番鸭细小病毒的研究进展

番鸭细小病毒(MPV)引起的雏鸭发病称为雏番鸭细小病毒病,主要侵害1～3周龄的雏番鸭,俗称"三周病".其发病率为26%～62%,病死率为22%～43%,病愈鸭大部分成为僵鸭,给养鸭业带来一定损失.该病具有高度传染性,多以腹泻、喘气和进行性消瘦及脚发软为主要症状.     

雏番鸭细小病毒和传染性法氏囊病病毒混合感染的检测

2017年12月,广西某番鸭养殖场的鸭群出现生长缓慢,临床表现软脚,拉白色粪便的腹泻并伴有一定的神经症状等;剖检可见肝脏肿大出血、腹腔有纤维素性渗出物、肠黏膜有不同程度的充血,法氏囊萎缩等病变;于20日龄开始发病,30日龄送检时发病率20%、死亡率10%。无菌取番鸭的病变组织,分别进行番鸭细小病毒(MDPV)和传染性法氏囊病病毒(IBDV)的核酸特异性检测,结果均呈阳性,并成功分离到一株番鸭细小病毒(命名为MDPV GX1712)和一株传染性法氏囊病病毒(命名为GL1712);基因序列分析结果显示,分离株GX1712与GX5、P 1988、 SAASSHNH等MDPV参考株的亲缘性最近,相似性高达98.5%～99.1%,在进化树中处同一个分支,并且与疫苗株FZ91-30亲缘性相距较远,表明该分离株为MDPV野毒株;基于VP1-b序列分析IBDV分离株GL1712与中国新型超强毒株HLJ0504同属HLJ0504-like cluster,而基于vVP2序列则与中等偏强毒力参考株同属C2-intermediate IBDV genotype,为基因重排毒株。根据发病日龄、临床症状、病理剖检和实验室诊断,确诊该临床发病番鸭群混合感染了番鸭细小病毒和传染性法氏囊病病毒。     

Evidence of Muscovy duck parvovirus in Muscovy ducklings in California

Muscovy duck parvovirus (MDPV) has been demonstrated in tissue samples from one- to four-week-old commercially reared Muscovy ducks that were weak, unable to walk and had a high mortality rate. On postmortem examination, the thigh and leg muscles, and the myocardium were found to be pale, and there was a fibrinous exudate on the capsule of the liver, and ascites. The parvovirus was isolated in embryonated Muscovy duck eggs and visualised by negative stain electron microscopy, detected by polymerase chain reaction (PCR) directly from the tissues, and antibodies to it were detected by immunoelectron microscopy, ELISA and immunofluorescence. In addition, the PCR products obtained that represented 1625 bp (74 per cent) of the capsid vP1 gene, including a hypervariable region between Derzsy's disease virus or goose parvovirus and MDPV, were sequenced and shown to be 100 per cent homologous with the MDPV 89384 reference strain, but only 82.3 per cent homologous with Derzsy's disease virus.     

番鸭饲养管理实用技术

番鸭又名瘤头鸭,俗称“红头鸭”、“木鸭”、“麝香鸭”,现社会上流传的“肉鸳鸯”就是番鸭.由于番鸭（包括杂交番鸭）瘦肉率较高和番味较浓而深受消费者的欢迎.市场的需求,促进了番鸭生产的迅速发展.番鸭可水养、陆养、圈养、牧养，生产设备简易，饲养管理方便，现将番鸭的饲养管理技术要点介绍如下：     

番鸭呼肠病毒病的病原学研究进展

番鸭呼肠病毒病是由番鸭呼肠病毒引起的一种急性传染病,主要发生于40日龄内的番鸭,临床上以软脚为主要症状,并伴有腹泻,发病率高,病情严重时可致全群死亡,给番鸭养殖业带来了巨大的损失。其病原为呼肠病毒科正呼肠病毒属番鸭呼肠病毒。文章综合了国内外对该病的病原学研究成果,从病毒的分类地位、生物学特性、基因组与编码蛋白、病原分布及流行特性、检测与防控等方面对该病的病原学进行了较全面的论述,以期对该病的深入研究和防控提供有用的资料。     

