Australian Journal of Basic and Applied Sciences, 5(9): 2274-2278, 2011 ISSN 1991-8178 Mıcrofungus Flora of Indoor and Outdoor Aır in Prımary Schools, Çorum, Turkey Adem İmalı, 1Ferudun Koçer, 2Burhanettin Yalçınkaya 1 1 Kilis 7 Aralik University, Faculty of Arts and Sciences, Department of Biology, 79000, Kilis, Turkey. 2 Ankara University, Faculty of Arts and Sciences, Department of Biology, Ankara, Turkey. Abstract: In the present study, it was aimed to determine the indoor and outdoor fungal flora of five different elementary schools using petri-plate method based on gravitation in Çorum city between the years of 2007-2008. 64 species belonging to the 29 genera from 80 petri plates were identified. The most prevalent genera were as follows Cladosporium spp. (%18.35), Aspergillus spp. (% 15.62), Penicillium spp. (%10.93), Ulocladium spp.(%9.37), Alternaria spp. (% 6.25), Scolecobasidium spp. (%3,12 ). Key words: Microfungi, indoor and outdoor air, Çorum, Turkey. INTRODUCTION Fungi represent about 10 % of the earth's biomass and capable of reproduction almost everywhere in indoor or outdoor air (Alexopoulos, 1979; Seltzer, 2007). Fungi can be prevalent in the atmosphere while optimum conditions for their reproduction are available and they cause many allergic diseases including asthma, allergic rhinosinusitis, and bronchopulmonary mycoses (Bıçakçı, 2001; Burrell, 1991; Celenk, 2007; Tatlıdil, 2001). There are over 80 species of fungi assocaited with allergic respiratory system, and especially the most important cause of allergic astma in children is a result of allergen inhalation (Kılıç, 2008; Horner, 1995). The most common genera identified in previous studies abroad are Cladosporium, Penicillium, Aspergillus and Alternaria (Petronella, 2005; Wan-Kuen, 2005). In our country, fungal spores belonging to the genera Cladosporium, Alternaria, Aspergillus, Penicillium are reported to be commonly found in many provinces in large part of the year (Ataygul, 2007; Celtik, 2011; Çeter, 2009). Children spend most of their time on the school and exposed to the fungi in school atmosphere without being aware. In addition, they are much more vulnerable and sensitive to environmental factors than adults (Wan-Kuen, 2005). For this reason, it has great importance to determine the fungal allergens at primary schools. MATERIAL AND METHODS Sampling Sites: The research materials are provided from five different elementary schools in the city center of Çorum, Turkey. Of these schools, Toprak Sanayii and Karşıyaka Elementary School were chosen due to being closer to the countryside and open to the wind. Albayrak Elementary school because of being in the center, where there is an intensive existence of the traffic and air pollution, Mehmet Akif Ersoy Elementary school because of having the high number of students and being open to the wind were preferred. Furthermore, Mustafa Kemal Elementary School was chosen due to the fact that there was a dense plant flora and less urbanization around school and the school was openly exposed to the wind. Some information about sampling sites are given in Table 1. Table 1: Some information about sampling sites. Sampling Site No Sampling Site Name 1 Toprak Sanayii Elemt. Sch. 2 Albayrak Elemt. Sch. 3 4 5 Karşıyaka Elemt. Sch. Mustafa Kemal Elemt. Sch. Mehmet Akif Elemt. Sch. Latitude 40 33 58 87 N 40 33 09 11 N Longitude 34 56 27 95 E 34 57 30 84 E Student Number 1180 1207 40 32 37 95 N 40 34 22 47 N 40 32 15 01 N 34 58 19 93 E 34 57 25 52 E 34 57 01 73 E 1295 1350 1860 Monthly-indoor and outdoor air samples were collected from the selected schools during the semesters of 2007-2008 education