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We determined nucleotide sequences and inferred amino acid sequences of viral protein (VP) 4, VP6, VP7, and nonstructural protein 4 genes of a porcine rotavirus strain (SKA-1) from Japan. The strain was closely related to a novel group of human rotavirus strains (B219 and J19). [ABSTRACT FROM AUTHOR]

Non-primate hepacivirus (NPHV) is a recently discovered homolog of the hepatitis C virus in horses. The frequency and distribution of NPHV infections among horses in Japan is unknown. In this study, serum samples from 453 horses across Japan were screened for NPHV RNA using real-time RT-PCR and anti-nonstructural 3 protein (NS3) antibodies using the Gaussia luciferase immunoprecipitation system assay. In order to monitor the course of NPHV infection in horses, we examined 31 stored samples (9 adult horses and 22 young horses) obtained one year ago and compared the results to the recent data. Stored sera from 7 mare-foal pairs were also examined. The NS3 region sequences of 14 NPHV strains from NPHV RNA positive serum samples were determined and analyzed phylogenically. Of the 453 serum samples tested, 33.55% were positive for anti-NS3 antibody and 13.68% were positive for NPHV RNA. We found a higher rate of NPHV RNA detection in serum obtained from young horses (1-2 years of age) than that of adults, in two geographically distinct areas. We observed higher variation in the course of infection over one year in young horses than in adult horses. The foals were infected with NPHV after the weaning period. Phylogenic analysis revealed that while NPHV NS3 genes isolated in Japan clustered with sequences previously classified as NPHV, but the genetic diversity of the Japanese NPHV strains we detected was not correlated with their geographic origin. In conclusion, Japanese horses exhibit a high prevalence of NPHV. Young age appears to be a risk factor for such viral infection in Japan, although the infectious route was not determined., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We evaluated the antiviral activity of a chlorine dioxide gas solution (CD) and sodium hypochlorite (SH) against feline calicivirus, human influenza virus, measles virus, canine distemper virus, human herpesvirus, human adenovirus, canine adenovirus and canine parvovirus. CD at concentrations ranging from 1 to 100 ppm produced potent antiviral activity, inactivating >or= 99.9% of the viruses with a 15 sec treatment for sensitization. The antiviral activity of CD was approximately 10 times higher than that of SH.

The efficacy of gaseous chlorine dioxide (ClO2) against feline calicivirus (FCV), a norovirus surrogate, in the dry and the wet states on a hard surface was evaluated. We demonstrated that low-concentration ClO2 gas (mean 0.08 ppm, 0.22 microg/I) could inactivate FCV in the wet state with 0.5% fetal bovine serum (FBS) within 6 h in 45 to 55% relative humidity (RH) (> 3 log10 reductions) and FCV in the dry state with 2% FBS (percentage of FBS in the viral suspension) within 10 h in 75 to 85% RH (> 3 log10 reductions) at 20 degrees C, respectively. Furthermore, a < 0.3 ppm concentration of ClO2 gas (mean 0.26 ppm, 0.73 microg/l) could inactivate (below the detection limit) FCV in the dry state with 5% FBS within 24 h in 75 to 85% RH at 20 degrees C. In contrast, in 45 to 55% RH at 20 degrees C, ClO2 gas had little effect even when the FCV in the dry state was exposed to high-concentration ClO2 (mean 8 ppm, 22.4 microg/l) for 24 h. These results suggest that humidity plays an important role in the inactivation by ClO2 gas of FCV in the dry state. According to the International Chemical Safety Card, threshold limit values for ClO2 gas are 0.1 ppm as an 8-h time-weighted average and 0.3 ppm as a 15 min short-term exposure limit. From these data, we propose that the treatment of wet areas of human activity such as kitchens, toilets, etc., with low-concentration ClO2 gas would be useful for reducing the risk of infection by noroviruses (NV) without adverse effects. In addition, we believe that the application of a combination of a < 0.3 ppm concentration of ClO2 gas and a humidifier in places without human activity may make it possible to inactivate NV in the dry state on any surface within a contaminated room without serious adverse effects.

