Isolation of azole-resistant Aspergillus spp. from food products.

The prevalence of azole-resistant Aspergillus fumigatus is increasing worldwide and is speculated to be related to the use of azole pesticides. Aspergillus spp., the causative agent of aspergillosis, could be brought into domestic dwellings through food. However, studies on azole-resistant Aspergillus spp. in food products are limited. Therefore, we aimed to isolate Aspergillus spp. from processed foods and commercial agricultural products and performed drug susceptibility tests for azoles. Among 692 food samples, we isolated 99 strains of Aspergillus spp. from 50 food samples, including vegetables (22.9%), citrus fruits (26.3%), cereals (25.5%), and processed foods (1.8%). The isolates belonged to 18 species across eight sections: Aspergillus, Candidi, Clavati, Flavi, Fumigati, Nidulantes, Nigri, and Terrei. The most frequently isolated section was Fumigati with 39 strains, followed by Nigri with 28 strains. Aspergillus fumigatus and A. welwitschiae were the predominant species. Ten A. fumigatus and four cryptic strains, four A. niger cryptic strains, two A. flavus, and four A. terreus strains exceeded epidemiological cutoff values for azoles. Aspergillus tubingensis, A. pseudoviridinutans, A. lentulus, A. terreus, and N. hiratsukae showed low susceptibility to multi-azoles. Foods containing agricultural products were found to be contaminated with Aspergillus spp., with 65.3% of isolates having minimal inhibitory concentrations below epidemiological cutoff values. Additionally, some samples harbored azole-resistant strains of Aspergillus spp. Our study serves as a basis for elucidating the relationship between food, environment, and clinically important Aspergillus spp.

ROS1 as a possible prognostic biomarker of cervical adenocarcinoma: An exploratory analysis with next-generation sequencing.

OBJECTIVES: Given the differences in clinical and biological characteristics between cervical adenocarcinoma and squamous cell carcinoma, this study aimed to conduct an exploratory analysis to examine the molecular characteristics of cervical adenocarcinoma in a Japanese population. METHODS: This study explored the simultaneous testing of multiple mutations targeting cervical adenocarcinoma using next-generation sequencing (NGS). The following genes were analyzed: BCAR4, CD274, PDCD1LG2, KRAS, ARID1A, PTEN, ALK, EGFR, ROS1, BRAF, PIK3CA, EP300, EBXW7, SHCBP1, TGFBR2, SMAD4, ERBB2, ERBB3, and KLF5. Tumor tissue and blood samples were obtained at the time of primary treatment. The NGS-based molecular profiles obtained from Tokai University (49 specimens) were compared with the registered data in The Cancer Genome Atlas (TCGA) database (133 specimens). RESULTS: The study cohort had higher rates of adenocarcinoma than the TCGA cohort (44.9% vs. 18.0%; P = 0.001). The adenocarcinomas in the study cohort had more alterations in ROS1, EGFR, EP300, SHCBP1, ALK, and PIK3CA than those in the TCGA cohort. Among them, ROS1 had the highest number of gene alterations (median, 7.00 ± 2.63). In the study cohort, patients with a high number of ROS1 alterations had a significantly higher recurrence rate (5-year recurrence rate, 48.8% vs. 14.6%; hazard ratio [HR], 4.32; 95% confidence interval [CI], 1.20-15.50; P = 0.014) and lower overall survival than those with low alterations (5-year survival rate, 70.7% vs. 93.1%; HR, 7.15; 95% CI, 1.08-58.22; P = 0.032). CONCLUSION: The current exploratory analysis suggests that ROS1 gene alteration may be a prognostic biomarker in cervical adenocarcinoma in Japanese patients.

A peer-led learning program about intimate and romantic relationships for persons with mental disorders (AIRIKI): co-creation pilot feasibility study.

