Distinct expression pattern of miRNAs in Marek's disease virus infected-chicken splenic tumors and non-tumorous spleen tissues

MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression. Emerging evidence suggests that differential miRNA expression is associated with viral infection and tumorigenesis. Recently discovered microRNAs in the Marek's disease virus (MDV) genome have been suggested to have regulatory roles during MDV oncogenesis. To gain more insight into the molecular mechanisms of the tumorigenesis of MDV, we used microarrays to screen host and viral miRNAs that were sensitive to infection by MDV. Microarray analysis showed significant differential expression of 79 miRNAs, which was confirmed by qRT-PCR analysis. These data suggest that differentially expressed miRNAs may have major roles in MDV-induced tumorigenesis. In addition, we found two clades of chicken miRNAs had increased expression in splenic tumors and non-tumorous spleen tissues from GA-infected chickens. Thus, the expression of these miRNAs can be considered signatures for MDV infection and tumorigenesis.     

马立克氏病病毒致弱毒株的致病性及致瘤相关基因序列分析

为培育及鉴定马立克氏病病毒(MDV)致弱病毒株,本研究将4株现地流行MDV强毒株(L-ZY、L-CZ、L-MS和L-SY株),在体外经CEF连续传代至110代,获得相应的4株高代次病毒株(L-ZYp 110C、L-CZp110C、L-MSp110C和L-SYp110C株).其中的L-MSp110C和L-SYp110C株对SPF鸡的致病性试验结果显示:以2×103 pfu和2×104 pfu的剂量接种1日龄SPF鸡,在试验期(12周)内试验鸡均没有引起马立克氏病病变;高代次病毒株在体外增殖能力增强,而在体内增殖能力显著低于亲本病毒株.与亲本病毒株基因序列比对发现:4株高代次病毒株病毒基因组中Meq和RLORF 12基因均有不同程度的缺失和插入;132 bpr序列拷贝数均显著增加;其中3株病毒株的RLORF4基因存在大片段缺失.以上结果表明,MDV强毒株体外传代至110代后,病毒增殖能力和主要致瘤相关基因的遗传特性均发生了改变,致弱毒株的毒力已完全弱化.该研究为进一步筛选MD弱毒疫苗提供了实验依据.     

Downregulation of cellular prion protein inhibited the proliferation and invasion and induced apoptosis of Marek's disease virus-transformed avian T cells

Cellular prion protein (PrP^C) is ubiquitously expressed in the cytomembrane of a considerable number of eukaryotic cells. Although several studies have investigated the functions of PrP^C in cell proliferation, cell apoptosis, and tumorigenesis of mammals, the correlated functions of chicken PrP^C (chPrP^C) remain unknown. In this study, stable chPrP^C-downregulated Marek''s disease (MD) virus-transformed avian T cells (MSB1-SiRNA-3) were established by introducing short interfering RNA (SiRNA) targeting chicken prion protein genes. We found that downregulation of chPrP^C inhibits proliferation, invasion, and migration, and induces G1 cell cycle phase arrest and apoptosis of MSB1-SiRNA-3 cells compared with Marek''s disease virus-transformed avian T cells (MSB1) and negative control cells. To the best of our knowledge, the present study provides the first evidence supporting the positive correlation between the expression level of chPrP^C and the proliferation, migration, and invasion ability of MSB1 cells, but appears to protect MSB1 cells from apoptosis, which suggests it functions in the formation and development of MD tumors. This evidence may contribute to future research into the specific molecular mechanisms of chPrP^C in the formation and development of MD tumors.     

