In developing countries where tuberculosis is still a health challenge, the prevalence of nontuberculous mycobacterial diseases is expected to rise as medical conditions that compromise the immune system become more widespread. In the current study, we aimed to determine the presence and diversity of nontuberculous mycobacteria (NTM) causing infections in Iranian patients. Sixty-seven clinical NTM isolates were identified using conventional and molecular methods, including PCR-restriction fragment length polymorphism analysis (PRA) and 16S rRNA sequencing. Out of 67 patients with confirmed mycobacterial infection, 29 had an associated immunosuppressive syndrome, including 9 who were HIV-infected. Forty-nine NTM isolates were identified using PRA, and the remaining 18 isolates were identified using 16S rRNA sequencing. We obtained the following results: Mycobacterium fortuitum, 30 isolates; M. kansasii, 12 isolates; M. gordonae, 8 isolates; M. porcinum, 3 isolates; M. conceptionense, 3 isolates; M. phlei, 2 isolates; and M. austroafricanum, M. avium, M. elephantis, M. intracellulare, M. lentiflavum, M. monacense, M. parascrofulaceum, and M. thermoresistibile, 1 isolate each; and 1 potentially novel mycobacterial species. With regard to the complexity of identification, it is recommended that laboratory diagnosis of NTM diseases be centralized by strengthening or setting up quality national and regional infrastructure.

The aim of the present study was to compare serological tests (Rose Bengal [RB]; standard agglutination test [SAT]; enzyme immunoassay [EIA] for detection of IgM, IgA, and IgG; and 2-mercaptoethanol [2-ME] test) that are routinely used in patients prediagnosed with different clinical types of brucellosis (acute, subacute, or chronic), and to evaluate the results of the IgG avidity test. Ninety-two patients having titers ≥1/160 as measured by SAT were included in the study. The IgG avidity test was performed in 78 patients who had positive EIA-IgG results. RB test results were positive in 88 (95.7%) patients. A statistically significant correlation was found between a positive EIA-IgM result and the diagnosis of acute brucellosis. When compared to the results of the SAT, the 2-ME test showed a lower titer in 55 (59.8%) patients, and the agreement between the 2-ME test and EIA-IgG was calculated as 84.8%. No statistical difference was found between the 40% avidity index used in the IgG avidity test and avidity maturation time (6 months). From our study, we concluded that (i) the RB and SAT tests are appropriate and reliable tests for the serological diagnosis of brucellosis; (ii) IgM can be used as a marker of acute brucellosis; (iii) the 2-ME test, similar to EIA, can be used to determine IgM levels; and (iv) the IgG avidity test should be standardized.

In anticipation of the imminent licensure of rotavirus vaccine, we evaluated the cost-effectiveness of rotavirus vaccine in Japan by taking into account the considerable variations in the incidence of rotavirus-associated hospitalizations previously reported in the literature. We assumed that the variation was due to local differences in healthcare utilization practices rather than a true difference in the incidence of severe rotavirus gastroenteritis. Thus, a Markov model was constructed such that the sum of rotavirus-associated hospitalizations and outpatient visits was set a constant value of 129 cases per 1,000 child-years. We calculated the direct medical cost, the indirect cost, and the quality-adjusted life year (QALY) loss in children aged less than 5 years. For the base case scenario, the incremental cost-effectiveness ratio (ICER) per QALY gained was 9.8 million Japanese yen from the healthcare perspective, but it was 900,000 Japanese yen from the societal perspective, making the program of universal immunization against rotavirus highly cost-effective. Furthermore, the universal immunization program was found to be cost-effective from the societal perspective for any of the previously reported incidence rates of rotavirus-associated hospitalization. Thus, the introduction of the rotavirus vaccine into the childhood immunization schedule and its co-administration with other childhood vaccines will be a cost-effective public health intervention in Japan.

Japanese encephalitis virus (JEV) and the four dengue viruses (DENV1–4) are co-distributed in Southeast and South Asia. Since JEV is antigenically cross-reactive with DENV1–4, the differentiation between these viruses using antibody assays may be difficult. Herein, we describe the development of an epitope-blocking enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for the nonstructural protein 1 (NS1) of JEV (JEV-NS1) to differentiate antibodies against JEV from those against DENV1–4. Hyperimmune mouse sera against JEV-NS1 showed >60% inhibition, whereas those against NS1 of DENV1–4 showed <30% inhibition. The present assay could therefore detect antibodies specific for JEV. For testing of human sera, a temporary cutoff value (30.8%) was calculated the average and standard deviation obtained for sera of control humans negative for JEV antibodies. Human sera positive for antibodies to any of DENV1–4 NS1 but negative for antibodies to JEV-NS1 showed a lower percentage inhibition than the cutoff value. On the other hand, sera with JEV-NS1 antibody levels of ≥0.400, as determined by the conventional ELISA (medially/strongly positive for JEV-NS1 antibodies), showed percentage inhibition greater than the cutoff. Although this blocking ELISA afforded false-negative results for most sera that were weakly positive for JEV-NS1 antibodies, it may be useful for investigating the seroepidemiology of JEV antibodies in dengue-endemic areas.

