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Atık Su Arıtma Sistemlerinde Mikrobiyal Kommunite

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Atık Su Arıtma Sistemlerinde Mikrobiyal Kommunite ATIK SU ARITMA SİSTEMLERİNDE MİKROBİYAL KOMMUNİTE ANALİZİ VE BİYODEGRADASYON POTANSİYELLERİNİN TESPİTİ Doktora Tezi Nilgün POYRAZ Eskişehir, 2018 ATIK SU ARITMA SİSTEMLERİNDE MİKROBİYAL KOMMUNİTE ANALİZİ VE BİYODEGRADASYON POTANSİYELLERİNİN TESPİTİ Nilgün POYRAZ DOKTORA TEZİ Biyoloji Anabilim Dalı Danışman: Doç. Dr. Mehmet Burçin MUTLU Eskişehir Anadolu Üniversitesi Fen Bilimleri Enstitüsü Nisan, 2018 Bu tez çalışması BAP Komisyonunca kabul edilen 1504F169 no.lu proje kapsamında desteklenmiştir JÜRİ VE ENSTİTÜ ONAYI Nilgün POYRAZ’ın “Atık Su Arıtma Sistemlerinde Mikrobiyal Kommunite Analizi ve Biyodegradasyon Potansiyellerinin Tespiti” başlıklı tezi .../.../2018 tarihinde aşağıdaki jüri tarafından değerlendirilerek “Anadolu Üniversitesi Lisansüstü Eğitim- Öğretim ve Sınav Yönetmeliği”nin ilgili maddeleri uyarınca, Biyoloji Anabilim dalında Doktora tezi olarak kabul edilmiştir. Unvanı Adı Soyadı İmza Üye (Tez Danışmanı) : ......................................................... ....................... Üye : ......................................................... ....................... Üye : ......................................................... ....................... Üye : ......................................................... ....................... Üye : ......................................................... ....................... ..................................... Enstitü Müdürü ÖZET ATIK SU ARITMA SİSTEMLERİNDE MİKROBİYAL KOMMUNİTE ANALİZİ VE BİYODEGRADASYON POTANSİYELLERİNİN TESPİTİ Nilgün POYRAZ Biyoloji Anabilim Dalı Anadolu Üniversitesi, Fen Bilimleri Enstitüsü, Nisan 2018 Danışman: Doç. Dr. Mehmet Burçin MUTLU Çalışma kapsamında belediyeye ait ve endüstriyel atık su arıtma tesisleri olmak üzere iki farklı tip atık su arıtma tesisinden mevsimsel olarak örnekleme yapılmış ve farklı operasyonel ünitelerden 40 örnek alınmıştır. Alınan örneklerden pH, biyolojik oksijen ihtiyacı gibi ölçümler, elementel analiz ve mezofilik bakteri sayımı yapılmıştır. Ortamdaki mikrobiyal çeşitliliği belirlemek amacıyla kültür bağımlı ve kültür bağımsız deneyler tasarlanmıştır. Kültür bağımlı çalışmalar kapsamında tesislerdeki fenol degradasyonu yapan mikroorganizmalar araştırılmıştır. Kültür bağımsız yöntemler kapsamında ise ortamın mikrobiyal çeşitliliğini ortaya çıkarmak için fluoresan in situ hibridizasyon (FISH) deneyleri yapılmış ve PCR ile 16S rRNA V3-V4 gen bölgeleri çoğaltılmış ve Illumina MiSeq ve ile Roche 454 GS FLX + / Junior platformları ile dizilenmiştir. Elde edilen veriler QIIME ve ChunLab CL Community programları kullanılarak analiz edilmiştir. Elementel analiz sonuçları, farklı ünitelerden alınan su örneklerinin elemental kompozisyonunun farklı olduğunu ortaya koymuş, pH değerlerinin ise havalandırma ünitelerinde sabit diğer ünitelerde ise değişken olduğu belirlenmiştir. Kültür bağımlı çalışmalar kapsamında izole edilen ve fenol degrade edebilme yeteneği bulunan izolatların genellikle Pseudomonas türleri olduğu tanımlanmıştır. Kültür bağımsız çalışmalar kapsamında taksonomik kompozisyonu incelemek için gerçekleştirilen 16S rRNA amplikon dizileme analizleri sonucu tesislerin kompozisyonun örnekleme dönemine göre değiştiği belirlenmiştir. Ayrıca belediyeye ait ve endüstriyel atık su arıtma tesislerinin taksonomik kompozisyonun da son derece farklı olduğu sonucuna ulaşılmıştır. FISH analizi sonucu da yoğun olarak tüm örneklerde Bacteria domaini üyeleri belirlenmiştir. Anahtar Kelimeler: Atık su, Fluoresan In Situ Hibridizasyon (FISH), biyodegradasyon, fenol, mikrobiyal çeşitlilik iii ABSTRACT MICROBIAL COMMUNITY ANALYSIS IN WASTE WATER TREATMENT SYSTEM AND DETERMINATION OF BIODEGRADATION POTENTIALS OF THESE COMMUNITIES Nilgun POYRAZ Department of Biology Programme Anadolu University, Graduate School of Science, April 2018 Supervisor: Associate Professor Mehmet Burcin MUTLU Within the scope of the study, two different types of wastewater treatment plants, namely municipal and industrial wastewater treatment plants, were seasonally sampled and 40 samples were taken from different operational units. Measurements such as pH, biological oxygen demand, elemental analysis and mesophilic bacteria counts were performed on the samples. Culture-dependent and culture-independent experiments were designed to determine the microbial diversity in the wastewater treatment plants. Culture- dependent studies have been carried out to investigate the microorganisms that degrade the phenol in the plants. As culture-independent methods,to elucidate the microbial diversity of the environment in detail, fluorescence in situ hybridization (FISH) experiments were performed and the 16S rRNA V3-V4 gene regions were amplified by PCR and sequenced with Illumina MiSeq and Roche 454 GS FLX + / Junior platforms. The obtained data were analyzed using QIIME and ChunLab CL Community programs. The results of the elemental analyzes indicated that the elemental values of the different units of the two plants were different and that the pH values are fixed in the aeration tanks and variable in the other units. It has been determined that isolates isolated in culture dependent studies and capable of phenol degrade are generally Pseudomonas species. 