Bacillus Calmette and Guérin (BCG) was introduced to Japan in 1924 by Kiyoshi Shiga and has been propagated for research purposes ever since propagation is accomplished using a glycerin-bile-potato mixture in the same manner used by Calmette and Guérin. To prepare a stable and safe freeze-dried BCG vaccine, several joint research projects were organized in 1949. At the National Institute of Infectious Diseases (formerly the National Institute of Health), the 172nd passage of BCG from the first culture was freeze-dried in 1961 and was used as the origin of the Japanese BCG strain, Tokyo-172. The Tokyo-172 was registered as an International Reference Strain in 1965 by the World Health Organization. In 1967, a multiple puncture method for BCG vaccination using a plastic cylinder implanted with nine fine needles at one end was introduced to Japan; there-after, percutaneous administration replaced intradermal injection. The efficacy and adverse reactions of BCG vaccines as well as recent knowledge on the genetic characterization of BCG is also discussed.

The prevalence of Mycoplasma pneumoniae among HIV-positive patients with community-acquired pneumonia (CAP) remains unclear. We investigated 300 HIV-positive adults (200 with CAP and 100 with no respiratory illness) and 75 HIV-negative adults with CAP for the prevalence of respiratory pathogens using culture and serology. A growth inhibition test was employed to confirm the isolates of M. pneumoniae using species-specific typing sera. The prevalence of M. pneumoniae in HIV-positive subjects was 17% by induced sputum and 11.3% by throat swab culture. The seroprevalence of anti-M. pneumoniae IgM was 11.7% by ELISA and 14.3% by the gelatin microparticle agglutination test. The prevalence of M. pneumoniae among HIV-negative cases was relatively low. Streptococcus pneumoniae was predominant (28%) among subjects with lower respiratory disease, whereas Staphylococcus aureus (15%) was common among upper respiratory symptomatic cases. Rales (P = 0.001), pharyngeal erythema (P = 0.02), cervical adenopathy (P = 0.004), skin rash (P = 0.001), and crepitations (P = 0.001) were each significantly related to M. pneumoniae positivity. Statistical significance was observed in relation to total lymphocyte count (P = 0.02) and erythrocyte sedimentation rate (P = 0.04), as well as to M. pneumoniae positivity. This study shows that the prevalence of M. pneumoniae in HIV-positive subjects is comparatively higher than in HIV-negative subjects with pulmonary symptoms, and concords with previous pilot studies carried out in Chennai, South India.

General theories on the inactivation of viruses in the presence of a concentrated protein, such as the allantoic fluid of chicken eggs, are not useful. That is, although sodium hypochlorite and sodium hydroxide are generally known as strong virucidal agents, they do not sufficiently inactivate viruses in allantoic fluid. We found that benzalkonium chloride (BC) is an effective virucidal agent against influenza, Newcastle disease, and avian infectious bronchitis viruses even in the presence of a concentrated protein. BC is easily biodegradable by activated sludge and is not very harmful to humans. We strongly recommend BC as a useful virucidal agent, especially in the manufacture of vaccines for these viruses.

To assess the importance of only IgA antibody positivity in the peptidebased ELISA (P-ELISA) examination of kinetic behaviors of antibodies (IgA, IgG) to Chlamydia trachomatis, 426 sera from 52 follow-up antigen-positive patients were assayed. In part, a microimmunofluorescence (MIF) test and an immunoblot (IB) assay were also used for confirmation. The results showed that the positivity rates of IgA and IgG antibodies were 82.7 and 96.2%, respectively, at the first testing. One patient had both IgA- and IgG-negative antibodies at the first testing, but this became only IgA-positive and then IgG-positive. The patient was co-infected with Candida albicans and C. trachomatis, and saw a gynecologist for the symptom of itching. Although the major outer membrane protein was negative in IB assay, the results of the MIF test and absorption experiments were positive. MIF titers for IgA and IgG antibodies to C. pneumoniae were <1:8 and 1:32, respectively, at the peak level of P-ELISA. These findings seem to suggest that when only the IgA antibody is detected by P-ELISA, C. trachomatis infection may be present at an early stage, so confirmation via testing for C. trachomatis is needed.