我国番鸭呼肠孤病毒病的研究现状

番鸭呼肠孤病毒病是我国近年来新出现的、以软脚为特征的高发病率、高致死率急性烈性病毒性传染病,由于肝脏表面的灰白点较为典型,因此俗称“花肝病”或“白点病”。初步认为其病原为呼肠孤病毒科正呼肠孤病毒属番鸭呼肠孤病毒。本文综合了国内近年来对该病的研究成果,从病原学、流行病学、临诊症状、病理学、区别诊断和防制措施等方面对该病进行了较全面的论述,以期对该病的深入研究和防制提供有益的资料。     

番鸭呼肠孤病毒p10.8蛋白致细胞凋亡功能

为确定番鸭呼肠孤病毒(DRV)p10.8蛋白的生物学功能,本研究采用RT-PCR方法扩增DRV S14株p10.8编码基因,将其克隆到真核表达载体中,构建了重组表达质粒pcDNA-p10.8;通过转染鸭胚成纤维细胞(DEF),首次对p10.8蛋白的凋亡功能进行了研究.细胞转染48 h后,Hoechest、DNA Ladder和TUNEL法的细胞凋亡检测结果显示:光镜下可见细胞形态学上出现的细胞皱缩,Hoecheg染色后可见细胞核固缩,染色质凝固成团块状;DNA Ladder法可检测到凋亡细胞DNA样品呈梯形条带;TUNEL法可观察到褐色调亡细胞的存在.以上结果均表明,p10.8在DEF中的表达具有诱导宿主细胞凋亡的作用,是番DRV的凋亡蛋白.     

番鸭呼肠病毒的分离与RT-PCR鉴定

从发病番鸭中分离到1株病毒,用该病毒接种番鸭胚,至第2代可引起鸭胚死亡,胚体出血,部分鸭胚肝脾有少量白点。分离毒经用RT-PCR方法进行检测,可与番鸭呼肠病毒标准株扩增出大小一致的特异条带,证实该分离毒为番鸭呼肠病毒。     

1株番鸭源新型鸭呼肠孤病毒(QY株)生物学鉴定

从广东清远地区临床病变为发病鸭软脚,肝、脾肿大,有点/块状出血、坏死,其他各脏器不同程度出血的病死雏番鸭肝、脾组织中分离到1株病毒.通过形态观察、核酸图谱鉴定、RT-PCR扩增、基因序列分析、动物回归试验结果表明,该病毒为一株不同于以往番鸭呼肠孤病毒的新型鸭呼肠孤病毒,并命名为DRV-QY株.     

广西番鸭细小病毒的分离和鉴定

198 5年以来在我国的福建、广东、浙江、江西等省先后发生了以腹泻、呼吸困难和脚软为主要症状的番鸭细小病毒病 ,对 1 -3周龄的雏番鸭危害极大。 1 989年法国西部也发生了致死率为 80 %的番鸭细小病毒病 ,并分离到了病毒。广西从 1 996年开始在番鸭中也出现了类似疾病 ,为了明确病因 ,我们从广西各地采集疑似番鸭细小病毒的病料 ,进行了病原分离和鉴定 ,结果如下。1　材料和方法1 .1　标本的采集和处理从南宁市郊、北流、博白、柳州等地采集 2 0余份病死雏番鸭的肝、脾、胰等脏器 ,用含双抗的Ｈａｎｋ’ｓ液按 1∶5的比例磨成组织混悬液 ,…     

1株重组型番鸭细小病毒的分子特征解析

为了对1株番鸭细小病毒(MDPV)分离株NY18株的分子特征予以准确揭示,通过设计重叠PCR 引物,扩增NY18株的完整基因组并进行了测序.结果显示,NY18株基因组由5 071个碱基组成,5'和3'端末端倒置重复序列(inverted terminal repeats,ITR)均由424个碱基组成,其外侧385个碱基...     

