Optimization of depth of filler media in horizontal flow constructed wetlands for maximizing removal rate coefficients of targeted pollutant(s).

Varying the depth of HFCW media causes differences in the redox status within the system, and hence the community structure and diversity of bacteria, affecting removal rates of different pollutants. The key functional microorganisms of CWs that remove contaminants belong to the phyla Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes. Secondary data of 111 HFCWs (1232 datasets) were analyzed to deduce the relationship between volumetric removal rate coefficients (KBOD, KTN, KTKN, and KTP) and depth. Equations of depth were derived in terms of rate coefficients using machine learning approach (MLR and SVR) (R2 = 0.85, 0.87 respectively). These equations were then used to find the optimum depth for pollutant(s) removal using Grey wolf optimization (GWO). The computed optimum depths were 1.48, 1.71, 1.91, 2.09, and 2.14 m for the removal of BOD, TKN, TN, TP, and combined nutrients, respectively, which were validated through primary data. This study would be helpful for optimal design of HFCWs.

Antibiotic resistance genes and bacterial diversity: A comparative molecular study of treated sewage from different origins and their impact on irrigated soils.

Present study aims to investigate how is soil affected following irrigation with treated effluents of different origins by analysing the bacterial diversity, metabolic diversity and antibiotic resistance genes (ARGs). Comparative analysis with previously reported ARGs in effluents was performed to understand the mobility of ARGs from treated wastewater to the irrigated soil with respect to the control soil regimen. Acinetobacter, Burkholderia and Pseudomonas were observed as the most abundant genera in all the samples. The metabolic gene abundance of all the samples suggests a prominent contribution to natural mineral recycling. Most abundant ARGs observed encode resistance for clindamycin, kanamycin A, macrolides, paromomycin, spectinomycin and tetracycline. Treated effluent reuse did not appear to enhance the ARG levels in soils in most cases except for institutional treatment site (M), where the ARGs for aminoglycosides, ß-lactams and sulfonamides were found to be abundantly present in both treated effluent and the irrigated soil. This study finds the importance of wastewater treatment from different origins and the impact of treated wastewater reuse in irrigation. This study also emphasises on the better understanding of ARGs mobility from water to soil.

A systematic review on chlorine tolerance among bacteria and standardization of their assessment protocol in wastewater.

Though chlorine is a cost-effective disinfectant for water and wastewaters, the bacteria surviving after chlorination pose serious public health and environmental problems. This review critically assesses the mechanism of chlorine disinfection as described by various researchers; factors affecting chlorination efficacy; and the re-growth potential of microbial contaminations in treated wastewater post chlorination to arrive at meaningful doses for ensuring health safety. Literature analysis shows procedural inconsistencies in the assessment of chlorine tolerant bacteria, making it extremely difficult to compare the tolerance characteristics of different reported tolerant bacteria. A comparison of logarithmic reduction after chlorination and the concentration-time values for prominent pathogens led to the generation of a standard protocol for the assessment of chlorine tolerance. The factors that need to be critically monitored include applied chlorine doses, contact time, determination of chlorine demands of the medium, and the consideration of bacterial counts immediately after chlorination and in post chlorinated samples (regrowth). The protocol devised here appropriately assesses the chlorine-tolerant bacteria and urges the scientific community to report the regrowth characteristics as well. This would increase the confidence in data interpretation that can provide a better understanding of chlorine tolerance in bacteria and aid in formulating strategies for effective chlorination.

Strategies for enhancing phosphorous removal in Vertical Flow Constructed Wetlands.

Constructed wetlands (CWs) are among the fastest emerging treatment methods for wastewater treatment. Unlike their organics and nitrogen removal capacities, the potential of CWs as a sink for phosphorous is still debatable. In this study, the secondary data from several CWs treating domestic sewage were compiled and assessed. Curves were plotted between orthophosphate-phosphorous (PO43--P) loading and the corresponding removal rates. Other factors affecting PO43--P removal like depth of the CW, surface area, organic loading rate etc. Were also analyzed. Removal rates of PO43--P were conforming to a linear positive relation with the loading rates. Pea gravel as a CW medium performed consistently well (60-80% removal) for a wide range of influent PO43--P loading (0.5-1.5 g/m3-d). The increased depth of the wetland appears to favor phosphate removal. PO43--P removal was found to be correlated with outlet dissolved oxygen, total Kjeldahl nitrogen removal and effluent nitrate. The study suggests that proper design, optimal organic loading and suitable pre-treatment may increase the applicability of CWs for phosphate removal from domestic wastewater. Larger area requirements can also be avoided by increasing their depth while keeping the volume of the filter media the same.