BACKGROUND: Intimate and romantic relationships are important in life for individuals, irrespective of mental health status. We developed a four-hour peer-led learning program for persons with mental disorders about intimate and romantic relationships through a co-creation process with service users and examined its preliminary effectiveness and feasibility of implementing the program. METHODS: A one-group pretest-posttest trial was conducted using a mixed-method design for 45 individuals with mental disorders in Japan. Outcome data were collected at three time points: baseline, post-intervention, and one month after program completion. Mixed models for repeated measures (MMRM) were used to examine changes over time in the Rosenberg Self-Esteem Scale (RSES), Recovery Assessment Scale (RAS), Herth Hope Index (HHI), and the original items. Group interviews were conducted for process evaluation. RESULTS: MMRM showed significant changes over time on RSES, RAS, HHI, and two original items "I am able to communicate well with others about myself" and "I am able to listen to others well." In multiple comparisons, RSES and HHI were significant one month after the program. Participants reported changes during the first month after attending the program in terms of their positive attitude toward romantic relationships (n = 14), taking romantic actions (n = 11), and feeling their overall communication improved (n = 11). Although two participants had an unscheduled psychiatric visit that could be attributed to attending the program, all recovered after one month. CONCLUSIONS: The program exhibited preliminary effectiveness to a moderate extent in improving recovery, particularly regarding self-esteem and hope. The program is feasible but requires further modifications regarding inclusion criteria for participants and the training of peer facilitators. TRIAL REGISTRATION: UMIN000041743;09/09/2020.

Video-based e-learning program for schoolteachers to support children of parents with mental illness: a cluster randomized trial.

BACKGROUND: Some children of parents with mental illness need support. This study aimed to develop and test the effectiveness of an e-learning program for training elementary schoolteachers to support children of parents with mental illness. METHODS: The program, which included a 30-min video-based e-learning program, aimed to help schoolteachers gain basic knowledge about mental illness and children of parents with mental illness, recognize children in need of support, and gain confidence in supporting them. A school-based cluster randomized controlled trial was conducted, and the schools were randomly divided into intervention and control groups. The teachers at these schools signed up for the program and participated individually. The outcome measures for the schoolteachers were evaluated at three time points: baseline (T1), post (T2), and one month later (T3). Along with the Sense of Coping Difficulty subscale (primary outcome measure), the following self-developed outcome measures were used: actual behaviors and attitude toward supporting children, knowledge, and self-assessment of program goals achievement. The Sense of Coping Difficulty subscale results at T3 were compared between the groups. Effectiveness over time was assessed for all the outcome measures. The interaction between baseline and intervention effects on the Sense of Coping Difficulty subscale was analyzed. As a part of the process evaluation, open-ended text responses were analyzed qualitatively. RESULTS: Baseline responses were collected from 87 participants in the intervention group and 84 in the control group. The total score of the Sense of Coping Difficulty subscale at T3 was significantly lower in the intervention group than in the control group (p = 0.007). Over time, a significant effect was observed on the Sense of Coping Difficulty subscale, actual behavior, knowledge of onset timing and probability of onset, and achievement of all program goals. Exploratory analysis was particularly effective for those who encountered a high level of difficulty in supporting children. The participants' text responses indicated that they planned to look carefully at children's backgrounds and stay close to them in the future. CONCLUSIONS: The program was effective for schoolteachers in supporting children of parents with mental illness. TRIAL REGISTRATION: UMIN000045483; 14/09/2021.

[Prevalence and Antimicrobial Resistance of Campylobacter jejuni/coli and Analysis of Macrolide Resistance Isolates from Retail Meat in Tokyo, Japan (2010-2019)].

This study aimed to investigate the prevalence and antimicrobial sensitivity of Campylobacter jejuni and Campylobacter coli in retail meat (chicken, beef, pork, venison, wild boar, horse, lamb and mutton) in Tokyo (Japan) from 2010 to 2019. Furthermore, the resistance mechanism of erythromycin (EM)-resistant strains was analysed. C. jejuni had a highly positive rate in domestic chicken meat (53.4%, 334/626 samples), domestic chicken offal (49.3%, 34/69 samples), and domestic beef offal (28.3%, 47/166 samples), while C. coli had a high positivity rate in domestic pork offal (31.7%, 44/139 samples). The positive rate of C. jejuni was significantly higher in offal than that in meat in domestic beef, while the positive rate of C. coli was significantly higher in offal than that in meat in domestic beef and domestic pork (p<0.05). In the isolates, 1.0% (6/631 strains) of C. jejuni and 36.2% (55/152 strains) of C. coli were EM resistant, with 41.5% (262/631 strains) of C. jejuni and 65.1% (99/152 strains) of C. coli being ciprofloxacin resistant. A2075G mutation of the 23S rRNA gene was confirmed in all EM-resistant strains.