Ascorbate (AsA) is a major antioxidant and free-radical scavenger in plants. Monodehydroascorbate reductase (MDAR; EC 1.6.5.4) is crucial for AsA regeneration and essential for maintaining a reduced pool of AsA. To examine whether an overexpressed level of MDAR could minimize the deleterious effects of environmental stresses, we developed transgenic tobacco plants overexpressing Arabidopsis thaliana MDAR gene (AtMDAR1) in the cytosol. Incorporation of the transgene in the genome of tobacco plants was confirmed by PCR and Southern-blot analysis and its expression was confirmed by Northern- and Western-blot analyses. These transgenic plants exhibited up to 2.1-fold higher MDAR activity and 2.2-fold higher level of reduced AsA compared to non-transformed control plants. The transgenic plants showed enhanced stress tolerance in term of significantly higher net photosynthesis rates under ozone, salt and polyethylene glycol (PEG) stresses and greater PSII effective quantum yield under ozone and salt stresses. Furthermore, these transgenic plants exhibited significantly lower hydrogen peroxide level when tested under salt stress. These results demonstrate that an overexpressed level of MDAR properly confers enhanced tolerance against ozone, salt and PEG stress.

Group B rotaviruses detected in Bangladesh in 2000 and 2001 were analyzed genetically to clarify relatedness to human group B rotaviruses reported previously in China and India, and to animal group B rotaviruses. VP7 gene sequences of the Bangladeshi group B rotaviruses (Bang373, Bang544, Bang334, and Bang402) were almost identical to each other and also showed high sequence identity to the Indian strain CAL-1 (98%) and Chinese strain adult diarrhea rotavirus (ADRV) (92%), while identities to bovine and murine viruses were considerably low (60-63%). Other genes of Bang373 and Bang544 encoding VP2, VP4, VP6, and NSP1 through NSP5 also showed much higher sequence identities to those of CAL-1 (97.7-99.4%) than to those of ADRV (89.9-93.9%). Characterization of nucleotide substitutions among Bang373, CAL-1, and ADRV suggested that all the gene segments might have evolved neutrally at similar mutation rates, while some of the gene segments (e.g., VP2 gene) were suggested to be more conserved than others. In conclusion, group B rotaviruses detected in Bangladesh represented by Bang373 and the Indian virus CAL-1 were considered as virtually identical viruses which are distinct genetically from ADRV, and it was suggested that Bang373 (CAL-1)-like group B rotavirus (Bengali strains) might be distributed primarily in an area around the Bay of Bengal., (Copyright 2004 Wiley-Liss, Inc.)

Human group B rotavirus was detected in 12 of 220 adult patients and 2 of 67 child patients with severe diarrhea in Bangladesh. Group B rotavirus may be virulent in both adults and children, and the virus may be an especially serious diarrheal agent in Bangladesh.

Canine parvovirus (CPV) was detected in faeces from dogs with diarrhoea by a specific slide agglutination test using latex particles coated with anti-CPV monoclonal antibody (LA-anti-CPV). The agglutination of LA-anti-CPV with CPV on a glass slide was evident macroscopically within 2 min. The sensitivity of the latex agglutination (LA) test was similar to that of the hemagglutination test. The LA test is available for the rapid diagnosis of CPV infection at an animal hospital.

Reoviruses designated as OS-320 to OS-324 were isolated from a total of 5 fecal specimens from 3 with diarrhea and 2 apparently healthy pigs aged 3 months. The serotype of these isolates was determined as reovirus type 2 by cross neutralizing tests. Furthermore, the results of hemagglutination test suggested the isolates were different from the other reoviruses. A survey of neutralizing antibody against reoviruses showed type 2 virus was widely prevalent among pigs in Okayama Prefecture. This paper is the first report of the isolation of reovirus type 2 from pigs.

