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The Goose/Guangdong-lineage (Gs/Gd) H5 high pathogenicity avian influenza viruses (HPAIVs) spread among poultry and wild birds worldwide; an association has been identified between the migration of wild birds and spread of HPAIVs. Every autumn-spring season, the mallard (Anas platyrhynchos) migrates to Japan in substantial numbers for overwintering; however, to the best of our knowledge, no virological studies have focused on mallards' susceptibility to the HPAIVs in Japan. To evaluate the susceptibility of mallards to infection with Gs/Gd H5 HPAIVs isolated during previous outbreaks in Japan, we experimentally infected the birds with various virus strains: A/chicken/Yamaguchi/7/2004 (H5N1) (clade 2.5), A/chicken/Miyazaki/K11/2007 (H5N1) (clade 2.2), A/whooper swan/Akita/1/2008 (H5N1) (clade 2.3.2), A/mandarin duck/Miyazaki/22M-765/2011 (H5N1) (clade 2.3.2.1c), A/duck/Chiba/26-372-48/2014 (H5N8) (clade 2.3.4.4c), A/duck/Hyogo/1/2016 (H5N6) (clade 2.3.4.4e) and A/mute swan/Shimane/3211A002/2017 (H5N6) (clade 2.3.4.4b). The birds exhibited high tracheal shedding for a prolonged period, particularly those infected with A/duck/Hyogo/1/2016 (H5N6). Various clinical manifestations ranging from asymptomatic to mild (corneal opacity) infections to neurological disorders accompanied by mortality were noted depending on the virus strain. Furthermore, virus-infected mallards contaminated both cohoused mallards and water in their surroundings. Thus, mallards may disseminate viruses in the environment, thereby influencing HPAI outbreaks in Japan. Therefore, mallards represent an important migratory bird species that spread HPAIVs in Japan.

We previously developed a multiplex PCR assay for the differentiation of serovar 1a, 1b, 2 and 5 strains of Erysipelothrix rhusiopathiae. In this study, we analyzed the serovar-defining chromosomal region of a serovar 2 swine isolate, which was PCR-positive for both serovars 1a and 2 by the multiplex PCR assay. Genetic analysis of the chromosomal region revealed that, as in serovar 1a strains, the ERH_1440 gene, which is usually truncated or missing in serovar 2 strains, was intact in this strain. This paper first shows an E. rhusiopathiae serovar 2 strain possessing an intact ERH_1440 gene and suggests that care may be needed when determining the serovar of such rare strains by PCR assay.

The first human case of zoonotic H7N9 avian influenza virus (AIV) infection was reported in March 2013 in China. This virus continues to circulate in poultry in China while mutating to highly pathogenic AIVs (HPAIVs). Through monitoring at airports in Japan, a novel H7N3 reassortant of the zoonotic H7N9 HPAIVs, A/duck/Japan/AQ-HE30-1/2018 (HE30-1), was detected in a poultry meat product illegally brought by a passenger from China into Japan. We analysed the genetic, pathogenic and antigenic characteristics of HE30-1 by comparing it with previous zoonotic H7N9 AIVs and their reassortants. Phylogenetic analysis of the entire HE30-1 genomic sequence revealed that it comprised at least three different sources; the HA (H7), PB1, PA, NP, M and NS segments of HE30-1 were directly derived from H7N9 AIVs, whereas the NA (N3) and PB2 segments of HE30-1 were unrelated to zoonotic H7N9. Experimental infection revealed that HE30-1 was lethal in chickens but not in domestic or mallard ducks. HE30-1 was shed from and replicated in domestic and mallard ducks and chickens, whereas previous zoonotic H7N9 AIVs have not adapted well to ducks. This finding suggests the possibility that HE30-1 may disseminate to remote area by wild bird migration once it establishes in wild bird population. A haemagglutination-inhibition assay indicated that antigenic drift has occurred among the reassortants of zoonotic H7N9 AIVs; HE30-1 showed similar antigenicity to some of those H7N9 AIVs, suggesting it might be prevented by the H5/H7 inactivated vaccine that was introduced in China in 2017. Our study reports the emergence of a new reassortant of zoonotic H7N9 AIVs with novel viral characteristics and warns of the challenge we still face to control the zoonotic H7N9 AIVs and their reassortants.

