We conducted descriptive analysis of available information regarding the epidemiology of cholera outbreaks in South and Southeast Asia during 2003–2012. Information from 58 articles, 8 reports, and World Health Organization databases were analyzed. Overall, 113 cholera outbreaks were studied in South and Southeast Asia during the past 10 years. The majority of the outbreaks (69%) occurred in Southeast Asia, including India (52%). The highest number of outbreaks was observed in 2004 (25.7%). The most commonly identified source was contaminated water: however, in some countries, the spread of cholera was facilitated via contaminated seafood (e.g., Myanmar, Thailand, and Singapore). Several genotypes and phenotypes of Vibrio cholerae, the causative agent of cholera, were identified in the outbreaks, including V. cholerae O1 El Tor (Ogawa and Inaba) and V. cholerae O139. The emergence of multidrug-resistant V. cholerae strains was a major concern. Cholera-related mortality was found to be low across the outbreaks, except in Orissa, India (currently Odisha) during 2007, where the case fatality rate was 8.6%. Potential limitations included underreporting, discrepancies, possible exclusion of nonindexed reports, and incomprehensive search terms. The provision of safe water and proper sanitation appear to be critical for the control of further spread of cholera in South Asian and Southeast Asian regions.

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We investigated the prevalence of non-typhoidal Salmonella (NTS) with “reduced susceptibility to ciprofloxacin” (RS-Cip) (minimum inhibitory concentration [MIC], 0.12–1.0 μg/mL) as well as their resistance genes in 75 NTS isolates (53 from stool, 21 from blood, and 1 from urine) from patients at a tertiary care Malaysian hospital between January and December 2009. RS-Cip was detected in 24/75 (32.0%) isolates. Using the ciprofloxacin MIC interpretive criteria for Salmonella in the Clinical and Laboratory Standards Institute 2013 guidelines, 51/75 (68.0%) isolates were found to be sensitive, 22/75 (29.3%) were intermediate, and 2/75 (2.7%) were resistant to ciprofloxacin. The 24 isolates that were intermediate or resistant to ciprofloxacin were the same isolates categorized as having RS-Cip. Among the 23 tested isolates with RS-Cip, the qnrS gene was detected in 17/23 (73.9%) and single gyrA mutations were detected in 6/23 (26.1%) (Asp87Tyr [n = 3], Asp87Asn [n = 2], and Ser83Phe [n = 1]). A parC (Thr57Ser) mutation was detected in 13/23 (56.5%) isolates, coexisting with either a qnrS gene or a gyrA mutation. The high incidence of the qnrS gene among isolates with RS-Cip needs to be monitored because qnr genes can spread via plasmids and aid in the emergence of increased resistance levels.

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The present study aimed to compare the main features of infection with pandemic influenza A virus in pregnant and nonpregnant women admitted to hospitals in Spain during the first waves of the 2009–2010 influenza pandemic. This was a prospective (November 2009 to June 2010), multicenter observational study. All cases were women of reproductive age who had not been vaccinated against seasonal or pandemic influenza A. Influenza infection was confirmed by reverse transcription-polymerase chain reaction (RT-PCR). The sociodemographic and clinical data of all cases were reviewed. A total of 219 inpatients, including 49 pregnant women and 170 nonpregnant women, were enrolled in the study upon admission to participating hospitals. The most substantially different symptoms between the groups were respiratory distress and unilobar consolidation, both of which were more frequent among nonpregnant women. Antibiotics and systemic corticosteroids were more frequently used in nonpregnant women; however, there were no differences in the rates of treatment with antivirals. Our findings indicated that the compared with nonpregnant women, pregnant women in this study did not have significantly different symptoms and were not at increased risk of complications from pandemic influenza virus infection.

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Biofilm production by microorganisms is critical for their pathogenicity. Serum promotes biofilm production by Aspergillus fumigatus; however, its effects on other Aspergillus spp. have not been reported. We analyzed biofilm formation by five Aspergillus spp., i.e., A. fumigatus, A. flavus, A. nidulans, A. niger, and A. terreus, and examined the effects of serum/serum proteins such as fetal bovine serum (FBS), fetuin A, and bovine serum albumin (BSA) on hyphal growth, hyphal branching, and extracellular matrix (ECM) formation. The antifungal susceptibility of A. fumigatus isolates that formed biofilms was also examined. All serum/serum proteins promoted the growth of all these fungal species; growth promotion was most evident with FBS, followed by fetuin A and BSA. This effect was most evident in case of A. fumigatus and least evident in case of A. terreus. Electron microscopy showed thick ECM layers surrounding fungal cell walls after culture with FBS, particularly in A. fumigatus. An increase in hyphal branching caused by fetuin A was the highest in case of A. fumigatus and A. nidulans. Biofilm-forming A. fumigatus showed resistance to most antifungal agents, although a synergism of micafungin and amphotericin B was suggested. Our results indicate that serum promotes biofilm formation, including thick ECM, by many Aspergillus spp., particularly A. fumigatus, and that this may be closely related to its virulence.