Bovine viral diarrhea virus: An updated American College of Veterinary Internal Medicine consensus statement with focus on virus biology,hosts, immunosuppression,and vaccination

Control of bovine viral diarrhea virus (BVDV) in cattle populations across most of the world has remained elusive in spite of advances in knowledge about this viral pathogen. A central feature of virus perseverance in cattle herds is the unique mechanism of persistent infection. Managing BVDV infection in herds involves controlling persistently infected carrier animals using a multidimensional approach of vaccination, biosecurity, and identification of BVDV reservoirs. A decade has passed since the original American College of Veterinary Internal Medicine consensus statement on BVDV. While much has remained the same with respect to clinical signs of disease, pathogenesis of infection including persistent infection, and diagnosis, scientific articles published since 2010 have led to a greater understanding of difficulties associated with control of BVDV. This consensus statement update on BVDV presents greater focus on topics currently relevant to the biology and control of this viral pathogen of cattle, including changes in virus subpopulations, infection in heterologous hosts, immunosuppression, and vaccination.     

人参皂苷及其衍生物抗马立克病病毒作用的超微结构和病理变化观察

为了探讨人参皂苷及其衍生物抗马立克氏病病毒的作用机制.采用马立克氏病毒感染的雏鸡作为模型,人参皂苷及其衍生物口服给药途径,动态观察不同时间段,试验鸡的器官病理变化和脾脏、法氏囊的组织超微结构改变.结果发现人参皂苷及其衍生物组的器官病变程度明显低于病毒对照组,法氏囊和脾脏的超微结构病变轻于病毒对照组,说明人参皂苷及其衍生物在体内可以通过保护机体免疫器官来发挥抗病毒作用.     

鸭病毒性肿头出血症病毒培养特性与形态观察

试验对鸭病毒性肿头出血症病毒在鸭胚上的培养特性、毒力、病毒的纯化及其形态进行了研究。结果表明:该病毒不能感染鸡胚,能在鸭胚上繁殖并传代,在鸭胚上的半数致死量为1×10~(-7.24)/0.2mL;氯化铯超速离心纯化的病毒效果优于蔗糖离心法,电镜观察病毒粒子呈球形或椭圆形,大小为60～80nm,无囊膜。     

Knowledge,attitudes and management of bovine viral diarrhoea virus among eastern Australian cattle producers: results from a 2013 cross-sectional study

Bovine viral diarrhoea virus (BVDV) is an economically significant disease affecting the Australian cattle industry, with losses stemming from decreased production and reproductive performance and control costs. However, these losses can be difficult to appreciate, particularly in endemic regions. Overall, there is a variable but high herd-level seroprevalence in Australia. Despite a potentially high financial burden of the disease, the onus for control ultimately falls on producers and strategies employed will vary between regions. A cross-sectional study, using a postal survey, was conducted in 2013 to evaluate the BVDV knowledge, attitudes and management practices utilised by Australian cattle producers. A total of 192 producers participated in the study, and results indicate that knowledge and attitudes towards disease risk are variable and can be improved. Producer knowledge of how persistently infected (PI) animals are produced was higher than that of disease outcomes or transmission pathways. Implementation of biosecurity practices was limited, with approximately half of respondents employing quarantine procedures for introduced stock and only 2% indicating they would antigen test introduced stock for BVDV. Approximately a third (36%) of producers reported engaging in BVDV control, with the majority of these using vaccination strategies over deliberate exposure to a PI. Knowledge of and engagement with BVDV control was positively influenced by the producer relationships with veterinarians. Findings from this study suggest that building on education and delivering a consistent message among stakeholders would likely improve producer awareness and understanding in relation to BVDV and support decision making in BVDV management.     

马立克氏病毒1.8-kb mRNA对其上游双向启动子活性的影响

构建了针对1.8-kb mRNA基因簇潜在阅读框的RNA干扰质粒pP(1.8-kb-RNAi).将该质粒与包含其上游双向启动子的质粒pP(pp38)-CAT和pP(1.8-kb)-CAT共转染鸡胚成纤维细胞(CEF)和MDV rMd5感染的CEF(rMd5-CEF),48 h后,通过测定转染细胞裂解液中氟霉素乙酰转移酶(CAT)的活性确定1.8-kb mRNA被干扰后对双向启动子活性的影响.结果显示,利用pP(1.8-kb-RNAi)质粒干扰1.8-kb mRNA,可以使其上游双向启动子两个方向的活性均显著下降(P<0.01),其中1.8-kb mRNA方向下跌29.5%,pp38方向下跌25.0%.本研究结果证明了1.8-kb mRNA对其上游双向启动子活性有增强作用.     