Since 2013, H5N6 highly pathogenic avian influenza viruses (HPAIVs) have been responsible for outbreaks in poultry and wild birds around Asia. H5N6 HPAIV is also a public concern due to sporadic human infections being reported in China. In the current study, we isolated an H5N6 HPAIV strain (A/Muscovy duck/Long An/AI470/2018; AI470) from an outbreak at a Muscovy duck farm in Long An Province in Southern Vietnam in July 2018 and genetically characterized it. Basic Local Alignment Search Tool (BLAST) analysis revealed that the eight genomic segments of AI470 were most closely related (99.6%-99.9%) to A/common gull/Saratov/1676/2018 (H5N6), which was isolated in October 2018 in Russia. Furthermore, AI470 also shared 99.4%-99.9% homology with A/Guangxi/32797/2018, an H5N6 HPAIV strain that infected humans in China in 2018. Phylogenetic analyses of the entire genome showed that AI470 was directly derived from H5N6 HPAIVs that were in South China from 2015 to 2018 and clustered with four H5N6 HPAIV strains of human origin in South China from 2017 to 2018. This indicated that AI470 was introduced into Vietnam from China. In addition, molecular characteristics related to mammalian adaptation among the recent human H5N6 HPAIV viruses, except PB2 E627K, were shared by AI470. These findings are cause for concern since H5N6 HPAIV strains that possess a risk of human infection have crossed the Chinese border.

An H5N6 highly pathogenic avian influenza virus (HPAIV) outbreak occurred in poultry in Japan during January 2018, and H5N6 HPAIVs killed several wild birds in 3 prefectures during Winter 2017-2018. Time-measured phylogenetic analyses demonstrated that the Hemagglutinin (HA) and internal genes of these isolates were genetically similar to clade 2.3.4.4.B H5N8 HPAIVs in Europe during Winter 2016-2017, and Neuraminidase (NA) genes of the poultry and wild bird isolates were gained through distinct reassortments with AIVs that were estimated to have circulated possibly in Siberia during Summer 2017 and Summer 2016, respectively. Lethal infectious dose to chickens was similar between the poultry and wild-bird isolates. H5N6 HPAIVs during Winter 2017-2018 in Japan had higher 50% chicken lethal doses and lower transmission efficiency than the H5Nx HPAIVs that caused previous outbreaks in Japan, thus explaining in part why cases during the 2017-2018 outbreak were sporadic.

Anti-O-antigen antibodies, such as anti-O4 antigen IgG, induce protective immunity against Salmonella enterica serovar Typhimurium (S. Typhimurium) infection. S. Typhimurium belongs to the group O4, which can be classified into two serological variants, namely factor O5 antigen positive (O5+) and factor O5 antigen negative (O5-). In this study, we determined the protective immunity induced by anti-O4 antigen IgG against O5+ and O5- S. Typhimurium infection in a mouse model. Unexpectedly, anti-O4 antigen IgG induced protection against O5- of S. Typhimurium, but not against O5+ of S. Typhimurium. We suggest that the affinity of the O4 antigen with anti-O4 antigen IgG is stronger in the O5- S. Typhimurium compared to the O5+ S. Typhimurium. Although anti-O4 antigen IgG has the potential to protect against S. Typhimurium infection, the effects of anti-O4 antigen IgG in protection against Salmonella infection differ depending on the presence or absence of the O5 antigen.

Prion diseases are fatal neurodegenerative disorders of humans and animals and no effective treatments are currently available. Allogenic transplantation of immortalized human mesenchymal stem cells (MSCs) can prolong the survival of mice infected with prions. However, autologous transplantation is an appropriate model for evaluating the effects of MSCs on prion diseases. Therefore, we isolated and purified MSCs from the femur and tibia of mice as compact bone-derived MSCs (CB-MSCs). Flow cytometric analysis showed that CB-MSCs were negative for myeloid stem cell-derived cell markers CD11b and CD45, but positive for molecules such as Sca-1, CD105 and CD90.2, which are reported to be expressed on MSCs. The ability of CB-MSCs to migrate to brain extracts from prion-infected mice was confirmed by an in vitro migration assay. Intra-hippocampus transplantation of CB-MSCs at 120 days post-inoculation marginally but significantly prolonged the survival of mice infected with the Chandler prion strain. The transplantation of CB-MSCs did not influence the accumulation of disease-specific prion protein. However, the CB-MSC transplantation enhanced microglial activation, which appeared to be polarized to the M2-type activation state. These results suggest that autologous MSC transplantation is a possible treatment for prion diseases, while the modification of microglial activation may be a therapeutic target for neurodegenerative diseases.