This study was designed to understand the seroprevalence of Mycoplasma pneumoniae infection in healthy Taiwanese adolescents. The study included 2,233 college freshmen (female:male = 1.29:1; mean age, 19.7 years). The percentages of subjects residing in northern, central, southern, and eastern Taiwan were 66.91, 15.89,9.0, and 8.2%, respectively. All enrolled subjects underwent a serologic agglutination test to detect serum concentrations of antibodies to M. pneumoniae. The test results showed that 19.84% of the subjects were infected and, of those, 6.0% were estimated to have subsequently acquired a current or acute infection during this study period. Moreover, the percentage of seropositive females (22.77%) was significantly higher than that of seropositive males (16.07%) (odds ratio, 1.54; 95% confidence interval, 1.241 - 1.911). Subjects residing in eastern counties were more likely to contract M. pneumoniae than those residing in other areas of Taiwan.

The genetic diagnosis of pathogenic agents using microarrays has the advantage of high-throughput detection, but a relatively large amount of DNA sample is required. To obtain a sufficient amount of DNA for molecular diagnoses, several whole genome amplification (WGA) methods have been proposed. In this study, using Francisella tularensis and Escherichia coli as models, we compared four WGA methods in terms of their efficiency of amplification of whole genomic DNA in order to identify the most suitable method for preparing DNA to be used for microarray analysis. It was possible to obtain more than 1.5 μg of products from 10 ng of F. tularensis and E. coli genomic DNA using four methods, but biases in the amplification of bacterial genes were least prominent in the multiple displacement amplification (MDA) or OmniPlex WGA. When the amplified DNAs were applied to microarray slides consisting of 32 different genes probes, DNAs amplified by Phi29 v2 of MDA and OmniPlex WGA showed high signal intensity as well as a high signal-to-noise ratio for all 32 genes. These results indicate that Phi29 v2 and OmniPlex WGA are useful methods for obtaining sufficient DNA from a limited amount of samples for the detection of microbes using microarrays.

The confirmed cases of human brucellosis in the area of Larissa in Central Greece from 2003 to 2005 were analyzed to assess the features of the affected population and to determine the factors influencing the acquisition of infection. Data of patients infected by Brucella spp. concerning age, gender, occupation, date of diagnosis and the observed symptoms were collected from regional hospitals, health centers and private practitioners. The incidence of human brucellosis in the area was 32.49 cases/100,000 inhabitants. Males, due to their professions, were affected more often by brucellosis than females. The majority of the cases were attributed to direct contact with animals or their products. Only in 8.49% of the cases was the infection attributed to the consumption of dairy products. The urban population is not at potential risk for acquiring brucellosis because all commercialized dairy products in Greece are produced from pasteurized milk. The occurrence of human brucellosis shows seasonality, with the majority of the cases diagnosed from December to May. Direct contact with animals, the season of the year and gender were the risk factors influencing the acquisition of infection. Fever, arthralgia, profuse sweating and anorexia were the symptoms most often observed.

In 106 inpatients with methicillin-resistant Staphylococcus aureus (MRSA)-positive body sites, MRSA contamination on their surrounding environmental surfaces and the palm of their dominant hand were detected and quantified. The association between MRSA-positive sites (palm of dominant hand, sputum, nasal discharge, pharyngeal mucosa, open pus, closed pus, urine, feces, and others) in patients and MRSA contamination in their surrounding environmental surfaces was evaluated by quantification theory II of multivariate analysis. The surrounding environmental surfaces were contaminated with MRSA in 54 (50.9%) of the 106 patients. The contamination of MRSA was 380.2 ± 2,198.0 colony forming units (cfu)/100 cm² in 41 (40.2%) of 102 bed linen samples, 15.2 ± 69.5 cfu/100 cm² in 19 (22.4%) of 85 overbed table samples, 12.8 ± 56.5 cfu/about 100 cm² in 18 (20.9%) of 86 bed side rail samples, and 0.2 ± 1.3 cfu/entire handle surface in 2 (2.7%) of 74 samples of room door handles on the inner side of the patients’ room doors. In the palm, 6,743.3 ± 65,446.5 cfu/palm MRSA was detected in 29 (29.6%) of 98 patients. MRSA in patients’ palms had the most marked influence on MRSA contamination of their surrounding environmental surfaces. When MRSA is detected in patients’ palms, the possibility of MRSA contamination of their surrounding environmental surfaces is high.

