Crimean-Congo hemorrhagic fever in European part of Turkey: genetic analysis of the virus strains from ticks and a seroepidemiological study in humans.

A survey of ticks from domestic ruminants, together with a serosurvey in humans was conducted in Thrace (northwestern Turkey) to evaluate the prevalence of Crimean-Congo hemorrhagic fever virus (CCHFV) in ticks and humans. More prevalent ticks were Hyalomma marginatum, Hyalomma aegyptium, Rhipicephalus bursa, and Rhipicephalus (Boophilus) annulatus, with low numbers of Dermacentor marginatus, Rhipicephalus sanguineus group, and Ixodes ricinus. No differences in the tick faunal composition were found among surveyed provinces. CCHFV was detected using specific primers for strains belonging to both Europe 1 and Europe 2 clades in a total of 15 pools of ticks collected in nine localities. The maximum likelihood estimate of infection rate was calculated as 0.72/100 ticks (95% CI = 0.42-1.16). Viral RNA was observed only in H. marginatum, R.(B.) annulatus, and R. bursa with overall maximum likelihood estimate infection rates being 0.93 (95% CI = 0.35-2.05), 0.74 (95% CI = 0.24-1.78), and 1.67 (95% CI = 0.69-3.46), respectively. The surveyed region is the only place where both viral strains are circulating together in nature in Turkey. Results from serosurvey on 193 samples from three localities in the region showed that immunoglobulin M and immunoglobulin G rates are compatible with an epidemiological situation in which the virus has been present for a long time and is not the result of a recent invasive event from the main epidemic center in Anatolia (north-central Turkey). Seropositivity rates cannot be compared against the tick faunal composition, because of the homogeneity in the results about tick surveys. The high rate of seropositivity, and the prevalence of CCHFV in both Europe 1 and 2 clades among the ticks, but few clinical cases suggest that the circulation of both viral strains may confer protection against the CCHFV infection.

[Detection of human bocavirus DNA by polymerase chain reaction in children and adults with acute respiratory tract infections]. / Akut solunum yolu enfeksiyonlu çocuk ve eriskinlerde insan bokavirus DNA'sinin polimeraz zincir reaksiyonu ile saptanmasi

Human bocavirus (HBoV) which was described in 2005 by molecular techniques, is a member of Parvoviridae. The role of HBoV is being questioned in acute respiratory diseases (ARD) in many recent studies. The aim of this study was to determine the presence of HBoV DNA in the respiratory specimens of patients with ARD. A total of 155 throat swab and/or washing specimens from 76 children and 79 adults with ARD were examined. HBoV DNA was investigated by single step in-house polymerase chain reaction (PCR) using NS1 primers (5-'TATGGCCAAGGCAATCGTCCAAG-3', 5'-GCC GCGTGAACATGAGAAA-CAG-3') which amplify the 290 base pair region of NS1 gene located between nucleotides 1545-1835 of prototype HBoV st1 strain. HBoV DNA was detected in 5 (6.5%) of 76 children and 2 (2.5%) of 79 adults. Three sequenced samples showed 100% homology with the reference sequences. This study in which HBoV DNA was detected in children and adults with ARD, is the first HBoV prevalence study in Turkey. Larger scale prospective clinical and molecular studies are required to explain the association between HBoV and respiratory disease.

A dark strain in the Fusarium solani species complex isolated from primary subcutaneous sporotrichioid lesions associated with traumatic inoculation via a rose bush thorn.

Fusarium species are hyaline hyphomycetes widely distributed in nature and documented agents of both superficial and systemic infections in humans. In this paper, we report a darkly-pigmented and initially non-sporulating isolate in the Fusarium solani species complex (FSSC) causing a post-traumatic sporotrichoid infection in an otherwise healthy, male patient. Sequencing of multiple loci showed that the isolate represented an otherwise unknown lineage, possibly corresponding to a separate species, within the multi-species F. solani complex. In prolonged culture, the non-sporulating isolate produced revertant wild-type subcultures with typical Fusarium conidiation. This suggests that the original dense, dark, non-sporulating isolate was a host-adapted form selected in vivo for characters compatible with human pathogenicity. The production of such forms by Fusarium species is increasingly recognized now that sequencing has allowed the identification of highly atypical isolates. In vitro antifungal susceptibility of the isolate was investigated against seven conventional and two newly approved antifungal agents. The isolate showed in vitro resistance to amphotericin B, but appeared susceptible to itraconazole and terbinafine. A cure was ultimately achieved with combined terbinafine/itraconazole therapy with prolonged itraconazole follow-up therapy.

