Tawan Limpiyakorn

amoA-encoding archaea in wastewater treatment plants: a review

Recent evidence from natural environments suggests that in addition to ammonia-oxidizing bacteria, ammonia-oxidizing archaea (AOA) affiliated with Thaumarcheota, a new phylum of the domain Archaea, also oxidize ammonia to nitrite and thus participate in the global nitrogen cycle. Besides natural environments, modern data indicate the presence of amoA-encoding archaea (AEA) in wastewater treatment plants (WWTPs). To further elucidate whether AEA in WWTPs are AOA and to clarify the role of AEA in WWTPs, this paper reviews the current knowledge on this matter for wastewater engineers and people in related fields. The initial section coveys a microbiological point of view and is particularly based upon data from AOA cultures. The later section summarizes what is currently known about AEA in relation to WWTPs. Based on the reviewed data, future research pathways are proposed in an effort to further what is known about AEA in wastewater treatment systems.

Atrazine degradation by stable mixed cultures enriched from agricultural soil and their characterization.

Aims:  The aim of this work was to enrich stable mixed cultures from atrazine‐contaminated soil. The cultures were examined for their atrazine biodegradation efficiencies in comparison with J14a, a known atrazine‐degrading strain of Agrobacterium radiobacter. The cultures were also characterized to identify community structure and bacterial species present.

Development and application of real-time PCR for quantification of specific ammonia-oxidizing bacteria in activated sludge of sewage treatment systems.

In this study, four real-time polymerase chain reaction (PCR) primer sets were developed for the 16S rRNA genes of specific ammonia-oxidizing bacteria (AOB) found in activated sludge of sewage treatment systems. The primer sets target two of several sequence types of the Nitrosomonas oligotropha cluster, members within the Nitrosomonas communis cluster, and all members of the Nitrosomonas europaea–Nitrosococcus mobilis cluster. The detection limit of each primer set was in the range of 3×101–6×102 genes reaction−1. Reliable quantification of the target AOB DNA was obtained when the target AOB DNA comprised more than 0.1% of total AOB DNA in the sample. The application of the primer sets to samples taken from five sewage treatment systems showed that, in all systems, the majority of the AOB population was comprised of one sequence type of the N. oligotropha cluster (3.9±1.5×109–1.7±0.5×1010 cell l−1) and, in most systems, followed by members within the N. communis cluster (2.8±0.3×109–1.0±0.1×1010 cell l−1) or/and another sequence type of the N. oligotropha cluster (1.5±0.6×108–5.5±0.5×108 cell l−1). N. europaea–N. mobilis cluster arose solely in small numbers (4.9±0.8×108 cell l−1) in one system. Real-time PCR-amplified products obtained from genomic DNA extracted from samples were verified using clone library, and it revealed that only the target AOB DNA were PCR amplified, without amplification of the nontarget sequences.

Effect of silver nanoparticles on Pseudomonas putida biofilms at different stages of maturity.

This study determined the effect of silver nanoparticles (AgNPs) on Pseudomonas putida KT2440 biofilms at different stages of maturity. Three biofilm stages (1-3, representing early to late stages of development) were identified from bacterial adenosine triphosphate (ATP) activity under static (96-well plate) and dynamic conditions (Center for Disease Control and Prevention biofilm reactor). Extracellular polymeric substance (EPS) levels, measured using crystal violet and total carbohydrate assays, and expression of the EPS-associated genes, csgA and alg8, supported the conclusion that biofilms at later stages were older than those at earlier stages. More mature biofilms (stages 2 and 3) showed little to no reduction in ATP activity following exposure to AgNPs. In contrast, the same treatment reduced ATP activity by more than 90% in the less mature stage 1 biofilms. Regardless of maturity, biofilms with EPS stripped off were more susceptible to AgNPs than controls with intact EPS, demonstrating that EPS is critical for biofilm tolerance of AgNPs. The findings from this study show that stage of maturity is an important factor to consider when studying effect of AgNPs on biofilms.

Fate of Estrogens and Estrogenic Potentials in Sewerage Systems

Release of steroid estrogens such as estrone (E1), 17β-estradiol (E2), estriol (E3), and 17α-ethinylestradiol (EE2) into the urban environment is of concern due to their prominent endocrine disrupting properties. This review focuses on the fate of estrogens as well as estrogenic potentials in the sewerage system starting from human excretion to municipal sewage treatment facilities. To examine the roles of different components in the municipal sewage treatment facilities in the removal of estrogens, data from 130 full-scale municipal sewage treatment plants (STPs) in 14 countries were summarized. Primary treatment systems removed less than 10% of the influent estrogens. Municipal STPs with the secondary biological treatment systems gave higher percentage removals of estrogens and estrogenic potentials than municipal STPs without secondary biological treatment systems. On average, municipal STPs with suspended growth systems gave better percentage removals of estrogens and estrogenic potentials than attached growth systems. Biological processes with nitrification had higher percentage removals than processes without nitrification. Microorganisms responsible for the degradation of estrogens under various conditions were also reviewed and presented.

Degradation of 17α-methyltestosterone by Rhodococcus sp. and Nocardioides sp. isolated from a masculinizing pond of Nile tilapia fry

17α-Methyltestosterone (MT), a synthetic anabolic androgenic steroid, is widely used in aquafarming for the production of an all male fish population such as Nile tilapia. This study isolated, identified and characterized MT-degrading bacteria in the sediment and water from a masculinizing pond of Nile tilapia fry. Based on the phylogeny, physiological properties and cell morphology, the three isolated MT-degrading bacteria were related closely to Rhodococcus equi, Nocardioides aromaticivorans, and Nocardioides nitrophenolicus. Growth of the three isolated strains was found to be inhibited for MT concentrations in the range of 1.0-10mg/L. The inhibition of cell growth was found to be modeled using the Haldanes substrate inhibition model. The kinetic constants ranged from 0.13 to 0.19h(-1) for μ(max), 0.7-24.8mg/L for K(s) and 19.6-76.2mg/L for K(i). Androgenic activity using β-galactosidase assay showed that all strains degraded MT to the products with no androgenic potency.

