A seroepidemiologic study of a measles outbreak, Yamagata Prefecture, Japan, 2017: The estimation of spreaders using serological assays in a measles elimination setting.

INTRODUCTION: In regions where the endemic measles virus has been eliminated, early detection of contagious patients is important for preventing the spread of measles and sustaining elimination. To investigate whether serological assays can be used for the estimation of highly infectious patients with measles, we performed a seroepidemiologic study of a measles outbreak in Yamagata Prefecture, Japan, in 2017. METHODS: We tested plaque reduction neutralization (PRN), IgG avidity, and gelatin particle agglutination (PA) assays in 31 patients with measles, subdivided into two super-spreaders, three spreaders, and 26 non-spreaders. Simultaneously, these results were compared with the cycle threshold (Ct) of a semi-quantitative real-time reverse transcription PCR for the measles virus from throat swab specimens. RESULTS: In the PRN assay, one super-spreader and two spreaders lacked protective antibodies. The IgG avidity assay showed that two super-spreaders and one spreader had low avidity. The PA assay indicated that two super-spreaders and two spreaders lacked protective antibodies. Comparison of the results of the three serological assays and Ct revealed that patients whose antibody titers were judged as low in the IgG avidity and PA assays showed low Ct (i.e., high viral load), whereas non-spreaders tended to show low viral load. CONCLUSIONS: Our preliminary seroepidemiologic analysis of a population of 31 patients with measles suggests that PA and IgG avidity assays may be used for the identification of super-spreader/spreader candidates. However, further investigations are necessary to validate the robustness of these serological assays in detecting contagious measles cases.

Longitudinal epidemiology of human coronavirus OC43 in Yamagata, Japan, 2010-2017: Two groups based on spike gene appear one after another.

Human coronavirus OC43 (HCoV-OC43) is divided into genotypes A to H based on genetic recombination including the spike (S) gene. To investigate the longitudinal transition of the phylogenetic feature of the HCoV-OC43 S gene in a community, phylogenetic analysis of the S1 region of the S gene was conducted using 208 strains detected in Yamagata during 2010 to 2017 with reference strains of the genotype. The S1 sequences were divisible into four groups: A to D. All Yamagata strains belonged to either group B or group D. In group B, 46 (90.2%) out of 51 Yamagata strains were clustered with those of genotype E reference strains (cluster E). In group D, 28 (17.8%) and 122 (77.7%) out of 157 Yamagata strains were clustered, respectively, with genotype F and genotype G reference strains. In cluster G, 28 strains formed a distinct cluster. Monthly distributions of HCoV-OC43 in Yamagata in 2010 to 2017 revealed that group B and group D appeared one after another. In group B, the cluster E strains were prevalent recurrently. In conclusion, epidemics of HCoV-OC43 in Yamagata, Japan might be attributable to two genetically different groups: group B showed a recurrent epidemic of strains belonging to a single phylogenetic cluster and group D showed epidemic strains belonging to multiple clusters.

Scrub typhus caused by Shimokoshi type Orientia tsutsugamushi showing variant 56-kDa type-specific antigen gene sequence in Tohoku region, Japan.

In 2018, a patient was diagnosed with Shimokoshi type scrub typhus in Yamagata Prefecture, Japan. The causative pathogen was likely a variant type because 43 (8.3%) of 521 deduced amino acid sequences of the 56-kDa type-specific antigen (TSA) were different from those of the Shimokoshi prototype strain. The patient's paired sera showed low antibody titers against the Shimokoshi prototype strain. Two cases of scrub typhus reported in the Tohoku region during 2011-2012 also involved the same 56-kDa TSA gene sequence. These findings suggest the presence of diversity in Shimokoshi type Orientia tsutsugamushi, which may impede the laboratory diagnosis of scrub typhus.

Detection of Saffold viruses from children with acute respiratory infections in Yamagata, Japan, between 2008 and 2015.