Four stains designated as OB94-1 to OB94-4 of group A bovine rotavirus (BRV) were isolated from 35 fecal samples of calves with diarrhea in sporadic outbreaks. In VP7 (G) and VP4 (P) serotyping of these isolates, OB94-1 to OB94-3 were determined as G6P5, G6P5 and G10P5, respectively, by cross neutralization (NT) test and the G- and P- serotyping polymerase chain reaction (PCR) analysis. OB94-4 showed a one-way antigenic relation with the Lincoln stain (G6P1) and a weak antigenic relationship with the KK3 strain (G10P11), and was determined as G6P11 by the PCR method. Thus, OB94-4 was shown to be a new G6 BRV with different antigenic properties from the others in the NT test.

We evaluated drug resistance and R plasmids of 554 stains of Escherichia coli isolated from feces of migratory watefowl, including whistling swans (Cygnus columbianus), pintails (Anas acuta) and black-tailed gulls (Larus crassirostris) collected from the San-in District, Japan, between each November and March, 1983 to 1984, 1984 to 1985, and 1985 to 1986. Seven antimicrobial agents were tested: dihydrostreptomycin (DSM), kanamycin, spectinomycin, ampicillin (ABPC), oxytetracycline (OTC), chloramphenicol, and sulfadimethoxine (SDMX). Many strains were resistant to several drugs; in particular, all strains were resistant to SDMX. Both multiple drug resistant strains and drug resistance patterns occurred most frequently in strains isolated from whistling swans, followed by black-tailed gulls, and pintails, respectively. Of 233 strains, 128 (55%) carried transmissible R plasmids. The drugs with the largest number of resistance patterns observed were, in descending order, OTC, DSM, ABPC, and SDXM.

Thirty strains of Yersinia pseudotuberculosis were isolated from rabbits (17 strains), wild rats (9 strains) and house rats (4 strains) in China between 1990 and 1993. The biochemical properties of these isolates were identical with those of Y. pseudotuberculosis and no special characteristics were found in these strains. Serologically, serogroups 4b and 5b were identical to isolates found in Japan, and a new serogroup 1c and unclassified strains have also been detected. The existence of virulence-associated properties were different among strains. The pYV plasmid was detected from 6 strains of 30 isolates. This report documents the presence of Y. pseudotuberculosis in China, providing important epidemiological information.

Chicks at 2, 4 or 6 weeks of age were experimentally infected individually with a nephrosis/nephritis-causing avian infectious bronchitis virus (IBV) strain Kagoshima-34. The susceptibility of chicks in each group to the infection was compared, based on the clinical signs, excretion of virus in the faeces and antibody titres in the serum. The results showed that although all chicks appeared to be susceptible to IBV infection, the most severe clinical response was observed following infection at 2-week-old. Likewise, whilst the virus was also recovered from the faeces of all the chicks infected, the duration of viral excretion was longest in the 2-week-old chicks. A high antibody titre was detected at 4 weeks post infection (PI) and was maintained for at least another 16 weeks in the 4- and 6-week-old chicks. In contrast, a low antibody titre was detected only between 8 to 12 weeks PI in the 2-week-old chicks. Thereafter, no antibody was detected despite the presence of clinical signs.

Yersinia pseudotuberculosis which were screened out depending on auto-agglutination and Ca2+ dependency, were examined for their production of hemagglutinin (HA), and its purification and characterization were performed. The HA with a broad reactivity with various mammalian erythrocytes was recovered from the culture supernatant of these strains grown at 37 C but not 25 C. HAs from two strains, R148R and T1040, were purified by salt precipitation, gel filtration and anion-exchange chromatography by HPLC. Both purified HAs were cysteine-deficient acidic protein with an apparent molecular weight in the range of 15,000 to 16,000. N-terminal amino acid sequences of the first 25 residues were found to share 12% identity with that of afimbrial adhesin from enterotoxigenic Escherichia coli 2230. Immunoelectron microscopy and immunodiffusion test with polyclonal antiserum raised against the purified R148RHA demonstrated that the HA was associated with the amorphous aggregates which were detached from bacteria. These results suggest that the HA of Y. pseudotuberculosis belongs to a third type of HA produced by the yersinial species.

