Cryptococcus neoformans Recovered From Olive Trees (Olea europaea) in Turkey Reveal Allopatry With African and South American Lineages.

Cryptococcus species are life-threatening human fungal pathogens that cause cryptococcal meningoencephalitis in both immunocompromised and healthy hosts. The natural environmental niches of Cryptococcus include pigeon (Columba livia) guano, soil, and a variety of tree species such as Eucalyptus camaldulensis, Ceratonia siliqua, Platanus orientalis, and Pinus spp. Genetic and genomic studies of extensive sample collections have provided insights into the population distribution and composition of different Cryptococcus species in geographic regions around the world. However, few such studies examined Cryptococcus in Turkey. We sampled 388 Olea europaea (olive) and 132 E. camaldulensis trees from seven locations in coastal and inland areas of the Aegean region of Anatolian Turkey in September 2016 to investigate the distribution and genetic diversity present in the natural Cryptococcus population. We isolated 84 Cryptococcus neoformans strains (83 MATα and 1 MATa) and 3 Cryptococcus deneoformans strains (all MATα) from 87 (22.4% of surveyed) O. europaea trees; a total of 32 C. neoformans strains were isolated from 32 (24.2%) of the E. camaldulensis trees, all of which were MATα. A statistically significant difference was observed in the frequency of C. neoformans isolation between coastal and inland areas (P < 0.05). Interestingly, the MATaC. neoformans isolate was fertile in laboratory crosses with VNI and VNB MATα tester strains and produced robust hyphae, basidia, and basidiospores, thus suggesting potential sexual reproduction in the natural population. Sequencing analyses of the URA5 gene identified at least five different genotypes among the isolates. Population genetics and genomic analyses revealed that most of the isolates in Turkey belong to the VNBII lineage of C. neoformans, which is predominantly found in southern Africa; these isolates are part of a distinct minor clade within VNBII that includes several isolates from Zambia and Brazil. Our study provides insights into the geographic distribution of different C. neoformans lineages in the Mediterranean region and highlights the need for wider geographic sampling to gain a better understanding of the natural habitats, migration, epidemiology, and evolution of this important human fungal pathogen.

[Prevalence and genotype distribution of human papillomavirus in patients attending to gynecology polyclinics]. / Jinekoloji poliklinigine basvuran hastalarda insan papillomavirüs prevalansi ve genotip dagilimi.

Human papillomavirus (HPV) infection is the most significant risk factor of the development of cervical cancer. The distribution of HPV prevalence and genotype varies widely between regions. In this study, it was aimed to investigate the prevalence and genotype distribution of HPV, retrospectively. One thousand one hundred and seventy patients who applied to the department of obstetrics and gynecology were included in this study. Samples were collected from patients for cervical HPV DNA and Pap smear. The Pap samples taken for Pap smear were fixed with alcohol and analyzed according to the modified Bethesda system. HPV identification and typing were performed using the "Linear Array HPV Genotyping Test (Roche Molecular System, USA)". Patients were divided into 5 groups due to their age. Total HPV ratio was most frequently found among the patients who were between 17-30 years old, while HR-HPV was most frequently found between 51-60 years. Nine hundred seventy-eight of 1170 (83.6%) patients had normal cytologic findings whereas 192 (16.4%) had abnormal cytologic findings. HPV was detected in 37.2% of the total patients. high-risk HPV (HR-HPV) rate was 21.2%, probable high risk (PR-HPV) rate was 6.4% and low risk HPV (LR-HPV) rate was 9.5%. When the relationship between cytologic findings and HPV was examined, normal cytology/HPV negative 67.8%; abnormal cytology/HPV negative 37.5%, normal cytology/HPV positive 32.2%, abnormal cytology/HPV positive 62.5% were detected. The highest prevalence of HPV was 8.9% with HPV 16, followed by 6, 53 and 52/53/35/58. A total of 354 patients were biopsied, 177 of whom were normal, 111 of whom were cervical intraepithelial neoplasia (CIN) 1, 66 of whom were CIN 2 and over. In the group with normal pathological findings, HR-HPV ratio was found as 15.8%, while in CIN 1 44.1% in CIN 2-3 63.6%. Sensitivity, specificity, positive predictive value, and negative predictive value of screening tests were examined in CIN 2 and more lesions. Sensitivity and specificity for HR-HPV were 63.6% and 73.3%, respectively, the same rates were 81.8% and 58.7% for HPV. The highest sensitivity was found in combination of HRHPV and Pap smear, the highest specificity in HPV. In conclusion, the HPV prevalence and genotype distribution in our study are similar to those reported in the world, but higher than previous studies in our country. These results may be due to our methodology and hospital based nature of our study group. We conclude that only smear or HR-HPV testing are not sufficient as a single pronged screening test, and that the participation of other genotypes of HPV in screening increases the sensitivity.

