Ramazan Gümral

Lack of Candida africana and Candida dubliniensis in Vaginal Candida albicans Isolates in Turkey Using HWP1 Gene Polymorphisms

Candida africana differs from the common strains of C. albicans and C. dubliniensis morphologically, physiologically, genetically, and, in particular, clinically. This fungal pathogen is primarily recovered from genital specimens, especially in vaginal specimens. In this investigation, we reexamined 195 vaginal C. albicans isolates for the presence of C. africana and C. dubliniensis by using hyphal wall protein 1 (HWP1) gene polymorphisms. All study isolates were confirmed to be C. albicans, and none were verified as either C. africana or C. dubliniensis. In conclusion, the HWP1 gene polymorphisms offer a useful tool in the discrimination of C. africana, C. albicans, and C. dubliniensis. Further studies may highlight the pathogenesis and importance of this yeast in vulvovaginal candidiasis.

Trichophyton tonsurans scalp carriage among wrestlers in a national competition in Turkey.

Trichophyton tonsurans tinea gladiatorum is an emerging epidemic among combat-sport athletes across the globe. In the present study, we investigated the prevalence of symptomatic and asymptomatic dermatophytic infections among wrestlers in the National Greco-Roman Championship in Turkey. In total, 194 wrestlers from 32 provinces and 72 clubs were examined for scalp, trunk, groin, and toe web dermatophytic infections. We also administered a questionnaire to obtain information on the participants’ lifestyles, wrestling characteristics, and risk factors for dermatophytic infections. The hairbrush method was used for scalp and trunk sampling, whereas a cotton swab was used for groin, toe web, and mat sampling. Three wrestling mats in the gymnasium were surveyed for dermatophytes using the touch preparation method. A total of 17 (8.8%) wrestlers harbored dermatophytes, and 22 strains were isolated: 13 (59.1%) T. tonsurans and 9 (40.9%) T. rubrum. These isolates were found on the scalp (8), trunk (2), forearm (1), hand (1), groin (3), and feet (7). In addition, we recovered 8 dermatophyte strains from the 150 mat samples (5.3%): T. rubrum in 6 samples (75%) and T. tonsurans in two samples (25%). T. tonsurans was only recovered from 11 out of 194 (5.7%) wrestlers. Scalp carriage represents the predominant (72.7%) clinical picture of a T. tonsurans infection in these Greco-Roman wrestlers in Turkey.

Detection of major HPVs by a new multiplex real-time PCR assay using type-specific primers.

In this study, we aimed to develop a cost-effective, practical, and sensitive method to be used for the diagnosis of HPV infections. The presence of HPV-DNA was investigated in cervical smear samples using three different methods: MY09/11 consensus PCR, TaqMan-based type-specific real-time PCR, and SYBR Green-based multiplex PCR. Of the 315 samples, 21.6% (68/315) were HPV-DNA positive by using at least one of the three methods. The relative sensitivities of MY09/11 PCR, type-specific PCR, and multiplex PCR were found to be 86.8% (59/68), 91.2% (62/68), and 91.2% (62/68), respectively. Genotyping analyses were successfully carried out in 62 of 68 HPV-DNA positive samples, and 77 isolates (8 low-risk and 69 high-risk HPV) were identified, while six samples were determined to be positive by consensus PCR only and could not be genotyped. The type distribution of the 69 high-risk HPV strains was as follows: 37.7% HPV 16, 13.0% HPV 52, 11.6% HPV 58, 7.2% HPV 18, 7.2% HPV 31, 7.2% HPV 68, 4.3% HPV 35, 4.3% HPV 39, 4.3% HPV 82, 1.4% HPV 33, and 1.4% HPV 45. Our data suggests that the diagnosis of HPV infections using only consensus PCR may lead to epidemiologically significant data loss, and that our multiplex PCR is more sensitive than consensus PCR and lower in cost than the type-specific PCR. We believe that the SYBR Green-based multiplex PCR may be useful and cost-effective for other microbiological fields. In addition, type-specific screening of HPV-DNA gives more reliable results, but it may also be used in combination with consensus PCR if the type spectrum of the test is not large enough.

Epidemiology of dermatophytosis in junior combat and non-combat sports participants.