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As the incidence of measles decreases, cases reported as suspected measles will increasingly involve rash associated with measles vaccination itself. In this study, we assessed vaccine-associated measles cases reported in Korea between 2002 and 2012 using a standardized assessment and following by the World Health Organization case definition criteria. We retrospectively analyzed data regarding (i) wild-type measles and (ii) vaccine-associated measles in patients aged 12–23 months. The presence or absence of fever, rash, cough, coryza, conjunctivitis, and Koplik spots were reviewed. Males were more likely to be reported with vaccine-associated measles than with wild-type measles (68% vs. 47%, P < 0.05). The number of patients with wild-type measles peaked between April and July, whereas that of patients with vaccine-associated measles remained relatively constant throughout the year. However, after excluding the cases reported during the 2007 outbreak in Korea, the trend was similar between the two groups. Cough, coryza, and conjunctivitis were more likely to be present in patients with wild-type measles (32–61% vs. 10–43%, P < 0.05); conversely, the absence of these symptoms was noted in most patients with vaccine-associated measles. We therefore conclude that cough, coryza, and conjunctivitis may be useful as key positive findings to distinguish between wild-type measles and vaccine-associated measles infection among 12–23-month-old patients in a country with a low incidence of measles.

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Infection of human with avian-like A (H1N1) swine influenza virus (SIV) occasionally occurs in China, suggesting a potential risk of cross-species transmission of the swine influenza H1N1 virus from pigs to humans, particularly to those having direct contact with pigs. A seroepidemiological study was conducted to assess the prevalence of antibodies against the avian-like A (H1N1) SIV among swine farm residents and pigs in southern China to evaluate the risk of infection to swine farm workers. Hemagglutination inhibition (HI) assays revealed that 11.17% (61/546) of the sera samples from swine farm residents in southern China were positive for antibodies against the avian-like A (H1N1) SIV. The difference in numbers of antibody-positive samples obtained from swine farm residents and a control group of healthy city residents was statistically significant (P = 0.031). In addition, 219 of the 1,180 serum samples from pigs were positive for the antibodies against an avian-like A (H1N1) SIV, A/swine/Guangdong/SS1/2013(H1N1), as assessed by HI. The data suggest that occupational exposure of swine farm residents and veterinarians in southern China to pigs may increase their risk of acquiring avian-like A (H1N1) SIV infection. According to a special pig farming model in southern China, the staff and residents are in close contact with infected pigs and may be among the first to become infected.

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Female sex workers (FSWs) and their clients are one of the major sources of spread of HIV/AIDS in Bangladesh. The purpose of this study was to determine the rate of unprotected sex among FSWs in Rajshahi City, Bangladesh. A total of 200 FSWs were randomly selected for the survey from February to September 2012. The age range of FSWs was 16–41 years (average age, 24.52 ± 6.26 years), and the majority of these women (84.4%) were married. More than 88% of FSWs reported practicing unprotected sex because of clients' insistence. Further analysis showed a significantly higher rate of protected sex among unmarried FSWs (P < 0.01). Approximately 32% of FSWs did not undergo voluntary counseling and testing (VCT) for HIV, and this factor was significantly associated with the education level (P < 0.01), age (P < 0.01), and economic status (P < 0.05). Furthermore, 89.5% respondents did not ask their new clients about VCT/HIV status, and this factor was associated with the resident area (P < 0.05), age (P < 0.05), and economic status (P < 0.01) of FSWs. Authorities in this country should focus their HIV/AIDS prevention efforts on illiterate FSWs, married FSWs, and FSWs aged below 21 years or above 30 years.

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Acute encephalitis syndrome (AES) is a major public health problem in eastern Uttar Pradesh, claiming thousands of lives every year. Here we report the common viral etiologic agents of AES and its epidemiology in the vicinity of Lucknow in Uttar Pradesh, North India. Cerebrospinal fluid (CSF) samples collected from patients with AES, who were referred to a viral diagnostic laboratory from January 2011 to December 2012, were tested for IgM antibodies against Japanese encephalitis virus (JEV), dengue virus (DV), herpes simplex virus (HSV), measles virus, mumps virus, varicella zoster virus (VZV), and enterovirus using commercial enzyme immuno-assays. Of the 1,578 enrolled patients, JEV was the most commonly detected (16.2%), followed by DV (10.8%), HSV (9.3%), measles virus (8.9%), mumps virus (8.7%), VZV (4.4%), and enterovirus (0%). Co-positivity with more than 1 virus was observed in 12 patients. The demographic distribution of patients pertaining to age, sex, and geographic and seasonal variation is discussed. Maximum mortality was caused by JEV infection, while patients with HSV infection had maximum residual neuro-psychiatric disability. JEV and DV are the chief causative agents of AES in North India, although other viruses should also be considered in a differential diagnosis.