[Situations in which individuals with mental disabilities perceive the need for advocacy support and how to deal with them].

Objectives　Policies promoting community integration of individuals with mental disabilities have increased the need for community advocacy. This study aimed to identify situations in which individuals with mental disabilities perceive the need for advocacy support and how to deal with them.Methods　In this qualitative descriptive study, group interviews were conducted with 13 peer advocates and 12 individuals with mental disabilities. A verbatim transcript of the interviews was created. Categories were generated by raising the level of abstraction from the perspective of "Situations where individuals with mental disabilities require advocacy support and how to address them."Results　Situations requiring advocacy support occurred in outpatient psychiatry settings, psychiatric hospitalizations, welfare facilities, schools, neighborhoods, and places of employment; among family and relatives; and at consultation services. In outpatient psychiatry, "difficulties in accessing medical care" were reported. In psychiatric hospitalizations, participants felt "pressured and unable to escape the environment." In welfare facilities, "romantic relationships between users were discouraged." Regarding familial difficulties, "limited understanding and acceptance of the disease," "relationship deterioration due to poor hospitalization conditions and forced hospitalization," and "marital difficulties due to mental illness" were prevalent. Participants in schools experienced "isolation due to their illness," and in the local community, there were "problems related to reasonable accommodation of individuals with disabilities in neighborhood association activities." Employed participants faced "inadequate consideration despite disclosing their illness to co-workers." At counseling institutions, participants felt "forced to endure without resolution when consulting." Individuals with disabilities coped with these situations by "transferring to a different clinic" or "changing facilities," but in the case of psychiatric hospitalization, they gave up and "did not go against the staff."Conclusion　Individuals with mental disabilities need advocacy support for psychiatric care and diverse situations such as family, school, and community engagement. Efforts should be made to introduce an advocacy system in psychiatric hospitals and to disseminate accurate information about mental illness to high-risk age groups. Moreover, it is necessary to disseminate knowledge of reasonable accommodation and appropriate responses to individuals with mental illness. Peer advocates should educate individuals with disabilities about their rights and encourage proactive measures.

A randomized controlled study of an e-learning program (YURAIKU-PRO) for public health nurses to support parents with severe and persistent mental illness and their family members.

BACKGROUND: Supporting parents with severe and persistent mental illness (SPMI) requires knowledge, skills, and a positive attitude toward parenthood. We developed a Japanese e-learning program for public health nurses (PHNs) to enable them to support parents with SPMI and their family members. This study aimed to evaluate the effectiveness of the program in improving the knowledge, skills, attitudes, and self-efficacy of PHNs in supporting them. METHODS: A three-hour video-based e-learning program was developed. A randomized controlled trial was conducted with 176 PHNs responsible for maternal and child health in Japan. The outcome measures included the Sense of Coping Difficulty/Possibility Scale, skills to support people with SPMI, and achievement of program goals. Outcome data were collected at three time points during the study: baseline (T1), post-intervention (T2), and one month after T2 (T3) using self-administered electronic questionnaires. Outcome measures were assessed by comparing the two groups at the endpoint (T3) using t-tests and ANOVA. Effectiveness over time was assessed using a mixed model for repeated measures, with group and time interactions as fixed effects. RESULTS: The study participants were randomly allocated to two groups:89 in the intervention group, and 87 in the control group. The total score and the scores in the two subscales of the Sense of Coping Difficulty/Possibility Scale in the intervention group at T3 were significantly higher than those in the control group, as shown by the t-test and ANOVA (all p<0.001). The Sense of Coping Difficulty subscale had a large effect size (Cohen's d=1.27). The analysis of the results of a mixed model for repeated measures showed that the group and time interactions on all outcome measures were not significantly different at T1 but were significantly different at T2 and T3. CONCLUSIONS: The program was effective one month after its completion, particularly in reducing PHNs' difficulties in supporting parents with SPMI. TRIAL REGISTRATION: UMIN000045765, November 1, 2021.

