Laboratuvar Ölçekli Anammox Reaktörlerde Mikrobiyal Topluluğun Moleküler Yöntemlerle Araştırılması
Laboratuvar Ölçekli Anammox Reaktörlerde Mikrobiyal Topluluğun Moleküler Yöntemlerle Araştırılması
Dosyalar
Tarih
2013-08-13
Yazarlar
Esen, Elif
Süreli Yayın başlığı
Süreli Yayın ISSN
Cilt Başlığı
Yayınevi
Fen Bilimleri Enstitüsü
Institute of Science and Technology
Institute of Science and Technology
Özet
Biyolojik atık su arıtımı organik ve inorganik bileşiklerin uzaklaştırılmasında kullanılan, yaklaşık yüzyıllık bir geçmişe sahip önemli biyoteknolojik proseslerden biridir. Nitrat, nitrit, amonyum gibi azotlu bileşiklerin toksik etkisinden dolayı nitrojen giderimi biyolojik atıksu arıtımında kritik bir yere sahiptir. Anammox (anaerobik amonyum oksidasyonu) prosesi ise konvensiyonel metodlara kıyasla korbon ve oksijen gerektirmediği için yeni ve az enerji harcayan bir alternatif.Atıksu arıtım sistemlerinde genel yaklaşım konvensiyonel parametreler ile çalışmaya meyilli olsa da, moleküler teknikler proses dizaynında, işletilmesinde ve kontrolünde bize daha geniş bir bakış açısı sağlayabilir. Moleküler teknikler proseste yer alan mikroorganizma türlerini ve miktarlarını belirlemede kullanılır. Böylece konvensiyonel parametrelerle moleküler parametreler karşılaştırılarak daha üst düzey arıtma sistemleri kurulabilir. Bu çalışma moleküler teknikler kullanılarak laboratuvar ölçekli Anammox reaktörde mikrobiyal komünitenin incelenmesini hedefemektedir. Bu amaçla floresan in situ hibridizasyon (FISH) ve 16S rDNA gen bazlı moleküler yöntemler anammox bakterisi, amonyağı oksitleyen bakteri (AOB), amonyağı oksitleyen arke (AOA) ve nitrit oksitleyen bakteri (NOB) gibi mikroorganizma çeşitlerinin varlığı araştırıldı. Örnekler Marmara Üniversitesi’nde kısmi nitrifikasyon reaktörü ve anammox reaktöründen elde edildi. FISH sonuçlarına göre kısmi nitrifikasyon reaktöründe NTSPA0662 ve NSO1225 probları kullanılarak NOB ve AOB türleri saptandı. Anammox reaktöründe ise PLA46, AMX368, BS820 ve AMX820 probları kullanılarak Planktomisetes ve anammox bakteri türlerinin olduğu belirlendi. DGGE sonuçlarına baktığımızda ise numuneler arası değişimin olduğunu biyonümerik analiz sonuçlarından görmekteyiz. Bu sonuçlara göre hem kısmi nitrifikasyon ve hem anammox reaktöründe hedeflenen türler aktif olarak yer almaktadır.