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The previously identified Shiga toxin (Stx) 2f-producing Escherichia coli O115:HNM strain F08/101-31, isolated from a symptomatic human, was confirmed to be E. albertii in the present study by whole genome DNA–DNA hybridizations, by sequencing (cpn60, dnaJ, and 16S rRNA genes), and by multi-locus sequence typing. The F08/101-31 strain was originally identified as E. coli rather than the relatively new bacterial species E. albertii, which was first described in 2003, because it did not display any of the biochemical characteristics of E. albertii. This new classification will impact public health management strategies in Japan because the present study showed that some E. albertii strains, which are often misidentified as E. coli, produce Stx and likely cause diarrhea in humans. Therefore, further guidelines for the management and identification of Stx-producing E. albertii are required in Japan.

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Replicon systems have been used for high-throughput screening of anti-dengue virus (anti-DENV) inhibitors and for understanding mechanisms of viral replication. In the present study, we constructed novel DENV-1 replicons encoding Gaussia luciferase that was secreted into the culture medium. Two types of constructs were generated: RNA-based and DNA-based. Each type was translated in an internal ribosome entry site (IRES)-dependent or IRES-independent manner. Among these constructs, the DNA-based replicon employing IRES-dependent translation (DGL2) produced the highest titer. Luciferase levels in the culture medium revealed that the DGL2 replicon was inhibited by ribavirin (a well-known DENV inhibitor) at levels similar to those measured for drug inhibition of multi-round DENV-1 infection. These results indicate that the DNA-based IRES-driven DENV-1 replicon may facilitate studies on viral replication and antiviral compound discovery.

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We encountered a 3.5-year-old girl with acute encephalopathy associated with human metapneumovirus (hMPV) infection. She had pyrexia and status epilepticus, followed by a coma. Cerebrospinal fluid analysis showed no pleocytosis or elevation of protein levels. hMPV RNA was detected in tracheal aspirate. Acute encephalopathy in the patient was probably related to the hMPV infection. Serum levels of interleukin-6 and matrix metalloproteinase-9 were elevated on admission, and these factors were presumed to be related to acute encephalopathy, associated with her viral infection, or due to status epilepticus. She was treated with dexamethasone pulse therapy, intravenous immunoglobulin, and continuous thiopental infusion. She recovered without neurological sequelae.

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We conducted a long-term follow-up study between December 2005 and February 2007 on 4 immunocompetent infants, who repeatedly presented with respiratory symptoms, using PCR-based techniques targeting 14 viruses related to acute respiratory tract infection. Of 38 specimens, 30 were collected from symptomatic infants and 8 were collected when respiratory symptoms were absent. Overall, one or more respiratory viruses were detected in 94.7% (36/38) of the specimens. Of the 36 PCR-positive specimens, 77.8% (28/36) were positive for more than one virus. Most of these co-infections were double infections (55.6% or 20/36). Of note, co-infections with 4 and 3 viruses were observed in 3 (8.3% or 3/36) and 5 (13.9% or 5/36) specimens, respectively. Of the 8 specimens collected from the 4 infants when apparent respiratory symptoms were absent, 7 (87.5%) were positive for respiratory viruses. Respiratory viral co-infections were also frequent and found in 5 of the specimens (62.5%). However, apparent correlation between disease severity and co-infection was undetectable due to the limit of the number of cases studied. Taken together, this longitudinal study revealed that respiratory viral co-infections were not infrequent in infants aged 0–2 years, regardless of the presence of respiratory symptoms (62.5–77.8%).

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An outbreak of cholera struck Bihar, an Indian state, in August 2008 following a massive flood. Here we report the phenotypic and genotypic characteristics of Vibrio cholerae strains isolated from patients with diarrhea. Rectal swabs were obtained from patients with diarrhea who were admitted to medical camps or the hospital, and the strains were biochemically and serologically characterized. V. cholerae was isolated from 21 (65.6%) of 32 rectal swabs. Serological studies revealed that all the 21 isolates belonged to V. cholerae O1 Ogawa. Mismatch amplification mutation assay (MAMA)-PCR showed that the isolates belonged to El Tor variant group, and pulsed-field gel electrophoresis (PFGE) proved that these isolates were of a different lineage than the conventional El Tor variant strains. These isolates were resistant to several drugs, including ampicillin, streptomycin, tetracycline, nalidixic acid, and furazolidone. The uniqueness of the current report arises from the fact that records of cholera in Bihar are availiable for the early 1960s but not for the next 4 decades. Moreover, the present study is the first to report a cholera outbreak in Bihar that was caused by an El Tor variant strain.

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