Molecular Evolution of the Capsid Gene in Norovirus Genogroup I.

We studied the molecular evolution of the capsid gene in all genotypes (genotypes 1-9) of human norovirus (NoV) genogroup I. The evolutionary time scale and rate were estimated by the Bayesian Markov chain Monte Carlo (MCMC) method. We also performed selective pressure analysis and B-cell linear epitope prediction in the deduced NoV GI capsid protein. Furthermore, we analysed the effective population size of the virus using Bayesian skyline plot (BSP) analysis. A phylogenetic tree by MCMC showed that NoV GI diverged from the common ancestor of NoV GII, GIII, and GIV approximately 2,800 years ago with rapid evolution (about 10(-3) substitutions/site/year). Some positive selection sites and over 400 negative selection sites were estimated in the deduced capsid protein. Many epitopes were estimated in the deduced virus capsid proteins. An epitope of GI.1 may be associated with histo-blood group antigen binding sites (Ser377, Pro378, and Ser380). Moreover, BSP suggested that the adaptation of NoV GI strains to humans was affected by natural selection. The results suggested that NoV GI strains evolved rapidly and date back to many years ago. Additionally, the virus may have undergone locally affected natural selection in the host resulting in its adaptation to humans.

Imported case of acute respiratory tract infection associated with a member of species nelson bay orthoreovirus.

A Japanese man suffered from acute respiratory tract infection after returning to Japan from Bali, Indonesia in 2007. Miyazaki-Bali/2007, a strain of the species of Nelson Bay orthoreovirus, was isolated from the patient's throat swab using Vero cells, in which syncytium formation was observed. This is the sixth report describing a patient with respiratory tract infection caused by an orthoreovirus classified to the species of Nelson Bay orthoreovirus. Given the possibility that all of the patients were infected in Malaysia and Indonesia, prospective surveillance on orthoreovirus infections should be carried out in Southeast Asia. Furthermore, contact surveillance study suggests that the risk of human-to-human infection of the species of Nelson Bay orthoreovirus would seem to be low.

Comparison of real-time reverse-transcription loop-mediated isothermal amplification and real-time reverse-transcription polymerase chain reaction for detection of noroviruses in municipal wastewater.

The monitoring of NVs in municipal wastewater by both real-time RT-LAMP and real-time RT-PCR, and the comparison of these two methods with respect to NV detection were carried out. The change in NVs detected by real-time RT-LAMP agreed well with that detected by real-time RT-PCR. In contrast, the correlation between the copy number determined by real-time RT-PCR and the threshold time (Tt) determined by real-time RT-LAMP obtained during monitoring was not significant (0.1

Quantitative analysis of fecal sapovirus shedding: identification of nucleotide substitutions in the capsid protein during prolonged excretion.

Sapovirus (SaV) is an important pathogen causing gastroenteritis in humans. Quantitative analysis of the viral loads in feces collected from two SaV outbreaks was performed. Our results showed that SaV excretion generally decreased to an undetectable level in 2 weeks; however, some individuals excrete SaV in feces at high concentrations for 2-4 weeks after the onset of illness. In addition, we identified for the first time nucleotide changes in the capsid region during prolonged excretion.

Investigation of reservoir animals of Leptospira in the northern part of Miyazaki Prefecture.

We surveyed reservoir animals of leptospires in the northern part of Miyazaki Prefecture, where a cluster of human leptospirosis had occurred during the summer of 2006. Leptospira was isolated from 6 of 57 large Japanese field mice (Apodemus speciosus). The serogroups of the isolates were Autumnalis (5 strains) and Hebdomadis (1 strain) and the partial nucleotide sequences of their flaB genes suggested that the isolates belonged to L. interrogans. The human patient sera reacted specifically with the Leptospira strain isolated from the mice captured around the area where each patient occurred, suggesting that mice are the source of human infection. We also detected leptospiral DNAs by flaB-polymerase chain reaction in the kidneys of large feral animals; wild boars (positive ratio 10.3%; 4 of 39) and deer (19.2%; 10 of 52). The Leptospira spp. harbored by these animals were deduced to be L. interrogans (in 5 animals) and L. borgpetersenii (in 9 animals) by the nucleotide sequences of the amplicons. Anti-Leptospira antibodies were also detected among symptomatic hound dogs. These results suggest that these feral animals may cause leptospirosis and pose a potential risk to hunters and workers in the meat processing industry.

[Pathogenicities of Mycobacterium intracellulare and M. avium strains to the mice which were isolated from non-tuberculous mycobactriosis patients].

The virulence of 5 strains of M. intracellulare and 6 strains of M. avium to mice were examined. The former bacteria were obtained from the patients with mycobacterial pulmonary disease and the latter were from AIDS patients respectively. C57BL/6 (NRAMP-1 susceptible) and its NRAMP-1 congenic mice (resistant) were used to evaluate the virulence of these bacteria. Three of the 5 strains of M. intracelllulare showed a relatively high virulence. They grew in the liver, spleen and lungs of susceptible mice and even in the lungs of resistant mice. On the other hand, none of 6 strains of M. avium could grow in either susceptible or resistant mice. In conclusion, our mouse model may be a useful tool for evaluating the virulence of bacteria isolated from mycobacteriosis patients.