Emine Gözde ÖZBAYRAM, Orhan İNCE, Sabine KLEINSTEUBER, Marcell NIKOLAUSZ

Pachnoda Marginata Larva'sının arka bağırsağından anaerobik lignoselülitik mikrobiyal kültür geliştirilmesi

Tarım atıkları, yüksek enerji içerikleri ve üretim miktarları ile düşük maliyetleri nedeniyle anaerobik çürütücüler için en önemli biyokütle olarak değerlendirilmesine rağmen karmaşık lignoselülozik yapıları bu maddelerin hidroliz aşamalarını sınırlandırmaktadır. Hidroliz verimini artırmak için etkin lignoselüloz degradasyon stratejileri geliştirilmektedir. Bu çalışmanın amacı, Pachnoda marginata larvalarının arka bağırsağından lignoselüloz parçalayabilen anaerobik mikrobiyal topluluğun zenginleştirilmesidir. Bu kapsamda, Pachnoda marginata larvaları 3 hafta süre ile lignoselülozik substrat ile beslenmiş, sonrasında disekte edilmiştir. Arka bağırsak kültür şişelerine transfer edilmiştir. Bakteriyel topluluk çeşitliliği 16S rRNA amplikon dizileme yöntemi ile Illumina MiSeq platformunda analiz edilmiş, metanojenik arkeal topluluk ise T-RFLP yöntemi ile incelenmiştir. Kültür şişelerinde biyogaz üretimi zaman ile artış göstermiş, tüm şişelerde metan üretimi gözlenmiştir. Zenginleştirme prosedürü sonucunda, kültürün bakteriyel topluluk profili değişiklik göstermiş, üç transfer sonrasında alınan örneklerde Porphyromonadaceae (phylum: Bacteroidetes) bolluğunun artarak baskıladığı görülmüştür. Metanojenik topluluk ise Methanobrevibacter ile baskılanmıştır. Çalışmada elde edilen sonuçlar, Pachnoda marginata larvasının sindirim sisteminden üretilen zenginleştirilmiş kültürün, lignoselülozca zengin kompleks biyokütleyi etkin bir şekide parçalayabildiğini göstermiştir.

Anahtar Kelimeler:

Bağırsak mikrobiyotası, Illumina Miseq, Lignoseüloz degredasyonu, Scarabaeidae, T-RFLP, Zenginleştirilmiş kültür

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