Although Saffold virus (SAFV) was reported as a novel human cardiovirus in 2007, no causative association between SAFV and clinical disease has been proven and the longitudinal epidemiology of SAFVs is not available. To establish the relationship between SAFVs and acute respiratory infections (ARIs) and to clarify the longitudinal epidemiology of SAFVs, 7258 nasopharyngeal specimens were collected from children with ARIs in Yamagata, Japan between 2008 and 2015. The specimens were inoculated on a microplate including six cell lines as part of routine surveillance, and molecular screening was performed for SAFVs using a reverse transcription (RT)-PCR method. Throughout the study period, 95 (1.3%) SAFV genotype 2 (SAFV2), and 28 (0.4%) SAFV3 were detected, mainly between September and November. There were two outbreaks of SAFV2 in 2009 and 2013, and one outbreak of SAFV3 in 2012 and the positive rates during these outbreaks were 12.1% (53/439), 11% (35/319), and 4.4% (20/453), respectively. Sixty-three SAFV2 and 28 SAFV3 strains were detected as a single virus from children with ARIs such as pharyngitis, herpangina, and tonsillitis. These results suggested that SAFV2 and SAFV3 are possible causative agents of ARIs among children and their infections occur mainly in the autumn season in Japan.

Seroepidemiology of human parechovirus types 1, 3, and 6 in Yamagata, Japan, in 2014.

To clarify the seroepidemiology of human parechovirus type 1 (HPeV1), 3 and 6, neutralizing antibodies (NT Abs) were measured in 214 serum specimens collected in 2014 in Yamagata, Japan. The seroprevalence against HPeV1 was 100% in all age groups, while that against HPeV3 and HPeV6 was 79.4% and 66.8%, respectively, overall. The geometric mean titers of NT Abs against HPeV1, 3 and 6 were 755.2, 255.0 and 55.9, respectively, overall. Our findings indicate that HPeV1 is the most prevalent HPeV circulating in Yamagata, followed by HPeV3 and HPeV6.

Genetic Reassortment in a Child Coinfected with Two Influenza B Viruses, B/Yamagata Lineage and B/Victoria-Lineage Strains.

We identified a child coinfected with influenza B viruses of B/Yamagata and B/Victoria lineages, in whom we analyzed the occurrence of genetic reassortment. Plaque purification was performed using a throat swab specimen from a 9-year-old child, resulting in 34 well-isolated plaques. The genomic composition of eight gene segments (HA, NA, PB1, PB2, PA, NP, M, and NS genes) for each plaque was determined at the lineage level. Of the 34 plaques, 21 (61.8%) had B/Phuket/3073/2013 (B/Yamagata)-like sequences in all gene segments, while the other 13 (38.2%) were reassortants with B/Texas/02/2013 (B/Victoria)-like sequences in 1-5 of the 8 segments. The PB1 segment had the most B/Victoria lineage genes (23.5%; 8 of 34 plaques), while PB2 and PA had the least (2.9%; 1 of 34 plaques). Reassortants with B/Victoria lineage genes in 2-5 segments showed the same level of growth as viruses with B/Yamagata lineage genes in all segments. However, reassortants with B/Victoria lineage genes only in the NA, PB1, NP, or NS segments exhibited reduced or undetectable growth. We demonstrated that various gene reassortments occurred in a child. These results suggest that simultaneous outbreaks of two influenza B virus lineages increase genetic diversity and could promote the emergence of new epidemic strains.

Detection of Tick-Borne Pathogens in Ticks from Dogs and Cats in the Yamagata Prefecture of Japan in 2018.

Companion animals can become infected with tick-borne diseases (TBDs) becoming a reservoir for human transfer, thereby damaging human health. To evaluate whether companion animals are infested with ticks harboring human TBD pathogens, we detected TBD pathogens in ticks collected from dogs and cats brought to animal hospitals in the Yamagata prefecture of Japan. An investigation of 164 adult ticks collected from 88 dogs and 41 cats between March and July 2018 revealed that this region was dominated by three tick species, Ixodes ovatus (n = 95, 57.9%), Ixodes nipponensis (n = 37, 22.6%) and Haemaphysalis flava (n = 10, 6.1%). To evaluate their pathogenic potential, we went on to test each tick for spotted fever group rickettsiae, Lyme disease borreliae, relapsing fever borreliae, tick-borne encephalitis virus, and Huaiyangshan banyangvirus (formerly SFTS virus). Our results identified two I. ovatus ticks infected with Borrelia miyamotoi, which causes emerging relapsing fever; several I. nipponensis ticks infected with Rickettsia monacensis, which cause rickettsiosis; and several Ixodes persulcatus ticks infected with Rickettsia helvetica, which can also cause rickettsiosis. These results suggest that dogs and cats, and veterinary professionals and pet owners, in the Yamagata prefecture have some risk of exposure to several TBDs. This means that there should be continuous monitoring and reporting of TBDs, even those known to be uncommon in Japan, in both companion animals and humans to ensure the health and safety of both humans and animals in Japan.