Lack of Chlamydophila pneumoniae and predominance of Alloiococcus otitidis in middle ear fluids of children with otitis media with effusion.

OBJECTIVE: To investigate the presence of Chlamydophila pneumoniae and other bacterial pathogens in middle ear effusion samples obtained from children with otitis media with effusion (OME). MATERIALS AND METHODS: Twenty-eight children (mean age 7.03; standard deviation 2.18) with OME unresponsive to medical therapy were included in the study. All of the children underwent ventilation tube insertion under general anesthesia. Eighteen patients were bilaterally affected whereas 10 children had unilateral disease. The middle ear fluids (46 samples in total) were collected during ventilation tube insertion, and were evaluated subsequently for the presence of C. pneumoniae and other bacterial pathogens using polymerase chain reaction (PCR). RESULTS: Although all samples were negative for C. pneumoniae, bacterial DNA was detected in 21 of 46 samples. Overall 40% of the patients (4/10) with unilateral involvement, and 61% of the patients (11/18) with bilateral involvement were positive for bacterial DNA. In 6 patients with bilateral OME bilateral samples were positive, whereas 5 patients with bilateral OME showed only unilateral positivity. According to the results of DNA sequencing analysis, all of the positive samples harbored only one bacterial species. In 12 of 46 samples Alloiococcus otitidis DNA (26%), in 7 Haemophilus influenzae DNA (15%), in one Streptococcus pneumoniae DNA (2%) and in one Moraxella catarrhalis DNA (2%) were present. CONCLUSIONS: Our findings support that C. pneumoniae does not seem to have a role in OME in children whereas A. otitidis was found to be more frequent than the other common pathogens. Further studies are required to elucidate the exact pathogenetic role of these microorganisms in OME.

Fatal Trichoderma harzianum infection in a leukemic pediatric patient.

We report the repeated isolation for Trichoderma.harzianum, a rare opportunistic pathogen from three sets of each of the following clinical samples; blood serum, skin lesions, sputum and throat of a pediatric ALL patient with neutropenia. The definition of invasive fungal infection requires evidence of the presence of fungal elements in tissue samples, in addition to the isolation of suspected etiologic agent in culture. However, invasive procedures are not always applicable due to several factors, as for example in our case, the poor general status of the individual patient or thrombocytopenia. The present paper also emphasizes the problems encountered in obtaining appropriate samples and diagnosing invasive fungal disease in immunocompromised patient populations, including those with hematological malignancy. Three cases involving T. harzianum, including this one, have been described thus far in the literature. All were fatal and the fungus was resistant to antifungal therapy. A critical review of the other two cases of Trichoderma infections in humans is provided.

Susceptibility testing of Cryptococcus diffluens against amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole and posaconazole.

Cryptococcus diffluens is a recently re-established species that shares several phenotypic features with Cryptococcus neoformans. We evaluated the application of the Clinical Laboratory Standards Institute (CLSI, formerly NCCLS) macro- and microbroth dilution methods and the E-test agar diffusion method to determine the in vitro susceptibilities of known strains of C. diffluens against amphotericin B (AMB), flucytosine (5-FC), fluconazole (FLC), itraconazole (ITC) and the novel triazoles, voriconazole (VRC) and posaconazole (PSC). Seven strains were found to be resistant in vitro to AMB (MICs >/=2 microg/ml), five were resistant to 5-FC (MICs of >/=32 microg/ml), four were resistant to FLC (MICs of FLC >/=32 microg/ml) and nine were resistant to ITC (MICs of ITC >1 microg/ml). In contrast, VRC and PSC showed good in vitro activity against C.diffluens strains, even those with elevated MICs to amphotericin B and/or established azoles. Most of the isolates were inhibited by 0.5 microg/ml of both VRC and PSC. A clinical isolate showing phenotypic switching exhibited elevated MICs to both agents, i.e., VRC (>16 microg/ml) and PSC (>8 microg/ml).

