Suzan Okten

Airborne Fungi and Actinomycetes Concentrations in the Air of Eskisehir City (Turkey)

The present study investigated the isolation and identification of airborne fungi from three different urban stations located in Eskisehir (Turkey). Air samples were taken by exposing a Petri dish with Rose-Bengal streptomycin agar medium for 15 min and after incubation the number of growing colonies was counted. The sampling procedure for fungi was performed 35 times at the research stations weekly between March and November 2001. A total of 2518 fungal and 465 actinomycetes colonies were counted on 420 Petri plates over a nine-month period. In total, some 20 mould species belonging to 12 genera were isolated. Alternaria alternata, Cladosporium cladosporioides and Scopulariopsis brevicaulis were the most abundant species in the study area (13.66, 5.80 and 5.50% of the total, respectively). Relationships between fungal spore numbers, aerosol air pollutants (that is the particulate matter in the air) and sulphur dioxide together with the meteorological conditions were examined using statistical analysis. Number of fungi and actinomycetes were tested by multivariate analysis (MANOVA) according to the areas and months. Fungal numbers were nonsignificant according to the areas and months (p > 0.05), but the number of actinomycetes recorded was significant (p < 0.01).

Airborne fungi and bacteria in indoor and outdoor environment of the Pediatric Unit of Edirne Government Hospital.

This study was performed between January 2004 and December 2004 in 13 stations in the Pediatric Unit of Edirne Government Hospital in order to determine the outdoor and indoor airborne microfungal and bacterial contents. The results of air samplings revealed that 1,376 microfungal and 2,429 bacterial colonies in total were isolated. The isolated microfungal specimens were identified and 65 species from 16 genera were determined. Among these, the most frequent genus was Cladosporium with 462 colonies (33.58%) followed by Alternaria with 310 (22.53%) and Penicillium with 280 (20.35%) colonies. The isolated bacterial samples were grouped based on their Gram-staining properties. The most frequent ones were Gram (+) cocci with 1,527 colonies (62.87%) followed by Gram (+) bacilli with 828 colonies (34.09%) and Gram (−) bacilli with 74 colonies (3.05%). Staphylococcus, Bacillus, Corynebacterium, and Microccus appeared to be the common genera isolated for all months. Statistical analyses were performed in order to see if there existed a relationship between meteorological conditions and the microfungal and bacterial species and their concentrations.

Aspergillus alliaceus, a new potential biological control of the root parasitic weed Orobanche

During extensive surveys in fields heavily infested by broomrape in the Trakya Region‐Turkey, a different new fungus, Aspergillus alliaceus, was isolated from the infected broomrape. It is aimed to investigate whether or not it is really a pathogen for Orobanche. The fungi was exposed to a greenhouse environment in order to assess its pathogenicity and virulence against Orobanche cernua. In addition, infection tests on Orobanche seeds were also performed under laboratory conditions. The fungus was subjected using two different methods, exposure to a liquid culture with conidial solution and a sclerotial solid culture with fungal mycelia. Cytological studies were carried out at light, TEM and SEM levels. The results show that the sclerotial solid culture with fungal mycelia quickly caused necrosis and was more effective than the other type. It also greatly diminished attachments, tubercles, and caused the emergence of shoots and an increase in the total shoot number of Orobanche. In addition, both when the fungi was exposed to both soil and used to contaminate sunflower seeds, its pathogenicity was more effective. Consequently, it was determined that A. alliaceus was an effective potential biological control of broomrape throughout its life cycle from dormant seed to mature plant.

Mycological contamination in dental unit waterlines in Istanbul, Turkey

Studies on dental units (DUs) are conducted either for the prevention or the reduction of the density of bacterial contamination in dental unit waterlines (DUWLs). However, the existence of fungi in the these systems requires more attention. During dental treatment, direct contact with water contaminated with fungi such as Candida, Aspergillus, or inhalation of aerosols from high-speed drill may cause various respiratory infections, such as asthma, allergies, and wounds on mucose membranes, especially on immunocompromised patients and dentists. The aims of this study are to investigate the number and colonization of fungi in DUWLs in the city of Istanbul, Turkey. Water samples were collected from air-water syringes, high-speed drills, and inlet waters from 41 DUs. The aerobic mesophilic fungi count in high- speed drills was higher than inlet waters and air-water syringes. Non-sporulating fungi were found in 7 DUs. The isolated fungi were identified as Penicillium waksmanii, Cladosporium spp., Penicillium spp., Candida famata, Cryptococcus laurentii, Candida guilliermondii, Penicillium verrucosum, Aspergillus pseudoglaucus, Penicillium decumbens, and Acremonium sp. Some of these fungal genera are known as opportunistic pathogens that led to respiratory diseases such as allergic rhinits. This study shows the importance of regular control of mycological contamination on water at DUs.