Longitudinal Epidemiology of Viral Infectious Diseases Combining Virus Isolation, Antigenic Analysis, and Phylogenetic Analysis as Well as Seroepidemiology in Yamagata, Japan, between 1999 and 2018.

We introduced a microplate method for virus isolation in the Department of Microbiology, Yamagata Prefectural Institute of Public Health (YPIPH) in 1999 in Yamagata, Japan. We have since carried out longitudinal epidemiological studies on viral infectious diseases, particularly respiratory viruses, combining traditional technologies such as virus isolation and serological techniques and newly developed molecular methods. Here, we provide an overview of our activities at YPIPH between 1999 and 2018. During the study period, we observed emerging and re-merging diseases such as those caused by echovirus type 13, enterovirus D68, parechovirus-A3 (PeV-A3), and Saffold virus. With regard to PeV-A3, we proposed a new disease concept, "PeV-A3-associated myalgia/myositis." We also revealed the longitudinal epidemiologies of several viruses such as enterovirus A71 and coxsackievirus A16. To perform longitudinal epidemiological studies at any time in Yamagata, we established a system for stocking clinical specimens, viral isolates, complementary DNAs, and serum specimens. We have also pursued collaboration works with virology laboratories across Japan. We hope our experiences, findings, and research materials will further contribute to the development of countermeasures against viral infectious diseases and improvement in public health strategies in Yamagata, Japan, Asia, and around the world.

Parechovirus A3 (PeV-A3)-associated myalgia/myositis occurs irrespective of its genetic cluster: a longitudinal molecular epidemiology of PeV-A3 in Yamagata, Japan between 2003 and 2016.

No longitudinal molecular epidemiology of parechovirus A3 (PeV-A3) over a decade is available and PeV-A3-associated myalgia/myositis has been reported only in Japan. Thus, we aimed to clarify the longitudinal molecular epidemiology of PeV-A3 with a major focus on the strains detected from PeV-A3-associated myalgia/myositis cases. We performed sequence and phylogenetic analysis for the VP1 region of PeV-A3 strains in Yamagata, Japan, between 2003 and 2016. The phylogenetic analysis indicated that PeV-A3 strains caused PeV-A3-associated myalgia/myositis as well as a variety of infectious diseases, ranging from mild to severe, in subjects ranging from neonates to adults, irrespective of genetic cluster or variations. PeV-A3 strains are causative agents of a variety of human diseases, irrespective of their genetic cluster. Furthermore, we consider that PeV-A3-associated myalgia/myositis may occur, not only in Japan, but also in other countries, as closely related PeV-A3 strains have been circulating around the world.

Phylogenetic and antigenic analyses of coxsackievirus A6 isolates in Yamagata, Japan between 2001 and 2017.

Although coxsackievirus A6 (CV-A6) is generally recognized as a causative agent of herpangina in children, CV-A6 infections globally emerged as a new and major cause of epidemic hand-foot-and-mouth-diseases (HFMDs) around 2008. To clarify the longitudinal epidemiology of CV-A6, we carried out sequence and phylogenetic analyses for the VP1 and partially for the VP4-3D regions as well as antigenic analysis using 115 CV-A6 isolates and 105 human sera in Yamagata, Japan between 2001 and 2017. Phylogenetic analysis revealed that CV-A6 isolates were clearly divided into two clusters; strains in circulation between 2001 and 2008 and those between 2010 and 2017. Neutralizing antibody titers of two rabbit antisera, which were immunized with Yamagata isolates in 2001 and 2015, respectively, against 28 Yamagata representative strains as well as the prototype Gdula strain were 1:2560-1:5120 and 1:160-1:640, respectively. The neutralizing antibody titers among residents in Yamagata against the above two strains were similar. Our analyses revealed that there were cross-antigenicities among all analyzed CV-A6 strains, although the newly emerged strains were introduced into Yamagata around 2010 and replaced the previous ones. With regard to control measures, these findings suggest that we can prevent CV-A6 infections through the development of a vaccine that effectively induces neutralizing antibodies against CV-A6, irrespective of genetic cluster.