Participation in competitive sports is popular and widely encouraged worldwide. Herein, we investigated 252 male and 67 female sports players, aged 16.4 ± 1.3 years, active in 15 different types of combat (n = 143) and non‐combat (n = 176) sports. Of the 319 participants in this study, 11 (3.5%) players, including six wrestlers, four football players and one handball player, all of whom were men, harboured dermatophytic fungi. Briefly, Trichophyton tonsurans was present in three athletes, who were scalp carriers of the fungus. Furthermore, T. rubrum (4), T. interdigitale (3) and Arthroderma simii (1) were recovered from eight participants with tinea inguinalis (4), tinea pedis (2) or both (1). One patient was a trunk carrier of concomitant tinea pedis. All dermatophytic fungi were identified using both direction sequence of the rDNA regions spanning the internal transcribed spacers (ITS1 and ITS2) and 5.8 rRNA gene. Although sports‐active individuals are active and sweat more, we observed a low prevalence of dermatophytosis, both in combat (5.2%) and non‐combat sports participants (3.4%) (P > 0.05). However, dermatophyte infections require more attention and appropriate management to eradicate the infection and to prevent possible outbreaks. This study also documents the first case of zoophilic A. simii in Turkey.

Species Distribution and in Vitro Susceptibility of Candida Bloodstream Isolates to Six New and Current Antifungal Agents in a Turkish Tertiary Care Military Hospital, Recovered Through 2001 and 2006

OBJECTIVE Species distribution and antifungal susceptibility of Candida bloodstream isolates. METHODS 173 Candida species recovered from fungemic patients admitted to the largest tertiary-care military hospital between 2001 and 2006 in Ankara, Turkey evaluated retrospectively. Antifungal susceptibility of 95 isolates (45 Candida parapsilosis, 35 C. albicans, 7 C. tropicalis, 4 C. krusei, 3 C. glabrata, and 1 C. kefyr) was determined against fluconazole, itraconazole, voriconazole, posaconazole, caspofungin, and amphotericin B. RESULTS C. albicans was the most common yeast (48.0%), followed by C. parapsilosis. Almost all strains showed low minimal inhibitory concentration (MIC) values to all six antifungals tested. Only 2 of 45 C. parapsilosis isolates were resistant to fluconazole, one was susceptible in a dose-dependent manner (SDD) to itraconazole, and 14 were nonsusceptible to caspofungin. CONCLUSIONS Increasing prevalence of C. parapsilosis emphasizes a need for better catheter care and continuous surveillance programs.

Coexistence of Epstein-Barr virus and Parvovirus B19 in tonsillar tissue samples: quantitative measurement by real-time PCR.

OBJECTIVE In this study, we aimed to investigate the presence and copy number of six different viruses in tonsillar tissue samples removed surgically because of chronic recurrent tonsillitis or chronic obstructive tonsillar hypertrophy. METHODS In total, 56 tissue samples (tonsillar core) collected from 44 children and 12 adults were included in this study. The presence of viruses was investigated using a new TaqMan-based quantitative real-time PCR assay. RESULTS Of the 56 tissue samples, 67.9% (38/56) were positive for at least one of the six viruses. Epstein-Barr virus was the most frequently detected virus, being found in 53.6% (30/56), followed by human Parvovirus B19 21.4% (12/56), human adenovirus 12.5% (7/56), human Cytomegalovirus 5.4% (3/56), BK polyomavirus 1.8% (1/56), and Herpes simplex virus 1.8% (1/56). Precancerous or cancerous changes were not detected in the tonsillar tissue samples by pathologic examination, whereas lymphoid hyperplasia was observed in 24 patients. In contrast to other viruses, B19 virus was present in high copy number in tonsillar tissues. The rates of EBV and B19 virus with high copy number (>500.000 copies/ml) were higher in children than in adults, and a positive relationship was also found between the presence of EBV and the presence of B19 virus with high copy number (P=0.037). CONCLUSIONS It is previously reported that some viral agents are associated with different chronic tonsillar pathologies. In the present study, the presence of B19 virus in tonsillar core samples was investigated quantitatively for the first time, and our data suggests that EBV infections could be associated with B19 virus infections or could facilitate B19 virus replication. However, further detailed studies are needed to clarify this observation.

Efficiency of MY09/11 consensus PCR in the detection of multiple HPV infections

Human papillomavirus (HPV) DNA testing has become an important component of cervical cancer screening programs. In this study, we aimed to evaluate the efficiency of MY09/11 consensus polymerase chain reaction (PCR) for the detection of multiple HPV infections. For this purpose, MY09/11 PCR was compared to an original TaqMan-based type-specific real-time PCR assay, which can detect 20 different HPV types. Of the 654 samples, 34.1% (223/654) were HPV DNA positive according to at least one method. The relative sensitivities of MY09/11 PCR and type-specific PCR were 80.7% (180/223) and 97.8% (218/223), respectively. In all, 352 different HPV isolates (66 low-risk and 286 high-risk or probable high-risk types) were identified in 218 samples, but 5 samples, which were positive by consensus PCR only, could not be genotyped. The distribution of the 286 high-risk or probable high-risk HPVs were as follows: 24.5% HPV-16, 8.4% HPV-52, 7.7% HPV-51, 6.3% HPV-39, 6.3% HPV-82, 5.6% HPV-35, 5.6% HPV-58, 5.6% HPV-66, 5.2% HPV-18, 5.2% HPV-68, and 19.6% the other 8 types. A single HPV type was detected in 57.3% (125/218) of the genotyped samples, and multiple HPV types were found in the remaining 42.7% (93/218). The false-negative rates of MY09/11 PCR were found to be 17.4% in single infections, 23.3% in multiple infections, and 34.6% in multiple infections that contained 3 or more HPV types, with the condition that the low-risk types HPV-6 and HPV-11 be considered as a monotype. These data suggest that broad-range PCR assays may lead to significant data loss and that type-specific PCR assays can provide accurate and reliable results during cervical cancer screening.