Mycoplasma pneumoniae is a common cause of community-acquired pneumonia (CAP) in children, but there has been no clinical report on M. pneumoniae infections in Vietnamese children. We investigated the clinical features of M. pneumoniae infection when the pathogen was detected in the respiratory tract in hospitalized children aged 1 - 15 years due to lower respiratory tract infections or CAP in Vietnamese children. Throat swabs from 47 patients (18.6%) of 252 patients with a clinical diagnosis of CAP were PCR positive (male, 34; female, 13), and 21 throat swabs (8.3%) showed culture positive for M. pneumoniae. The M. pneumoniae pathogen could be detected by PCR and/or culture in 52 patients (male, 36; female, 16). The major clinical signs in the 52 patients were fever (>38°C) in 100%, pharyngitis in 100%, tachypnea in 94%, dry cough in 86.5%, and rough breathing in 83% of patients. The average term of illness prior to hospitalization was 7.5 ± 4.1 days, and the average number of hospitalized days was 7.9 ± 3.5 days. β-lactam group antibiotics, which were ineffective against M. pneumoniae infection, were used in 37 cases (71%).

The objective of this study was to clarify the longitudinal epidemiology of Chlamydia trachomatis serovars in Japan. A total of 339 endocervical swab specimens obtained from female patients who attended the Department of Obstetrics and Gynecology, Saitama Medical School, were used. Positive specimens of either transport medium of IDEIA Chlamydia (1st group, from 1999 to 2001), or DNA extract of Cobas Amplicor STD-1 Chlamydia trachomatis (2nd group, from 2003 to 2005) were used for serotyping. Typing of C. trachomatis serovars in DNA extracts was performed by polymerase chain reaction-restriction fragment length polymorphism. Ten serovars, A, B, D, E, F, G, H, I, J and K, were identified in the 1st group, and serovar E was most frequently identified (27.6%). In the 2nd group, nine serovars, B, C, D, E, F, G, H, I and K, were identified, and serovar D was most frequently identified (24.7%). Serovars B and Ba were significantly more common around 2000 and the mid-1990s (from 1993 to 1996), respectively. Numbers of serovar I increased significantly during the research period. In addition, serovar I was more frequent in the 2nd group than in the 1st group in women aged 20 - 29 years. There were no significant differences of serovar distribution between pregnant and non-pregnant women.

Hepatitis C virus (HCV) infection has been associated with hepatic steatosis. However, the role of hepatic steatosis in the pathogenesis of HCV infection remains controversial. In our study, 425 consecutive HCV-viremic patients with biopsy-proven chronic hepatitis C (male, 264; mean age, 49.0 years) were enrolled. Scoring of hepatic steatosis was based on the method described by Kleiner and on histopathology performed using the Knodell and Scheuer systems. HCV RNA level and genotypes were determined at the time of biopsy. Hepatic steatosis was observed in 30.8% of patients, including 113 mild, 16 moderate, and 3 with severe hepatic steatosis. Patients with a body mass index (BMI) <23 kg/m² had a significantly lower rate (18.9%) of hepatic steatosis (P < 0.001). Hepatic steatosis did not correlate with the hepatic necroinflammatory activity, but was related to hepatic fibrosis (P = 0.035). Hepatic steatosis was also not associated with HCV RNA level, and the distribution was similar between patients with HCV genotype 1 and genotype 2 infection. According to multivariate analysis, BMI is the strongest risk factor associated with hepatic steatosis, followed by hepatic fibrosis and triglyceride level with odds ratios (95% confidence intervals) of 2.51 (1.49 - 4.23), 2.06 (1.14 - 3.70), and 1.02 (1.01 - 1.03), respectively. Hepatic steatosis was associated with being overweight, hepatic fibrosis, and triglyceride level in chronic hepatitis C.

