机体识别病毒核酸的几种分子模式及途径

近些年机体病原相关模式受体及其识别分子机制的研究,极大地促进了先天性分子免疫学的发展,并成为现代免疫学研究的重点和热点领域。作者通过机体识别病毒核酸的Toll样受体(Toll-like receptors,TLRs)、RIG-I样受体(RIG-I like receptors,RLRs)、NOD样受体(NOD-like receptors,NLRs)和DAI(DNA-dependent activatorof interferon-regulatory factors,DAI)等模式识别分子的细胞定位、分子结构及其识别病毒核酸途径的介绍,系统、全面地探讨了宿主机体如何全方位识别和消除入侵病毒的分子免疫防御途径,为抗病毒药物和疫苗的设计以及抗病育种提供了理论依据。     

未知病毒检测方法在动物病毒病诊断中的应用

近年来,新发动物疫病多以病毒病为主,往往具有不可预测性且对畜牧业生产造成重大损失,因此如何快速诊断其主要病原成为防控新发疫病的关键因素。对新发疫情的传统诊断,往往需要依靠经验,逐步排除已知病原后才能确定疫情的主要因素,随着分子生物学及测序技术的发展,临床上出现了很多直接从病料中检测未知病毒的方法,这些方法逐渐成为确诊未知疫情的主要手段。作者主要阐述近年来应用于发现未知病毒的检测方法。     

玉树部分地区牦牛病毒性腹泻血清学检测报告

为摸清牛病毒性腹泻/黏膜病(BVD/MD)在玉树地区流行情况,应用酶联免疫吸附试验,对采自囊谦、治多两县的155份(囊谦40份,治多115分)牦牛血清进行牛病毒性腹泻抗体检测。检测结果表明:两县的155份样品中阳性率囊谦县100%、治多县87.83%,玉树地区牦牛群中BVDV感染非常严重。     

云南省某腹泻仔猪群猪流行性腹泻病毒核酸检测及其N基因序列分析

猪流行性腹泻是猪场新生仔猪腹泻的主要病原之一.为确定云南省曲靖市某猪场新生仔猪出现呕吐、腹泻和高死亡率的病因,采集病死仔猪肠道、肠系膜淋巴结、脾脏等组织病料,处理后提取病原核酸,通过RT-PCR进行导致仔猪腹泻的猪流行性腹泻病毒(PEDV)、猪传染性胃肠炎病毒、猪轮状病毒、猪伪狂犬病病毒、猪瘟病毒、猪繁殖与呼吸综合征病...     

Detection of Bovine Viral Diarrhoea Virus Infected Cattle – Testing Tissue Samples Derived from Ear Tagging Using an Erns Capture ELISA

A new diagnostic approach testing tissue samples derived from cattle ear tagging for bovine viral diarrhoea virus (BVDV) antigen in a commercially available antigen capture enzyme‐linked immunosorbent assay (ACE) was developed. To validate this method, 99 positive and 469 negative samples were tested. With those samples the assay yielded a sensitivity of 100% and specificity of ≥99.6%. Serum and ear tissue samples from 11 persistently infected (PI) BVDV calves were tested. While serum samples were negative after intake of colostrum, the ear tissue samples could be detected positive for BVDV all the time. Testing multiple samples derived from the same ear from PI cattle yielded positive results and low variation. Using cattle ear tags combining the ear tag application with sampling of a small ear tissue plug and testing those tissue samples with an ACE could be a reliable and economic way of BVDV testing.     

云南省部分养殖场鸡传染性贫血病毒核酸检测及VP2基因序列分析

为了解云南省鸡传染性贫血病毒（chicken anemia virus,CAV）的流行及其VP2基因变异情况,2017—2020年,在昆明市、楚雄州、大理州、红河州、玉溪市等家禽养殖密集区的89个鸡场,采集422份疑似病鸡肝脏样品,通过PCR方法检测CAV核酸。结果显示,检出阳性245份,平均阳性率为58.06%;不同年份的CAV核酸阳性率为53.61%~60.91%,不同区域核酸阳性率为51.68%~63.52%。对3份CAV阳性样品进行VP2基因序列分析,发现核苷酸序列同源性为99.6%~99.8%,与参考毒株1852TW和N7的核苷酸同源性为99.6%~99.8%,均属于GroupA分支。结果表明：云南省普遍存在CAV感染,且感染较严重;VP2基因仍可作为PCR病毒核酸检测的首选目标基因。通过加强鸡群CAV监测,淘汰阳性鸡群和结合疫苗免疫,可减少该病造成的损失。     

