Impact of Long-Term Fertilization on Community Structure of Ammonia Oxidizing and Denitrifying Bacteria Based on amoA and nirK Genes in a Rice Paddy from Tai Lake Region,China

Ammonia oxidizing （AOB） and denitrifying bacteria （DNB） play an important role in soil nitrogen transformation in natural and agricultural ecosystems. Effects of long-term fertilization on abundance and community composition of AOB and DNB were studied with targeting ammonia monooxygenase （amoA） and nitrite reductase （nirK） genes using polymerase chain reaction- denaturing gradient gel electrophoresis （PCR-DGGE） and real-time PCR, respectively. A field trial with different fertilization treatments in a rice paddy from Tai Lake region, centre East China was used in this study, including no fertilizer application （NF）, balanced chemical fertilizers （CF）, combined organic/inorganic fertilizer of balanced chemical fertilizers plus pig manure （CFM）, and plus rice straw return （CFS）. The abundances and riehnesses of amoA and nirK were increased in CF, CFM and CFS compared to NF. Principle component analysis of DGGE profiles showed significant difference in nirK and amoA genes composition between organic amended （CFS and CFM） and the non-organic amended （CF and NF） plots. Number of amoA copies was significantly positively correlated with normalized soil nutrient richness （NSNR） of soil organic carbon （SOC） and total nitrogen （T-N）, and that of nirK copies was with NSNR of SOC, T-N plus total phosphorus. Moreover, nitrification potential showed a positive correlation with SOC content, while a significantly lower denitrification potential was found under CFM compared to under CFS. Therefore, SOC accumulation accompanied with soil nutrient richness under long-term balanced and organic/inorganic combined fertilization promoted abundance and diversity of AOB and DNB in the rice paddy.

Applicability of the Arrhenius model for Ammonia Oxidizing Bacteria subjected to temperature time gradients

The aim of this work is to identify the range of applicability of Arrhenius type temperature dependence for Ammonia Oxidizing Bacteria （AOB） subjected to tem- perature time gradients through continuous titrimetric tests. An innovative online differential titrimetric technique was used to continuously monitor the maximum biologic ammonia oxidation rate of the biomass selected in a pilot scale membrane bioreactor, as a function of temperature time gradients. The monitoring technique is based on the measurement of alkalinity and hydrogen peroxide con- sumption rates in two parallel reactors operated in non- limiting substrate conditions for AOB; both reactors were continuously fed with mixed liquor and in one of them AOB were inhibited with allylthiourea. The effects of temperature decrease rates in the range 1 to 4℃h^-1 were evaluated by controlling the titrimetric reactor in the temperature range 10℃-20℃. The dependence of growth kinetics on temperature time gradients and the range of applicability of Arrhenius model for temperature depen- dency of AOB growth kinetics were assessed. The Arrhenius model was found to be accurate only with temperature gradients lower than 2℃·h^-1. The estimated Arrhenius coefficients （θ） were shown to increase from 1.07 to 1.6 when the temperature decrease rate reached 4℃.h^-1.

Simultaneous removal of COD and nitrogen using a novel carbon-membrane aerated biofilm reactor

A membrane aerated biofilm reactor is a promising technology for wastewater treatment. In this study, a carbon-membrane aerated biofilm reactor （CMABR） has been developed, to remove carbon organics and nitrogen simultaneously from one reactor. The results showed that CMABR has a high chemical oxygen demand （COD） and nitrogen removal efficiency, as it is operated with a hydraulic retention time （HRT） of 20 h, and it also showed a perfect performance, even if the HRT was shortened to 12 h. In this period, the removal efficiencies of COD, ammonia nitrogen （NH4^＋-N）, and total nitrogen （TN） reached 86%, 94%, and 84%, respectively. However, the removal efficiencies of NH4^＋-N and TN declined rapidly as the HRT was shortened to 8 h. This is because of the excessive growth of biomass on the nonwoven fiber and very high organic loading rate. The fluorescence in situ hybridization （FISH） analysis indicated that the ammonia oxidizing bacteria （AOB） were mainly distributed in the inner layer of the biofilm. The coexistence of AOB and eubacteria in one biofilm can enhance the simultaneous removal of COD and nitrogen.

Feasibility of low-intensity ultrasound treatment with hydroxylamine to accelerate the initiation of partial nitrification and allow operation under intermittent aeration

Partial nitrification is a key aspect of efficient nitrogen removal,although practically it suf-fers from long start-up cycles and unstable long-term operational performance.To address these drawbacks,this study investigated the effect of low intensity ultrasound treatment combined with hydroxylamine(NH2OH)on the performance of partial nitrification.Results showthat compared with the control group,low-intensity ultrasound treatment(0.10W/mL,15 min)combined with NH2OH(5 mg/L)reduced the time required for partial nitrification initiation by 6 days,increasing the nitrite accumulation rate(NAR)and ammonia nitro-gen removal rate(NRR)by 20.4% and 6.7%,respectively,achieving 96.48% NRR.Mechanis-tic analysis showed that NH2OH enhanced ammonia oxidation,inhibited nitrite-oxidizing bacteria(NOB)activity and shortened the time required for partial nitrification initiation.Furthermore,ultrasonication combined with NH2OH dosing stimulated EPS(extracellular polymeric substances)secretion,increased carbonyl,hydroxyl and amine functional group abundances and enhanced mass transfer.In addition,16S rRNA gene sequencing results showed that ultrasonication-sensitive Nitrospira disappeared from the ultrasound+NH_(2)OH system,while Nitrosomonas gradually became the dominant group.Collectively,the results of this study provide valuable insight into the enhancement of partial nitrification start-up during the process of wastewater nitrogen removal.

Overlooked nitrogen-cycling microorganisms in biological wastewater treatment

Nitrogen-cycling microorganisms play key roles at the intersection of microbiology and wastewater engineering.In addition to the well-studied ammonia oxidizing bacteria,nitrite oxidizing bacteria,heterotrophic denitrifiers,and anammox bacteria,there are some other N-cycling microorganisms that are less abundant but functionally important in wastewater nitrogen removal.These microbes include,but not limited to ammonia oxidizing archaea(AOA),complete ammonia oxidation(comammox)bacteria,dissimilatory nitrate reduction to ammonia(DNRA)bacteria,and nitrate/nitrite-dependent anaerobic methane oxidizing(NO_(x)-DAMO)microorganisms.In the past decade,the development of high-throughput molecular technologies has enabled the detection,quantification,and characterization of these minor populations.The aim of this review is therefore to synthesize the current knowledge on the distribution,ecological niche,and kinetic properties of these“overlooked”N-cycling microbes at wastewater treatment plants.Their potential applications in novel wastewater nitrogen removal processes are also discussed.A comprehensive understanding of these overlooked N-cycling microbes from microbiology,ecology,and engineering perspectives will facilitate the design and operation of more efficient and sustainable biological nitrogen removal processes.