Intensive care unit (ICU)-acquired infections can bring some degree of morbidity and mortality to patients staying in the ICU. In this study, over a 6-month period, the prevalence of infections and mortality rates among ICU patients, the predominant organisms and their resistance patterns were determined in a cross-sectional study. Samples of all the patients hospitalized longer than 48 h in two Shiraz university teaching hospitals in Shiraz, Iran, were microbiologically cultured, and antimicrobial susceptibility was determined using the disk diffusion method. We studied 123 specimens from 89 patients aged 1 month to 80 years (38.3 ± 13.4), and among them 46 patients (51.7%; 95% CI, 41.3-62.1%) showed infection based on culture and clinical findings. Of these, 37 patients (41.6%) had more than one ICU-related nosocomial infection and 9 patients (10.1%) had only one ICU-related nosocomial infection. The overall mortality rate for ICU-acquired infections was 10.9% (5 patients). Gram-negative bacteria were significantly more involved in infections than were Gram-positive bacteria (P < 0.05). The most frequently reported infections were urinary tract infections. The most frequently isolated bacteria were Pseudomonas (39.1%), which was mainly sensitive to amikacin and ceftazidime. Our study results underscore the importance of specific measures for infection control in critically ill patients in ICUs in Shiraz, Iran, especially when using urinary catheters.

We characterized the genetic diversity of the complete VP1 region of coxsackievirus A16 (CA16) and enterovirus 71 (EV71) isolated from patients with hand, foot, and mouth disease in Toyama from 1981 to 2007 to evaluate the relationship between epidemics and genetic changes. The predominant genogroups of CA16 changed from B to C in 1995-1998, and genogroup C further changed from subgenogroup C1 to C2 around 2002, and to C3 in 2005-2007. The subgenogroups of the EV71 isolates were classified into B1, B4, C1, and C3 in 1983-1994, and into C4 in 1997-2006. However, changes of the amino acid sequences of the VP1 regions of CA16 were restored, and those of the EV71 isolates were not observed among the same subgenogroups during this survey period, indicating that the prevalence that occurred at intervals of several years seemed to depend on an accumulating number of immunologically naive children, not viral antigenic changes.

In the present study, we tried to reduce the non-specific reactions for measuring anti-Chlamydia pneumoniae IgM antibodies by the ELISA kit of HITAZYME C. pneumoniae Ab-IgM (HITAZYME IgM) by using a new absorbent. We also tried to reset the IgM cut-off index (ID) of HITAZYME IgM by testing serum samples from healthy children and healthy adults with no respiratory symptoms. The results suggest that the use of the new absorbent (anti-human IgG antibodies) may reduce the non-specific reactions by rheumatoid factor and anti-nuclear antibodies, and that the setting of the higher cut-off ID (2.00), calculated as the mean ID + 3SD of the serum samples from healthy children and healthy adults, respectively, would improve the specificity of IgM during the measurement by HITAZYME IgM.

Reports from studies conducted in several countries indicate a high incidence of bacterial contamination of donor blood. The prevalence of bacterial contamination of blood and its products in Ghana is not known. This study was conducted to determine the prevalence of bacterial contamination of blood and its products at the three major blood transfusion centers in the Greater Accra Region of Ghana. Stored whole blood and its products were cultured on different media, and isolates were identified using standard biochemical and bacteriological methods. The susceptibility of the isolates to selected antimicrobial agents was also determined by the disc diffusion method. The overall prevalence rate was 9% (28/303; whole blood, 13% [24/192]; plasma, 3% [2/79]; platelet, 9% [2/22]). The Gram-positive bacteria isolated were coagulase-negative Staphylococcus, S. aureus, and Bacillus spp., and the Gram-negative organisms were Yersinia enterocolitica, Citrobacter freundii, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae. The Gram-positive bacteria were sensitive to cloxacillin, erythromycin, tetracycline, and gentamicin but resistant to penicillin, ampicillin, cefuroxime, and cotrimoxazole, while the Gram-negative bacteria were sensitive to amikacin and gentamicin but resistant to chloramphenicol, tetracycline, ampicillin, cefuroxime, cefotaxime (except Y. enterocolitica), and cotrimoxazole. Our results suggest that bacterial contamination of blood and its products is prevalent in Ghana.

