Klebsiella pneumoniae (Kp) associated with hospital acquired infections are extensively-drug resistant (XDR), making treatment problematic. Understanding the genetic epidemiology of XDR -Kp can determine their potential to be hypervirulent (hv) through the presence of siderophores. We characterized genomes of 18 colistin-resistant XDR-Kpisolated from 14 patients with complicated urinary tract infection in an Indian healthcare facility. 18 organisms comprised STs: ST14 (9/18), ST147 (5/18), ST231 (2/18), ST2096 (1/18), and ST25 (1/18). Many patients in one ward were infected with the same ST, indicating a common infection source. Some patients had recurrent infections with multiple STs that were circulating in that ward, providing evidence for hospital transmission. Beta lactamase genes (blaCTX-M-1, blaSHV, and blaampH) were present in all isolates. blaNDM-1 was present in isolates 15/18, blaOXA-1 was present in isolates 16/18, blaTEM-1D was present in 13/18 isolates, and blaOXA-48 was present in isolates 14,19 and 30. Disruption of mgrB with various IS elements was responsible for colistin resistance in 6 isolates. The most common K type among these isolates was K2 (10/18). One XDR convergent hv-Kp ST2096 was associated with prolonged hospitalisation (iuc+ybt+blaOXA-1+blaOXA-48).Convergent XDR-hv-Kp detected has outbreak potential, warranting effective antimicrobial stewardship and infection control.
Equine botulinum antitoxin is one of the most popular countermeasures for human botulism. The unitage of the antitoxin product is defined according to national minimum requirement or pharmacopoeia in each country by referring to national standard antitoxins for four types (A, B, E, and F). With the expected depletion of the national standard antitoxins, replacement national standard antitoxins are produced and standardized through collaboration of the National Control Laboratory and other participants, including manufacturer(s). Therefore, Japanese National Standard Botulinum Antitoxin Type A, Equine, was replaced according to the results of a collaborative study involving the National Institute of Infectious Diseases and KM Biologics Co., Ltd. The unitage of the replacement material was determined through mouse neutralization tests, which involved toxin-antitoxin mixture injection at pH 7.0. Potency value of 440 units/vial was obtained. However, the Japanese Minimum Requirement for Biological Products was revised, and the neutralization reactions were repeated at pH 6.0, for which considerably different potency value (656 units/vial) and survival profile of mice were obtained. In September 2021, the replacement material, Japanese National Standard Botulinum Antitoxin Type A, Equine, lot 2, was established with potency value of 656 Units/vial. The impact of pH-dependent change in potency on antitoxin quality control is discussed.
This study investigated the role of genetic variant rs8177374 in MAL/TIRAP gene in mediating the cytokine levels of IFN-γ, TNF-α, IL-10 and TGF-β in malaria patients due to P. falciparum or P. vivax infection. The study included human blood samples of malaria patients (n=228) and healthy controls (n=226). P. falciparum and P. vivax groups were established based on the causative species of Plasmodium. Malaria samples were divided into mild and severe malaria groups based on symptoms appeared in patients as per WHO criteria. In previous study, we genotyped rs8177374 via allele specific PCR based strategy. In this study, cytokines levels were estimated from blood plasma of samples of genotypes of rs8177374 via Sandwich Enzyme Linked Immunosorbent Assay kits. Increased levels of IFN-γ and TNF-α in presence of CC genotype indicates the role of CC genotype in both severe malaria and mild malaria groups. Enhanced levels of IL-10 in CT genotype and mild malaria group suggest the role of CT genotype and IL-10 in mild clinical outcomes of malaria. rs8177374 polymorphism of MAL/TIRAP gene has important role in malaria pathogenesis.
To demonstrate the transmission cycle of Shimokoshi type Orientia tsutsugamushi in Shimane prefecture, field rodents were captured from areas where four human infections caused by the pathogen were reported. The rodents were investigated for the transmission cycle of the pathogen based on the pathogen’s genome, antibodies against the pathogen, and vector of the pathogen (Leptotrombidium palpale). In addition, the vector was captured from soil of the study areas. A total of 44 rodents was captured. No DNA of O. tsutsugamushi were detected in the blood or spleen samples using real-time polymerase chain reaction. However, the specific antibody against the pathogen was detected in 2 out of 44 (4.5%) rodents by the indirect immunoperoxidase method, indicating the presence of the pathogen in the study areas. Although 29 L. palpale were identified, DNA detection was not performed, because of insufficient number of vectors according to the DNA detection rate in previous studies. However, the identification of the vector as well as the specific antibody in rodents suggest the presence of the transmission cycle of Shimokoshi type O. tsutsugamushi in Shimane prefecture.