[Previous experiences of Japanese children with parents who have a mental illness, and their consultation situation at school: A survey of grown-up children].

Objectives　The purpose of this study was to understand the experience of grown-up children with parents suffering from mental illness during their elementary, junior high, and high school years, and the consultation situations at their schools.Methods　A web-based questionnaire survey was conducted on 240 grown-up children who had participated in groups for children with parents suffering from mental illness. The analysis was a simple tabulation and compared the consultation history inside and outside the school environment, according to the respondent's age. Free-response descriptions were categorized according to the content.Results　A total of 120 respondents answered, ranging in age from 20 to over 50 years, with the majority being women (85.8%). Their mothers were the most likely parent to suffer from a mental illness (67.5%), and their estimated age at illness onset was before entering elementary school (73.1%).　During their elementary, junior high, and high school years, 57.8-61.5% of the respondents provided emotional care, and 29.7-32.1% did household chores. When they were in elementary school, most recognized adult fights (62.4%) and experienced attacks by parents (51.4%). Signs that others could have noticed included the fact that parents did not visit the school or talk to teachers, being bullied, forgetfulness, absenteeism, arriving late for school, and academic deterioration. Nearly half of the respondents (43.2-55.0%) hid their challenges during their elementary, junior high, and high school years. They were aware of teachers' prejudice towards mental illness, discriminatory behavior, and lack of consideration for privacy. At the same time, many respondents wanted teachers to be concerned about their circumstances and listen to their stories.　More than half of the respondents (62.7-80.7%) did not approach school staff, who did not notice their problems. When they were high school students, they had an inadequate consultation environment or were unprepared to consult. The easiest person to approach was the teacher in charge.Conclusion　Children with parents suffering from a mental illness were those most in need of support, while being the least connected to sources of support. Schools need to support children to become aware of their problems at home, and teachers need to pay more attention to their students, and listen to them carefully, including to what they may not say.

MicroRNA profiling of gastric content from breast-fed and formula-fed infants to estimate last feeding: a pilot study.

Recently, we were consulted about a challenging case, where an infant died by poisoning and the drug-dependent mother insisted that she unintentionally gave the toxic drug through breast milk. Accordingly, we investigated the utility of immunoblotting and microRNA (miRNA) profiling of the infant's gastric content (GC) to differentiate between breast-feeding and formula-feeding. As a pilot study, we sampled the GC from breast-fed (GCB) and formula-fed (GCF) infants, as well as gastric juice (GJ) from fasted adults at autopsy. Breast milk (BM) samples were collected from volunteers within 1 year post-delivery. By immunoblotting, lactoferrin and gross cystic disease fluid protein (GCDEP) were clearly detected in BM, but could not be detected in GCB. Similarly, ß-lactoglobulin was detected in formula milk, but could not be detected in GCF. Meanwhile, miRNA sequencing revealed that the miRNA expression profile of GCB was closer to BM than GCF and GJ. Especially, miR-151a and miR-186 were more abundant in BM and GCB than in GCF and GJ. Our study is the first to elucidate the human GJ miRNA profile and demonstrate the possibility that miR-151a and miR-186 in GC may be the biomarker of breast-feeding.

Pilot Study of a Video-Based Educational Program to Reduce Family Violence for Parents of Adult Children with Schizophrenia.

This pilot study evaluated a video-based educational program for improving communication skills and reducing family violence between parents and their adult children with schizophrenia. We used a one group pretest-posttest design. The program included a main 90-min video and six stories, each 20-30 min long. We made assessments at baseline and program completion (three months after baseline). Sixty-six parent participants completed the intervention. The average frequency of acts of family violence significantly decreased from 11.4 (SD = 26.2) at pretest to 5.1 (SD = 13.2) at posttest (p = 0.016). Our findings showed significant improvements regarding expressed emotion, psychological distress, family empowerment, and hope, demonstrating preliminary positive results for this video-based educational program. The program was shown to be feasible for support/educational groups of family members of adults with mental disorders to deliver and may also be useful for practitioner-led educational groups for families in public health centers or medical settings to offer.

