The natural occurrence of vertical transmission of dengue viruses in Aedes albopictus (Skuse) mosquitoes was examined in the state of Kerala in southern India. Adults and larvae of Ae. albopictus collected from Kerala were screened for dengue viruses by using enzyme-linked immunosorbent assay with dengue-specific monoclonal antibodies. The possibility of the vertical transmission of dengue virus in Ae. albopictus was further evidenced by the detection of the virus in field-collected adult males as well as females emerged from field-collected larvae. Two pools, one pool from the adult males and one pool from the emerged females derived from field-collected larvae, were collected in the relatively hot months of June and March, respectively, and found to be positive for dengue virus antigen. Dengue serotype 2 virus was isolated from field-collected male adults in Kerala. These findings suggest that dengue virus is maintained in Ae. albopictus mosquitoes during the dry season by vertical transmission in nature.

The objective of this study was to characterize the gene cassettes of class 1 integrons and anti-microbial resistance among CTX-M-3-producing Serratia marcescens isolates from different specimens in southern Taiwan. One hundred and twenty-two isolates (70.5%) of 173 CTX-M-3-producing S. marcescens isolates were positive for class 1 integrons, including 53.3% of blood isolates, 94.1% of urine isolates, and 87.2% of sputum isolates. No class 2 or class 3 integrons were detected in this study. By PCR with primers 5 ́-CS and 3 ́-CS for the amplification of gene cassettes regions, amplicons ranging from 0.7 to 3.0 kb in length were found in 108 (88.5%) of the 122 class 1 integron-containing isolates of CTX-M-3-producing S. marcescens isolates. Ten different types by pattern of amplicons for class 1 integrons were obtained. The Type I amplicon (46.3%) harbors two different class 1 integrons containing the gene cassettes of aadA2 and aadB-catB3, respectively, and was most prevalent in the gene cassette region-positive S. marcescens isolates, followed by the Type II amplicon, which harbors one class 1 integron containing the gene cassette dfrA12-orfF-aadA2 (28.7%). Most of the S. marcescens isolates (66.7%, 8/12) harboring three different class 1 integrons (Type IV amplicon) were found in blood isolates. Class 1 integrons were conjugally transferred to recipients in 92.0% of S. marcescens harboring two different class 1 integrons containing the gene cassettes aadA2 and aadB-catB3, respectively. The transfer rate of class 1 integron carrying dfrA12-orfF-aadA2 was detected in 77.4% of S. marcescens isolates. The results showed that all those isolates with conjugative transfer of integrons carried their class 1 integrons on the conjugative plasmids.

The aim of this study was to examine the prevalence rates and risk factors for reproductive tract infections (RTIs) among married women of reproductive age in a rural area of Shandong Province in China. A population-based cohort of 4,039 married women of reproductive age was cluster-randomly selected from the local birth control registry. All subjects underwent clinical and microbiological tests and an interview in the form of a standardized questionnaire. The prevalences of trichomoniasis, bacterial vaginosis (BV), and candidiasis as diagnosed by clinical tests were 2.8, 5.9, and 3.1%, respectively. The infection rates of Trichomonas, BV, and Candida were 2.9, 6.6, and 3.9%, respectively. The infection rates of gonorrhea and syphilis were low and no cases of HIV infection were found. After adjustment for confounding factors the risk factors for trichomoniasis were income higher than $200, lack of knowledge about sexually transmitted diseases, and marriage to a businessmen. For candidiasis the risk factors were three or more abortions, income higher than $200, age of 30 - 39 years, and women with extramarital sex partner(s). For BV the risk factors were three or more abortions and age of 30 - 39 years. The prevalence of RTI/sexually transmitted infection (STI) and the risk behavior observed in this study indicate a need for primary programs to prevent the increase of RTI/STI and HIV infections in rural areas.

The clinical and laboratory characteristics of 38 staphylococcal adult bacterial meningitis (ABM) cases (19 Staphylococcus aureus infections and 19 coagulase-negative staphylococcal [CoNS] infections), collected over a period of 6.5 years (July 1999 - December 2005; total ABM cases = 181) were analyzed. The results were compared with those of our previous study (January 1986 - June 1999; total ABM cases = 202: monomicrobial infection = 180, mixed infection = 22, staphylococcal infection = 27). The 38 staphylococcal meningitis cases were 21 men and 17 women. Fever and altered consciousness were the leading clinical manifestations. A preceding postneurosurgical state was noted in 12 of the 19 S. aureus infections and all 19 CoNS infections. The ages of onset and mortality rates of S. aureus infection and CoNS infection were 58.21 ± 13.05 years and 36.8% (7/19), and 44.16 ± 17.57 and 5.3% (1/19), respectively. Eleven of the 19 implicated S. aureus strains and 18 of the 19 implicated CoNS strains were mecA gene-positive and methicillin-resistant; all the strains retained their susceptibility to linezolid. The therapeutic results showed a mortality rate of 21% (8/38). This study revealed an increase of methicillin-resistant, postneurosurgical staphylococcal infection in ABM. Patients with CoNS infection had a younger age at onset and a lower mortality rate.