番鸭白细胞介素2基因的克隆及其原核表达

为原核表达番鸭白细胞介素2 (MdIL-2),本研究利用ConA刺激诱导番鸭脾淋巴细胞,采用RT-PCR技术扩增MdIL-2基因的编码序列,并将其克隆至pGEX-6P-1中,构建重组质粒pGEX-MdIL-2,转化E.coli BL21受体菌.测序结果表明,MdIL-2基因ORF全长420 bp,编码140个氨基酸.重组菌经IPTG诱导表达后,SDS-PAGE分析显示,重组蛋白约为40 ku.Western blot分析表明,重组蛋白能够被抗GST单克隆抗体特异性识别.MdIL-2基因的克隆及其原核表达为进一步进行MdIL-2的活性检测以及应用研究奠定了基础.     

番鸭细小病毒VP3基因和鸭IL-2基因真核融合表达载体的构建

将番鸭细小病毒(MDPV)主要结构蛋白VP3基因和免疫刺激基序(CpG)依次克隆至质粒pIRES-dIL2上,构建了重组质粒pIRES-dIL2-VP3-CpG。核酸疫苗重组质粒转染BHK-21细胞进行间接免疫荧光试验,可在细胞浆中检测到绿色荧光,表明重组质粒可以成功地在真核细胞内表达。     

番鸭呼肠病毒活疫苗免疫途径及其效果研究

1日龄雏番鸭经肌肉注射、口服、点眼 和喷雾等途径免疫番鸭呼肠病毒活疫苗, 7d后攻毒。结果表明,肌肉注射免疫组保护 率为100%,口服免疫组保护率为85.7%,点 眼组保护率为0%,喷雾组保护率为0%;血液 淋巴细胞转化能力测定结果显示肌肉注射免 疫组淋巴细胞活性最高,口服免疫组次之,点 眼组、喷雾组最差。与对照组相比,35d后无 显著差异,与攻毒保护结果类似。番鸭呼肠病 毒活疫苗以肌肉注射免疫效果最好,口服免疫 也能提供有效保护,但点眼和喷雾途径不能提 供有效保护;口服免疫有望成为比肌肉免疫更 安全、方便、快捷的免疫途径。     

番鸭呼肠孤病毒S基因组的克隆与序列分析

对番鸭呼肠孤病毒S12和S14毒株S基因组（S1-4）中σA、σB、σNS和σC蛋白基因进行了克隆和测序。序列分析表明：鸭呼肠孤病毒（DRV）与禽呼肠孤病毒（ARV）的σA和σNS基因的核苷酸同源性分别为76．0％～77．1％和78．4％～79．6％，氨基酸同源性分别为89．5％～91．2％和91.6％～92．7％；而具有诱导群特异性和型特异性中和抗体的σB和σC基因的核苷酸同源性分别为60．3％～64．4%和2．7%～9．9％，氨基酸同源性分别为61．4％～62．0％和22．6％～26．7％；DRV和ARV抗原性存在差异。而DRVS12／S14与法国89026株σA、σB、σNS和σC基因的核苷酸同源性分别为90．0％、93．6％、87．9％～88．0％和93．1%，氨基酸同源性分别为97．1%、94．3%、95．7%～95．9%和93．7％。进化树分析表明，DRV与ARV形成不同的分支，DRV是正呼肠孤病毒属中不同于ARV的一种新的呼肠孤病毒。     

Indigenous Muscovy ducks in Congo-Brazzaville. 1. A survey of indigenous Muscovy duck management in households in Dolisie City