The aim of this study was to characterize 160 clinical Mycobacterium tuberculosis isolates from Guangdong with respect to their drug susceptibility phenotypes to three common anti-tuberculosis drugs, isoniazid (INH), rifampin (RIF) and streptomycin (SM), and with respect to genetic mutations in the most commonly corresponding resistance genes (katG, rpoB and rpsL). The drug susceptibility profiles were evaluated by the absolute concentration method, and genetic mutations in the corresponding resistance genes were identified by DNA sequencing. Among these isolates, 33.1% (53/160) were drug-resistant. The percentages of isolates resistant to INH, RIF and SM were 21.9% (35/160), 16.9% (27/160) and 15.6% (25/160), respectively. Twenty-five of 35 (71%) INH-resistant isolates, 22 of 27 (81.5%) RIF-resistant isolates and 19 of 25 (76%) SM-resistant isolates were found to have mutations in the analyzed katG, rpoB and rpsL gene fragments. The most frequent mutation patterns for the three drugs were as follows: INH, Ser315Thr (68.6%) in katG; RIF, Ser531Leu (55.6%) in rpoB; and SM, Lys43Arg (72%) in rpsL. These findings provide useful data on the mutation types of drug-resistant genes in M. tuberculosis isolates from Guangdong province in China.

The aim of this study was to evaluate the seroprevalence and correlates of diphtheria and tetanus in Edirne, Turkey. Tetanus and diphtheria antitoxin levels were determined by enzyme-linked immunosorbent assay. Among 99 participants, a diphtheria antitoxin level of ≥0.1 IU/mL was found in 97 (98%), while 2 (2%) had antitoxin levels of 0.011-0.099 IU/mL. The geometric mean titres (GMTs) in men were statistically higher. Among 295 participants, a tetanus antitoxin level of ≥0.1 IU/mL was found in 291 (98.6%), while 4 (1.4%) had antitoxin levels of 0.011-0.099 IU/mL. Participants who had completed secondary school or higher education showed higher GMT values. Additionally, participants vaccinated within the previous 5 years had higher GMT values and the percentage of participants who had completed secondary school or higher education was higher among them. GMTs decrease with increasing age and increase as the poverty index increases. The average socioeconomic status index of the participants was high for both diphtheria and tetanus seroepidemiology. In this community-based study, antitoxin levels of diphtheria and tetanus were high. However, revaccination of adults with tetanus-diphtheria toxoids at every opportunity (military service, pregnancy, post-injury prophylaxis, etc.) together with a single booster every 10 years should be considered as an immunization policy.

Group A rotavirus (GARV) genes (the VP7 and NSP3 genes) in acute-phase cerebrospinal fluid (CSF), sera and stool samples from 6 children with convulsions accompanied by GARV gastroenteritis were investigated by reverse transcription-polymerase chain reaction (RT-PCR). When the VP7 gene was amplified from the samples, the G genotype (G type) of GARV was determined by RT-PCR. GARV genes were detected in the CSF samples of all 6 children, in 2 of the 3 blood samples, and in all of 4 stool samples. The G typing of GARV from 12 of a total of 13 samples indicated that G3 was the predominant G type in all samples. GARV antigens were detected by enzyme-linked immunosorbent assay in all of the 3 tested sera samples, while no GARV antigens were detected in any of the 5 tested CSF samples. We confirmed the presence of GARV genomes in the CSF samples from all of the children with rotavirus-associated seizures, including encephalopathy. However, the relationship between convulsions and the existence of GARV RNA in CSF remains unclear and further study is required.

The purpose of this study was to characterize the frequency, activation, and coreceptor expression of lymphocytes in Chinese human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infected patients, and to study the impact of HCV on immune status and disease progression of HIV infection. Flow cytometry was used to analyze the numbers of T cells and NK cells, the level of activation and the coreceptors of T lymphocytes in different disease stages among 110 HIV/HCV co-infected and 101 HIV mono-infected patients. With disease progression, co-infected patients expressed lower numbers of CD4 T-cells and NK cells, and higher activation levels and coreceptor expression of T lymphocytes. Compared to the counts of HIV mono-infected patients, the NK cell counts of co-infected patients were significantly lower in the asymptomatic HIV-infected and AIDS groups, and the levels of HLA-DR and CXCR4 were significantly elevated in the AIDS group. The viral load of HIV and HCV in the co-infected group increased gradually with the progression of disease. With disease progression, the immune status of HIV/HCV co-infected patients decreased gradually, and the HIV viral load increased. HCV appears to accelerate the natural course of the HIV disease by damaging the innate immune function and aggravating the levels of activating markers and coreceptors on T lymphocytes in co-infected patients.