Acute and convalescent plasma samples were obtained from 101 confirmed primary dengue cases: 48 cases infected with dengue virus type 1, 10 cases with type 2, 42 cases with type 3 and one case with type 4. The hemagglutination inhibition (HI) titers of individual samples were at levels similar to each of the 4 dengue viruses at both the acute and convalescent stages, irrespective of the dengue virus that infected the patients. The results indicate that HI antibodies to dengue viruses are cross-reactive. When an HI test is used as a diagnostic test for dengue virus infection, the cross-reactive nature needs to be considered when interpreting the results.

This paper reports the strategy and effectiveness of an emergency control program conducted in Pingtung City, Taiwan in response to dengue outbreaks. In our control strategy, we carried out 3 insecticide space sprays with an interval of 6 - 7 days and 2 source reductions to cover the entire duration of dengue virus exposure in humans and mosquito vectors. The control effect was demonstrated by a significant reduction in the Breteau (51.1%) and larval (80.0%) indices, but no such effect was demonstrated by alterations in the adult index (54.9%), house index (45.0%), container index (33.8%), or by indoor (15.8%), outdoor (31.2%), or total water-filled containers (22.7%) per 100 premises examined. The contribution made by the reduction in the number of positive containers was primarily in the outdoor (77.2%), and not the indoor containers (–6.0%). This reduction attributed to an overall reduction of 96.0% Aedes albopictus larvae and 71.0% Aedes aegypti. Therefore, 4 weeks after this extensive emergency control measure, the number of dengue cases dropped to one. Finally, due to both the decrease in temperature resulting from the upcoming winter, and to the sustained effort toward source reduction, the transmission cycle of DENV-2 in Pingtung City was interrupted at the beginning of 2003, and no additional cases were identified in late 2003.

The aim of the present study was to determine in vitro phospholipase and protease activities in 122 Candida spp. isolated from several anatomically distinct sites of healthy adults. C. albicans (66.4%) was the most frequently isolated Candida spp. C. glabrata (7.3%), C. tropicalis (6.3%) and C. kefyr (4.9%) were the most frequently isolated non-C. albicans Candida spp. Fifty (40.9%) of the isolates examined were phospholipase positive and 64 (52.4%) were protease positive. Forty-three (53.8%) of the C. albicans isolates tested were phospholipase producers-however, only a few strains of non-C. albicans Candida spp. behaved in the same way. Protease activity was detected in 46 (56.7%) of the C. albicans strains tested and in a few strains of non-C. albicans Candida spp. The levels of phospholipase and protease activities in commensal isolates were found to be lower than the levels of other enzyme activities previously reported in clinical Candida spp. isolates. The phospholipase activity of Candida spp. was found to be higher in oral (59.0%) and fecal (42.8%) isolates. The protease activity of Candida spp. was found to be higher in urogenital (55.1%) and skin (58.8%) isolates. We conclude that further investigations will be needed on the phospholipase and protease activity of Candida spp. in healthy subjects in order to clarify their contribution to fungal virulence.

The aims of this study were to illustrate the prevalence and determinants of mutations associated with antiretroviral drug resistance in a group of antiretroviral-naïve and treatment-experienced patients in Thailand, where antiretroviral drugs are widely used. One hundred and thirteen treatment-naïve (92 CRF01_AE and 21 subtype B patients) and 1,709 treatment-experienced patients were recruited. Genotypic resistance to antiretroviral drugs was studied by sequencing the isolated viruses. Mutation frequencies in treatment-naïve patients were reported along with those for treatment-experienced patients. The results showed that all of the patients with treatment experience showed the same pattern of genotypic resistance. The results also showed that only 14 drug-naïve patients (12.4%) carried HIV-1, with at least one drug-resistant mutation. Moreover, four drug-naïve patients were found to carry the marker mutations for transmission of drug resistance. The most commonly found marker in drug-naïve patients was M36I/V/L (n = 90, 81.1%), which is a common natural polymorphism among HIV-1 subtype CRF01_AE individuals. In order to prevent the rapid emergence of resistant virus strains, a national program to monitor antiretroviral drug resistance should be established. We also recommend routine genotypic testing in treatment-naïve patients before starting antiretroviral therapy to prevent subtherapeutic response and viral failure.