[Seroprevalence of visceral leishmaniasis in stray dogs in Kocaeli]. / Kocaeli Sokak Köpeklerinde Visseral Leishmaniasis Seroprevalansi.

Dogs are the most important reservoir of visceral leishmaniasis (VL). A male child who lives in Köseköy in Kocaeli was diagnosed with VL. Since this child had never been outside Kocaeli, serum samples of 65 stray dogs were analyzed for canine VL using the indirect fluorescence antibody test (IFAT) and ELISA. Two dogs (3.07%) tested positive with both ELISA and IFAT. Leishmania amastigotes were observed in the lymph aspiration material from one of them. Growth was observed in NNN medium inoculated with lymph aspiration material from the other dog. This was the first study investigating canine VL prevalence in our city and plans were made for control of the disease.

Detection of HHV-8 and HPV in laryngeal carcinoma.

OBJECTIVE: In this study we aimed to investigate the presence of human herpesvirus 8 (HHV-8) and human papillomavirus (HPV) in laryngeal carcinoma. MATERIALS AND METHODS: Fifty patients operated on because of laryngeal carcinoma were included in the study. Forty-seven had squamous cell carcinoma (SCC) whereas three had verrucous carcinoma. Fresh tumoral tissues, or tumoral tissues obtained from archival paraffin-embedded blocks, were examined. HHV-8 DNA and HPV DNA were detected using polymerase chain reaction (PCR) and viral genotypes of HPV were determined via the hybrid capture method. The presence of HHV-8 DNA and HPV DNA were also investigated in normal appearing laryngeal tissue collected from 50 cadavers at autopsy. RESULTS: HPV DNA was detected in seven patients (7/50; 14%) (5 out of 47 patients with SCC (5/47; 10.6%) and two out of three patients with verrucous carcinoma). HHV-8 DNA was detected in five patients and they all had SCC (5/47; 10.6%). One case had both HHV-8 and HPV DNA. None of the control samples from cadavers harbored HHV-8 DNA, or HPV DNA. There was a statistically significant correlation between HHV-8 DNA and HPV DNA positivity and laryngeal SCC (Fisher exact test; p=0.023 for each). No statistically significant correlations were found between the presence of HHV-8 and/or HPV and age, gender, tumor stage, differentiation, the site of the tumor, smoking and alcohol use. CONCLUSIONS: The findings of the present study suggest that beside HPV, HHV-8 might have a role in laryngeal carcinogenesis. Further investigations are necessary to clarify the exact role of these viruses in laryngeal carcinoma.

Subcutaneous cryptococcosis due to Cryptococcus diffluens in a patient with sporotrichoid lesions case report, features of the case isolate and in vitro antifungal susceptibilities.

Environmental fungi, in particular primary pathogens and Cryptococcus spp. can be responsible for skin lesions mimicking sporotrichosis. In this paper, we report a case of subcutaneous cryptococcosis in an apparently healthy, young male patient due to a non-C. neoformans Cryptococcus species, C. diffluens. The isolate showed in vitro phenotypic switching that may affect virulence and host inflammatory and immune responses, and in vitro resistance to amphotericin B and 5-flucytosin. This species shares several phenotypic traits with C. neoformans, and, therefore, decisive diagnosis should be based on biopsy and culturing results followed by molecular identification.

A Rhizopus oryzae strain isolated from resected bone and soft tissue specimens from a sinonasal and palatal mucormycosis case. Report of a case and in vitro experiments of yeastlike cell development.

We report a histologically and mycologically proven sinonasal mucormycosis case causing palatal necrosis in a nondiabetic patient with renal failure. Mycological examination of Giemsa stained imprinted tissue preparations revealed abundant yeast-like cells besides the typical mucoraceous hyphae. The fungus was isolated from surgical specimens and identified as Rhizopus oryzae by phenotypic and genotypic tests. Laboratory studies were performed to investigate the association of the yeast-like cells observed in tissue specimens and the fungus recovered in culture. In vitro induced yeast-like cell development of the case isolate was found under certain growth conditions and documented by photomicrographs.

Cryptococcal parotid involvement: an uncommon localization of Cryptococcus neoformans.

