Experimental infection of domestic pigs with the virus of peste des petits ruminants.

Pigs could be subclinically infected with PPRV by inoculation or contact with infected goats. There was no evidence that the virus could spread to uninfected pigs or goats and pigs are not considered important in the epidemiology of PPRV.     

小反刍兽疫的研究进展

概述了小反刍兽疫的易感动物、临床症状及病原的理化特征和基因组结构，重点阐述了病毒6种结构蛋白的大小和组成、小反刍兽疫病毒的分离培养、血清学及分子生物学诊断技术，并对其传统疫苗以及新型基因工程疫苗的研制进行了详细论述，以为该病的流行病学研究、诊断和免疫防制提供参考。     

Experimental infection of alpine goats with a Moroccan strain of peste des petits ruminants virus (PPRV)

Peste des petits ruminants virus (PPRV) recently caused a serious outbreak of disease in Moroccan sheep and goats. Alpine goats were highly susceptible to PPRV with mortality rates approaching 100%, as opposed to local breeds of sheep which were less susceptible to the disease. The relative susceptibility of alpine goats was investigated through an experimental infection study with the Moroccan strain of PPRV. Severe clinical signs were observed in the alpine goats with virus being excreted through ocular, nasal and oral routes. No difference in the severity of the disease in goats was observed with different inoculation routes and transmission of the virus by direct contact was confirmed. This study confirmed the susceptibility of the alpine goat to PPRV infection and describes a challenge protocol that effectively and consistently reproduced severe clinical signs of PPR in experimentally infected goats.     

小反刍兽疫病毒H基因的原核表达与鉴定

根据GenBank中已发表的小反刍兽疫病毒(PPRV)Nigeria 75/1株的H基因序列,设计上下游引物并添加BamH I酶切位点,以含有小反刍兽疫病毒H基因的Topo-PPRVH质粒为模板进行PCR扩增,扩增产物克隆于pEASY-T载体中,用BamH I单酶切后将目的片段连接到原核表达载体pET-32a(+)中,核酸序列分析证明.成功构建了PPRV H原核表达质粒pET-32a-H.将pET-32a-H重组质粒转化大肠杆茵 BL21(DE3)进行融合表达.经SDS-PAGE电泳,可见H蛋白获得了高效表达,融合蛋白的分子量约为87 Ku,表达产物以包涵体的形式存在,其表达量达到茵体总蛋白的38%,占包涵体蛋白的80%以上.Western blot鉴定表明,所表达的重组蛋白能被抗组氨酸单抗、抗PPRV标准阳性羊血清及抗PPRV疫苗的羊血清所识别,具有良好的免疫原性.     

Pathological and immunohistochemical study of experimental peste des petits ruminants virus infection in goats

Peste des petits ruminants (PPR) is an emerging, economically important viral disease of goats and sheep in the Indian subcontinent. In the present investigation, 15 hill goats were experimentally infected with 2 ml of 10% splenic suspension of a virulent isolate of PPR virus (PPR/Izatnagar/94) that had caused heavy mortality (>75%) in goats during 1994 outbreaks in northern India. More than 86% (13 of 15) animals died between 9 and 13 days post inoculation at the height of temperature or when temperatures were declining. Necropsy findings included congestion of gastrointestinal tract (GIT), nasal sinuses, consolidation of antero-ventral lobes of lungs, engorged spleen, and occasionally oedematous lymph nodes. Histopathological examination of major organs of GIT revealed degeneration and necrosis of labial mucosa, severe mucosal and submucosal congestion, degeneration and necrosis of intestinal epithelium and lymphoid cell depletion from Peyer's patches along with presence of syncytia at times. Lungs showed broncho-interstitial changes and presence of intracytoplasmic and intranuclear eosinophilic inclusions in alveolar macrophages and syncytial cells. These changes in lungs were frequently complicated with serofibrinous pneumonia (57%, eight of 14). Lymphocytolysis and occasional syncytia formation were evident in the lymphoid tissues. Immunohistochemical (IHC) findings included presence of PPR virus antigen in the labial, intestinal, and bronchiolar epithelial cells, pneumocytes, macrophages and syncytial cells in lungs, and lymphoid (intact and necrotic) and reticular cells in lymphoid organs. The findings of the study indicated the highly virulent nature of the PPR virus isolate (PPR/Izatnagar/94), causing 100% mortality and characteristic pathological changes in the target organs such as lungs, intestines and lymphoid tissues. The results of the IHC study suggested that indirect immunoperoxidase could be an alternative method in the absence of more sophisticated methods of laboratory diagnosis of PPR virus infection in goats.     