Polio vaccination coverage and seroprevalence of poliovirus antibodies after the introduction of inactivated poliovirus vaccines for routine immunization in Japan.

In Japan, the oral poliovirus vaccine (OPV) was changed to 2 types of inactivated poliovirus vaccine (IPV), the standalone conventional IPV (cIPV) and the Sabin-derived IPV combined with diphtheria-tetanus-acellular pertussis vaccine (DTaP-sIPV), for routine immunization in 2012. We evaluated polio vaccination coverage and the seroprevalence of poliovirus antibodies using data from the National Epidemiological Surveillance of Vaccine-Preventable Diseases (NESVPD) from 2011 to 2015. Several years before the introduction of IPV in 2012, OPV administration for children was refused by some parents because of concerns about the risk of vaccine-associated paralytic poliomyelitis. Consequently, in children aged <1 years who were surveyed in 2011-2012, polio vaccination coverage (45.0-48.8%) and seropositivity rates for poliovirus (type 1: 51.7-65.9%, type 2: 48.3-53.7%, and type 3: 15.0-29.3%) were decreased compared to those surveyed in 2009. However, after IPV introduction, the vaccination coverage (95.5-100%) and seropositivity rates (type 1: 93.2-96.6%, type 2: 93.1-100%, and type 3: 88.6-93.9%) increased among children aged <1 years in 2013-2015. In particular, seropositivity rates and geometric mean titers (GMTs) for poliovirus type 3 in <5-year-old children who received 4 doses of IPV (98.5% and 247.4, respectively) were significantly higher than in those who received 2 doses of OPV (72.5% and 22.9, respectively). Furthermore, in <5-year-old children who received 4 doses of either DTaP-sIPV or cIPV, the seropositivity rates and the GMTs for all 3 types of poliovirus were similarly high (96.5-100% and 170.3-368.8, respectively). Our findings from the NESVPD demonstrate that both the vaccination coverage and seropositivity rates for polio remained high in children after IPV introduction.

Clinical characteristics of children infected with enterovirus D68 in an outpatient clinic and the association with bronchial asthma.

BACKGROUND: All reports of increases in severe respiratory disease associated with human enterovirus D68 (EV-D68) are from hospital settings. However, there are few reports describing clinical characteristics in less severely affected populations. METHODS: We conducted a retrospective observational study from January 2010 to December 2015 in Yamagata, Japan. Using regional passive surveillance, 5794 respiratory specimens were collected from children who initially presented to an outpatient clinic with acute respiratory symptoms. The collected samples were tested for EV-D68 by reverse transcription PCR. RESULTS: EV-D68 was detected in 79 specimens mainly during the two epidemic periods in August-October 2010 and August-October 2015, when detection rates were 10.2% (31 of 304 specimens) and 16.3% (46 of 282 specimens), respectively. Among the 69 EV-D68-positive children, excluding those with viral coinfection, 39 (57%) had upper respiratory tract infections, 23 (33%) bronchiolitis or asthma attack, 5 (7%) bronchitis, 1 (1%) meningitis and 1 (1%) acute flaccid paralysis. In 23 children with wheezing, retraction was observed in 10 (43%), and six (26%) were diagnosed with asthma exacerbation. Six children required hospital admission, five (83%) because of asthma exacerbation. A history of asthma or wheezing was the most significant risk factor for the development of wheezing (odds ratio, 8.23; 95% CI, 2.65-25.50; p < .001). CONCLUSIONS: The low rate of hospitalization (9%, 6 of 69) indicates that most cases with EV-D68 infection were managed as outpatients. A history of asthma or wheezing was a potential risk factor for wheezing, resulting in hospitalization due to a severe asthma attack.

The Largest Measles Outbreak, Including 38 Modified Measles and 22 Typical Measles Cases in Its Elimination Era in Yamagata, Japan, 2017.