用PCR技术从同一兔出血症病料扩增和检出2种病毒核酸

应用 Ｐ Ｃ Ｒ 和 Ｒ Ｔ Ｐ Ｃ Ｒ 技术,对兔出血症病毒核酸作了鉴定。分别设计合成 ２ 对 Ｐ Ｃ Ｒ 扩增特异性引物,即按兔出血症病毒中国分离的 Ｎ Ｊ株核酸序列合成 １ 对引物（ Ｎ Ｊ引物）,按德国分离的 Ｆ Ｒ Ｇ 株核酸序列合成另 １ 对引物（ Ｆ Ｒ Ｇ 引物）,进行 Ｐ Ｃ Ｒ 和 Ｒ Ｔ Ｐ Ｃ Ｒ。从纯化的病毒核酸样品和感染 Ｄ Ｊ Ｒ Ｋ 细胞的病毒核酸样品中,均扩增出 ２ 对引物范围内的特异性核酸片段, Ｎ Ｊ引物扩增产物为 ３６４ ｂｐ, Ｆ Ｒ Ｇ 引物扩增产物为 ５１３ｂｐ。模板用 Ｒ Ｎａｓｅ 消化后,仍出现 ３６４ ｂｐ 带,用 Ｄ Ｎａｓｅ Ｓ１ 消化,则此带消失;反之,用 Ｄ Ｎａｓｅ Ｓ１ 消化,出现５１３ ｂｐ 带,用 Ｒ Ｎａｓｅ 消化,则此带消失,证实前者模板为 Ｄ Ｎ Ａ,后者为 Ｒ Ｎ Ａ。 Ｓｏｕｔｈｅｒｎ 杂交试验证实, Ｐ Ｃ Ｒ扩增出的核酸片段与各自的地高辛标记特异性探针发生强阳性杂交反应。由此认为,在兔出血症病毒感染中,同时存在着 ２ 种不同核酸型的病毒。     

Informative Value of an Indirect Enzyme‐Linked Immunosorbent Assay (ELISA) for the Detection of Bovine Viral Diarrhoea Virus (BVDV) Antibodies in Milk

Bulk and individual milk samples from 117 herds located in Brittany (west France) were used to assess: (i) the performance characteristics of an indirect enzyme‐linked immunosorbent assay (ELISA) applied to individual milk for the detection of antibodies to bovine viral diarrhoea virus (BVDV); and (ii) the relationship between the bulk milk result obtained from this test and the within‐herd prevalence of antibody‐positive lactating cows. This ELISA test was based on a monoclonal antibody directed against non‐structural protein NS2‐3 of pestiviruses. At the individual level, based on 1113 matched milk/serum samples, the sensitivity and specificity of this test applied to milk, compared with the virus neutralization test on serum, were 95.0 and 97.7%, respectively. At the herd level, the relationship between the optical density percentage (OD%) of bulk milk and the within‐herd prevalence of antibody‐positive lactating cows was assessed using the receiver operating characteristics (ROC) analysis. Classes of OD% of bulk milk were determined so that they were associated with minimum intraclass and maximum between‐class variances of within‐herd prevalence of antibody‐positive cows. The ROC analysis resulted in two classes of bulk milk results corresponding to different expected levels of within‐herd prevalence. Herds with an OD% of bulk milk <75% and ≥75% had a mean observed prevalence of antibody‐positive cows of 8.9 and 60.6%, respectively. Herds with a bulk milk result <75% were expected to be BVDV free, whereas large variations in prevalence of antibody‐positive cows existed in the herds with OD% ≥75%. The test described in this study is suitable to identify herds likely to have a low prevalence of BVDV antibody‐positive cows.     

Experimental Infection of Cows with Bovine Viral Diarrhoea Virus in Early Pregnancy – Findings in Serum and Foetal Fluids

Nineteen pregnant cows were experimentally infected with bovine viral diarrhoea virus (BVDV) between day 74 and 81 of pregnancy. All cows became infected and developed serum antibodies. Sixteen of the cows delivered persistently infected (PI) offspring, whereas the remaining three gave birth to calves with detectable serum antibodies and free from BVDV. The 16 cows with PI foetuses developed higher levels of antibodies in serum during pregnancy than did their three peers carrying non‐PI calves. Multivariate analysis showed that the antibody levels in these two groups of cows were significantly different from day 135 of pregnancy. Foetal fluid was successfully collected from 18 of the 19 infected cows and from five uninfected control cows between 10 and 24 days before delivery by use of a percutaneous, blind puncture technique. No negative effects were observed in the cows or their offspring. BVDV was isolated and detected with an immunoperoxidase test in foetal fluid from 13 of the 16 cows carrying PI foetuses, and from 15 of the cows when a quantitative fluorescent polymerase chain reaction (PCR) technique was used. The negative sample in the PCR assay was positive for BVDV antibodies. The number of viral copies per microlitre in foetal fluids varied between 103 and 1080 in the positive samples. All samples taken from the cows carrying non‐PI foetuses were negative for BVDV in both assays. In this experiment, examination of either serum or foetal fluids could identify the cows carrying a PI foetus. Examination of serum for BVDV antibodies was a reliable indicator of a PI foetus if the serum was collected during the last 2 months of pregnancy. For examination of foetal fluids, both viral and serological analyses should be performed. For viral analysis, PCR should be the test of choice. High levels of BVDV antibodies in conjunction with a negative result in the PCR may be indicative of a false‐negative virus result. Further experience with the method of collection of foetal fluids is necessary for evaluation of its safety. Investigation of pregnant cows in order to discover a PI offspring before it is born could be a useful tool in control and eradication of BVDV.     