To document the association of pathogenic Escherichia coli with diarrhea in Romanian children, 250 E. coli fecal isolates from children under 5 years of age were PCR-screened for well-recognized virulence determinants, as well as for their phylogenetic background. The putative diarrheagenic E. coli (DEC) were investigated for susceptibility to various antibiotics. Overall, 61 E. coli isolates were classified as enteroaggregative E. coli (29 isolates), atypical enteropathogenic E. coli (22 isolates), enterotoxigenic E. coli (8 isolates), and verotoxin-producing E. coli (1 isolate), and one isolate was categorized as unconventional DEC. Only 8 of the PCR-positive isolates would have been assumed to be pathogenic based on their O antigenicity, which highlights the limited effectiveness of serotyping. More than a half (51%) of the pathogenic isolates expressed a multidrug-resistant phenotype, which raises concerns about the therapeutic pediatric approach. The DEC isolates were heterogeneous phylogenetically, deriving from all four major groups: A (31 isolates), B2 (14 isolates), B1 (10 isolates), and D (6 isolates), respectively. Thus, the phylogenetic descent was less significant than the virulence gene content. Our findings document the importance of DEC as a cause of childhood diarrhea in Romania, providing evidence that efforts should be made to estimate the burden of infections by etiology for a better medical approach.

A mark-release-recapture study of the dispersal ability of blow flies, Calliphora nigribarbis, was conducted in Ikumo-Naka, Ato Town, Yamaguchi Prefecture, Japan in December 2004. A location where a fatal avian influenza outbreak had occurred 1 year previously was selected for the present study. A total of 3,884 C. nigribarbis were collected, 1,915 of which were marked and released from 4 different collection sites during 2 successive days. The recapture rate of the released C. nigribarbis ranged from 0.014 to 0.029 among the collection sites, and the overall recapture rate was calculated as 0.022. Based on the distance between the released site and the recaptured site, the dispersal rate of C. nigribarbis was estimated as 256 m/h on the 1st day and 179 m/h on the 2nd day of the experiment, and the maximum dispersal rate observed in this study was estimated as 500 m/h. Taking into account the active period of C. nigribarbis on a fine day (7 h/day), the distance traveled by C. nigribarbis within a day was estimated as 1,789 and 1,250 m/day on average for the 1st and 2nd days, respectively, and the maximum distance was 3,500 m/day. These results suggest that C. nigribarbis could play a role in the mechanical dissemination of avian influenza virus and spread of the outbreak in Japan.

The reporting of vancomycin resistance in Staphylococcus spp. has enormous therapeutic and epidemiological consequences. We evaluated Vitek software version 2.01 by the CLSI-recommended broth dilution method as well as the CDC-recommended vancomycin screen agar and Etest for vancomycin susceptibility testing of Staphylococcus spp. Of the 105 isolates of Staphylococci tested by the above methods, Vitek 2.01 gave 16 (15%) a false vancomycin intermediate/resistant phenotype. Laboratories using automated systems for routine microbiological susceptibility testing must confirm such resistance by validated methods.

Ten of 140 patients (7.1%) showed negative results on a QFT-2G test. The 10 patients consisted of seven males and three females (mean age: 66.0 years old). Three of the 10 patients had no clinical symptoms and no underlying diseases. They showed negative results on the QFT-2G test repeatedly within 1 month and a negative response on a tuberculin skin test (TST). Six of the seven remaining patients were of advanced age, and all of them had underlying diseases. Although all seven patients showed negative results on the QFT-2G test repeatedly within 1 month, one of them showed a negative response on TST. The lymphocyte count was within the normal range in three patients with nutritional conditions. Although five of nine patients with pulmonary tuberculosis had cavitary lesions, six had lesions within one-third of the unilateral lung field. The clinical effect in all patients was good. We suspect that there are two patterns by which false negative results are shown on QFT-2G tests; one pattern involves the decrease of interferon-gamma production due to advanced patient age or lymphocytopenia, and in the other lymphocytes cannot produce interferon-gamma for Mycobacterium tuberculosis-specific antigen in young patients without underlying disease.