Adenovirus (Ad) type 40 and 41 DNAs were directly extracted from stool specimens of children with gastroenteritis. Two new strains of Ad41, Sanekata and Ehime strain, were cloned and their restriction maps were constructed. The left terminal end of the cloned Ad41 genome, EcoRI-E fragment of the Sanekata strain and EcoRI-F fragment of the Ehime strain, had transforming ability in rat 3Y1 cells. Only one of the 35 isolates of Ad40 tested showed a different restriction profile, while three different restriction profiles were found in DNAs from Ad41 isolates.

We developed a simple agglutination test for the detection of porcine rotavirus in stools from pigs with diarrhea. The virus was detected with high sensitivity and specificity by a slide agglutination test using latex particles coated with antibody against the porcine rotavirus strain OSU (LA-antiOSU). The agglutination of LA-antiOSU with OSU on a glass slide was evident macroscopically within 2 min. The sensitivity of this latex agglutination (LA) test was four times higher than that of the electron microscope method. The LA test is available for the rapid diagnosis of porcine rotavirus infections.

Avian infectious bronchitis virus strain Kagoshima-34 isolated from the kidneys of a chicken that died of nephrosis/nephritis lost its nephropathogenicity during intratracheal passage in SPF chickens. The resultant virus acquired stronger respirotropism but reduced tropism for kidneys. On the other hand strain Tottori-2 isolated from the trachea of a chicken suffering from severe respiratory disease did not lose its respirotropism after serial intravenous passage in SPF chickens. The serological properties of the passaged virus were investigated by virus neutralisation test. The antibody titres of both strains of virus fluctuated with progressive passage. The serological properties of the virus isolated from respiratory organs were not necessarily the same as those of the isolates made from the kidneys.

Human rotavirus (HRV) in faeces of patients may be readily detected with high sensitivity and specificity using latex agglutination (LA) on a glass slide by making use of the cross-reactivity of anti-calf rotavirus (CRV) antibody. Latex particles were coated with anti-CRV immunoglobin. The antibody coated particles (AC-L) are specifically agglutinated by both CRV and HRV, and the agglutination is evident macroscopically within a minute. To examine the sensitivity and reliability of the LA method compared to other methods, HRV in faecal extracts of 48 infants with acute gastroenteritis was sought by the LA, reversed passive haemagglutination (RPHA) and electron microscope (EM) methods. Samples positive by the EM method were all positive by the LA method, and samples negative by EM were all negative by LA. The LA method is suitable for application as a simple clinical diagnostic test.

We sensitized sheep erythrocytes (SRBC) with antibodies against human rotavirus strain Wa (SRBC-antiWa) and antibodies against calf rotavirus strain NCDV (SRBC-antiNCDV). These were readily agglutinated in the presence of homologous antigens, i.e., human rotavirus and calf rotavirus. By the hemagglutination of SRBC-antiWa and SRBC-antiNCDV (reverse passive hemagglutination [RPHA]), titration of rotavirus in extracts from feces of children suffering from diarrhea (61 specimens) was carried out. We found that the ratio of titers determined with SRBC-antiWa and SRBC-antiNCDV varied remarkably from specimen to specimen. This indicated that the antigenic determinants on human rotavirus in patients feces cross-react with antibodies against NCDV to varying extents. To express the cross-reactivity of human rotavirus with antibodies to NCDV, we propose a Wa/NCDV rotavirus index which can be calculated from the RPHA titer with SRBC-antiWa and SRBC-antiNCDV as follows: Wa/NCDV rotavirus index = (antiWa-RPHA titer of specimen/antiWa-RPHA titer of NCDV)/(antiNCDV-RPHA titer of specimen/antiNCDV-RPHA titer of NCDV).