Ubiquity and Diversity of Cold Adapted Denitrifying Bacteria Isolated From Diverse Antarctic Ecosystems.

Nitrogen cycle has been poorly investigated in Antarctic ecosystems. In particular, how extreme conditions of low temperature, dryness, and high radiation select the microorganisms involved in the cycle is not yet understood. Denitrification is an important step in the nitrogen cycle in which nitrate is reduced stepwise to the gases NO, N2O, and N2. Denitrification is carried out by a wide group of microorganisms spread in the phylogenetic tree. The aim of this work was to isolate and characterize denitrifying bacteria present in different cold environments from Antarctica. Bacterial isolates were obtained from lake, meltwater, sea, glacier ice, ornithogenic soil, and penguin feces samples from King George Island, Fildes peninsula in the Antarctic. Samples were taken during the deicing season in five sampling campaigns. From all the samples we were able to isolate denitrifying strains. A total of 199 bacterial isolates with the capacity to grow in anaerobic mineral media reducing nitrate at 4°C were obtained. The characterization of the isolates by 16S rRNA gene sequence analysis showed a high predominance of the genus Pseudomonas, followed by Janthinobacterium, Flavobacterium, Psychrobacter, and Yersinia. Other minor genera detected were Cryobacterium, Iodobacter, Kaistella, and Carnobacterium. The capacity to denitrify was not previously described for most of the bacteria related to our isolates and in many of them denitrifying genes were not present suggesting the presence of new genes in this extreme environment. Our work demonstrates the ubiquity of denitrification in the Maritime Antarctica and gives important information linking denitrification at cold temperature with taxa in an unequivocal way.

Evaluation of nitrogen removal and the microbial community in a submerged aerated biological filter (SABF), secondary decanters (SD), and horizontal subsurface flow constructed wetlands (HSSF-CW) for the treatment of kennel effluent.

To ensure microbial activity and a reaction equilibrium with efficiency and energy saving, it is important to know the factors that influence microbiological nitrogen removal in wastewater. Thus, it was investigated the microorganisms and their products involved in the treatment of kennel effluents operated with different aeration times, phase 1 (7 h of continuous daily aeration), phase 2 (5 h of continuous daily aeration), and phase 3 (intermittent aeration every 2 h), monitoring chemical and physical parameters weekly, monthly microbiological, and qualitative and quantitative microbiological analyzes at the end of each applied aeration phase. The results showed a higher mean growth of nitrifying bacteria (NB) (106) and denitrifying bacteria (DB) (1022) in phase with intermittent aeration, in which better total nitrogen (TN) removal performance, with 33%, was achieved, against 21% in phase 1 and 17% in phase 2, due to the longer aeration time and lower carbon/nitrogen ratio (15.7), compared with the other phases. The presence of ammonia-oxidizing bacteria (AOB), the genus Nitrobacter nitrite-oxidizing bacteria (NOB), and DB were detected by PCR with specific primers at all phases. The analysis performed by 16S-rRNA DGGE revealed the genres Thauera at all phases; Betaproteobacteria and Acidovorax in phase 3; Azoarcus in phases 2 and 3; Clostridium, Bacillus, Lactobacillus, Turicibacter, Rhodopseudomonas, and Saccharibacteria in phase 1, which are related to the nitrogen removal, most of them by denitrifying. It is concluded that, with the characterization of the microbial community and the analysis of nitrogen compounds, it was determined, consistently, that the studied treatment system has microbiological capacity to remove TN, with the phase 3 aeration strategy, by simultaneous nitrification and denitrification (SND). Due to the high density of DB, most of the nitrification occurred by heterotrophic nitrification-aerobic. And denitrification occurred by heterotrophic and autotrophic forms, since the higher rate of oxygen application did not harm the DB. Therefore, the aeration and carbon conditions in phase 3 favored the activity of the microorganisms involved in these different routes. It is considered that, in order to increase autotrophic nitrification-aerobic, it is necessary to exhaust the volume of sludge in the secondary settlers (SD), further reducing the carbon/nitrogen ratio, through more frequent cleaning, whose periodicity should be the object of further studies. Graphical abstract.

Estimate of denitrifying microbiota in tertiary sewage treatment and kinetics of the denitrification process using different sources of carbon

A study of the kinetics of denitrification was carried out in the laboratory based on the quantification of N2O, the final product of the activity of denitrifying microorganisms, when the enzymatic reduction of N2O to N2 was blocked by acetylene. Concentrated mixed liquor (sludge from a reactor with intermittent aeration used for sewage treatment) was used as the inoculum, while methanol, acetic acid, glucose, effluent sewage from an anaerobic fluidized bed reactor and synthetic substrate simulating domestic sewage were used as carbon sources. The mean concentration of nitrate was 20 mg/L. Maxima of N2O production and NO3- consumption occurred between 0.5h and 2.0h of incubation using all the carbon sources, which characterized the denitrification process. Acetic acid and methanol were responsible for the highest rates of N2O production. The estimated number of denitrifying microorganisms in the reactor with intermittent aeration, using the MPN technique, varied from 10(9) to 10(10) MPN/g VSS, indicating a high potential for the occurrence of denitrification.

O estudo da cinética do processo de desnitrificação foi realizado em laboratório a partir da quantificação de N2O, produto final da atividade de organismos desnitrificantes, quando acetileno bloqueou a redução enzimática de N2O para N2. Foram utilizados, como inóculo, licor misto concentrado (lodo proveniente de reator com aeração intermitente tratando esgoto sanitário) e, como fontes de carbono: metanol, ácido acético, glicose, esgoto efluente de reator anaeróbio de leito expandido/fluidificado e substrato sintético simulando esgoto sanitário. A concentração média de nitrato foi de 20 mg/L. Entre 0,5h e 2,0h de incubação, ocorreram máximos de produção de N2O e de consumo de NO3-, com todas as fontes de carbono usadas, caracterizando o processo de desnitrificação. Ácido acético e metanol foram responsáveis pelas maiores velocidades de produção de N2O. Os números estimados de microrganismos desnitrificantes no reator com aeração intermitente, empregando a técnica de NMP, variaram entre 10(9) a 10(10) NMP/g.SSV, indicando elevado potencial para a ocorrência da desnitrificação.