Equine placenta – A clinician's perspective. Part 1: Normal placenta – Physiology and evaluation

A complex organ, the equine placenta is responsible for fetal nourishment, protection from external and internal insults, and the production and/or metabolisation of various hormones. The endometrial cups are unique structures of the equine placenta that are responsible for producing an essential hormone for equine pregnancy, equine chorionic gonadotropin. Since mares have epitheliochorial placentae, with 6 layers of tissue separating maternal from fetal circulation, almost the entire surface of the chorioallantois must be attached to the maternal endometrium in order to support a developing fetus adequately. The only avillous areas of the normal chorioallantois are: the cervical star, sites of the endometrial cups, areas facing oviductal papillae, folds overlying major allantoic vessels, and fetal foot PADs (placental areas of degeneration). There are characteristic differences between the gravid and the nongravid horn of the chorioallantois. Allantoic vesicles, allantoic pouches, hippomanes, amniotic plaques and yolk sac remnants are normal features of the equine placenta. The clinician should thoroughly examine the entire placenta immediately after its expulsion. The most important aspect of this evaluation is to check for completeness of the chorioallantois, along with identifying any pathological lesions on the fetal membranes or the umbilical cord.     

Multistate Outbreak of Human Salmonella Typhimurium Infections Linked to Pet Hedgehogs – United States, 2011–2013

Zoonotic Salmonella infections cause approximately 130 000 illnesses annually in the United States. Of 72.9 million US households owning at least one pet, five million own small mammals; 3000 hedgehogs were documented by USDA in USDA‐licensed breeding facilities and pet stores in 2012. State health department collaborators and PulseNet, the national bacterial subtyping network, identified human infections of a Salmonella Typhimurium outbreak strain, which were investigated by CDC, USDA‐APHIS and state public and animal health officials. A case was defined as an illness in a person infected with the outbreak strain identified between 1 December 2011 and 3 June 2013. Investigators collected information on patient exposures, cultured animal and environmental specimens for Salmonella, and conducted traceback investigations of USDA‐licensed hedgehog facilities. There were 26 cases in 12 states. Illness onset dates ranged from 26 December 2011 to 8 April 2013. The median patient age was 15 years (range = <1–91 years); 58% were female. Among 23 persons with available information, 8 (35%) were hospitalized and one outbreak strain‐associated death was reported. Of 25 patients with available information, 20 (80%) reported pet hedgehog contact in the week before illness onset. The outbreak strain was isolated from animal and environmental samples collected from three ill persons’ homes in three states. Hedgehogs were purchased in geographically distant states from USDA‐licensed breeders (10/17, 59%); a USDA‐licensed pet store (1/17, 6%); unlicensed or unknown status breeders (3/17, 18%); and private individuals (3/17, 18%). Traceback investigations of USDA‐licensed facilities did not reveal a single source of infection. Public and animal health collaboration linked pet hedgehog contact to human infections of Salmonella Typhimurium, highlighting the importance of a One Health investigative approach to zoonotic salmonellosis outbreaks. More efforts are needed to increase awareness among multiple stakeholders on the risk of illness associated with pet hedgehogs.     

Priapism Secondary to Perineal Abscess in a Dog – A Case Report

A 7‐year‐old intact male Boxer was referred to our services at the Veterinary Teaching Hospital of the University of Trás‐os‐Montes and Alto Douro, suffering from a persistently erect penis (including the bulbus glandis) that had been exposed for several days. Radiographic and ultrasonographic examinations detected a 5.0 × 3.5 cm mass located dorso‐laterally to the urinary bladder. The microbial culture of the mass revealed Staphylococcus spp. At that time, we suspected the involvement of an abscess in the origin of the priapism. Medical and surgical treatments were promptly instituted, which allowed for penile withdrawal into the prepuce; however, the resolution of the penile erection was not accomplished in the following days and penile amputation was required. Histological evaluation of the excised penis revealed extensive infarction of the erectile tissue of the pars longa and bulbus glandis, and also of the blood vessels of the penis. Following penile amputation and antimicrobial therapy, the animal fully recovered. Ultimately, the animal died as a consequence of gastric torsion. At necropsy, some lesions compatible with a previous perforation of the intestinal wall were recorded. The data gathered from the anamnesis, the physical and imaging examinations, along with the post‐mortem findings, allowed us to conclude that in this clinical case the primary cause of priapism was a perineal abscess due to bowel perforation.