The combination of lactoferrin with fluconazole has been reported to synergistically enhance the antifungal activity of fluconazole against Candida spp. and inhibit the hyphal formation in fluconazole-resistant strains of Candida albicans. In this study, we investigated the association between the therapeutic effects of this combination and the pharmacological characteristics of fluconazole and itraconazole and the variation in these effects with differences among the strains in terms of the susceptibility and resistance mechanisms. Lactoferrin enhanced the growth-inhibitory activity of fluconazole against two different ergosterol mutants but not againt pump mutants or an azole-susceptible strain; but increased the activity of itraconazole against all the strains tested in this study. Exogenous iron cancelled the synergistic effect, which suggests that the iron-chelating function of lactoferrin may contribute to the synergism. Besides, radiolabeled fluconazole assays revealed that lactoferrin did not affect the intracellular concentrations of fluconazole, thereby indicating that these synergistic effects were not due to the alteration of the intracellular uptake of the drug. The development of new clinical treatments and therapeutic method against resistant Candida will depend on our understanding of the resistance mechanisms and methods to overcome them by the application of suitable drug combinations with synergistic effects. The results of this study might contribute to the improvement of our understand of the mechanisms underlying the resistance of Candida strains.

Surveillance of vaccine safety is one of the public health interventions used to investigate the causal relationship between vaccines and adverse events. Using active surveillance data, we aimed to compile a detailed summary describing the safety of the pandemic influenza A (H1N1) vaccine. Computer-assisted telephone interview was used to investigate adverse events for 9,000 subjects who had received non-adjuvanted vaccines between November 2009 and January 2010, and for 19,000 adults who received adjuvanted vaccines from January through March 2010. The participants were interviewed to obtain information about local and systemic adverse events. Among subjects who received the non-adjuvanted vaccine, 5.5% (n = 492) reported adverse events after vaccination, while 6.7% of those who received the adjuvanted vaccine reported adverse events. In the group receiving the adjuvanted vaccine, the highest reported rate of adverse events was among persons aged 19–49 years (9.1%, 577/6,329), followed by persons aged 50–64 years (7.2%, 485/6,718), and elderly persons aged 65 years and over (3.4%, 204/5,953). The implementation of this active surveillance study demonstrated the safety of both the adjuvanted and non-adjuvanted H1N1 vaccines.

Staphylococcus epidermidis is a cause of orthopedic device-related infection, and to treat such infection, biofilms should be controlled. Polysaccharide intercellular adhesin (PIA) is associated with the biofilm-forming ability of staphylococcal strains. PIA in biofilm-positive staphylococcal strains can be detected by the Congo red agar (CRA) method. In this study, we used the CRA method to examine the effects of subminimal inhibitory concentrations (sub-MICs) of 11 antibacterial agents on PIA production by S. epidermidis. We found that the PIA-negative variants were selected only by sub-MICs of gentamicin (GM). This PIA-negative phenotype was maintained over several generations in the absence of GM. Such selection occurred in six of eight clinical isolates, as well as in the biofilm-positive control strain. No such selection occurred with aminoglycoside antibiotics except for GM. Most of the PIA-negative variants that were selected by GM showed a markedly lower biofilm-forming ability on stainless steel washers than their untreated parent strains. In conclusion, variants with lower biofilm-forming ability may be selected by a sub-MIC of GM. Investigation of the reason why variants with reduced biofilm-forming ability can be selected in the presence of sub-MICs of GM may contribute to strategies against biofilm-related infections.

In 2007–2008, a measles outbreak occurred among children above the age of 10 years in Akita Prefecture, northeastern Japan (population, approximately 1,120,000 at the time). Our group controlled the outbreak by (i) implementing a publically financed urgent vaccination program and (ii) prohibiting non-vaccinated and non-infected students from attending school as per regulations of the school public health law. We encouraged high-risk students to undergo a vaccination program, which resulted in the successful containment of the outbreak without the development of any severe cases. After the outbreak, the Akita Prefectural Government began an annual“Akita measles elimination month” every April, and no measles case found in Akita Prefecture during 2009–2010 subsequently. Our outbreak response initiative can be applied nationally for the complete elimination of measles throughout Japan.