Pesta granule trials with Aspergillus alliaceus for the biocontrol of Orobanche spp.

The pathogenic fungus Aspergillus alliaceus has been shown to have potential for the biocontrol of Orobanche spp. (broomrape), a root parasitic plant. The effectiveness of A. alliaceus in reducing Orobanche infection was analysed using pesta granules prepared with different food formulations. The results showed that pesta granules comprising of fungal mycelia/spore mixtures from liquid and solid culture, sclerotia and fungal mycelia reduced Orobanche infection to a greater extent in below ground conditions when applied early and at high doses before crop sowing. In addition, pesta granules eliminated the risk of broomrape contamination within a 0.2–0.3 cm diameter of the granules. The sclerotial pathogenicity of A. alliaceus was compared with those of other fungi reported in other studies. In addition, some morphological and histological studies on the fungal pathogenicity on broomrape plants after infection are presented. The present study reveals the potential of sclerotial A. alliaceus pesta granule applications for long-term broomrape biocontrol under field conditions.

Screening Brucella spp. in bovine raw milk by real-time quantitative PCR and conventional methods in a pilot region of vaccination, Edirne, Turkey

Brucellosis is a worldwide zoonotic disease transmitted to humans by consumption of contaminated milk and milk products. Brucellosis is endemic in Turkey, and Edirne has a high Brucella prevalence. Brucellosis is prevented by live-attenuated vaccines for animals and the vaccination program has been in place since 1984 in Turkey. Thrace is the pilot region for this vaccination program. The gold standard diagnostic technique for brucellosis is still the isolation of suspicious bacterial colonies followed by bacteriological identification, but it is very time consuming and laborious. In many studies, Brucella has been investigated by PCR techniques. However, PCR-based methods cannot differentiate between the vaccine strain and the virulent strain; thus, the vaccine strain may interfere with the virulent strain and causes false-positive reactions. To monitor brucellosis control programs effectively, it is important to distinguish vaccine and field strains of Brucella spp. In this study, raw milk samples were collected from 99 cows at 12 different barns in 5 villages of Edirne (Turkey). Bacteriological analyses and real-time quantitative (q)PCR experiments were applied to all samples. The DNA was isolated using Biospeedy DNA-Tricky Purification Kit (Bioeksen, Istanbul, Turkey). For all reactions, Roche Light Cycler Nano (Roche Diagnostics, Mannheim, Germany) instrument and Biospeedy EvaGreen qPCR Pre-Mix (Bioeksen) were used. The data were analyzed using Roche LightCycler NanoSoftware 1.0. For samples that were negative by bacteriological analyses and positive by qPCR, we developed a novel qPCR-based method to differentiate the virulent B. abortus strains and B. abortus S19 vaccine strain. We designed qPCR primers targeting the outer membrane protein of B. abortus. The qPCR products were sequenced using the ABI Prism Big Dye Terminator Cycle Sequencing Ready Reaction Kit on an ABI Prism 377 DNA sequencer (Applied Biosystems, Foster City, CA). In total, 2.02% of the samples were Brucella positive, by both bacteriological method and the novel qPCR method. We concluded that, to obtain true-positive results in Brucella spp. screening studies for milk, differentiating the virulent and vaccine strain should not be disregarded.

A NOVEL BIOSENSOR BASED ON Lactobacillus acidophilus FOR DETERMINATION OF PHENOLIC COMPOUNDS IN MILK PRODUCTS AND WASTEWATER