Seroprevalence and associated risk factors of important pig viral diseases in Bhutan

A cross-sectional serological study was conducted in Bhutan between October 2011 and February 2012 to determine the prevalence of antibodies to classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), swine influenza virus (SIV) subtype H1N1 and Aujeszky's disease virus (ADV). Furthermore, risk factors for the seropositive status were investigated.     

Characterisation of genotype VII Newcastle disease virus (NDV) isolated from NDV vaccinated chickens,and the efficacy of LaSota and recombinant genotype VII vaccines against challenge with velogenic NDV

A Newcastle disease virus (NDV) isolate designated IBS002 was isolated from a commercial broiler farm in Malaysia. The virus was characterised as a virulent strain based on the multiple basic amino acid motif of the fusion (F) cleavage site ¹¹²RRRKGF¹¹⁷ and length of the C-terminus extension of the hemagglutinin-neuraminidase (HN) gene. Furthermore, IBS002 was classified as a velogenic NDV with mean death time (MDT) of 51.2 h and intracerebral pathogenicity index (ICPI) of 1.76. A genetic distance analysis based on the full-length F and HN genes showed that both velogenic viruses used in this study, genotype VII NDV isolate IBS002 and genotype VIII NDV isolate AF2240-I, had high genetic variations with genotype II LaSota vaccine. In this study, the protection efficacy of the recombinant genotype VII NDV inactivated vaccine was also evaluated when added to an existing commercial vaccination program against challenge with velogenic NDV IBS002 and NDV AF2240-I in commercial broilers. The results indicated that both LaSota and recombinant genotype VII vaccines offered full protection against challenge with AF2240-I. However, the LaSota vaccine only conferred partial protection against IBS002. In addition, significantly reduced viral shedding was observed in the recombinant genotype VII-vaccinated chickens compared to LaSota-vaccinated chickens.     

甘草多糖和甘草酸对NDV感染鸡胚成纤维细胞能力的影响

从甘草中提取甘草多糖和甘草酸,进行抗病毒活性比较。用MTT法比较了它们对鸡胚成纤维细胞(CEF)生长及以三种加药方式(先药后毒、先毒后药、药毒同时)抵抗新城疫病毒(NDV)感染细胞的影响,并用平均病毒抑制率比较甘草多糖和甘草酸的抗病毒活性。结果表明,先药后毒,甘草多糖能显著抑制NDV感染细胞(P0.05),平均病毒抑制率为60.6%;先毒后药,甘草多糖和甘草酸都能显著抑制NDV感染细胞(P0.05),其中甘草多糖的平均病毒抑制率为74.28%,甘草酸为62.3%。通过平均病毒抑制率比较,表明甘草多糖的抗病毒活性优于甘草酸。     

Associations between genetics, farm characteristics and clinical disease in field outbreaks of porcine reproductive and respiratory syndrome virus

Porcine reproductive and respiratory syndrome (PRRS) is a disease of domestic swine characterized by exceptionally high clinical variability. This study addresses the question of whether clinical variability in PRRS results from (a) genetic variation among viral isolates and/or (b) variation in management practices among farms on which isolates are found. Genetic data (open reading frame 5 gene sequences) and data on farm characteristics and associated clinical disease signs were collected for 62 PRRS virus (PRRSV) field isolates, representing 52 farms. Clinical disease signs were interrelated — confirming that a true reproductive syndrome exists (involving abortions, infertility in sows, deaths of sows and preweaning mortality).