Designing the vertical flow constructed wetland based on targeted limiting pollutant.

The requirement of large land area limits the adoption of constructed wetlands (CWs) in urban settings with limited land availability. The area calculations for CW design are commonly carried out following Kikuth approach where the removal rate constant (K) is derived from literature. Investigation of secondary data of 82 vertical flow CWs, performed in this study, yielded wide variations (0.0003 - 0.822 md-1) in the calculated K values for different pollutants under different environmental and operational conditions indicating that it is important to incorporate the desired levels of pollutant removal to arrive at customized design of CWs. The results indicated that the relative standard deviation of K values could be narrowed by classifying the datasets based on design parameters like depth, hydraulic loading rates and substrate loading rates. These calculations can help arrive at more scientific design of CW to achieve the prevailing standards for the discharge or reuse of sewage.

Impact of combined chlorine and UV technology on the bacterial diversity, antibiotic resistance genes and disinfection by-products in treated sewage.

This paper explores the effect of hybrid chlorine and UV disinfection treatment against their individual usage on microbial community, functional genes, antibiotic resistant genes (ARGs) and disinfection by-products (DBPs) formation. The disinfectant doses of 2.5 mg L-1 chlorine and 41 mJ cm-2 UV were selected based on the coliform counts to be attained in treated sewage. The highest bacterial diversity was observed in control (secondary treated) sample followed by UV, chlorine and hybrid disinfection. The highest elimination of bacterial species (296) was achieved in hybrid treatment, which was far better than the standalone treatments. The disinfection with all the disinfectants used resulted in increased abundance of ARGs. Motility genes were found to be enriched in hybrid disinfected samples. DBP concentrations were within the stipulated norms for all the disinfectant treatments used. Hybrid disinfection was observed to be more effective in alleviating the risks associated with the reuse of treated sewage.

Novel microbial nitrogen transformation processes in constructed wetlands treating municipal sewage: a mini-review.

Traditionally nitrogen transformation in constructed wetlands (CWs) has been attributed to the activities of aerobic autotrophic nitrifiers followed by anoxic heterotrophic denitrifiers. However, the nitrogen balances in such systems are far from being explained as a large fraction of the losses remain unaccounted for. The classical nitrification-denitrification theory has been successfully employed in certain unit processes by culturing fast-growing bacteria, but the CWs offer an ideal environment for slow-growing bacteria that may be beneficially exploited to achieve enhanced nitrogen removal by manipulating the environmental conditions in their favor. In the last three decades, many novel microorganisms have been isolated from CWs that have led to the discovery of some other routes that have made researchers believe could play a significant role in nitrogen transformation processes. The increased understanding of novel discerned pathways like anaerobic ammonium oxidation (ANAMMOX), heterotrophic nitrification and aerobic denitrification, which are mediated by specialized bacteria has indicated that these microorganisms could be enriched by applying selection pressures within CWs for achieving high rates of nitrogen removal. Understanding these novel nitrogen transformation processes along with the associated microbial population can provide new dimensions to the design of CWs for enhanced nitrogen removal.

Investigation of chlorine tolerance profile of dominant gram negative bacteria recovered from secondary treated wastewater in Jaipur, India.

Management of bacterial survival post chlorine disinfection is vital for safe wastewater reuse for irrigation, as the presence of microorganisms in large numbers may lead to subsequent contamination of the surface and ground water reservoirs. Even after satisfying the current norms of coliform counts after disinfection (less than 1000 MPN per 100 mL), chlorine tolerant bacteria surviving in inadequately treated wastewater may pose a public health threat as many of these bacteria are able to re-grow upon storage. The current study is aimed to assess the magnitude of the problem posed by chlorine tolerant bacteria during chlorine disinfection and attempts to derive a strategy for safe disinfection. The chlorine tolerance was examined in the dominant gram negative bacteria (GNB) recovered from secondary treated sewage from a treatment plant located at Jaipur, India. Bacterial survival and re-growth (after 24 h) studies on test species (n = 11) with reference to E.coli ATCC 25922 reveal that, while the lethal doses of isolates ranged from 0.5 to 1.25 mgL-1,the chlorine doses for complete inhibition of re-growth were much higher (0.75-1.75 mgL-1).The isolates showing highest lethal dose and re-growth inhibition dose, identified as Citrobacter freundii, Klebsiella sp. and Stenotrophomonas maltophilia also exhibited very low log effective reduction (0.72-1.90) values and were selected as chlorine tolerant bacteria. Results of inactivation kinetics experiments on chlorine tolerant bacteria reveal a strong correlation (R2 > 0.89-0.99) between log reduction values and contact time. In re-growth kinetics experiments, maximum re-growth was observed after 6 h exposure following which, only marginal increase was registered up to 24 h. The study indicates that the existing approach of bacterial elimination post chlorine treatment may be grossly inadequate to assess the performance of the disinfection process adopted for drinking water treatment. It further brings out a novel approach to arrive at meaningful chlorine doses that take bacterial re-growth into account for achieving safe disinfection.