There has been no report on the prevalence of Campylobacter spp. in farm animals in Mongolia. To uncover the prevalence of Campylobacter spp. in chickens in Mongolia and their antimicrobial resistance, in this study, we isolated and characterized Campylobacter spp. from chickens in Mongolia. We collected 71 cloacal swabs of chickens from 5 farms including 4 layer farms and one broiler farm near Ulaanbaatar city and isolated 25 Campylobacter jejuni and 6 Campylobacter coli isolates. All isolates were resistant to tetracycline, and 3 C. coli isolates were resistant to erythromycin. The C. coli isolates possessed either the erm(B) gene or nucleotide substitution at nt 2,075 of 23S rDNA, both of which are known to be associated with erythromycin resistance. Sixteen of the 31 C. jejuni/C. coli isolates (51.6%) were resistant to nalidixic acid and fluoroquinolones. All the fluoroquinolone-resistant isolates possessed amino acid substitution from threonine to isoleucine at codon 86 (nucleotide substitution: ACA to ATA). Multilocus sequence typing and phylogenetic analyses showed a variation in C. jejuni/C. coli in chickens in Mongolia. In addition, some of the C. jejuni isolates seemed to be phylogenetically close to isolates in Asian and Oceanian countries. This is the first report on the characterization of antimicrobial resistance of Campylobacter spp. in farm animals in Mongolia and is valuable for implementation of measures for a prudent use of antimicrobials in farm animals.

High pathogenic avian influenza viruses (HPAIVs) of the H5 subtype have spread in poultry and wild birds worldwide. Current studies have highlighted the association between the migration of wild birds and the spread of HPAIVs. However, virological studies examining responsible species of migratory birds to spread HPAIVs are limited. In Japan, the common teal (Anas crecca) arrives in great numbers for overwintering every autumn-spring season; therefore, we performed experimental infection using six H5 HPAIVs isolated in past outbreaks in Japan (A/chicken/Yamaguchi/7/2004 (H5N1), A/whooper swan/Akita/1/2008 (H5N1), A/mandarin duck/Miyazaki/22M-765/2011 (H5N1), A/duck/Chiba/26-372-48/2014 (H5N8), A/duck/Hyogo/1/2016 (H5N6) and A/mute swan/Shimane/3211A002/2017 (H5N6)) to evaluate the susceptibility of the species to HPAIV infection. The results illustrated that most birds in all experimental groups were infected by the strains, and they shed viruses for a prolonged period, in trachea than cloaca, without displaying distinctive clinical signs. In addition, comparative analysis using calculation value of total viral shedding during the experiment revealed that the birds shed viruses at above a certain level regardless of the differences of strains. These results suggested that the common teal could be a migratory bird species that disseminates viruses in the environment, thereby influencing HPAI outbreaks in wild birds in Japan.

To examine the behaviour of ozone (O 3 ) in the seasonal snowpack, measurements were taken of O 3 and CO 2 in the interstitial air on Rishiri Island, which is located in northern Japan, during the 2010/11 winter season. Exhibiting variation on timescales ranging from several minutes to several days, the atmospheric O 3 in the surface air generally increased from December (38 ppb) to April (52 ppb). The ozone mixing ratio sharply decreased below the snow surface. Whereas the CO 2 data in the interstitial air indicated that a rapid exchange between the snow and the atmosphere occurred intermittently, the O 3 mixing ratio remained low and constant (

To investigate the dynamics and environmental drivers of CO 2 flux through the winter snowpack in a dwarf bamboo ecosystem (Hokkaido, northeast Japan), we constructed an automated sampling system to measured CO 2 concentrations at five different levels in the snowpack, from the base to the upper snow surface. Using a gas diffusion approach, we estimated an average apparent soil CO 2 flux of 0.26 μmol m −2 s −1 during the snow season (December–April); temporally, the CO 2 flux increased until mid-snow season, but showed no clear trend thereafter; late-season snow-melting events resulted in rapid decreases in apparent CO 2 flux values. Air temperature and subnivean CO 2 flux exhibited a positive linear relationship. After eliminating the effects of wind pumping, we estimated the actual soil CO 2 flux (0.41 μmol m −2 s −1 ) to be 54% larger than the apparent flux. This study provides new constraints on snow-season carbon emissions in a dwarf bamboo ecosystem in northeast Asia.