[Comparative analysis of residual neurovirulence of vaccine and low attenuated rubella virus (Matonaviridae: Rubivirus: Rubella virus) strains in the experiments on the macaque rhesus (Macaca mulatta) monkeys].

INTRODUCTION: Rubella is currently an infection controlled by specific prophylaxis. Not only the right vaccine prophylaxis strategy and tactics, but also the use of effective and safe vaccine preparations is crucial for the elimination of this disease.The aim of the investigation was to study the morphological and pathogenetic patterns of changes developing in the central nervous system (CNS) and internal organs of monkeys (Haplorhini) during intracerebral inoculation with 2 strains of rubella virus (Matonaviridae: Rubivirus: Rubella virus) (RV): highly attenuated Orlov-B, and low attenuated Orlov-14. MATERIAL AND METHODS: In the experiments, seronegative rhesus macaque monkeys (Macaca mulatta) weighing 3.3-5.1 kg (n = 7) were used. Neurovirulence of the strains was determined by a complex of clinical, pathomorphological, and virological methods. RESULTS AND DISCUSSION: It was found that during attenuation, the Orlov-B strain lost the ability to replicate in CNS cells and induce moderate/expressed specific changes in them, as well as to overcome the blood-brain barrier and cause the damage of sensitive organs and tissues. This fact indicates a low level of residual neurovirulence of the vaccine strain. CONCLUSION: The results obtained in this study regarding the clinical symptoms of CNS lesions and the nature of the pathological process in its tissues in experimental animals can be significant for the improvement of safety control of live rubella vaccines. These data indicate that the Orlov-B strain can be considered as a candidate strain for further study on the development of a rubella vaccine based on the domestic vaccine strain.

Revisiting Rustrela Virus: New Cases of Encephalitis and a Solution to the Capsid Enigma.

Rustrela virus (RusV; species Rubivirus strelense) is a recently discovered relative of rubella virus (RuV) that has been detected in cases of encephalitis in diverse mammals. Here, we diagnosed two additional cases of fatal RusV-associated meningoencephalitis in a South American coati (Nasua nasua) and a Eurasian or European otter (Lutra lutra) that were detected in a zoological garden with history of prior RusV infections. Both animals showed abnormal movement or unusual behavior and their brains tested positive for RusV using specific reverse transcription quantitative PCR (RT-qPCR) and RNA in situ hybridization. As previous sequencing of the RusV genome proved to be very challenging, we employed a sophisticated target-specific capture enrichment with specifically designed RNA baits to generate complete RusV genome sequences from both detected encephalitic animals and apparently healthy wild yellow-necked field mice (Apodemus flavicollis). Furthermore, the technique was used to revise three previously published RusV genomes from two encephalitic animals and a wild yellow-necked field mouse. When comparing the newly generated RusV sequences to the previously published RusV genomes, we identified a previously undetected stretch of 309 nucleotides predicted to represent the intergenic region and the sequence encoding the N terminus of the capsid protein. This indicated that the original RusV sequence was likely incomplete due to misassembly of the genome at a region with an exceptionally high G+C content of >80 mol%. The new sequence data indicate that RusV has an overall genome length of 9,631 nucleotides with the longest intergenic region (290 nucleotides) and capsid protein-encoding sequence (331 codons) within the genus Rubivirus. IMPORTANCE The detection of rustrela virus (RusV)-associated encephalitis in two carnivoran mammal species further extends the knowledge on susceptible species. Furthermore, we provide clinical and pathological data for the two new RusV cases, which were until now limited to the initial description of this fatal encephalitis. Using a sophisticated enrichment method prior to sequencing of the viral genome, we markedly improved the virus-to-background sequence ratio compared to that of standard procedures. Consequently, we were able to resolve and update the intergenic region and the coding region for the N terminus of the capsid protein of the initial RusV genome sequence. The updated putative capsid protein now resembles those of rubella and ruhugu virus in size and harbors a predicted RNA-binding domain that had not been identified in the initial RusV genome version. The newly determined complete RusV genomes strongly improve our knowledge of the genome structure of this novel rubivirus.

A Novel Rubi-Like Virus in the Pacific Electric Ray (Tetronarce californica) Reveals the Complex Evolutionary History of the Matonaviridae.

Rubella virus (RuV) is the causative agent of rubella ("German measles") and remains a global health concern. Until recently, RuV was the only known member of the genus Rubivirus and the only virus species classified within the Matonaviridae family of positive-sense RNA viruses. Recently, two new rubella-like matonaviruses, Rustrela virus and Ruhugu virus, have been identified in several mammalian species, along with more divergent viruses in fish and reptiles. To screen for the presence of additional novel rubella-like viruses, we mined published transcriptome data using genome sequences from Rubella, Rustrela, and Ruhugu viruses as baits. From this, we identified a novel rubella-like virus in a transcriptome of Tetronarce californica-order Torpediniformes (Pacific electric ray)-that is more closely related to mammalian Rustrela virus than to the divergent fish matonavirus and indicative of a complex pattern of cross-species virus transmission. Analysis of host reads confirmed that the sample analysed was indeed from a Pacific electric ray, and two other viruses identified in this animal, from the Arenaviridae and Reoviridae, grouped with other fish viruses. These findings indicate that the evolutionary history of the Matonaviridae is more complex than previously thought and highlights the vast number of viruses that remain undiscovered.