year. Gravity- Based Petri Plate Method was used for the samplings. This sampling method was preferred because of adhesion of air particles with the effect of gravity and its easiness was another reason for preference of this method. Samplings were taken during school hours and at the same time in all Coresponding Author: Adem İmalı, Kilis 7 Aralik University, Faculty of Arts and Sciences, Department of Biology, 79000, Kilis, Turkey Tel: +90 348 822 23 50-1434 / Fax:+90 348 822 23 51, E-mail: adem672002@gmail.com. 2274 Aust. J. Basic & Appl. Sci., 5(9): 2274-2278, 2011 stations from a height of 1.5 m (Sarıca, 2007; Srikanth, 2008). Two petri plates containing Rose-bengalstreptomycin inside and outside air were left open for 15 minutes. Closed plates have been covered with strech film to prevent contamination until incubation. Then, the plates were periodically incubated for 8-12 days at 25°C. Penicillium and Aspergillus isolates obtained after incubation were inoculated into the Czapek Solution agar and the other species were inoculated into the Malt agar (Domsch, 1980). Isolates were inoculated into agar slant tubes for long term storage (Imalı, 2005). For identification of isolated fungi, Samson and Pitt (2000), Klich (2002), Pitt and Hocking (2009), Hasenekoğlu (1991) were referred, respectively. Results: The method used here is a common method applied by many researchers (Çolakoğlu, 2004; Fischer, 2009; Ramachandran, 2005) to determine the fungal flora of air. In the present study, species belonging to the genera Acremonium, Alternaria, Arthrobotrys, Arthroderma, Aspergillus, Botriyotricum, Botrytis, Ceratocystis, Chrysosporium, Cladosporium, Curvularia, Fusarium, Geotrichum, Monilia, Mucor, Mycelia, Nectria, Penicillium, Phialophora, Phoma, Phytophthora, Pleospora, Rhizopus, Scolecobasidium, Stachybotrys, Stemphylium, Torula, Trichosporan, Ulocladium were determined (Table 5). The most common determined genera in the study and their percentages (%) are as follows Aspergillus spp. (% 15.62), Cladosporium spp. (% 18.35), Penicillium spp. (% 10.93), Ulocladium spp. (% 9.37), Alternaria spp. (% 6.25), Scolecobasidium spp. (% 3.12). 64 species belonging to the 29 genera from 734 colonies including 212 indoor and 522 outdoor colonies were identified. While the highest number of colony with a number of 234 (31.88%) was found in October, the lowest number of colony with a number of 18 (2.45%) was determined in February.The lowest number of colony at stations was counted in February whereas the highest number of colony at stations was found in March. The highest colony number was determined at Station 4 (Mustafa Kemal Elemt. Sch.). However the lowest colony number was found at station 1 (Toprak Sanayii Elemt. Sch.) (Table 2; Figure 1). Table 2: Monthly-dependent number and percentages (%) of colonies at stations Monthly-dependent number and percentages (%) of colonies at stations Month 1 2 3 4 Oct. 2007 25 (3,41 %) 60 (8,17 %) 30 (4,09 %) 89 (12,12 %) Nov. 2007 5 (0,68%) 16 (2,18 %) 7 (0,95 %) 24 (3,27 %) Dec. 2007 6 (0,82 %) 4 (0,55 %) 6 (0,82 %) 18 (2,45 %) Jan. 2008 5 (0,68 %) 5 (0,68 %) 5 (0,68 %) 4 (0,55 %) Feb. 2008 4 (0,55 %) 4 (0,55 %) 3 (0,41 %) 3 (0,41 %) March2008 38 (5,18 %) 39 (5,31 %) 39 (5,31 %) 34 (4,63 %) Apr. 2008 15 (2,04 %) 14 (1,91 %) 16 (2,18 %) 16 (2,18 %) May.2008 18 (2,45 %) 11 (1,50 %) 15 (2,04 %) 8 (1,09 %) Total 116(15,80%) 153(20,84%) 121(16,48%) 196(26,70%) 5 30 (4,09 %) 8 (1,09 %) 7 (0,95 %) 5 (0,68%) 4 (0,55 %) 47 (6,40 %) 23 (3,13 %) 24 (3,27 %) 148(20,16%) Total - % 234 (31,88 %) 60 ( 8,17%) 41 (5,58 %) 24 (3,27 %) 18 (2,45 %) 197 (26,84 %) 84 (11,44 %) 76 (10,35 %) 734 (100 %) Fig. 1:Total colony number in indoor and outdoor air according to the months. Dıscussıon: A large number of studies have been conducted, reviewed and discussed to determine the fungal flora of indoor and outdoor of elsewhere (Aydoğdu, 2008; Çolakoğlu, 2004; Fischer, 2009; Ramachandran, 2005). In a study in determination of the indoor and outdoor fungal flora of kindergartens by Aydoğdu and Asan (4), while lowest number of colony was found to be in the months of January and Februry the highest number was determined in October. Similarly, the report on the atmosphere of Porto and Amares by Olivera et al. [23], the lowest spore concentration was found to be in the months of January and February the highest spore concentration was recorded in October. 2275 Aust. J. Basic & Appl. Sci., 5(9): 2274-2278, 2011 Table 3: Statistical analyses. 1 Mean 14,50 Maximum 38 Minimum 4 Median 10,50 St. deviation 12,18 F- test 148,29 2 19,13 60 4 12,50 20,09 403,55 3 15,13 39 3 11,00 13,04 170,13 4 24,50 89 3 17,00 28,08 788,57 5 18,60 47 4 15,50 15,26 232,86 Total 91,75 234 18 68,00 80,39 6.462 In the present study, lowest number of colony was also determined in January and February, which is in a good agreement with previous studies (Aydoğdu, 2008; Oliveira, 2009), the highest number of colony, in addition to the months of October, was also observed in March. The decline number of colony or spore concentration in the months of January and February was shown to be the results of climatic fluctuations such as precipitation, humidity, temperature and wind speed by many reports (Bıyık, 2002; Burge, H.A., 1986; Celtik, 2011; Imalı, 2002; Wan-Kuen, 2005). As shown in table 5 obtained from State Meteorological Agency (2007-2008), the reason of lowest number of colony to be found in the months of January and February may be attributed to the lower value of monthly total precipitation (10,2- 22,5 mm) and temperature [(-)5,8- (-) 2,9 0C] and wind speed (1,1- 1,4 m/sec) as well as the higher value of meteorological data of monthly mean pressure (932,2-933,1 hPa) and relative humidity (82,5-85, 5 %). The higher number of colony in October and March may be attributed to the optimum conditions for fungal reproduction, which were moderate high temperature and lower precipitation. The fungal concentration and compositions varying with various meteorological factors we obtained in here are in a good agreement with previous reports (Asan, 2002; Bıyık, 2002; Celtik, 2011; Imalı, 2002; Wan-Kuen, 2005). Outdoor fungal concentration, which is statistically significant factor on asthma and rhinitis patients, appears to be the one of important agent of biological pollutants. Inal et al., (2007) indicated that there was a positive correlation on fungal allergic concentration in stimulating asthma and rhinitis symptoms with a reports on effect of indoor mold concentrations on daily symptom severity of children with asthma and/or rhinitis. Aspergillus and Penicillium spores were reported to be found in humid buildings. Because of the fact that the microbial concentration of open and close areas are dependent on seasonal variations, while the microbial concentration is higher in open areas especially in summer, the higher number of concentration close area was reported to be especially found in winter (Asan, 2002; Bıyık, 2002; Celtik, 2011; Imalı, 2002; Wan-Kuen, 2005). The fungal concentration intensity was found to be higher in indoor air in fall and higher in outdoor air during spring season (Table 4; 5). These findings are parallel with previous works. Since the cold weather in indoor during winter is unfavorable for fungal reproduction, hot