Salmonella enterica subsp. enterica serovar Typhimurium has recently emerged worldwide as a producer of extended-spectrum β-lactamase (ESBLs). However, such resistant clinical isolates are still rare in Japan. The common types of ESBLs found are the CTX-M type β-lactamases, including novel ones such as CTX-M-64. CTX-M-64 has a chimeric structure that is a combination of the CTX-M-1 and CTX-M-9 groups. In 2017, S. Typhimurium was isolated from cultures of stool, blood, and urine of an 82-year-old man. Here, we describe the discovery of a clinical isolate of S. Typhimurium in Japan. Antimicrobial susceptibility testing revealed that S. Typhimurium was resistant to third- and fourth-generation cephalosporins, including ceftazidime and monobactam. The MICs of ceftazidime and ceftriaxone were restored by clavulanic acid. Whole-genome sequencing analysis revealed that S. Typhimurium had blaCTX-M-64 gene on an IncHI2/IncHI2A-type plasmid with an assembly length of 174,477 bp. The genetic structure of the region surrounding the blaCTX-M-64 gene, ISKpn26-ΔISEcp1-blaCTX-M-64-orf477, was only shared with the chromosome sequence of S. Typhimurium detected from food-producing chicken in Guangdong, China. Although rare, S. Typhimurium can induce bloodstream infections and produces ESBLs. This is the first report of a CTX-M-64-producing Enterobacterales clinical isolate of domestic origin in Japan.
In the present study, 26 Staphylococcus aureus strains, including 16 methicillin-resistant S. aureus (MRSA) and 10 methicillin-susceptible S. aureus (MSSA), which were collected from eight medical institutions in Chiba Prefecture that requested toxin gene analysis from 2015 to 2021, were analyzed. A total of 14 Panton-Valentine leukocidin (PVL) positive strains were identified, including one of MSSA. PVL-positive strains were classified into seven types based on polymerase chain reaction-based open reading frame typing (POT). Of these, three POT types of MRSA strains were not previously reported, and one had been previously reported as PVL-negative. Some strains were positive for both PVL and toxic shock syndrome toxin 1 (TSST-1). One POT type was identified in both PVL-positive and PVL-negative strains. This is the first report on the regional spread of highly pathogenic S. aureus strains in children from multiple medical institutions based on the POT method. This method is useful for estimating the spread of toxin gene-carrying strains in the community because of its association with toxin genes. As the number of PVL-positive strains is increasing in Japan, it is important to analyze the isolates of severe S. aureus infections in children by combining toxin gene analysis and the POT method.
The aim of this study was to determine influencing factors including regular financial social support program on tuberculosis (TB) treatment outcomes and success rates. TB patients registered 2018-2019 were included for this retrospective cohort study. We classified into two groups as those who received financial support for at least one month and those who did not. A total of 22867 patients were included in the study, 5033 of whom received financial social support and 17834 of whom did not. The success rate was 11.9% higher among the group of patients received financial social support compared to the those who did not receive (97.34% vs. 85.40%). When other factors in the model were controlled, the success rate among all patients was 1.3 times higher for female, 2.6 times higher for under 50 years of age, 1.5 times higher for extra-pulmonary TB, 1.5 times higher for new case, 5.9 times higher for drug susceptible TB and 5.8 times higher for those who received financial support. This is the first study from Türkiye evaluating the effect of regular financial social support program on TB treatment outcomes since the program started in 2018. We recommend regular financial support to TB patients for all countries.
Mycobacterium tuberculosis (M.tb) infection caused communicable disease tuberculosis (TB) is a major cause of ill health and one of the leading causes of death worldwide. Protein encoded by region of deletion (RD) of M.tb mediated the pathogenic properties of M.tb through inducing inflammation response or disrupting host cell metabolism. We cloned and purified Rv2653 protein from RD13 to explore its regulatory effect on host macrophages. We found Rv2653 protein promoted the glycolysis level and upregulated the expression of glycolytic key enzymes HK2 and LDHA of THP1 cells. Furthermore, Rv2653 induced glycolysis contributes to the activation of NLRP3 inflammasome. Rv2653 activated the mTORC1 signaling pathway, and the mTORC1 inhibitor NR1 blocked Rv2653-induced HK2, LDHA and NLRP3 expression. siRNA interfering HK2 or LDHA significantly inhibited the activation of NLRP3 inflammasome by Rv2653, blocked Rv2653 triggered inflammatory factors IL-1β, IL-6, TNF-α, ROS and NO, and promoted the survival of BCG in THP1 cells. Altogether, Rv2653 promotes the glycolysis process by activating mTORC1 signaling pathway, activates NLRP3 inflammasome and the release of inflammatory factors, ultimately inhibits the intracellular survival of BCG in THP1 cells. Therefore, we revealed Rv2653-induced anti-M.tb immune mechanisms which contributes to the development of new anti-TB strategy.