[Isolation of Cryptococcus neoformans from a chesnut tree (Castanea sativa), Denizli, Turkey]. / Denizli bölgesinde kestane (Castanea sativa) agacindan Cryptococcus neoformans izolasyonu.

Cryptococcus neoformans is a basidiomycetous encapsulated yeast that can cause life-threatening infections in immunosuppressed humans and animals. C.neoformans/Cryptococcus gattii infections are considered to be acquired via inhalation of aerosolized particles from the environment. Avian guano, decaying tree hollows and soil are known as environmental niches. In recent years, colonization of the woody structures of different trees such as Eucalyptus camaldulensis, Tamarix hispida, Platanus orientalis and Punica granatum has been reported in the environmental study of the western Anatolian region. Based on the results of previous studies, our country may have intensive Cryptococcus colonization niches in the western regions. The aim of this study was to investigate the presence of the colonization of C.neoformans niche in chestnut (Castanea spp.) trees on higher altitudes. In the study, the colonization of C.neoformans was screened on chestnut trees (Castanea spp.) in Aydin-Ödemis-Denizli geographical area. This area consists of mountainous terrain between the fertile plain formed by two major rivers.This region is one of the widespreading areas of chestnut farming in Anatolia. Two hundred and fourteen chestnut trees that had deep fissures or trunk hollows were screened during mid-summer 2017. A swabbing technique was used, and all samples were cultured on Staib agar medium containing biphenyl and antibiotics. Cultures were checked for ten days for suspicious brown colonies. Suspicious yeast colonies were tested for the identification of pathogenic Cryptococcus by conventional methods and canavanine-glycine-bromothymol agar reactions. ITS 1-4 primers were used for strain PCR tests. We determined the mating type and serotypes by PCR analysis of the STE20 genes using STE20 (Aa), STE20 (Aα), STE20 (Da), and STE20 (Dα) primers. V8 agar medium was used for mating cultivation. Only one (0.47%) strain of C.neoformans was isolated from 214 screened trees. This strain was confirmed by ITS 1-4 sequencing. The serotype A MATα mating type was observed. Basidium, basidiospores and clamp connections in hyphal structure were noted with MATα mating on V8 agar medium. In this study, the first C.neoformans isolate from a chestnut tree (Castanea sativa) was determined from Denizli region. Further studies of distribution of human pathogenic Cryptococcus will be helpful to determine the risk areas for the living organisms in our region.

[A comparison of biosurfactant production between clinical and environmental Exophiala isolates]. / Klinik ve çevresel Exophiala izolatlarinda biyosurfaktan üretiminin karsilastirilmasi.

Black yeast in the genus Exophiala are able to grow in hydrocarbon-contaminated environments and are pathogenic in immunosuppressed hosts. The biosurfactant produced by Exophiala species may be associated with strain pathogenicity by changing the hydrophobicity. The aim of this study was to prove the hypothesis that biosurfactant production in Exophilia strains isolated from clinical samples is lower than the strains isolates from toxic (dishwasher and railway sleepers) environments. A total of 122 Exophiala isolates 108 environmental (isolated from 82 dishwashers and 36 railway sleepers) and 14 clinical isolates confirmed by molecular tests were included in the study. Biosurfactant activity was tested by the drop collapse method, in which the surface of a microtiter plate well was evaluated for the presence of a biosurfactant, and by the oil spreading technique on crude oil. An open source analyses program, ImageJ®, was used for crude oil spreading technique data. A clear surface zone that differs more than two standard deviations from the mean size was accepted as a positive result. Among the 122 Exophiala species, 11 (9.0%) and 10 (8.2%) strains showed biosurfactant activity by the drop collapse test and oil spreading method, respectively. An acceptable relation was found between the drop collapse test and oil spreading method (Cohen κ coefficient= 0.30). Despite the presence of isolates showing biosurfactant activity, no statistically significant difference was detected between Exophiala strains (p= 0.72). The biosurfactant levels of environmental isolates were higher than the isolates obtained from the patients (p= 0.03). The highest biosurfactant level was observed in one Exophiala phaeomuriformis strain isolated from a dishwasher. There was no difference between the biosurfactant levels of the dishwasher and railway sleeper isolates (p= 0.66). Biosurfactant production may be a more important determinant of virulence in Exophiala species than expected. In this study, biosurfactant activity was higher in environmental isolates compared to the clinical isolates. Consensus of multiple biosurfactant screening protocols may clarify why environmental Exophiala species are less virulent. Further studies should evaluate biosurfactant activity in additional clinical Exophiala isolates. The biosurfactant activity of more Exophiala isolates obtained from patients should be investigated with further planned studies.