Serological studies with the virus of peste des petits ruminants in Nigeria.

It was possible to distinguish separate serological responses when experimental goats were inoculated with either rinderpest virus or peste des petits ruminants virus. Examination of field samples established that peste des petits ruminants occur commonly in Nigerian sheep and goats although some villages have escaped recent infection. There was no evidence of infection with rinderpest in these animals.     

Peste des petits ruminants infection in domestic ruminants in Sudan

The existence of peste des petits ruminants (PPR) in domestic ruminants and camels in Sudan during 2008–2012 was investigated. Lung tissues and serum samples were randomly collected from sheep, goats, cattle, and camels at different areas of Sudan. A total of 12,384 serum samples were collected from clinically healthy 7413 sheep, 1988 camels, 1501 cattle, 1459 goats, and 23 gazelles at different areas in the Sudan. They were examined for PPR antibodies using competitive ELISA (cELISA). The overall detected seroprevalence of PPR in tested sera was 49.4%; seroprevalence values within species were 67.1, 48.2, 25.8, 2.1, and 21.7% in sheep, goat, cattle, camels, and gazelles, respectively. The highest seroprevalence (68.1%) was observed in sera collected from Darfur states, then the central states (54.3%). A total of 1276 lung tissue samples (623 sheep, 324 cattle, 220 camels, and 109 goats) were collected. The majority of lung samples were collected from clinically healthy animals that showed lesions on PM in slaughterhouses (95%) and during PPR outbreaks; samples were tested for PPR antigen using immunocapture ELISA (IcELISA). PPR antigen was detected in 233 out of the 1276 tested samples (18.3%). Positive results were observed in samples collected from clinically healthy and diseased animals. The observed prevalence values in each species were 33.6, 21.1, 15.4, and 12.3% in camel, goat, sheep, and cattle, respectively. PPR antigen was detected in samples from different areas; however, the highest prevalence (63.9%) was found in samples collected from the eastern states, then Khartoum state (28%). Trials for virus isolation were done in different cell cultures. Out of 30 IcELISA-positive samples inoculated in primary bovine and ovine kidney cells, Vero cells, the PPR virus was successfully isolated from 15 (eight sheep, five camels, and two goats) samples in the three cell culture types. Using RT-PCR, PPRV nucleic acid was detected in all 25 IcELISA-positive tested samples.     

Pathogenicity of attenuated peste des petits ruminants virus in sheep and goats

Progressive loss of virulence for goat kids was noticed when peste des petits ruminants (PPR) virus was passaged in Vero cells. While goats inoculated with the 60th passage suffered from the clinical PPR disease and mortality, goats inoculated with the 80th passage did not show any sign of the disease. If the progressive loss of virulence of the virus with passage continues, it will not be long before a homologous PPR vaccine will be obtained at the National Veterinary Institute, Vom.     

Sample type is vital for diagnosing infection with peste des petits ruminants virus by reverse transcription PCR

Peste des petits ruminants (PPR) diagnosis from suspected samples from sheep and goats was carried out. Buffy coat, tissues, and oculo-nasal swabs were analyzed using nucleoprotein (NP3/NP4) and fusion protein (F1/F2) gene primers, respectively. Analysis of the sample types and primer set revealed that buffy coat are the best type of samples for PPR diagnosis and the use of two set of primers will increase the number of positives.     