Cholera has been a prevalent disease worldwide since the early 19th century. Vibrio cholerae O1 and O139 are the two serogroups that have been mainly implicated in causing cholera. This study reports the results of biotyping, serotyping and phage typing of V. cholerae O1 and O139 (1998–2007) strains received from different parts of India for the identification of the trends in the occurrence and spread of cholera in the country. However, there has been a notable steep decline in the occurrence of V. cholerae O139 strains over the past few years resulting in no strain of V. cholerae O139 being received from any part of India in 2007 and 2008. Of the total strains received, 79.1% were serotyped as Ogawa and the remaining 20.9% were found to be Inaba, which indicates that Ogawa was the predominant serotype. Almost 100% typeability was observed with the new scheme of V. cholerae O1, with type 27 being the dominant phage type and V. cholerae O139 strains were clustered into the predominant phage type T-1. From the phage typing and serotyping results, it can be concluded that V. cholerae O1 (T-27) and O139 (T-1) strains circulate throughout the country at any given time.

The aim of this study was to evaluate lymph node tuberculosis (LNT) cases in two provinces in Turkey with different demographic and socioeconomic characteristics. A total of 109 LNT cases were reviewed retrospectively. The cases were analyzed and compared for symptoms, findings, age, vaccination status, and diagnostic procedures. Socioeconomic conditions were also assessed for the two provinces. A palpable cervical node was considered a significant predictor for all LNT. Mediastinal lymph node involvement was found to be common in cases of pulmonary manifestation of LNT. Female patients were predominantly from the Van Province, while older patients were found to be from Izmir Province. LNT should be suspected in lymphadenitis patients of all age-groups especially in young adolescents with cervical lymph node enlargements. In the presence of mediastinal lymphadenopathy, pulmonary tuberculosis should be investigated.

A parasitological survey of Schistosoma haematobium infection among primary schoolchildren in the remote areas of Hhohho and Manzini Provinces in northwestern Swaziland was undertaken. Presence of infection in subjects was confirmed on detection of S. haematobium ova in urine or the presence of hematuria. The intensity of the infection was estimated by calculating the total number of S. haematobium ova present in a 10-ml urine specimen and was expressed in terms of geometric mean intensity (GMI). The prevalence of S. haematobium infection in these populations was 5.3% (21/395) with a GMI of 46.5. Boys had higher prevalence (7.1%, 13/182) and GMI (50.4) than girls (3.8%, 8/213; 40.0) did (P > 0.05). Geographically, the prevalence in Manzini schoolchildren (14.6%, 12/82) was significantly higher than that in Hhohho schoolchildren (2.9%, 9/313; P < 0.001); however, Hhohho schoolchildren had a higher GMI (70.2) than that observed in Manzini schoolchildren (21.9). Children from schools located in Lowveld had a significantly higher prevalence (11.4%, 19/166) than that in children from schools located in Highveld (0.6%, 1/162) (P < 0.0001).

We report a new case of Cardiobacterium hominis endocarditis identified during an acute coronary syndrome. The positivity of the blood cultures was confirmed rapidly (50 h) as a result of improvements to the automated detection system, whereby it is no longer necessary to incubate the vials for long periods of time when Aggregatibacter-Cardiobacterium-Eikenella-Kingella infections is suspected. The phenotype-based VITEK 2 NH identification system is not able to distinguish between the two species of Cardiobacterium, as it does not contain C. valvarum in its library. The method for 16S rRNA gene sequence analysis is able to separate the two species but is not available in all laboratories. We used MALDI-TOF mass spectrometry, as an alternative, to rapidly distinguish between C. hominis and C. valvarum, because both species are contained in the system library.

Dengue is currently the most important arboviral disease in the world, particularly in tropical countries in which the environmental conditions favor the development and proliferation of the mosquito vector. Dengue hemorrhagic fever presents in two phases: an initial phase, which is characterized by sudden onset of fever and a variety of nonspecific signs and symptoms, and a critical phase, which is characterized by the recovery from fever and development of hemorrhagic symptoms and circulatory insufficiency. This report documents a case of splenic rupture in a patient with dengue hemorrhagic fever who developed hypovolemic shock and subsequently died. Although splenic rupture is a known complication of other acute infections, it is a rare complication of dengue; therefore, it may be misdiagnosed. In the case described here, the poor outcome mainly resulted from the sudden onset of complications; the patient died of splenic rupture less than 24 h after admission, and the cause of death was confirmed at necropsy.