Biotransformation of 17α-methyltestosterone in sediment under different electron acceptor conditions.

17α-Methyltestosterone (MT), an anabolic androgenic steroid, is used widely in inducing an all male population in aquaculture farming of fish, such as Nile tilapia (Oreochromis niloticus). Current understanding of the occurrence and fate of MT in the sediments and the surrounding areas of the aquaculture ponds are very limited. Bioassay tests showed that MT was biotransformed under aerobic and sulfate-reducing conditions with a half-life of 3.8d and 5.3d, respectively, with complete disappearance of androgenic activity. However, under methanogenic condition, MT was found to biotransform but the androgenic activity continued to persist even after 45 d of incubation. In contrast, MT was found to transform slowly under iron(III)-reducing condition and was hardly transformed under nitrate-reducing condition. A possible reason for the lack of transformation of MT under nitrate-reducing condition is the presence of the methyl group at the C-17 position. The results of this study suggest that MT and its degradation products with androgenic activity may potentially accumulate in the sediments of fish farming ponds under iron(III)-reducing, nitrate-reducing and methanogenic conditions.

Communities of ammonia-oxidizing bacteria, ammonia-oxidizing archaea and nitrite-oxidizing bacteria in shrimp ponds

The communities of nitrifying microorganisms were identified in samples taken from six shrimp ponds in Thailand (five outdoor-earthen ponds and one indoor pond). The sequences of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA), and nitrite-oxidizing bacteria (NOB) were analyzed after specific PCR amplification of the 16S rRNA or amoA genes. Among the ammonia-oxidizing microorganisms, AOB appeared to be the most dispersed throughout the ponds, while AOA sequences were only retrieved from three ponds. The AOB found in the shrimp ponds belonged to only two AOB clusters, the Nitrosomonas sp. Nm143 cluster and Nitrosomonas marina cluster, which are reported with salt requirements. The majority of AOA sequences fell closer to group I.1a Thaumarcheota rather than group I.1b Thaumarcheota. For NOB, Nitrospira, but not Nitrobacter, were detected. NOB of sublineage II and IV Nitrospira, which were previously reported as salt tolerant and salt requirement NOB, were found to be common in the shrimp ponds. Insight into ammonia-oxidizing microorganisms, numbers of AOB and AOA amoA genes in two selected ponds (one outdoor-earthen ponds and one indoor pond) were quantified using qPCR. High numbers of AOA amoA genes were found in both ponds. The information obtained from this study clearly identifies the microorganisms responsible for nitrification in shrimp ponds.

Effects of inoculum type and bulk dissolved oxygen concentration on achieving partial nitrification by entrapped-cell-based reactors.

An entrapment of nitrifiers into gel matrix is employed as a tool to fulfill partial nitrification under non-limiting dissolved oxygen (DO) concentrations in bulk solutions. This study aims to clarify which of these two attributes, inoculum type and DO concentration in bulk solutions, is the decisive factor for partial nitrification in an entrapped-cell based system. Four polyvinyl alcohol entrapped inocula were prepared to have different proportions of nitrite-oxidizing bacteria (NOB) and nitrite-oxidizing activity. At a DO concentration of 3 mg l(-1), the number of active NOB cells in an inoculum was the decisive factor for partial nitrification enhancement. However, when the DO concentration was reduced to 2 mg l(-1), all entrapped cell inocula showed similar degrees of partial nitrification. The results suggested that with the lower bulk DO concentration, the preparation of entrapped cell inocula is not useful as the DO level becomes the decisive factor for achieving partial nitrification.

Survey of Microbial Diversity in Flood Areas during Thailand 2011 Flood Crisis Using High-Throughput Tagged Amplicon Pyrosequencing

The Thailand flood crisis in 2011 was one of the largest recorded floods in modern history, causing enormous damage to the economy and ecological habitats of the country. In this study, bacterial and fungal diversity in sediments and waters collected from ten flood areas in Bangkok and its suburbs, covering residential and agricultural areas, were analyzed using high-throughput 454 pyrosequencing of 16S rRNA gene and internal transcribed spacer sequences. Analysis of microbial community showed differences in taxa distribution in water and sediment with variations in the diversity of saprophytic microbes and sulfate/nitrate reducers among sampling locations, suggesting differences in microbial activity in the habitats. Overall, Proteobacteria represented a major bacterial group in waters, while this group co-existed with Firmicutes, Bacteroidetes, and Actinobacteria in sediments. Anaeromyxobacter, Steroidobacter, and Geobacter were the dominant bacterial genera in sediments, while Sulfuricurvum, Thiovirga, and Hydrogenophaga predominated in waters. For fungi in sediments, Ascomycota, Glomeromycota, and Basidiomycota, particularly in genera Philipsia, Rozella, and Acaulospora, were most frequently detected. Chytridiomycota and Ascomycota were the major fungal phyla, and Rhizophlyctis and Mortierella were the most frequently detected fungal genera in water. Diversity of sulfate-reducing bacteria, related to odor problems, was further investigated using analysis of the dsrB gene which indicated the presence of sulfate-reducing bacteria of families Desulfobacteraceae, Desulfobulbaceae, Syntrobacteraceae, and Desulfoarculaceae in the flood sediments. The work provides an insight into the diversity and function of microbes related to biological processes in flood areas.