Candida vaginitis during contraceptive use: the influence of methods, antifungal susceptibility and virulence patterns.

No consensus exists about whether contraceptives cause an increased risk of vaginitis, including vulvovaginal candidosis (VVC). We investigated 495 women (252 who used contraceptives; 243 who did not) for the presence of VVC. Antifungal susceptibility testing was performed for five antifungal agents and for boric acid, and three virulence factors were also examined. We recovered 129 (26.1%) monofungal populations from vaginal samples of women with acute VVC (AVVC, n = 18), symptomatic recurrent VVC (RVVC, n = 22) and asymptomatic RVVC (n = 28), as well as of other contraceptive users who carried Candida in their vaginas (n = 61). It is important to note that the women who had VVC used the same contraceptive methods (p > 0.05). Candida albicans was the most common species isolated (45%), followed by C. glabrata (40.3%). Most of the vaginal yeast isolates exhibited low minimum inhibitory concentration levels for the five antifungals tested. However, this was not the case for boric acid. In addition, the yeast fungi that was derived from the AVVC and RVVC patients showed higher amounts of haemolytic activity than the yeast fungi found among the controls (p < 0.05). The use of contraception does not predispose women to VVC (p > 0.05). Also, both host- and organism-related factors were required to achieve optimal clinical treatment for VVC.

A case of rhinoorbital mucormycosis in a leukemic patient with a literature review from Turkey.

Mucormycosis (Zygomycosis) is a rare, invasive, opportunistic fungal infection of the paranasal sinuses, caused by a fungus of the order Mucorales. We report a case of rhinoorbital mucormycosis caused by Rhizopusoryzae in an acute lymphoblastic leukemia patient and review the 79 Mucormycosis cases reported in the last decade from Turkey. In our case, the diagnosis was made with endoscopic appearance, computerized tomography of the paranasal sinuses, and culture of the surgical materials. Following aggressive surgical debridement and parenteral amphotericin B therapy, the patient recovered completely. In Turkish literature, rhinocerebral manifestations were the most common form of the mucormycosis (64 cases), followed by pulmonary form (6 cases). The most common risk factor was hematologic malignancies (32 cases) and diabetes mellitus (32 cases), similar to those reported from the rest of the world. The etiologic agents responsible for the review cases were Rhizopus sp., Mucor spp., Rhizomucor spp., Rhizopusoryzae, Mucorcircinelloides, and Lichtheimia corymbifera. Although various treatment modalities were used, amphotericin B was the mainstay of therapy. Mortality rate was found to be 49.4% in review cases. It seems that strong clinical suspicion and early diagnosis, along with aggressive antifungal therapy and endoscopic sinus surgery, have great importance for better prognosis in mucormycosis.

Haemolytic and co-haemolytic (CAMP-like) activity in dermatophytes

Dermatophytes are some of the most common fungal pathogens in both humans and animals. These fungi release enzymes (e.g., keratinases) that play roles in their pathogenesis. Little is known about their haemolytic and co‐haemolytic (CAMP‐like) activities; however, in bacteria, these components play significant roles in pathogenesis. This study characterised these two factors in 45 dermatophyte strains (representing the genera Arthroderma, Epidermophyton, Microsporum and Trichophyton) using Columbia agar (CA) supplemented with 5% bovine, ovine and equine erythrocytes. Haemolysis was best observed on CA supplemented with ovine erythrocytes followed by equine and bovine erythrocytes, while CAMP‐like reactions occurred using bovine and ovine but not equine erythrocytes. Haemolytic and CAMP‐like activities were best observed using ovine and bovine erythrocytes in CA in 44 and 38 strains at 7 and 3 days respectively. Most dermatophytes recovered from both symptomatic and asymptomatic lesions had haemolytic and CAMP‐like activities. We suggest that the haemolytic and CAMP‐like activities are not correlated with ecological characteristics, isolation sites or clinical manifestations of dermatophytic fungi. We also believe that this study has the potential to contribute to the existing literature on dermatophytes and dermatophyte pathogenesis.