A cross-sectional study by means of a questionnaire with open-ended questions and multiple-choice questions was used to collect data on the profile of duck keepers, husbandry practices, and performances, opportunities and constraints of Muscovy duck breeding in households (n = 88) in Dolisie city (Congo-Brazzaville). The study confirmed the common observations on traditional poultry keeping such as scavenging during the day and housing overnight. The flock size (7.7 ± 3 ducks per unit) showed no specialization of husbandry (100% of surveyed flocks were kept for simultaneous production of ducklings, meat and eggs) and a high drake-to-duck ratio (1:3). The hatchability was close to 80.5% ± 13%, whereas the average number of eggs was 13.2 ± 5 per clutch. In addition, a high mortality (80%) was observed in ducklings, which was due to poor feeding, lack of veterinary care and housing conditions. Eggs and live ducks were sold by duck farmers in response to the family needs rather than market price. The three most important findings were as follows: (1) duck keepers were mainly men (80% versus 20% of women); (2) there was no evidence of taboo; and (3) the duck as an exotic bird was not proscribed by cultural beliefs, and therefore development of the Muscovy duck in Congo Brazzaville should be unhindered.     

商品代黑番鸭日粮适宜蛋白质水平的试验

本试验探讨了商品代黑番鸭日粮适宜的CP水平。试验设计1～30日龄和31～70日龄两独立阶段。1．30日龄日粮ME为11．50MJ儿g,CP水平设21％、18％、15％3种：31～70日龄日粮ME为11．91MJ／kg．CP水平设18％、15％、12％3种。结果表明：以增重、饲料转化率和胴体性能为指标．在本试验条件下．商品代黑番鸭1～30日龄日粮以ME11．50MJ／kg和CP21％为最适宜：31～70日龄日粮营养水平以ME11．91MJ／kg和CP18％为最适宜。     

Cloning and expression analysis of PRL and PRLR genes in black Muscovy duck

ABSTRACT

1. Prolactin hormone (governed by the PRL gene) is secreted by the anterior pituitary of animals, which combines with its receptor (prolactin receptor, PRLR) to act on target cells. Both PRL and PRLR are mainly associated with reproductive performance. The genetic mechanism of nesting in poultry is not yet clear, and so the aim of the current study was to determine expression patterns of PRL and PRLR at different times across the breeding stages of black Muscovy ducks.

2. In this study, the CDS regions of PRL and PRLR were determined by RACE sequencing. The expression levels of PRL and PRLR in the pituitary, ovary and uterus from the black Muscovy duck were compared and analysed during the pre-laying, laying and nesting periods.

3. The results showed that PRL and PRLR are highly homologous in a variety of poultry species. The expression of the PRL gene in the pituitary was the highest, which was significantly higher than seen in the ovary and uterus. This trend ran through the entire prenatal period, i.e. the laying period and the nesting period. The expression level of the PRLR gene in the pituitary and ovary was generally low, and expression in the uterus was the highest. There was no significant difference in expression of the PRLR gene between pituitary and ovary during different periods, but the expression level of the PRLR gene in the uterus reached its highest level during the nesting stage, which was significantly higher than seen in the early laying period.     

番鸭肠道乳酸菌的分离鉴定及抑菌性能

通过革兰染色、菌落形态观察、生化分析鉴定、牛津杯抑菌试验、Caco-2细胞黏附试验对分离到的6株乳酸产生菌进行研究。结果显示:经16SrDNA序列测定,比对分析6株乳酸产生菌与鼠李糖乳杆菌、乳酸乳球菌乳亚种、唾液乳杆菌、乳链球菌、哥伦比亚肠球菌和鼠乳杆菌高度同源。6株乳酸菌抑菌试验检测显示:对大肠杆菌、沙门菌、蜡样芽胞杆菌和金黄色葡萄球菌具有很强的抑菌效应,以CM3菌株的抑菌效果最强。同时,这6株乳酸菌对Caco-2细胞均有一定的黏附能力。结果表明:6株番鸭肠道乳酸产生菌能够黏附于肠道模型Caco-2细胞上并具有较强的抑菌活性,但其黏附性能及保护机制还有待进一步研究。