The use of the novel inflammatory indicator—systemic immune-inflammatory index (SII)—in pediatric patients with bronchiolitis remains unreported. Therefore, this study investigated the relationship between the SII and neutrophil-to-lymphocyte ratio (NLR) in patients with respiratory syncytial virus (RSV) and non-RSV bronchiolitis and clinical severity prediction.This study analyzed the data of 155 patients aged <2 years hospitalized with viral bronchiolitis due to RSV and/or other viral pathogens. The SII [platelet count×(neutrophil/lymphocyte)] value was calculated and compared among patients with RSV monoinfection, other viral monoinfections, and RSV coinfections. Severity was defined according to the number of days of supplemental oxygen treatment. The NLR and SII values were significantly higher in the non-RSV monoinfection group than in the RSV monoinfection group; the number of days of supplemental oxygen therapy was significantly increased in the RSV monoinfection group. No significant differences in the NLR, SII, and days of supplemental oxygen therapy were found between the RSV monoinfection and RSV coinfection groups. Although patients with RSV monoinfection showed more severe clinical findings than those with non-RSV monoinfection, the NLR and SII values were significantly higher in the non-RSV monoinfection group. Therefore, the NLR and SII do not appear to be very useful measurements for determining the severity of acute bronchiolitis attacks, but lower NLR and SII values in RSV group compared to non RSV group may be potential biomarker for RSV infection. Additional studies are required.
In 2019, the Japanese government set up a scheme for rubella antibody testing during workplace health check-ups for men born between fiscal years 1962 and 1978. However, use of vouchers for rubella antibody testing remains low. Health check-up data analyses are needed to determine the reason rubella antibody testing is not widely used. In this research, we aimed to clarify how rubella antibody test-taking behaviour during health check-ups has changed over the first 3 years of the rubella catch-up campaign in Japan. The vouchers were sent in 2019, 2020, and 2021 (2020 in some areas) to men born in fiscal years 1972–1978, 1966–1971, and 1962–1965, respectively. We calculated the prevalence of men born between 1962 and 1978 having received rubella antibody testing during mandatory health check-ups under the Industrial Health and Safety Act. It was relatively high (approximately 15%) soon after the distribution of vouchers in all three age groups, and then declined to below 2% over the second and third years. A further population approach with continuous public engagement is required in workplaces to effectively promote and expand the rubella vaccination program in Japan.
Myroides species, are reported more frequently as outbreaks in clinics and intensive care units (ICUs). In this study, it is aimed to investigate the epidemic potential, antibiotic resistance profile and risk factors of the M. odoratimimus isolates, are increasingly isolated in the intensive care units (ICUs) of our hospital. Data of patients, whose Myroides spp. were isolated in their clinical specimens, over a five-year period (September 2016-January 2022) were retrospectively analyzed. Identification of bacteria were performed with the Matrix: Assisted Laser Desorption Ionization/Time of Flight-Mass Spectrometer (MALDI-TOF MS) System. The presence of antibiotic resistance genes were analyzed by polymerase chain reaction (PCR) method. Possible clonal association between isolates was investigated the Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR method. 66 isolates were identified as M. odoratimimus and one isolate as M. odoratus. The presence of the blaMUS resistance gene was detected in all M. odoratimimus isolates, while the presence of sul2 in 10 isolates and tetX in 11 isolates was detected. Other resistance genes, including blaTUS, were not detected. Also 2 different clonal association patterns were found in 24 selected isolates by (ERIC)-PCR method.The increase in the immunosuppressive patient population point to the possibility of encountering this agent and other opportunistic pathogens more frequently in the future.
Reverse-transcriptase polymerase chain reaction (RT-PCR)-confirmed Enterovirus (EV) meningitis without pleocytosis has reported in only children. We examined the frequency of EV meningitis without pleocytosis and compared clinical features in adults. We retrospectively analyzed the data of adult patients with EV meningitis confirmed by cerebrospinal fluid (CSF) RT-PCR. Among 17 patients finally included, 58.8% showed no pleocytosis. The median age and clinical symptoms were not different between the pleocytosis and the non-pleocytosis group. There were no statistically significant differences in terms of seasonal variation or time from the onset of meningitis symptoms to undergoing lumbar puncture. The peripheral white blood cell (WBC) count of the pleocytosis was significantly higher than in patients without pleocytosis. The median CSF pressure showed higher trend in the non-pleocytosis group. Patients with higher CSF pressure than the normal level were more common in the non-pleocytosis group. The median CSF protein values were higher than the normal values in both groups. We confirmed a high frequency of EV meningitis without pleocytosis in adults. An accurate diagnosis using RT-PCR is necessary when meningitis symptoms are prominent during an EV epidemic and CSF protein levels and pressure are high, even if the CSF WBC count is normal.
Hepatitis E infection is an important problem in developing countries. Hepatitis E vaccination is important for prevention but it influenced by the resident’s knowledge. The Qingdao residents’ knowledge of hepatitis E is still unknown. This study used online survey on the Wechat platform for investigation. Chi-square test was used to compare the hepatitis E influencing factors between subgroups. Binary logistic regression was used for multiple factor analysis to explore the hepatitis E influencing factors. We have found the total awareness rate of hepatitis E was 60.51%. Females, aged between 51 and 60, aged 61 and above, worked in government-affiliated departments were found to have higher awareness rate than other subgroups. The participants who have family members infected with hepatitis E were found to have lower awareness rate. The government and relevant departments should focus on the education of the hepatitis E vaccination and the disease process.