16S rRNA amplicon sequencing assays performed to study microbial populations within culture-independent studies have determined that the composition of the resulting plants changes with the sampling period. Furthermore, the taxonomic composition of municipal and industrial wastewater treatment plants is also very different. The result of FISH analysis was also extensively determined for Bacteria domain members in all samples. Keywords: Wastewater, FISH (Fluorescence In Situ Hybridization), biodegradation, phenol, microbial diversity iv TEŞEKKÜR Doktora tezi olarak hazırlamış olduğum bu çalışma, atık su arıtma sistemlerindeki farklı operasyonel ünitelerin mikrobiyal çeşitliliğinin mevsimsel olarak değerlendirilmesini kapsamaktadır. Bu çalışma çerçevesinde ayrıca biyodegradasyon üzerine de çeşitli çalışmalar yapılmıştır. Bu çalışmanın ortaya çıkmasında bana öncülük eden, deneysel çalışmalar sırasında yönlendirmeleriyle bana yol gösteren, ilgi ve yardımlarını esirgemeyen Sayın Danışmanım Doç. Dr. Mehmet Burçin MUTLU’ya sonsuz teşekkürlerimi sunarım. Deneysel çalışmalarım sırasında atık su örneklerinin sağlanmasında bana destek olan atık su arıtma tesislerinin yetkililerine sağladıkları yardımlar ve verdikleri bilgiler için teşekkür ederim. Doktora eğitimim süresince ilgi ve desteklerini benden esirgemeyen Anadolu Üniveristesi Fen Fakültesi Biyoloji Bölümü’ndeki tüm hocalarıma, desteklerini esirgemeyen arkadaşlarıma teşekkür ederim Doktora çalışmam süresince desteğini hiçbir zaman esirgemeyen, her an yanımda hissettiğim değerli eşime ve beni bugünlere getiren aileme sonsuz teşekkürlerimi sunarım. Nilgün POYRAZ v 17/04/2018 ETİK İLKE VE KURALLARA UYGUNLUK BEYANNAMESİ Bu tezin bana ait, özgün bir çalışma olduğunu; çalışmamın hazırlık, veri toplama, analiz ve bilgilerin sunumu olmak üzere tüm aşamalarında bilimsel etik ilke ve kurallara uygun davrandığımı; bu çalışma kapsamında elde edilen tüm veri ve bilgiler için kaynak gösterdiğimi ve bu kaynaklara kaynakçada yer verdiğimi; bu çalışmanın Anadolu Üniversitesi tarafından kullanılan “bilimsel intihal tespit programı”yla tarandığını ve hiçbir şekilde “intihal içermediğini” beyan ederim. Herhangi bir zamanda, çalışmamla ilgili yaptığım bu beyana aykırı bir durumun saptanması durumunda, ortaya çıkacak tüm ahlaki ve hukuki sonuçları kabul ettiğimi bildiririm. Nilgün POYRAZ vi İÇİNDEKİLER Sayfa BAŞLIK SAYFASI .......................................................................................................... i JÜRİ VE ENSTİTÜ ONAYI .......................................................................................... ii ÖZET .............................................................................................................................. iii ABSTRACT .................................................................................................................... iv TEŞEKKÜR .................................................................................................................... v ETİK İLKE VE KURALLARA UYGUNLUK BEYANNAMESİ ............................ vi İÇİNDEKİLER .............................................................................................................. ix TABLOLAR DİZİNİ ..................................................................................................... ix ŞEKİLLER DİZİNİ ....................................................................................................... ix SİMGELER ve KISALTMALAR DİZİNİ ................................................................... x 1. GİRİŞ ........................................................................................................................ 1 1.1. Atık su Kavramı ............................................................................................. 2 1.1.1. Evsel ve endüstriyel atık su ............................................................... 3 1.2. Atık Su Arıtımı Önemi ve Tarihi ................................................................. 4 1.3. Atık Su Arıtım Yöntemleri ........................................................................... 5 1.3.1. Ön arıtım ........................................................................................... 6 1.3.1.1. Eleme .................................................................................. 6 1.3.1.2. Kum tutucu ........................................................................
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