Biological wastewater treatment (WWT) is undouptedly one of the most important biotechnological process, which have been used for over a century to remove organic and inorganic compounds. Nitrogen removal has a very crucial role in these biological treatment processes because of toxicity of nitrate, nitrite and ammonium. Conventional methods for nitrogen removal involve two different steps namely nitrification and denitrification. Anammox (Anaerobic ammonium oxidation) process is a new low-cost alternate to conventional biological nitrogen removal, since it does not require any carbon and oxygen sources. Even though the general approach embrace to use conventional parameters in WWT systems, practising culture-independent molecular techniques are expanding our understanding process design, operation and control. These techniques determine the identities of microorganisms involved in the process. Thus comparing the conventional datas with molecular results gain us a new aspect to set up advanced WWT plants. This study aims to highlight investigation of microbial communities in laboratory scale anammox reactor and partial nitrification reactor. Both reactor were started up using a mixed activated sludge seed obtained from a local domestic wastewater treatment plant. Molecular approaches that have proven to be particularly useful for study of microorganisms in WWT systems will be focused on in this study. For this purpose, Flourescent in situ hybridization (FISH) and 16S rDNA gene based molecular methods such as PCR (Polymerase Chain Reaction) and DGGE (Denaturing Gradient Gel Electrophoresis) was used to show presence of anammox bacteria, ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA). Samples are collected from a partial nitrification sequencing batch reactor (PN-SBR) and an Anammox sequencing batch reactor (AN-SBR) in Marmara University. According to FISH results in partial nitrification reactor, nitrospira-like nitrite oxidiing bacteria and ammonia-oxidiing bacteria were detected by using NTSPA0662 and NSO1225 probes, prespectively. Moreover, Planctomycetes and anammox species were observed in Anammox reactor bu using PLA46 Planctomycetes species specific probe and AMX368, BS820 and AMX820 anammox species specific probes. When we look at the DGGE results, we can see the microbial diversity change in samples by bionumerics cluster analysis. These result revealed that there are active anammox bacteria, ammonia oxidiing bacteria and ammonia oxidiing archaea with in these reactor system for ammonium treatment.
Biological wastewater treatment (WWT) is undouptedly one of the most important biotechnological process, which have been used for over a century to remove organic and inorganic compounds. Nitrogen removal has a very crucial role in these biological treatment processes because of toxicity of nitrate, nitrite and ammonium. Conventional methods for nitrogen removal involve two different steps namely nitrification and denitrification. Anammox (Anaerobic ammonium oxidation) process is a new low-cost alternate to conventional biological nitrogen removal, since it does not require any carbon and oxygen sources. Even though the general approach embrace to use conventional parameters in WWT systems, practising culture-independent molecular techniques are expanding our understanding process design, operation and control. These techniques determine the identities of microorganisms involved in the process. Thus comparing the conventional datas with molecular results gain us a new aspect to set up advanced WWT plants. This study aims to highlight investigation of microbial communities in laboratory scale anammox reactor and partial nitrification reactor. Both reactor were started up using a mixed activated sludge seed obtained from a local domestic wastewater treatment plant. Molecular approaches that have proven to be particularly useful for study of microorganisms in WWT systems will be focused on in this study. For this purpose, Flourescent in situ hybridization (FISH) and 16S rDNA gene based molecular methods such as PCR (Polymerase Chain Reaction) and DGGE (Denaturing Gradient Gel Electrophoresis) was used to show presence of anammox bacteria, ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA). Samples are collected from a partial nitrification sequencing batch reactor (PN-SBR) and an Anammox sequencing batch reactor (AN-SBR) in Marmara University. According to FISH results in partial nitrification reactor, nitrospira-like nitrite oxidiing bacteria and ammonia-oxidiing bacteria were detected by using NTSPA0662 and NSO1225 probes, prespectively. Moreover, Planctomycetes and anammox species were observed in Anammox reactor bu using PLA46 Planctomycetes species specific probe and AMX368, BS820 and AMX820 anammox species specific probes. When we look at the DGGE results, we can see the microbial diversity change in samples by bionumerics cluster analysis. These result revealed that there are active anammox bacteria, ammonia oxidiing bacteria and ammonia oxidiing archaea with in these reactor system for ammonium treatment.
Açıklama
Tez (Yüksek Lisans) -- İstanbul Teknik Üniversitesi, Fen Bilimleri Enstitüsü, 2013
Thesis (M.Sc.) -- İstanbul Technical University, Institute of Science and Technology, 2013
Thesis (M.Sc.) -- İstanbul Technical University, Institute of Science and Technology, 2013
Anahtar kelimeler
Anammox,
Moleküler yöntemler,
Anammox,
Molecular methods