Pairs of farms experiencing deaths in their sow populations also tended to share viral isolates which were more similar to one another than expected by chance alone. This implies that sow death (one of the more-severe manifestations of PRRS) is under genetic influence. Large herd size was a significant risk factor for the death of sows and for respiratory disease in nursery pigs. All-in–all-out management practices in the nursery were protective against reproductive signs in the sow herd. All-in–all-out management practices in the finishing stages of production were protective against respiratory disease in nursery pigs — but were paradoxically associated with an increased risk of infertility in sows. These results suggest that farm-management practices can also influence which PRRS clinical signs are manifested during an outbreak. In general, signs associated with PRRS appear to result from a combination of genetic factors and herd-management characteristics. The relative contributions of these two influences differ depending on the specific clinical sign in question.     

Effects of vaccine dose, virus challenge dose and interval from vaccination to challenge on protection of broiler chickens against Marek's disease virus challenge

OBJECTIVES: To examine the effects of varying the doses of turkey herpesvirus (HVT) vaccine and Marek's disease virus (MDV) challenge at two intervals after vaccination on the protection of chickens against challenge with MDV. DESIGN AND PROCEDURE: Experiment 1, a dose response study, consisted of 11 doses of HVT vaccine administered at hatch followed by challenge with 100 plaque forming units (pfu) of MDV 5 days post vaccination. Experiment 2, a 2 x 6 x 2 factorial design, included two HVT vaccine types, six different doses of HVT vaccine and 50 pfu and 200 pfu of MDV challenge 2 days post vaccination. All chickens were reared up to day 56 post challenge when all survivors were killed humanely. Dead and killed chickens were examined for gross MD tumours. RESULTS: Experiment 1 showed a significant positive linear relationship between dose of HVT vaccine and protective index in chickens challenged 5 days post vaccination. However the range of protective index observed was limited. In Experiment 2 neither HVT vaccine provided significant protection at any dose. There was no significant effect of vaccine type or MDV challenge dose on overall protection against challenge. Chickens challenged with 200 pfu of MDV had significantly higher mortality and MD incidence than those with 50 pfu. CONCLUSIONS: HVT vaccine dose had a significant impact on protective index, but vaccination to challenge interval appeared to have greater impact on the protective efficacy of vaccination. A fourfold increase in challenge dose increased mortality rate and incidence of MD.     

Characterisation of a SAT-1 outbreak of foot-and-mouth disease in captive African buffalo (Syncerus caffer): clinical symptoms, genetic characterisation and phylogenetic comparison of outbreak isolates

African buffalo (Syncerus caffer) play an important role in the maintenance of the SAT types of foot-and-mouth disease (FMD) in southern Africa. These long-term carriers mostly become sub-clinically infected, maintaining the disease and posing a threat to other susceptible wildlife and domestic species. During an unrelated bovine tuberculosis experiment using captive buffalo in the Kruger National Park (KNP), an outbreak of SAT-1 occurred and was further investigated. The clinical signs were recorded and all animals demonstrated significant weight loss and lymphopenia that lasted 100 days. In addition, the mean cell volume and mean cell haemoglobin values were significantly higher than before the outbreak started. Virus was isolated from several buffalo over a period of 167 days post infection and the molecular clock estimated to be 3 × 10⁻⁵ nucleotide substitutions per site per day. Seven amino acid changes occurred of which four occurred in hypervariable regions previously described for SAT-1. The genetic relationship of the outbreak virus was compared to buffalo viruses previously obtained from the KNP but the phylogeny was largely unresolved, therefore the relationship of this outbreak strain to others isolated from the KNP remains unclear.     

Studies on the role of humoral and cell-mediated immunity in pigs after vaccination with Aujeszky's disease virus

Humoral and cellular immunity in pigs vaccinated twice with Aujeszky's disease virus (ADV) was studied by seroneutralizing test and direct leucocyte migration inhibition technique. Significant migration inhibition of leucocytes (LMI) was found on the fifth day, whereas specific antibodies began to appear at that time only in very low titers. Anamnestic reaction due to the second injection of ADV did not bring about a significant increase of migration inhibition of leucocytes, instead the level of antibodies elevated markedly.