microbial community and functional characteristics

to analyze microbial community in landfill leachate

1. MICROBIAL COMMUNITY AND FUNCTIONAL CHARACTERISTIC OF LEACHATE FROM JERAM AND JABOR LANDFILL USING 16S rRNA AMPLICON SEQUENCING APPROACH Supervisor : Assoc. Prof. Dr. Noor Faizul Hadry Nordin Co-supervisor : Prof. Dr. Hamzah Mohd Salleh Assoc. Prof. Dr. Husna Ahmad Tajuddin Student : Siti Marhamah Drahaman Institution : INHART, International Islamic University Malaysia

2. INTRODUCTION

3. Illustration for 16S rRNA amplicon sequencing Beneficial for microorganism that has specific growth requirements, slow-growing organisms and out-competed microorganisms. P rovide a broad-spectrum identification of different microbes along with a high resolution phylogenetic microbial community profiling (Mukherjee et al., 2016). Effective method for the characterization of bacterial populations, taxonomical analysis, and species identification.

4. OBJECTIVES

5. OBJECTIVE METHODOLOGY OUTCOME 1. Microbial community in leachate sample Sample collection & processing Using sampling device Leachate samples stored at 4 ℃ eDNA extraction Using CTAB based extraction method with simple modification (Healey et al., 2014) Microbial community analysis Using 16s rRNA amplicon sequencing Microbial diversity Taxonomy profile

6. OBJECTIVE METHODOLOGY OUTCOME 2. Metabolic function prediction PICRUSt2 analysis To predict the functional composition of the microbial communities based on 16s rRNA marker gene profiles (Douglas et al., 2020). Pathway prediction

7. RESULT 1: Microbial diversity in landfill leachate 538 ASVs; 240 ASVs for Jabor , 232 ASVs for Jeram Reach the saturation plateau Majority of species have been detected Read depth is likely sufficient 2. Rarefaction curve for Jabor & Jeram leachate 1. Venn diagram of ASVs in Jabor & Jeram leachate

8. RESULT 1: Microbial diversity in landfill leachate Chao-1 index The community richness in Jabor & Jeram slightly similar; 273 & 265 Shannon index 4.663 for Jabor & 4.822 for Jeram reflect the species richness & diversity Simpson index - Jabor and Jeram have high Simpson value, 0.968 and 0.973 respectively, indicate all samples randomly selected 3. The α -diversity indices

9. RESULT 2: Microbial community composition 17 bacterial phyla identified Firmicutes (40%), Bacteroidetes (25%) and Proteobacteria (22.5%) among the most abundant phyla in the Jabor leachate Jeram leachate dominated by Firmicutes (60%), Proteobacteria (20%) and Synergistetes (15%) Bacteroidetes and Firmicutes abundantly found in anaerobic environments: To degrade carbohydrates, proteins and lipids (Semrau, 2011; Xu et al., 2020) Proteobacteria involved in the fermentation of soluble substrates ( Bareither et al., 2013; Xu et al., 2017). Synergistetes in landfill leachate (Song et al., 2015) play important roles in refuse decomposition 1. Taxonomy profile at phylum level

10. RESULT 2: Microbial community composition Anaerobic digester metagenome , Taibaiella , Pusillimonas , uncultured bacterium , Caldicoprobacter , Rubellimicrobium , Syntrophaceticus , and uncultured bacteria were the 8 most abundant bacterial genera shared in almost all samples genus Taibaiella belonging to Bacteroidetes phylum, was the most abundant in the Jabor leachate (30%), while Anaerobic digester metagenome (35%), belonging to Firmicutes phylum, had the highest relative abundance in the Jeram leachate. Taibaiella genus  can remove aniline, other than been found in phenol-degrading microbial communities (Hou et al., 2021). A naerobic digester metagenome genus reduce metabolic intermediates and act as major fermentative bacteria ( Seo et al., 2019). 2. Taxonomy profile at genus level

11. RESULT 3: Predicted metabolic function using PICRUSt2 The NSTI values of Jabor leachate sample is 0.395 , whereas Jeram leachate sample is 0.483 . C arbohydrate metabolism useful in degrading cellulose and hemicellulose for carbon-related metabolism (Toledo et al., 2017) A mino acid metabolism acts as energy and carbon sources during landfilling (Lopez et al., 2015). E nergy metabolism related to methanogenesis, sulfur cycle, and nitrogen cycle in landfill leachate. The diverse microbial communities with various metabolic pathways highlighted the potential roles of microorganisms in the degradation of landfill. Metabolism pathway predicted by PICRUSt2

12. CONCLUSION

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