Different branches of industry need to use phenolic compounds (PCs) in their production, so determination of PCs sensitively, accurately, rapidly, and economically is very important. For the sensitive determination of PCs, some biosensors based on pure polyphenol oxidase, plant tissu,e and microorganisms were developed before. But there has been no study to develop a microbial phenolic compounds biosensor based on Lactobacillus species, which contain polyphenol oxidase enzyme. In this study, we used different forms of Lactobacillus species as enzyme sources of biosensor and compared biosensor performances of these forms for determination of PCs. For this purpose, we used lyophilized Lactobacillus cells (containing L. bulgaricus, L. acidophilus, Streptococcus thermophilus), pure L. acidophilus, pure L. bulgaricus, and L. acidophilus- and L. bulgaricus adapted to catechol in Lactobacilli MRS Broth. The most suitable form was determined and optimization studies of the biosensor were carried out by using this form. For preparing the bioactive layer of the biosensor, the Lactobacillus cells were immobilized in gelatin by using glutaraldehyde. In the study, we used catechol as a substrate. Phenolic compound determination is based on the assay of the differences on the respiration activity of the cells on the oxygen meter in the absence and the presence of catechol. The microbial biosensor response depends directly on catechol concentration between 0.5 and 5.0 mM with 18 min response time. In the optimization studies of the microbial biosensor the most suitable microorganism amount was found to be 10 mg, and also phosphate buffer (pH 8.0; 50 mM) and 37.5°C were obtained as the optimum working conditions. In the characterization studies of the microbial biosensor some parameters such as substrate specificity on the biosensor response and operational and storage stability were examine. Furthermore, the determination of PC levels in synthetic wastewater, industrial wastewater, and milk products was investigated by using the developed biosensor under optimum conditions.

Indoor airborne fungal pollution in newborn units in Turkey

Pathogenic and/or opportunistic fungal species are major causes of nosocomial infections, especially in controlled environments where immunocompromised patients are hospitalized. Indoor fungal contamination in hospital air is associated with a wide range of adverse health effects. Regular determination of fungal spore counts in controlled hospital environments may help reduce the risk of fungal infections. Because infants have inchoate immune systems, they are given immunocompromised patient status. The aim of the present study was to evaluate culturable airborne fungi in the air of hospital newborn units in the Thrace, Marmara, Aegean, and Central Anatolia regions of Turkey. A total of 108 air samples were collected seasonally from newborn units in July 2012, October 2012, January 2013, and April 2013 by using an air sampler and dichloran 18% glycerol agar (DG18) as isolation media. We obtained 2593 fungal colonies comprising 370 fungal isolates representing 109 species of 28 genera, which were identified through multi-loci gene sequencing. Penicillium, Aspergillus, Cladosporium, Talaromyces, and Alternaria were the most abundant genera identified (35.14, 25.40, 17.57, 2.70, and 6.22% of the total, respectively).

Synthesis and antimicrobial evaluation of some novel sulfonylamido-benzoxazoles

A series of 2-(p-substituted phenyl)-5-[(4-substituted phenyl) sulfonylamido]-benzoxazoles were synthesized and tested for their antimicrobial activities. The structures of the new derivatives were elucidated by spectral techniques. The minimum inhibitory concentrations (MIC) of the new benzoxazoles were determined against standard bacterial and fungal strains and drug-resistant isolates and compared to those of several reference drugs.

Edirne İli Söğütlük Ormanı Toprağından İzole Edilen Aspergillus Türlerinin Biyoçeşitliliği

Calismamizda, Edirne ili orman topraklarindan izole edilen Aspergillus turlerinin morfolojik-koloniyal ve molekuler yontemlerle teshis edilmesi amaclanmistir. Bu amacla, Edirne Sogutluk Ormanindan, 2016 yili Mart ayinda Brown’un metodu kullanilarak toprak ornekleri alinmistir. Topraktan fungus izolasyonu icin “Topragi Sulandirma Metodu” kullanilmistir. Morfolojik calismalar icin CZ, CYA, CY20S, MEA besiyerlerine yapilan ekimlerden sonra, mikroskobik ve makroskobik karakterler incelenmistir. Molekuler calismalar icin ise sirasiyla DNA izolasyonu, calmodulin gen bolgesini hedefleyen PCR islemleri, PCR urununun saflastirilmasi, dizi analizi ve filogenetik analiz asamalari uygulanmistir. Calismalar sonucunda 7 adet Aspergillus turu tespit edilmistir. Bu turler, Aspergillus affinis (Turkiye icin yeni kayit), Aspergillus awamori , Aspergillus carbonarius , Aspergillus dimorphicus , Aspergillus europaeus (Turkiye icin yeni kayit), Aspergillus spelaeus (Turkiye icin yeni kayit) ve Aspergillus fischeri ’dir.