On 1,374 food-producing animal farms in 2001 - 2004, we investigated the influence of antimicrobial usage within 6 months before a survey conducted on antimicrobial resistance in Campylobacter. Tetracyclines and fluoroquinolones were used for therapeutic purposes on 107 and 21 farms, respectively. Oxytetracycline-resistant C. jejuni and C. coli were found in 57.1% (8/14) and 92.5% (37/40) of the farms, respectively, when the animals were treated with tetracyclines. However, they were found in 43.2% (112/259) and 74.3% (122/164) of the farms, respectively, when the animals were not treated with tetracyclines. Enrofloxacin-resistant C. jejuni and C. coli were found in 66.7% (2/3) and 16.7% (1/6) of the farms, respectively, when the animals were treated with fluoroquinolones. However, they were found in 15.5% (42/270) and 28.8% (57/198) of the farms, respectively, when the animals were not treated with fluoroquinolones. The percentage of oxytetracycline resistance in C. coli was significantly elevated when the animals were treated with tetracyclines (P < 0.05). The farms on which antimicrobials included by their resistance pattern were used accounted for 9.1% (4/44) and 24.1% (14/58) of the farms where enrofloxacin-resistant C. jejuni and C. coli were isolated, respectively. It is likely that several factors account for the presence of fluoroquinolone-resistant Campylobacter.

We analyzed the resistance to expanded-spectrum cephalosporins of an Enterobacter cloacae clinical isolate, EC002, by transconjugation, isoelectric-focusing analysis, and cloning experiments. It produced two β-lactamases with isoelectric point values of 5.4 and 8.7, corresponding to TEM-141, a novel variant of TEM-1, and CTX-M-22, encoded by a transferable plasmid.

The examination of archival samples by molecular techniques has been found to be a valuable tool in providing retrospective and epidemiological data. In contrast to DNA, there are fewer comprehensive studies quantitatively addressing the feasibility of assessing RNA from archival samples. In tropical countries like India, such studies are even more rare. One problem is that the warm climate and the fluctuations in the ambient temperature during transport and storage of tissues lead to early decomposition of the tissues. The present study was carried out to evaluate the usefulness of reverse transcriptase-polymerase chain reaction (RT-PCR) in unfixed archival samples to assess whether a retrospective diagnosis of human rabies could be made from archival brain samples from patients suspected to have died of rabies. A nested RT-PCR was performed on 6 rabies confirmed and 8 rabies negative brain tissue samples. Rabies viral RNA could be detected in all 6 positive brain tissue samples and was absent in all mouse inoculation test negative samples. None of the control samples revealed cross-reaction with rabies primers, indicating its specificity. Our results demonstrate the importance of RT-PCR in the detection of rabies virus RNA in 5- to 6-year-old preserved samples without substantial loss. The study proves the role of retrospective diagnosis by RT-PCR in archival autopsy material and decomposed tissues and paves the path for further molecular epidemiologic and phylogenetic studies.

Geotrichosis is an uncommon fungal infection. Geotrichum capitatum is commonly acknowledged as an opportunistic fungal pathogen that causes systemic geotrichosis in immunocompromised patients, especially patients with acute leukemia and severe neutropenia. Here, we report a case of oral geotrichosis caused by G. capitatum in an old patient with no hematological malignancies. Fungal cells were detected in clinical specimens obtained with oral swabs using the KOH technique. Yeast colonies with peripheral hairs were exclusively isolated as fungi from the oral mucosa and feces of the patient. The isolates were identified as G. capitatum by morphological findings, sugar-assimilation tests, and the nucleotide sequences of the ITS regions of the rDNA. Effective treatment of the patient was achieved with amphotericin B syrup in accord with the results of in vitro susceptibility tests. G. capitatum should be recognized as a fungal pathogen involved in superficial infections of older persons, as should Candida spp., even in the absence of hematological malignancies.

We evaluated the in vitro activity of ketoconazole (KET), fluconazole (FLU), amphotericin B (AmpB), and flucytosine (FCU) in comparison to voriconazole (VOR) as a triazole derivative and caspofungin (CAS) as an echinocandin against 114 Candida spp. isolated from different cultures (blood, urine, sputum). The most common species of identified Candida were C. albicans (88), followed by C. parapsilosis (8), C. glabrata (7), C. tropicalis (6), C. famata (2), C. kefyr (2), and C. sake (1). The Clinical and Laboratory Standards Institute M 27-A method was used to evaluate the activity of antifungal agents. The minimal inhibitory concentrations of the strains were evaluated by RPMI 1640 medium using a microdilution method. Of 114 isolates, 100% were sensitive to AmpB, VOR, and CAS, 1.75% showed intermediate resistant to FCU also 0.87% showed intermediate resistant to FLU, and 2.63% were fully resistant to FLU and FCU. These results suggest that KET, AmpB, CAS, and VOR demonstrated excellent activity against all Candida spp. Taken together; these antifungal agents should be effective in the treatment of a broad range of Candida infections.

