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.

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.