clinical manifestation of a previously unreported herpesvirus in the GHOW. Identification of the herpes virus was critical to administration of appropriate therapy and resolution of the conjunctivitis, and corneal epithelial debridement promoted resolution of the chronic corneal epithelial defect.     

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.     

Conjunctival xanthoma in a blue and gold macaw (Ara ararauna)

A 17-year-old female blue and gold macaw ( Ara ararauna ) presented for evaluation of a discreet, conjunctival mass of the OD. No other abnormalities were found on ophthalmic or physical examination. A heterophilic leukocytosis was present on the complete blood count, and elevated aspartate aminotransferase activity, creatinine kinase activity and cholesterol were present on the plasma biochemistry panel. Surgical removal of the mass was complete and no recurrence has occurred by six months after excision. Reduction of dietary fats was recommended to reduce serum cholesterol levels and reduce the likelihood of future occurrence of xanthomas in this bird.     

Acute pancreatitis in parrots

Acute pancreatitis was diagnosed in three parrots. Antemortem diagnosis of pancreatitis in the avian patient is rarely documented, and should be suspected in birds showing clinical signs attributable to abdominal pain or gastro-intestinal dysfunction. However, not all birds with confirmed pancreatitis display these signs. A serum amylase level greater than 1,500 U/L is suggestive of pancreatitis and pancreatic biopsy can be used to confirm a diagnosis. The aetiology of acute pancreatitis in parrots is discussed and a suggested treatment protocol is described.     

Trade-off expression of melatonin receptor subtypes (Mel1a and Mel1b) and androgen receptor in lung of a tropical bird,Perdicula asiatica

The role of circulatory steroid hormone along with melatonin in lung of any seasonally breeding bird has never been explored so far. This could be interesting because steroid hormones are immunosuppressive while melatonin is immunostimulatory in nature. In our present study, we report the effect of exogenous melatonin and testosterone on expression of melatonin receptor subtypes (Mel_1a and Mel_1b) and androgen receptor in lung of a tropical bird Perdicula asiatica. Birds were collected from vicinity of Varanasi and acclimatized in laboratory with sufficient food and water. The birds were treated with melatonin and testosterone at dose of 25 µg/100 g B.wt./day and 1 mg/100 g B.wt./day, respectively, for 28 days. At the end of the experiment, the birds were sacrificed and lung tissue and blood sample were collected for immunohistochemistry, Western blot analysis and hormonal assay. Testosterone treatment increased circulatory testosterone and upregulated expression of androgen receptors whereas downregulated expression of melatonin receptor subtypes Mel_1a and Mel_1b. Melatonin administration increased peripheral melatonin and upregulated expression of melatonin receptor subtypes Mel_1a and Mel_1b while downregulated androgen receptor. Thus, our results suggest that a trade-off relationship between melatonin and testosterone exists in regulation of their receptors in lung of Perdicula asiatica.     

番鸭呼肠孤病毒和H9亚型禽流感病毒共感染对番鸭法氏囊和抗体产生的影响

为探讨番鸭呼肠孤病毒(MDRV)和H9亚型禽流感病毒(H9 AIV)共感染对番鸭法氏囊免疫应答能力的影响,本研究将MDRV或/和H9 AIV人工感染8日龄番鸭,观察法氏囊病理组织学变化,检测法氏囊B细胞增殖能力及RE-5 AIV疫苗免疫后抗体变化规律.结果显示:H9 AIV感染组番鸭发病率低(10％),无死亡,法氏囊无病理变化,显著抑制番鸭法氏囊细胞增殖反应;MDRV感染番鸭发病率70％,死亡率40％,生长迟缓,法氏囊病理变化为萎缩,淋巴细胞减少,局部出现范围较小的坏死灶,番鸭法氏囊细胞增殖反应下降;共感染组番鸭发病率100％,死亡率80％,番鸭生长迟缓,法氏囊萎缩和淋巴细胞增殖反应下降程度均比单一病毒感染组严重.病毒感染使番鸭对RE-5 AIV疫苗免疫应答能力明显下降,其共感染组抑制抗体应答程度最严重;共感染组的病毒检出时间早于并且检出率大于单一病毒感染组.表明MDRV与H9 AIV共感染在番鸭免疫应答抑制方面有协同作用.