The recent identification of several atypical cases of bovine spongiform encephalopathy (BSE) has raised the possibility of the existence of distinct strains of BSE agents, arguing against the previous notion that BSE is caused by a single strain. To date, at least two atypical types (L and H) of agent have been reported based on the molecular sizes of the proteinase K-resistant forms of prion protein (PrP^Sc). These atypical agents were identified first in Japan, Italy, France, and Germany, and later in other European countries. Here, we have identified a case of BSE in a 169-month-old cow (designated as BSE/JP24), in which predominant deposition of the mono-glycosylated form of PrP^Sc was observed by Western-blot analysis, and plaques of PrP^Sc were detected in the brain by immunohistochemical analysis. The glycoform ratio of PrP^Sc was different from that of the typical BSE agent, in which the di-glycosylated form is dominant; instead, the ratio resembled that of type-2 human sporadic Creutzfeldt-Jakob disease and that reported for the L-type BSE. The characteristic glycoform ratio and plaques of PrP^Sc suggested that the agent in BSE/JP24 was relevant, if not identical, to the agent in bovine amyloidotic spongiform encephalopathy (BASE), an L-type BSE identified in Italy. It was of interest that at the level of the obex, the medulla oblongata was devoid of plaques of PrP^Sc, and a pathological phenotype similar to that of typical BSE specimens with vacuolations and coarse granular/linear deposition of PrP^Sc were observed.

Serum specimens were collected from 99 wild boars in the Northern area of the main Okinawa Island and from 27 wild boars on Iriomote Island in Okinawa Prefecture from 1997 to 2005. Sera were tested for Japanese encephalitis virus (JEV) antibody by hemagglutination inhibition assay and IgG enzyme-linked immunosorbent assay. Sixty-four samples (64.6%) in the Northern area and 1 sample (3.7%) from Iriomote Island were positive for the JEV antibody. The difference in seroprevalence between the Northern area and Iriomote Island was statistically significant (P < 0.01, χ² test). This difference may be due to the lack of a pig farm on Iriomote Island, whereas wild boars in the Northern area may be infected with JEV, amplified on pig farms. It is likely that there has recently been an increase in the number of wild boars living close to humans in certain areas of Japan. This in turn increases the possibility that wild boars are infected with JEV, which is amplified on pig farms, and these infected animals may play a role in carrying JEV to other regions of the country.

Staphylococcus lugdunensis is an unusually virulent coagulase-negative staphylococcus (CoNS) that can cause many types of infection. All culture specimens were collected from patients at Inje University Busan Paik Hospital between October 2005 and March 2006. S. lugdunensis was identified using the phenotypic biochemical tests and 16S rDNA sequencing. Among 358 CoNS, three strains were identified as S. lugdunensis. All three isolates showed positive results in the clumping factor test, but the L-pyrrolidonyl-beta-naphthylamide hydrolysis test was positive in only one and the ornithine decarboxylase test in two. Two of the three isolates were correctly identified by API Staph, but none of them was identified correctly by the Vitek I system. All three strains were penicillin resistant secondary to beta-lactamase production. S. lugdunensis was an unrecognized but infrequent cause of infection.

Although shigellosis is a potentially fatal disease that may cause a number of extra-intestinal manifestations, intractable septic shock is an unusual complication. Here we describe a 6-month-old infant who developed severe septic shock and convulsions during an episode of dysentery caused by multidrug-resistant Shigella dysenteriae. The case presentation demonstrates how shigellosis can lead to rare and potentially misleading complications such as septic shock when not treated adequately and promptly.

Bovine spongiform encephalopathy (BSE) is caused by a prion that primarily consists of an abnormal isoform of the prion protein (PrP^Sc). Since PrP^Sc is partially resistant to proteolytic digestion, the routine diagnosis of BSE is based on the immunological detection of the proteinase K (PK)-resistant moiety of PrP^Sc (PrP^core). However, transmission studies are indispensable in order to demonstrate prion infectivity and to analyze prion characteristics. Transmission experiments were accordingly performed on 2 young BSE cases (BSE/JP8, BSE/JP9) and 1 suspected BSE case (Suspended-1) that were detected by the BSE screening program in Japan. In this study, we attempted to transmit the prion from these 3 animals by using transgenic mice overexpressing bovine PrP (TgBoPrP). In spite of the use of BSE-sensitive transgenic mice, none of the mice developed neurological signs nor accumulated PrP^Sc in their brains for more than 600 days post-inoculation, even with subsequent blind passages. The results of a dilution experiment using the classical BSE prion indicated that prion infectivity in these 3 cattle was below the detection limit of 10^3.0 LD₅₀/g.