How Training Tools Physically Linking Soccer Players Improve Interpersonal Coordination.

Interpersonal coordination is an important skill for promoting collective behavior in team sports. This study tested the impact of two types of tools in facilitating triadic coordination. 16 males aged under 12 years were divided into four groups with similar skill levels and average ages. Each group performed a three-versus-one ball passing task under three conditions: a one-elastic-band tool linking the three players, a three-elastic-bands tool linking the three players, and without a tool linking the three players. The dependent variables were ball passing frequency, frequency and amplitude of inner angles of the triangle formed by the players, and duration of the synchronized patterns of the inner angles. The results show that neither tool increased ball-passing frequency or the duration of synchronized patterns. However, both tools increased the frequency of inner angles, and the three-elastic-bands tool decreased the amplitude of inner angles. From these results, we conclude that elastic-band tools affect spatial and temporal triadic formation by means of haptic and visual information. Specifically, compared with the one-elastic-band tool, the three-elastic-bands tool stabilizes the triadic spatial formation. We also explore the implications for how these tools can be used in practice.

Effect of spraying of fine water particles on facial skin moisture and viscoelasticity in adult women.

BACKGROUND/PURPOSE: It is known that the elderly and adult women with reduction in sebum secretion have reduced skin barrier function, drying of the skin in a low humidity environment is accompanied by physiological distress. As our hypothesis, when fine water particles are sprayed on the skin, the water content of the corneal layer is significantly increased. In the present study, we examined the ability of fine water particles to improve facial skin moisture levels in adult women. METHODS: We examined skin conductance, transepidermal water loss (TEWL), and skin elasticity as an index of skin barrier function at the cheek in 17 healthy adult women in the spraying of fine water particles, in the environment temperature at 24°C and 34.5% relative humidity. RESULTS: The skin conductance of stratum corneum after 120 minute of spraying, A condition (peak particle size below 0.5 µm) was 119.7 ± 25.1%, B condition (peak particle size 1.8 µm) was 100.4 ± 31.7%, C condition (peak particle size 5.4 µm) was 110.1 ± 25.0%, and the A condition was significantly higher than the B condition. Also, skin elasticity in the A condition tended to be higher value than in the other conditions. Transepidermal water loss (TEWL) after 120 minute of spraying showed a lower value in the A condition than in the other conditions. In the A condition, the skin conductance steadily maintained their initial levels up to 360 minute after spraying. CONCLUSION: Especially, by spraying smallest fine water particles, skin barrier function at the cheek was improved. These data indicated that non-charged fine water particles played an important role on moisten skin in a low humidity environment.

Acute Effects of Transdermal Administration of Jojoba Oil on Lipid Metabolism in Mice.

Background and objectives: Aroma therapy is a complementary therapy using essential oils diluted with carrier oils. Jojoba oils have been widely used as carrier oils. However, limited information is available regarding their effects on blood biochemical parameters. This study aimed to investigate the effect of transdermal administration of jojoba oil on blood biochemical parameters in mice. Materials and Methods: Eight-week-old male hairless mice were randomly divided into naïve control and treatment groups. In the treatment group, mice were topically administered 4 µL of jojoba oil, per gram of body weight, on the dorsa 30 min before euthanasia. Thereafter, serum biochemical parameters were assayed, and gene expression was analyzed in various tissues via a real-time polymerase chain reaction. Results: Serum non-esterified fatty acid (NEFA) levels increased significantly 30 min after topical application of jojoba oil (p < 0.05). Atgl was significantly upregulated in the liver (p < 0.05), and Atgl upregulation in the liver was positively correlated with serum NEFA levels (r = 0.592, p < 0.05). Furthermore, a trend of decreasing fatty acid trafficking-related gene (FABPpm, FATP-1, FATP-3, and FATP-4) expression in the skin after topical application of jojoba oil (p = 0.067, 0.074, 0.076, and 0.082, respectively) was observed. Conclusions: Serum NEFA levels were elevated 30 min after transdermal administration of jojoba oil. The mechanisms of elevated serum NEFA levels might be related to both enhanced lipolysis in the liver and reduced fatty acid trafficking in the skin.