Defluoridation of water using autochthonous bacterial isolates.

Prolonged consumption of fluoride-contaminated water poses health problems like dental and skeletal fluorosis in many parts of the world including India. In regions with acute water scarcity, it demands immediate intervention like de-fluoridation of water before consumption. In the current study, fluoride-resistant bacteria were isolated from fluoride-contaminated groundwater and soil samples collected from Dungarpur district, India, for their potential use in defluoridation. Out of a total of 53 bacterial isolates that were recovered and screened for fluoride resistance, three highly fluoride-resistant isolates DWC1, DWC2 and DWB5, resistant to up to 9200 mg L-1, 7200 mg L-1 and 5200 mg L-1 fluoride respectively, were characterized and identified as Aeromonas sp., Brevibacterium sp. and Paenibacillus sp. respectively. The fluoride removal capacity of isolates DWC1, DWC2, DWB5 and a consortium of all the three isolates was found to be 68.7%, 73.4%, 76.7 % and 70.1% respectively on nutrient broth supplemented with NaF (2000 mg L-1) after 8 days of incubation. Defluoridation conditions for the strain showing the best result (Paenibacillus sp.) were optimized for real fluoride-rich water collected from Ajmer District, India, using the Taguchi design of experiment. A defluoridation of up to 73.3% was observed at 40 °C temperature and pH 8 with inoculum: water ratio of 2:1 after 8 days of incubation. To the best of our knowledge, the defluoridation capacity of Paenibacillus sp. is the highest reported in literature to date for real water samples and could be investigated in further detail for commercial application.

Correction: ExtremeDB: A Unified Web Repository of Extremophilic Archaea and Bacteria.

[This corrects the article DOI: 10.1371/journal.pone.0063083.].

ExtremeDB: a unified web repository of extremophilic archaea and bacteria.

Extremophiles are the microorganisms which can survive under extreme conditions of temperature, pressure, pH, salinity etc. They have gained much attention for their potential role in biotechnological and industrial applications. The large amount of experimental data in the literature is so diverse, that it becomes difficult and time consuming for the researcher to implement it in various areas of research. Therefore, a systematic arrangement of data and redirection in a similar fashion through web interface can assist researchers in analyzing the data as per their requirement. ExtremeDB is a freely available web based relational database which integrates general characteristics, genome-proteome information, industrial applications and recent scientific investigations of the seven major groups of 865 extremophillic microorganisms. The search options are user friendly and analyses tools such as Compare and Extreme BLAST have been incorporated for comparative analysis of two or more extremophiles and determining the sequence similarity of a given protein/nucleotide in relation to other extremophiles respectively. The effort put forth herein in the form of database, would open up new avenues on the potential utility of extremophiles in applied research. ExtremeDB is freely accessible via http://extrem.igib.res.in.

Usefulness of organic acid produced by Exiguobacterium sp. 12/1 on neutralization of alkaline wastewater.

The aim of this study was to investigate the role of organic acids produced by Exiguobacterium sp. strain 12/1 (DSM 21148) in neutralization of alkaline wastewater emanated from beverage industry. This bacterium is known to be able to grow in medium of pH as high as pH 12.0 and to neutralize alkaline industrial wastewater from pH 12.0 to pH 7.5. The initial investigation on the type of functional groups present in medium, carried out using FT-IR spectroscopy, revealed the presence of peaks corresponding to carbonyl group and hydroxyl group, suggesting the release of carboxylic acid or related metabolic product(s). The identification of specific carboxylic group, carried out using RP-HPLC, revealed the presence of a single peak in the culture supernatant with retention time most similar to formic acid. The concentration of acid produced on different carbon sources was studied as a function of time. Although acid was present in same final concentration, the rate of acid production was highest in case of medium supplemented with sucrose followed by fructose and glucose. The knowledge of metabolic products of the bacterium can be considered as a first step towards realization of its potential for large-scale bioremediation of alkaline wastewater from beverage industry.