Osteonecrosis, a disabling complication associated with antiretroviral therapy (ART) and human immunodeficiency virus (HIV) infection, has rarely been reported in an Asian population. After an observation of 3,250 person-years (PY), 11 of 967 (1.1%) HIV-infected patients at a median age of 34 years developed osteonecrosis involving the hip joints (incidence, 3.4 per 1,000 PY). Their median CD4+ lymphocyte count had increased from 35 cells/μL at the diagnosis of HIV infection to 297 cells/μL at the diagnosis of osteonecrosis. The crude rate of osteonecrosis increased from 0% in patients without exposure to ART to 2.6 and 1.7% in patients with exposure to nucleoside reverse transcriptase inhibitors (NRTIs) and who had undergone highly active antiretroviral therapy (HAART) for 5 years or longer, respectively (P = 0.18 and 0.09, respectively). Among the patients receiving HAART, the estimated incidence of osteonecrosis was 4.2 per 1,000 PY. Patients with osteonecrosis had a longer duration of exposure to NRTIs (1,641 versus 1,264 days, P = 0.26) and to HAART (1,603 versus 1,251 days, P = 0.42), a higher serum triglyceride (median, 1,130 versus 351 mg/dl; P = 0.09), and a higher proportion of lipodystrophy (81.8 versus 15.0%, P < 0.0001). Our report suggests that osteonecrosis is a rare complication in HIV-infected patients with prolonged exposure to ART with resultant metabolic complications.

Despite the demonstrated benefits of influenza vaccinations, the coverage is lower than expected among health-care personnel (HCP). In this study we surveyed the attitudes of pediatric residents regarding influenza immunization and adverse reactions. Forty-five female and 35 male pediatric residents with ages ranging from 24 to 28 years were vaccinated with an influenza vaccine on 2 days in the 3rd week of September 2005 by the same nurse. Among our residents, 27 (33.7%) thought the vaccine unnecessary; their vaccine coverage was only 12% in the previous year. Thirteen residents (16%) had soreness at the vaccination site; 7 (8%) had other local reactions that did not interfere with everyday activities, and 16 (20%) had any systemic side effects. The overall rate of side effects from the vaccination was 36.5% (n = 29). Twenty of the 29 vaccinees who experienced side effects stated they did not want to receive the vaccine the following year because of the side effects, while 13% in the group without side effects stated the same thing, mainly because of the cost of vaccination. We would like to recommend an influenza vaccination campaign for HCP by employers, but first we must plan to take steps to improve the acceptability of the influenza vaccine among HCP.

Transfusion transmitted infections (TTIs) are a major problem associated with blood transfusion practices. A 4-year retrospective study from 2002 to 2005 was conducted at the blood bank of Lady Hardinge Medical College and associated hospitals in New Delhi, India. Donors were evaluated for the prevalence of HCV, HBsAg and HIV. A total of 28,956 healthy blood donors were tested, out of which 28,805 (99.48%) were replacement donors and 151 (0.52%) were voluntary donors. The proportion of voluntary donors was significantly low (P < 0.001). Males formed the bulk of the donor population (97.24%). The prevalence of HCV, HIV and HBsAg was 0.66% (ranging from 1.01% in 2002 to 0.29% in 2005), 0.56% (ranging from 0.70% in 2002 to 0.44% in 2005) and 2.23% (ranging from 2.42% in 2002 to 1.97% in 2005), respectively. For all three major TTIs, we found a decreasing trend in the prevalence over the past 4 years. The decreasing trend of HCV prevalence was significant (P < 0.001), but the same was not true for HIV and HBsAg. We suggest the need to stress more stringent donor selection criteria with emphasis on non-remunerated voluntary donations to ensure a safer blood supply.

Disseminated mycobacterial infection after Bacille Calmette-Guérin (BCG) vaccination is a rare disorder, usually presenting with fever, weight loss, anemia, lymphadenopathy and hepatosplenomegaly. We report a case of disseminated BCG infection in a 28-month-old girl with prolonged fever and abdominal mass due to retroperitoneal abscess. Appropriate therapy resulted in a good response. This is the second reported case of retroperitoneal abscess complicating BCG vaccination.