FRI-4 carbapenemase-producing Enterobacter cloacae complex isolated in Tokyo, Japan.

Objectives: A carbapenem-resistant Enterobacter cloacae complex isolated in Tokyo, Japan, produced a carbapenemase that was detected by a Carba NP test and a modified carbapenem inactivation method, but none of the 'Big Five' carbapenemase genes was detected by PCR. This study aimed to identify the carbapenemase. Methods: Carbapenemase genes were screened by WGS. Next, we generated a recombinant plasmid in which the carbapenemase gene was inserted. We also extracted the carbapenemase gene-carrying plasmid from the E. cloacae complex. The effects of both plasmids on the antibiotic susceptibility of Escherichia coli were then tested. The carbapenemase gene-carrying plasmid in the E. cloacae complex was completely sequenced. Results: A novel carbapenemase gene, blaFRI-4, encoded an amino acid sequence that was 93.2% identical to French imipenemase (FRI-1). E. coli transformed with blaFRI-4 showed reduced carbapenem susceptibility. A complete sequence of the blaFRI-4-carrying 98 508 bp IncFII/IncR plasmid (pTMTA61661) showed that blaFRI-4 and the surrounding region (18.7 kb) were duplicated. Conclusions: The FRI-4-producing E. cloacae complex was isolated in Japan, whereas all other FRI variants have been found in Europe, suggesting that the spread of FRI carbapenemases is global.

The coping process of Japanese parents who experience violence from adult children with schizophrenia.

With the acceleration of deinstitutionalization might increase families' chances of suffering violence by patients. This study clarified parents' coping processes with violence experienced from patients with schizophrenia. The grounded theory approach was used, and 26 parents were interviewed. We identified a four-stage coping process: (1) hope for treatment, (2) living with violence, (3) trying to solve violence, and (4) last solution for violence. This coping process had two illness-related characteristics: (1) a process of coping with two main stressful events (the illness and violence), and (2) the need for long-term appraisal of violence because of its unclear causes.

Violence Towards Family Caregivers by Their Relative with Schizophrenia in Japan.

There have been several violence-related deaths in Japan due to family violence by persons with severe mental illness against their caregivers. However, it is not often acknowledged that these violent acts are mainly directed at family members. This study aimed to clarify what acts of violence family caregivers experienced from their relative with schizophrenia, and how frequently these violent incidents occurred in their lifetime. We also examined caregivers' thoughts of death about themselves and their relatives, as well as their consultation efforts and escape from the violence perpetrated by their relative. Of the 277 caregivers, 87.7% had experienced psychological violence and 75.8% had experienced physical violence perpetrated by their relative. Of 210 caregivers who had experienced physical violence, 26.7% had thought of murder-suicide and 31.0% had wished for their relative's death. Family violence by persons with schizophrenia is not rare but a common occurrence in Japan and may have fatal consequences.

Changes in Families' Caregiving Experiences through Involvement as Participants then Facilitators in a Family Peer-Education Program for Mental Disorders in Japan.

A family peer-education program for mental disorders was developed in Japan, similar to existing programs in the United States and Hong Kong. Families that serve as facilitators in such programs may enhance their caregiving processes and, thereby, their well-being. This study's aim was to describe how families' caregiving experiences change, beginning with the onset of a family member's mental illness, through their involvement in a family group or peer-education program as participants then facilitators. Thus, this study was conducted in a family peer-education program for mental disorders in Japan. Group interviews were conducted with 27 facilitators from seven program sites about their experiences before, during, and after becoming facilitators. Interview data were coded and categorized into five stages of caregiving processes: (1) withdrawing and suppressing negative experiences with difficulty and regret; (2) finding comfort through being listened to about negative experiences; (3) supporting participants' sharing as facilitators; (4) understanding and affirming oneself through repeated sharing of experiences; and (5) finding value and social roles in one's experiences. The third, fourth, and fifth stages were experienced by the facilitators. The value that the facilitators placed on their caregiving experiences changed from negative to positive, which participants regarded as helpful and supportive. We conclude that serving as facilitators may improve families' caregiving processes.