weather in indoor provides optimum conditions for fungi, increasing the fungal concentration. Density of spore concentration of Aspergillus and Penicillium vary disproportionally regarding the seasonal fluctuations in indoor and outdoor air. Therefore, human populations should be more aware of and sensitive against those invisible microorganisms significantly affected by changing environmental factor. The students in range of 6-18 years should be informed about fungi and fungal allergy, which may lead to significant problems in the ahead. The present study may be a promising work for the further investigations. Table 4: The meteorological data of Çorum province in 2007 and 2008. Meteorological Data Fall / Spring Season Oct. Nov. Dec. Jan. Feb. Month/Year 2007 2007 2007 2008 2008 March 2008 Apr. 2008 May. Mean 2008 2007 Mean 2008 Monthly Total Precipitation (mm) 28,8 60,3 34,5 22,5 10,2 44 33,2 55,2 33.17 32.18 Monthly Mean Pressure (hPa) 929,6 927,5 930 932,2 933,1 921,3 923,4 925 925.67 926.97 Monthly Mean Temperature (° C) 13,5 5,2 1,2 -5,8 -2,9 9,3 12,8 14,1 11.46 10.80 Monthly Mean Wind Speed (m_sec) 2,8 3,3 1,3 1,1 1,4 2,3 2 1,8 3.29 1.82 69,3 82,3 84 85,5 82,5 67,3 68,9 63,8 67.26 71.62 Monthly Mean Relative Humidity (%) Table 5: Identified Microfungus species and their references. Species and their references Acremonium butryi (van Beyma) W. Gams 1971 Cladosporium variabile (Cooke) De Vries 1952 Alternaria alternata Keisler 1912 Alternaria raphani Groves & Skolko 1944 Alternaria brassicicola Wiltshire 1947 Alternaria pluriseptata Jorstad 1945 Arthrobotrys arthrobotryoides Lindau 1907 Curvularia pallescens Boedijn 1933* Fusarium sp. Link ex Fr. 1821 Geotrichum spp. Link ex Lmon 1821 Monilia sp. Pers ex. Fr. 1832 Mucor hiemalis Wehmer 1903 2276 Aust. J. Basic & Appl. Sci., 5(9): 2274-2278, 2011 Arthroderma guadrifidum Dawson&Gentles 1961 Aspergillus flavus Link ex.Gray. 1821 Aspergillus niger Van. Tieghem 1867 Aspergillus parasiticus Speare 1912 Aspergillus japonicus Saito 1906 Aspergillus clavatus Desm. 1834 Aspergillus cervinus Masse 1914 Aspergillus fumigatus Fresenius 1863 Aspergillus ornatus Raper, Fennell &Tresner 1953 Aspergillus flavipes Bain & Sartory 1926 Aspergillus clavato-flavus Raper&Fennell 1965 Botriyotricum sp. Sacc.& March. 1885 Botrytis sp. Mich. Ex Fr.1821 Ceratocystis moniliformis (Hedgcock) C. Moreau 1952* Chrysosporium georgii Van Oorschot 1980* Cladosporium herbarum (Pers) Link ex. Gray 1821 Cladosporium tenuissimum Cooke 1878 Cladosporium cladosporiodes (Fres) De Vries 1952 Cladosporium obtectum Rabel 1889 Cladosporium gallicola Sutton 1973 Cladosporium orchidis E.A.& M.B. Ellis 1972 Cladosporium apsicale Berk&Br.1873 Cladosporium brassicae M.B.Ellis 1976 Cladosporium aecidiicola (Thüm) 1876 Cladosporium ornithogali (Klotzsch) De Vries 1952 Cladosporium oxysporum Berk& M.A. Curtis 1868 *It is thought to be a new record for Microflora of Turkey Mycelia sterilia Nectria inventa Pethybr 1919 Penicillium citrinum Thom 1910 Penicillium rolfsii var. Sclerotiale Novobr 1974 Penicillium soppii W. Zalesky 1927 Penicillium turbatum Westling 1911 Penicillium yarmokense Baghd 1968 Penicillium rolfsii Thom 1910 Penicillium thomi Maire 1917 Phialophora melini (Nonnf) Conant 1937* Phoma glomerata (Corda) Wolenw&Hochapfel 1936 Phytophthora citricola Sawada 1927 Pleospora herbarum (Fr.) Rabenh.1857 Rhizopus oryzae Went&Prinsen Geerlings 1895 Stachybotrys sp. Corda 1837 Stemphylium sarciniforme (Cav.)Wiltshire 1938 Scolecobasidium constrictum Abbott 1927 Scolecobasidium dendroides Pirozynski& Horlges 1973* Trichosporan cutaneum (Beum et al.) 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