An immunohistochemical study of the pneumonia caused by peste des petits ruminants virus

Ten goats were inoculated with peste des petits ruminants virus, a paramyxovirus closely related to rinderpest virus. All goats developed severe clinical disease, 8/10 having coughing or dyspnea as prominent clinical signs. In addition, all of the goats had stomatitis and diarrhea. Histopathologic and immunohistochemical studies were done only on the respiratory tracts. Pathologic changes ranged from mild multifocal bronchiolitis and bronchitis to severe bronchointerstitial pneumonia. Lesions were more severe in anteroventral than caudal lobes. The histologic nature of the viral process in the goat lungs had many features in common with the processes of pneumonia in dogs, due to canine distemper, or pneumonia in human beings, due to measles virus. Immunohistochemical staining of formalin-fixed, paraffin-embedded respiratory tract tissue was performed using an indirect system with rabbit anti-rinderpest virus serum, biotinylated anti-rabbit antibody, streptavidin-alkaline phosphatase, and nitroblue tetrazolium chromogen. Staining was sensitive, highlighting the presence of viral antigen in both lung and trachea of all goats. Viral antigen was found in both cytoplasm and nucleus of tracheal, bronchial, and bronchiolar epithelial cells, type II pneumocytes, syncytial cells, and alveolar macrophages. In general, the amount of staining correlated directly with the severity of the inflammatory process.     

Experimental studies on immunosuppressive effects of peste des petits ruminants (PPR) virus in goats

Effect of virulent and attenuated peste des petits ruminants (PPR) virus on the immune response to nonspecific antigen (ovalbumin) was investigated. Clinical and serological responses were monitored in goats administered with ovalbumin concurrently with either PPR vaccine or virulent virus. Study showed that PPR virulent virus causes marked immunosuppression as evidenced by leukopenia, lymphopenia, and reduced early antibody response to both specific and nonspecific antigen. These observations were predominant particularly during acute phase of disease (4-10 days post-infection). On the other hand, the vaccine virus induced only a transient lymphopenia without significantly affecting the immune response to nonspecific antigen or to itself during this period. Further, the antibody levels to ovalbumin in the group administered with virulent PPRV increased significantly between days 28 and 35 post-infection in comparison to the titers in other two groups given with either ovalbumin alone or in combination with vaccine.     

小反刍兽疫活疫苗效力评价

以新疆天康畜牧生物技术股份有限公司生产的6批小反刍兽疫活疫苗为材料,对其进行最小免疫剂量、疫苗免疫期和疫苗保存期的研究。结果显示,该疫苗最小免疫剂量为103 TCID50/头份;免疫期至少为26个月,保存期为-20℃保存12个月。结果表明,该疫苗免疫原性好,对山羊和绵羊均可提供良好保护。     

The IMPDH inhibitors,ribavirin and mycophenolic acid,inhibit peste des petits ruminants virus infection

Peste des petits ruminants virus (PPRV) causes highly contagious diseases in domestic and particular wild small ruminants, leading to substantial economic loss. The development of effective and cheap antiviral medications shall help to circumvent this emerging burden. In this study, we found that ribavirin, a competitive inosine-5’-monophosphate dehydrogenase (IMPDH) inhibitor, significantly inhibits the replication of PPRV. As IMPDH is a key enzyme in purine nucleotide synthesis, supplementation of exogenous guanosine attenuate the anti-PPRV effect of ribavirin. Interestingly, an uncompetitive IMPDH inhibitor, mycophenolic acid (MPA), exerted more potent antiviral effect again PPRV. Similarly, this effect was largely restored upon supplementation of guanosine. Thus, we have demonstrated that the IMPDH inhibitors ribavirin and MPA combat PPRV infection through purine nucleotide depletion. Because both regimens have been widely used in the clinic for treating viral infection or organ rejection in transplantation patients for decades, respectively, repurposing these existing safe and cheap medications may provide a new avenue for combating PPRV infection.