Psychological Distress and Violence Towards Parents of Patients with Schizophrenia.

The present study investigated the relationship between violence and psychological distress experienced by parents of patients with schizophrenia. Questionnaire data from 379 parents were analyzed. A total of 151 parents (39.8%) had not experienced violence in the past year, whereas 96 (25.3%) and 132 (34.8%) had experienced psychological violence only or physical violence, respectively. A total of 216 (57.0%) of parents reported being psychologically distressed. Multiple logistic regression revealed that the risk of psychological distress significantly increased with the experience of psychological and physical violence, lower household income, greater family stigma, and the increasing age of patients.

[Two Outbreaks of Yersinia enterocolitica O:8 Infections in Tokyo and the Characterization of Isolates].

Although the number of outbreaks caused by Yersinia enterocolitica has been very small in Japan, 4 outbreaks were occurred during the 2 years between 2012 and 2013. We describe herein 2 outbreaks which were examined in Tokyo in the present study. Outbreak 1: A total of 39 people (37 high school students and 2 staff) stayed at a hotel in mountain area in Japan had experienced abdominal pain, diarrhea and fever in August, 2012. The Y. enterocolitica serogroup O:8 was isolated from 18 (64.3%) out of 28 fecal specimens of 28 patients. The infection roots could not be revealed because Y. enterocolitica was not detected from any meals at the hotel or its environment. Outbreak 2: A total of 52 students at a dormitory had diarrhea and fever in April, 2013. The results of the bacteriological and virological examinations of fecal specimens of patients showed that the Y. enterocolitica serogroup O:8 was isolated from 24 fecal specimens of 21 patients and 3 kitchen staff. We performed bacteriological and virological examination of the stored and preserved foods at the kitchen of the dormitory to reveal the suspect food. For the detection of Y. enterocolitica, food samples. together with phosphate buffered saline (PBS) were incubated at 4 degrees C for 21 days. Then, a screening test for Y. enterocolitica using realtime-PCR targeting the ail gene was performed against the PBS culture. One sample (fresh vegetable salad) tested was positive on realtime-PCR. No Y. enterocolitica was isolated on CIN agar from the PBS culture because many bacteria colonies other than Y. enterocolitica appeared on the CIN agar. After the alkaline-treatments of the culture broth or the immunomagnetic beads concentration method using anti-Y. enterocolitica O:8 antibodies, Y. enterocolitica O:8 which was the same serogroup as the patients' isolates was successfully isolated from the PBS culture. The fresh vegetable salad was confirmed as the incrimination food of this outbreak.

Four foodborne disease outbreaks caused by a new type of enterotoxin-producing Clostridium perfringens.

The epidemiological and bacteriological investigations on four foodborne outbreaks caused by a new type of enterotoxin-producing Clostridium perfringens are described. C. perfringens isolated from patients of these outbreaks did not produce any known enterotoxin and did not carry the C. perfringens enterotoxin gene. However, the culture filtrates of these isolates induced the accumulation of fluid in rabbit ileal loop tests. The molecular weight of the new enterotoxin may be between 50,000 and 100,000, although the known C. perfringens enterotoxin is ca. 35,000. This new enterotoxin was heat labile, and its biological activities were inactivated by heating for 5 min at 60°C. The new enterotoxin was sensitive to pH values higher than 11.0 and protease treatment but was resistant to trypsin treatment. These results suggest that the new enterotoxin may be a protein. Although C. perfringens enterotoxin induced morphological changes in Vero cells, the changes induced by the new enterotoxin differed from those by the known C. perfringens enterotoxin. The new enterotoxin also induced morphological changes in L929 cells, whereas the known C. perfringens enterotoxin did not, because L929 cells lacked an appropriate enterotoxin receptor. Although C. perfringens enterotoxin is recognized as the only diarrheagenic toxin responsible for C. perfringens foodborne outbreaks, the results of the present study indicate that C. perfringens isolated from these four outbreaks produced a new type of enterotoxin.