The incidence of modified measles (M-Me), characterized by milder symptoms than those of typical measles (T-Me), has been increasing in Japan. However, the outbreak dominated by M-Me cases has not been thoroughly investigated worldwide. The largest importation-related outbreak of measles with genotype D8 occurred in Yamagata Prefecture, Japan, from March to April 2017. This phenomenon was observed after Japan had achieved measles elimination in 2015. We confirmed 60 cases by detecting the genome of the measles virus (MeV). Among the cases, 38 were M-Me and 22 were T-Me. Thirty-nine (65.0%) patients were 20-39 years of age. Three out of 7 primary cases produced 50 transmissions, of which each patient caused 9-25 transmissions. These patients were 22-31 years old and were not vaccinated. Moreover, they developed T-Me and kept contact with the public during their symptomatic periods. Considering that M-Me is generally caused by vaccine failure, some individuals in Japan may have insufficient immunity for MeV. Accordingly, additional doses of measles vaccine may be necessary in preventing measles importation and endemicity among individuals aged 20-39 years. Furthermore, to accurately and promptly diagnose individuals with measles, particularly those who can be considered as primary cases, efforts must be exerted to detect all measles cases using epidemiological and genetic approaches in countries where measles elimination had been achieved.

Emotional relationships between child patients and their mothers during dental treatments.

BACKGROUND/PURPOSE: The behavioral control of child patients is an important issue in pediatric dentistry. The emotional states of the mothers of patients may influence the attitudes of their children. The aim of this study was to investigate the emotional states estimated from physiological responses of child patients and the subjective anxieties of their mothers during dental treatments and discuss the emotional relationships between children and their mothers. MATERIALS AND METHODS: To assess physiological responses associated with emotional changes induced by dental treatments in child patients aged 3-6 years, activity in the autonomic nervous were analyzed from variations in inter-beat intervals in electrocardiogram. Anxiety levels of accompanying mothers were examined using the State Trait Anxiety Inventory, which was filled out during the treatment of their child. RESULTS: Regarding the stress of child patients from the aspect of autonomic nervous activities during dental treatments, comparison between the cooperative and uncooperative patient groups showed that the uncooperative group demonstrated significantly higher sympathetic nervous activity and significantly lower parasympathetic nervous activity relative to the cooperative group, and their accompanying mothers showed significantly higher state anxiety scores relative to the mothers of cooperative children. Moreover, positive correlation between state anxiety scores of mothers and sympathetic nervous activities of their children was observed. CONCLUSION: These results indicated that uncooperative child patients undergo more stress and their mothers feel more anxiety from dental treatments, resulting in an emotional relationship between children and their mothers, which requires dental professionals to make special considerations to calm the anxiety of the mother, as well as the stress of the child patient.

Chronological changes of mumps virus genotypes in Japan between 1999-2013.

BACKGROUND: The molecular epidemiology of mumps virus (MuV) has been carried out worldwide based on genotyping proposed by the World Health Organisation. However, longitudinal molecular epidemiological studies of MuV are still limited. METHODS: This study carried out genotyping of MuVs isolated in Yamagata prefecture, which is located in northern Japan, between 1999-2013, using standard nomenclature based on the sequence analysis of the entire 316 nucleotides of the small hydrophobic (SH) gene. RESULTS: During this 15-year period, 249 MuVs were isolated, with the majority of them belonging to genotype G. Phylogenetic analysis revealed that genotype G strains were divided into two distinct clusters 1 and 2, consisting of 178 and 47 strains, respectively. The cluster 1 strains were isolated every year since 2001, except for 2012. The cluster 2 strains first appeared in 2011 and were dominant in 2011 and 2012. The epidemic pattern of genotype G strains observed in Yamagata was similar to those in Kanagawa and Hyogo prefectures located in eastern and western Japan, respectively. Only one L, three H and one F genotype strains were isolated in 2001, 2004 and 2010, respectively. Almost every year several genotype B strains related to Japanese vaccine strains were isolated. CONCLUSIONS: These data demonstrated that the genotype G strains have been endemically perpetuating as the major type over a wide area of Japan since 2001, although the genotype G strains that emerged after 2011 differed from the earlier strains.