This study was conducted to determine the prevalence and geographic distribution of Anaplasma phagocytophilum, the causative agent of human granulocytic anaplasmosis, in wild deer and boars in Japan. We analyzed the blood samples of the study animals using PCR-targeting the 16S rDNA followed by DNA sequencing. Wild deer infected with Anaplasma spp., including Anaplasma bovis and Anaplasma centrale were detected in the region from Hokkaido to Kyushu. The infection rates of A. phagocytophilum, A. bovis, and A. centrale in deer were 15.6, 21.9, and 37.5%, respectively, and the corresponding infection rates in wild boar were 3.6, 17.9, and 3.6%, respectively. However, p44/msp2 genes specific to A. phagocytophilum were not detected among the 16S rDNA-positive samples on PCR analysis. In addition, the 16S rDNA sequences of A. phagocytophilum were 100% similar to those detected previously in the deer in Japan and 98.6% similar to those of A. phagocytophilum detected in the United States and Europe, and from the tick, Ixodes ovatus and Ixodes persulcatus in Japan. These findings suggested that the A. phagocytophilum-related sequences detected from deer and wild boars in Japan were different from those of typical A. phagocytophilum strains.

The resistance phenotypes and genomic diversity of 185 Acinetobacter baumannii isolates obtained from the intensive care unit (ICU) of a local teaching hospital in Kuala Lumpur from 2006 to 2009 were determined using antimicrobial susceptibility testing and pulsed-field gel electrophoresis (PFGE). Antibiogram analyses showed that the isolates were fully resistant to β-lactam antimicrobials and had high resistance rates to the other antimicrobial agents tested. However, the isolates were susceptible to polymyxin B. Resistance to cefoperazone/sulbactam was only detected in strains isolated from 2007 to 2009. Some environmental isolates and an isolate from the hands of a healthcare worker (HCW) had identical resistance profiles and PFGE profiles that were closely related to patient isolates. Cluster analyses based on the PFGE profiles showed there was a persistent clone of endemic isolates in the ICU environment. The transmission route from HCWs to fomites to patients, which caused a long-term infection in the ICU of the University Malaya Medical Centre, was observed in this study. These data provide a better understanding of A. baumannii epidemiology within the hospital and the possible transmission routes. Knowledge of changes in the resistance rates of A. baumannii in our local hospital will improve antimicrobial therapy.

In Japan, the 7-valent pneumococcal conjugate vaccine (PCV7) was introduced in 2010. To assess the effects of PCV7 on invasive pneumococcal infection in children, a population-based prospective survey has been conducted in 10 prefectures. As a part of the study, blood and nasopharyngeal isolates from children admitted to the Shibata Hospital, Niigata Prefecture, were analyzed for determining the serotypes, their susceptibilities to antimicrobial agents, and multilocus sequence types. Sixteen blood isolates were obtained from October 2007 to December 2009. Sixty-three nasopharyngeal isolates were obtained from the posterior nares of 118 children with pneumonia from April to September 2008. The coverage rates of the blood and nasopharyngeal isolates for PCV7 were 81.3% and 57.1%, respectively. Although none of these children had received PCV7, serotype 19A isolates were recovered from 12.5% (2/16) of the blood samples and 12.7% (8/63) of the nasopharyngeal samples. The sequence type of a nasopharyngeal isolate of serotype 19A was ST320, and the minimum inhibitory concentration of penicillin G was 4 μg/mL. In addition to the continuous prospective survey of pneumococcal infection, early introduction of the 13-valent conjugate vaccine, in which the 19A conjugate is included, will be necessary in Japan.

The incidence of anthrax, which is caused by Bacillus anthracis, in the human and animal population of Mongolia has increased recently, and control of this infection is a nationwide concern. In this study, 29 isolates obtained from animals and various regions in Mongolia from 2001 to 2007 were analyzed by performing multiple-locus variable-number tandem-repeat analysis for 8 loci (MLVA-8) to understand the genetic relationship between the Mongolian B. anthracis isolates. We found that all the Mongolian isolates can be classified into A3 cluster along with the Japanese and the Chinese B. anthracis isolates. Our data revealed that MLVA-8 is useful for studying the molecular epidemiology of the Mongolian B. anthracis isolates and would help characterize B. anthracis infections in Mongolia.