Nosocomial Sphingomonas paucimobilis infections can arise from contaminated water and the contaminated hands of hospital staff. Within a 1-month period, we isolated six S. paucimobilis strains, including four from blood cultures of four patients and two from hospital environment specimens including tap water and a bathtub in a hemato/oncology unit. We described here these strains’ molecular epidemiological analyses by pulsed-field gel electrophoresis (PFGE) and antibiotic susceptibilities by E-test. Although clinical and environmental isolates yielded three different antibiotic resistances and PFGE patterns, all four clinical strains had an identical pattern by both methods. Thus, the isolated clinical strain clone could be traced neither to health care workers nor to environmental samples. It was concluded that S. paucimobilis strains can cause outbreaks in hemato/oncology units. We did not demonstrate genetic relatedness between clinical and environmental isolates by PFGE, but did find PFGE a useful identification technique for epidemiological investigation.

We analyzed the clinical and laboratory characteristics, therapeutic outcome and prognostic factors of 25 cases of cerebrospinal fluid (CSF) culture-proven Pseudomonas aeruginosa adult bacterial meningitis (ABM). Twelve P. aeruginosa strains, isolated from clinical CSF specimens, were tested for antibiotic susceptibility. The 25 cases included 17 men and 8 women, aged 17 to 86 years (median = 51). Of the 25 cases of P. aeruginosa ABM, 18 were the result of postneurosurgical infection and the other 7 were spontaneous infections. The latter 7 cases had serious underlying medical conditions. The antibiotic susceptibility rates of the 12 strains were as follows: ceftriaxone 16.7% (2/12), ceftazidime 91.7% (11/12), cefepime 83.3% (10/12), imipenem 83.3% (10/12), meropenem 83.3% (10/12) and ciprofloxacin 66.7% (8/12). The therapeutic results showed an overall mortality rate of 40% (10/25). The emergence of third-generation cephalosporin-resistant P. aeruginosa strains cultured from clinical CSF specimens in recent years has resulted in a therapeutic challenge in the treatment of ABM.

Prostate infection caused by Aeromonas sobria has not been reported in the literature. We presented a 44-year-old healthy man with a habit of alcoholic consumption who was admitted because of primary A. sobria prostatitis with septicemia. The patient was successfully treated with intravenous ofloxacin and was uneventful after a half-year follow-up. Based on our rare case, we suggest to clinical physicians that prostatitis might be considered in healthy male patients with A. sobria bacteremia and a vague primary infection focus.

In 2007, relatively large outbreaks of measles occurred in the Kanto region of Japan, including Gunma Prefecture. We performed sequence and phylogenetic analysis of the nucleoprotein gene (N gene) of measles viruses from 3 measles patients in this area in May 2007. The N gene sequences of the present strains were identical to each other, and phylogenetic analysis showed these viruses were classified into genotype D5. The results suggest that highly homologous measles viruses may be associated with outbreaks of measles in Gunma, Japan.

On October 17, 2006, the Sakai City Public Health Center received a report of acute gastroenteritis among 4 members from the same company who had eaten raw meat dish called “Yukke: Korean-style beef tartar” and raw liver at a rotisserie in Sakai City on October 7. Based on information from interviews, the median incubation period was 5.5 (range, 5 - 7 days), and the median length of illness was 7 days (range, 4 - 10 days). The illness was characterized by a prolonged incubation period, non-bloody watery diarrhea, reduced vomiting, and light fever, which led us to suspect an enteric protozoan infection. Stool specimens obtained from 3 of the 4 symptomatic patients were positive for Cryptosporidium oocysts. They, along with 2 food workers, were negative for food poisoning bacteria or Norovirus. Genotyping of the Cryptosporidium isolates by direct sequencing of PCR products revealed that all the isolates were the C. parvum genotype II (bovine) and the subgenotype of IIa with 100 % homology with respective 18S rRNA and Cpgp40/15 genes. Positive implementation of tests for enteric protozoa including Cryptosporidium is necessary in the differential diagnosis of suspected foodborne gastroenteritis, particularly when it is characterized by a prolonged incubation period and severe watery diarrhea. In fact, we were able to diagnose the illness as cryptosporidiosis without waiting for the results of bacteriological and virological examinations, and thus prevented the possible occurrence of a secondary infection through an ill patient who works as cooking personnel in the company.