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

Pachnoda Marginata Larva'sının arka bağırsağından anaerobiklignoselülitik 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.

Anaerobic lignocellulolytic microbial community derived from hindgut of Pachnoda Marginata Larva

Agricultural wastes are considered as the most important biomass foranaerobic digesters due to the high energy content, high abundance,and low costs. However, complex lignocellulosic structure limits thehydrolysis of these materials. Thus, effective lignocellulose degradationstrategies are developing to enhance the hydrolysis rate. The aim of thisstudy is to enrich microbial consortia that degrade lignocellulosicbiomass from the hindgut of Pachnoda marginata larvae. Within thisscope, Pachnoda marginata larvae kept on a lignocellulose rich diet for3 weeks and then dissected. The hindgut compartment was transferredto culture bottles. Bacterial community compositions were examined byamplicon sequencing of the 16S rRNA gene on the Illumina MiSeqplatform, methanogenic archaeal communities were determined by theT-RFLP method. Biogas production was increased over time andmethane production was observed in all bottles. The bacterialcommunity profile of the enrichment culture was shifted as a result ofthe enrichment procedure with particularly the abundance ofPorphyromonadaceae (phylum: Bacteroidetes) increasing duringprolonged subcultivation. The methanogenic community wasdominated by Methanobrevibacter. Our results show the successfulestablishment of an enrichment culture from the gut system ofPachnoda marginata larvae that effectively degrades complexlignocellulose rich biomass.

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