We describe a cryptococcal infection localized in the parotid gland of an otherwise healthy 72-year-old woman. The patient presented with a painful, approximately 4.5 cm diameter mass in the anterior region of her right ear. Her symptoms were mild and uncharacteristic. The patient had previously fallen on her face in her garden, causing the loss and breakage of her dentures. Since the soil of the garden contained chicken droppings, it is quite likely that the oral prothesis became contaminated on contacting the soil. The fungus probably entered the parotid gland through the traumatization of the posterior lateral wall of her oral cavity by her broken denture. Numerous intra- and extracellular cryptococcal yeast cells were observed in both histopathological and mycological slide preparations. The yeastlike fungus was recovered in cultures inoculated with tissue collected through three biopsies of her parotid region. The isolates were identified as Cryptococcus neoformans by classical mycology methods and found to be susceptible, in vitro, to fluconazole, amphotericin B and flucytosine. Fluconazole treatment (400 mg/d, for 6 months) was started and the patients facial swelling resolved and the pain significantly reduced within 5 weeks of the initiation of treatment. While fungal infection of the parotid gland have been reported, to our knowledge, this is the first description of a non-disseminated primary parotid infection due to C. neoformans.

Genetic subtypes of human immunodeficiency virus type 1 (HIV-1) in Istanbul, Turkey.

BACKGROUND: Epidemiological surveillance of HIV-1 subtypes is an important and ongoing element of preparation for global antiviral interventions. OBJECTIVE: To assess the molecular epidemiology of HIV-1 in Istanbul, Turkey. STUDY DESIGN: 27 HIV/AIDS patients were investigated. Data on age, sex, country of birth, and HIV acquisition route were collected. Following amplification with PCR the sequences of the gp41 region of the env gene were determined using a 310 DNA sequencer (ABI prism, Foster City, USA) and phylogenetically analyzed. RESULTS: Among the 27 patients (26 adults and 1 infant), 22 were male, born in Turkey, and 20 infected through heterosexual contact. Two patients acquired the virus through blood and/or blood transfusion and one infant by vertical transmission. The distribution of the subtypes was as follows: four were subtype A, 19 subtype B, one subtype C, one subtype D, and two subtype F1. According to our results, although the B subtype is still predominant, non-B subtypes are also present, even though the number of registered HIV/AIDS patients is low. CONCLUSION: These are the first subtyped HIV-1 strains in Turkey where a low level of HIV prevalence has been observed since the first reported case in 1985. These findings and Turkey's specific geographic localization indicate the need for a nationwide surveillance to detect all subtypes including the new recombinant ones.

The value of Gram staining of catheter segments for rapid detection of peripheral venous catheter infections.

Catheter-related bloodstream infections are common in hospitalized patients. Peripheral venous catheters (PVCs) are the most frequently used vascular access devices with usually low reported infection rates. We conducted a study to find the rate of PVC-related infections using semiquantitative (roll plate) and quantitative catheter culture techniques. We found significant growth in 9.5% of the PVCs by quantitative culture, which was predictive of a BSI in 43% of cases, which seems to be the major limitation of the method. We conclude that Gram stain is a fast and reliable diagnostic tool for rapid detection of PVC infection.

Assessment of antibiotic susceptibility of Ureaplasma urealyticum from prostitutes and outpatient clinic patients using the E-test and agar dilution method.

In this study, a total of 647 vaginal discharge samples were examined. Ureaplasma urealyticum growth was seen in 68 samples (10.5%). The antibiotic sensitivity of 30 types of U.urealyticum was determined with the E-test and agar dilution method. With the agar dilution method, all types were sensitive to ciprofloxacin and ofloxacin (MIC 0.94 microg/ml), tetracycline (MIC 0.125 microg/ml) and doxycycline (MIC 0.125 and 0.190 microg/ml). Furthermore, with the agar dilution method, 18 types (60%) were resistant to roxithromycin and 12 (40%) were sensitive (MIC 12 microg/ml); 3 types (10%) were resistant to erythromycin and 27 (90%) were sensitive (MIC 12 microg/ml); 9 types (30%) were resistant to clarithromycin and 21 (70%) were sensitive (MIC 12 microg/ml), and all types were sensitive to azithromycin (MIC 14 microg/ml).