In 2008, an outbreak of chikungunya virus (CHIKV) occurred in Narathiwat province, south Thailand. To determine the clinical significance, molecular epidemiology and evolutionary origin of the CHIKV causing this outbreak, 47 patients who had been admitted to Narathiwatratchanakharin provincial hospital due to acute febrile illness were enrolled in this study. Sera were tested for IgM antibodies, and RT-PCR was performed for CHIKV and dengue virus. We diagnosed 10 patients with CHIKV infection and 5 with dengue virus infection. Joint pain is a significant symptom of chikungunya fever. Five strains of CHIKV were isolated. Their genome sequences were different from those isolated from the previous outbreaks in Thailand (1988, 1995-1996) but similar to the sequences isolated from the 2008 Singapore outbreak. We speculated that the outbreak was caused by a group of viruses different from the previous outbreaks. RT-PCR, serology to detect IgM antibodies or paired sera for IgG for CHIKV should be performed in all patients with presumed hemorrhagic fever to promptly detect outbreaks of CHIKV. This precaution would help control global epidemics of this virus.

Drug susceptibility tests were performed with a series of Candida spp. in media supplemented with serum or bile. The azole susceptibilities of several medically important Candida spp., including C. albicans but not C. parapsilosis, were significantly reduced in supplemented media. These findings have important implications for the mechanisms of acquired azole resistance in pathogenic fungi.

A total of 52 methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates were collected from patients attending the teaching hospital of Tehran University of Medical Sciences. Disks containing antibiotics were used to determine the susceptibility of MRSA isolates. Analysis of SmaI macrorestriction profiles of the 52 MRSA isolates were grouped into three PFGE types. The majority of isolates (n = 49) were clustered into only one major PFGE type, designated as pulsotype A; these belonged to SCCmec type III or IIIA and showed resistance to ampicillin, ciprofloxacin, cotrimoxazole, erythromycin, gentamicin, and tetracycline. The remaining isolates fell into pulsotypes B and C, both belonging to SCCmec-type IV. All MRSA isolates were susceptible to vancomycin, teicoplanin, quinupristin-dalfopristin, linezolid, and tigecycline. The present study shows that a MRSA clone similar to the Brazilian clone (ST 239) of MRSA, which is a multiresistant MRSA clone with a high level of methicillin resistance, is very common in this teaching hospital in Tehran.

Twenty-eight of the 302 Salmonella strains isolated from patients with overseas travelers' diarrhea who were examined at Kansai Airport Department Station during the 6-year period between 2001 and 2007 showed decreased susceptibility to both nalidixic acid (NA) and ciprofloxacin (CPFX) (MIC of NA, 16-64 μg/mL; MIC of CPFX, 0.064-2.0 μg/mL). These 28 strains revealed no variations in the quinolone resistance-determining region; however, 25 of the strains showed retention of the qnr gene. The qnr-retaining Salmonella belonged to 6 serotypes, and 21 and 4 of the 25 strains showed qnrS1 and qnrS2, respectively. The most common serovar was S. Corvallis (17 strains).

Shiga toxin 2f-producing Escherichia coli (O115:HNM) with eae was isolated from a symptomatic patient in Fukuoka Prefecture, Japan. The patient was a 23-year-old male and his symptoms were diarrhea, abdominal pain, headaches and a fever (37.7ºC). He had eaten raw chicken meat, raw chicken eggs, cooked chicken meat and raw vegetables about 13 h prior to the onset of the symptoms. The patient's specimen was examined, and no diarrheagenic agents were detected except for Shiga toxin 2f-producing E. coli (STEC_2f) with eae. This is the first report of the serotype O115:HNM possessing stx_2f. We discuss the necessity of routinely using stx_2f-detecting PCR primers for detection of this enteric pathogen.

The etiological roles of diarrheagenic Escherichia coli (DEC), including enteroaggregative E. coli (EAggEC), diffusely adherent E. coli (DAEC) and EAST1EC—a strain of E. coli that possesses no diarrheagenic characteristics other than the EAggEC heat-stable toxin 1 (EAST1) gene—remain controversial. To clarify the prevalence of DEC among healthy individuals in Osaka City, Japan, and to compare the virulence properties of strains previously isolated from diarrheal patients, fecal specimens were examined for DEC. Isolation rates of Shiga toxin-producing E. coli, enterotoxigenic E. coli, and EAggEC were significantly lower among healthy adults than sporadic adult patients. There were no differences in enteropathogenic E. coli (EPEC), DAEC and EAST1EC between patients and healthy carriers. Subtyping of the intimin gene (eae) of EPEC, and measuring the IL-8 inductivity of DAEC on epithelial cells could provide criteria to distinguish strains in diarrheal patients from those in healthy carriers. Proper criteria should be established in order to diagnose subtypes of DEC as causative agents.