Congenital rubella syndrome in child of woman without known risk factors, New Jersey, USA.

We report a case of congenital rubella syndrome in a child born to a vaccinated New Jersey woman who had not traveled internationally. Although rubella and congenital rubella syndrome have been eliminated from the United States, clinicians should remain vigilant and immediately notify public health authorities when either is suspected.

[Preliminary study on ToRCH epidemic laws in Wuhan region].

OBJECTIVE: To investigate the ToRCH epidemic law in Wuhan region. METHODS: The ToRCH infection in the normal populations of Wuhan region was inspected through a whole year (from Mar. 1997 to Feb. 1998) by detecting the specific IgM antibodies to ToRCH in sera. RESULTS: The results showed that in the whole year, the average positive rates of IgM aitibodies to Toxo, RuV, CMV and HSV were 5.8%, 7.5%, 4.2% and 4.9% respectively. Three characteristics of the ToRCH epidemic in the region were concluded: (1) The epidemic of RuV obviously changed with seasons. Its infection rates were the highest (8.5%, 19.5%, 14.0%) during spring (March, April and May). However, the changes of Toxo, CMV and HSV were not obvious with seasons; (2). The adult infections were mainly recessive, which had no apparent clinical symptoms; (3) The cross infections of the four pathogens were common, the cross-reaction rate of the surveyed year was 2.6%. CONCLUSIONS: RuV infection of pregnant woman may result in congenital malformation of the emlbryo. Accoroding to the seasonal epidemic characteristics of RuV, women in pregnancy had better not to go to the public places during the sping seasons (March, April, May) as far as possible.

Inquérito sorológico de rubéola na era pré-vacinaçäo, em creches, escolas e maternidades de Fortaleza / Serologic survey of rubella in the pre-vacine era in child-care centers, schools and maternity units of Fortaleza

Objetivo: identificar a prevalência de rubéola em diversas idades e grupos populacionais e a suscetibilidade de mulheres gestantes e puérpuras segundo idade, número de filhos e aborto espoatâneo. Métodos: Estudo transversal de tipo inquérito sorológico. Pré-escolares e escolare foram selecionados em creches e escolas públicas distribuídas por distritos sanitários de Fortaleza. As gestantes e puérperas saudáveis foram recrutadas em duas grandes maternidades e três ambulatórios públicos de pré-natal. Indivíduos previamente vacinados e portadores de doenças crônicas ou agudas foram excluídos. Foi obtido consentimento escrito de todos os participantes ou responsáveis. Para detecçäo qualitativa de IgG sérica anti-rubéola utilizou-se método Elisa-indireto. Resultados: As soroprevalências médias por idade de 999 amostras foram 2 a 5 anos=59 por cento (136/231); 6 a 9 anos=53 porcento (109/204); 10 a 19 anos=56 por cento (243/432); e 20 a 39 anos=80 por cento (106/132). A idade média de 187 gestantes e puérperas foi de 23 anos (10-39) com soroprevalência de 76 por cento (142/187), 62 por cento no grupo de 15-19 anos e 83 por cento no grupo de 26-39 anos. Uma maior soroprevalência esteve associada à maior idade materna (p<0,001), a história de aborto e espotâneo (p=0,03) e a 2 ou mais filhos (p=0,01). Conclusöes: A elevada soroprevalência em pré-escolares revela a intensa transmissäo do vírus nas creches. A alta uscetibilidade nos adolescentes (45 por cento), entre os quais a gravidez é freqüênte, enfatiza a necessidade de se introduzir a vacina precocemente e manter elevada cobertura nos jovens, visando erradicar a rubéola congênita. Igualmente, a vacinaçäo rotineira das puérperas nesse grupo etário é particularmente benéfica.

Polymerase chain reaction-based detection of bacteria in semen.

OBJECTIVE: To determine if the presently used bacterial detection techniques provide accurate and complete profiles of microorganisms found in human semen. DESIGN: Routine bacterial cultures and molecular biology techniques using polymerase chain reaction (PCR), with a universal eubacterial primer, cloning, then sequence analysis were used to detect bacteria (culturable or nonculturable) in the semen. SETTING: University and hospital-based research laboratory. PATIENTS: Thirty infertile men and nine semen donors, all with no symptoms of a urinary tract infection, donated semen for the study. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Detection of bacteria using routine cultures and molecular biology techniques. RESULTS: Using PCR, we found > 10(4) bacteria/mL in the semen of 66% of the infertile asymptomatic men and 66% of the semen donors. This contrasts with our routine culture results which detected "significant" bacteriospermia in only 27% of the infertile men and in none of the preselected semen donors. From four of these semen specimens, DNA sequence analysis identified an average of nine different bacterial species per specimen, with close to 90% of the species being anaerobes. CONCLUSIONS: These data indicate that the present microbiologic detection methods underestimate the incidence of significant bacteriospermia, particularly anaerobic bacteria. The molecular biologic methods should help researchers confirm or refute the role of infection in male infertility.