Enrichment of Anaerobic Ammonium Oxidation (Anammox) Bacteria for Biological Nitrogen Removal of Wastewater

Abstract

Anaerobic ammonium oxidation (anammox) bacteria enrichment was explored for the potential application of ammonium rich wastewater removal. Samples of sludge from mature and young landfill leachate treatment plants were screened and used as inocula for anammox enrichment cultures. Enrichments were monitored for N-NH3, N-NO2-and N-NO3-to detect anammox potential activity. Six of the twelve enrichment cultures showed anammox activity after more than five months of enrichment period. All enrichment cultures that gave positive results were obtained from bottom part of sequencing batch reactor (SBR) lagoon indicating localization of anammox bacteria in anaerobic condition. Polymerase Chain Reaction (PCR) with specific primers targeting anammox and planctomycete were able to amplify the 16S rRNA sequence for anammox bacteria under PCR optimum condition. However, only three of six positive samples were successfully sequenced. DNA sequence analysis using NCBI (BLAST) and RDP showed that the anammox bacterial sequences of the investigated samples were identified as Candidatus Kuenenia stuttgartiensis with similarity of 100% (NCBI) and 99.3% (RDP). Keywords: Anammox; biological nitrogen removal AbstrakPengayaan bakteria anaerobic ammonium oxidation (anammox) dikaji untuk potensi aplikasi bagi rawatan air buangan yang mempunyai kandungan ammonium yang tinggi. Sampel enapcemar daripada loji rawatan cecair larut resap tapak kambusan sisa pepejal disaring dan digunakan sebagai inokulum untuk kultur pengayaan. Pemantauan N-NH3, N-NO2-and N-NO3-semasa pengayaan dilakukan untuk mengesan potensi aktiviti anammox. Enam daripada dua belas kultur pengayaan menunjukkan aktiviti anammox selepas lebih lima bulan tempoh pengayaan. Semua kultur pengayaan yang menunjukkan hasil yang positif adalah diperolehi dari bahagian bawah sequencing batch reactor (SBR) menggambarkan penempatan bakteria anammox di dalam keadaan anaerobik. Polymerase Chain Reaction (PCR) menggunakan primer spesifik mensasarkan bakteria anammox dan planctomycete mampu menggandakan jujukan 16S rRNA di bawah keadaan optima PCR. Walaubagaimanapun, hanya tiga daripada enam sampel yang positif berjaya dijujukkan. Analisis jujukan DNA menggunakan NCBI (BLAST) dan RDP menunjukkan jujukan bakteria anammox bagi sampel yang dikaji dikenalpasti sebagai Candidatus Kuenenia stuttgartiensis dengan 100% persamaan (NCBI) dan 99.3% (RDP).Kata kunci: Anammox; penyingkiran nitrogen biologi

Enrichment of anaerobic ammonium oxidation (anammox) bacteria for short start-up of the anammox process: a review

“…Typically, anammox batch assays are performed during inhibition studies of anammox activity [71][72][73][74] and to determine specific anammox activity (SAA) [75]. However, the batch system is also applicable for anammox culture because 'Candidatus Kuenenia stuttgartiensis' was reported to have been successfully enriched using the batch method [2]. It is recommended that a batch assay be performed for seeding sludge to test several parameters that indicate the existence of anammox activity.…”

“…To avoid oxygen contamination at the start, the inoculation work can be performed in an anaerobic chamber and the medium should be sparged using nitrogen or argon gas in advance. The technique of sparging anammox cultures using argon or nitrogen at the start of the culture process has been applied to remove oxygen from batch experiments [2,71,74,75]. The initial concentration of nitrite that is added to the culture needs to be below the inhibition level because bacterial activity is inhibited at nitrite concentrations ≥15.1 mM [72].…”

“…landfill leachate, wastewater from seafood processing industries, slaughterhouse wastewater, fertilizer manufacturing wastewater and tannery wastewater [1][2][3]) is a potential hazard to the environment. The discharge of untreated wastewater with a high ammonium concentration into the receiving waters can harm aquatic life because the presence of the substance may lead to serious problems, including oxygen depletion and eutrophication [4,5].…”

Penyisihan Nitrogen Melalui Proses Anammox dengan Inokulum Lumpur Instalasi Pengolahan Air Limbah Pabrik Pupuk

“…Berdasarkan karakteristik bakteri anammox, proses penyisihan nitrogen dari air limbah mencapai kondisi yang optimal pada rentang suhu 35-40 o C (Yusof et al 2013). Proses anammox masih memungkinkan terjadi pada suhu di bawah 20 o C namun akan mengalami penurunan kinerja.…”

Microbial Assisted Phytoremediation of Palm Oil Mill Final Discharge (Pomfd) Wastewater

“…Pure isolates were subjected to DNA extraction using Ultraclean™ soil DNA kit (MO Bio Laboratories, Inc., Carlsbad, CA, USA) according to manufacturer's instructions. Partial 16S rRNA fragments of the entire bacterial community were amplified by PCR from the extracted genomic DNA using the general bacteria primers, 27F (AGA GTT TGA TCC TGG CTC AG) and 1492R (ACG GTT ACC TTG TTA CGA CTT) (Yusof et al, 2013). The PCR programme consisted of an initial 4 min denaturation step at 95°C followed by 35 cycles of repeated denaturation at 95°C for 45 s, annealing at 56°C for 45 s, and an extension at 70°C for 1 min followed by a final extension at 72°C for 3 min.…”