Illumina sequencing-based analysis of a microbial community enriched under anaerobic methane oxidation condition coupled to denitrification revealed coexistence of aerobic and anaerobic methanotrophs

Methane is produced in anaerobic environments, such as reactors used to treat wastewaters, and can be consumed by methanotrophs. The composition and structure of a microbial community enriched from anaerobic sewage sludge under methane-oxidation condition coupled to denitrification were investigated. Denaturing gradient gel electrophoresis (DGGE) analysis retrieved sequences of Methylocaldum and Chloroflexi. Deep sequencing analysis revealed a complex community that changed over time and was affected by methane concentration. Methylocaldum (8.2%), Methylosinus (2.3%), Methylomonas (0.02%), Methylacidiphilales (0.45%), Nitrospirales (0.18%), and Methanosarcinales (0.3%) were detected. Despite denitrifying conditions provided, Nitrospirales and Methanosarcinales, known to perform anaerobic methane oxidation coupled to denitrification (DAMO) process, were in very low abundance. Results demonstrated that aerobic and anaerobic methanotrophs coexisted in the reactor together with heterotrophic microorganisms, suggesting that a diverse microbial community was important to sustain methanotrophic activity. The methanogenic sludge was a good inoculum to enrich methanotrophs, and cultivation conditions play a selective role in determining community composition.     

Start-up of sequencing batch reactor with <Emphasis Type="Italic">Thiosphaera pantotropha</Emphasis> for treatment of high-strength nitrogenous wastewater and sludge characterization

Biological treatment of high-strength nitrogenous wastewater is challenging due to low growth rate of autotrophic nitrifiers. This study reports bioaugmentation of Thiosphaera pantotropha capable of simultaneously performing heterotrophic nitrification and aerobic denitrification (SND) in sequencing batch reactors (SBRs). SBRs fed with 1:1 organic-nitrogen (N) and NH₄ ⁺-N were started up with activated sludge and T. pantotropha by gradual increase in N concentration. Sludge bulking problems initially observed could be overcome through improved aeration and mixing and change in carbon source. N removal decreased with increase in initial nitrogen concentration, and only 50–60 % removal could be achieved at the highest N concentration of 1000 mg L^?1 at 12-h cycle time. SND accounted for 28 % nitrogen loss. Reducing the settling time to 5–10 min and addition of divalent metal ions gradually improved the settling characteristics of sludge. Sludge aggregates of 0.05–0.2 mm diameter, much smaller than typical aerobic granules, were formed and progressive increase in settling velocity, specific gravity, Ca²⁺, Mg²⁺, protein, and polysaccharides was observed over time. Granulation facilitated total nitrogen (TN) removal at a constant rate over the entire 12-h cycle and thus increased TN removal up to 70 %. Concentrations of NO₂ ^?-N and NO₃ ^?-N were consistently low indicating effective denitrification. Nitrogen removal was possibly limited by urea hydrolysis/nitrification. Presence of T. pantotropha in the SBRs was confirmed through biochemical tests and 16S rDNA analysis.     

Evaluation of the giant reed (<Emphasis Type="Italic">Arundo donax</Emphasis>) in horizontal subsurface flow wetlands for the treatment of recirculating aquaculture system effluent

Introduction

Two emergent macrophytes, Arundo donax and Phragmites australis, were established in experimental subsurface flow, gravel-based constructed wetlands (CWs) receiving untreated recirculating aquaculture system wastewater.

Materials and methods

The hydraulic loading rate was 3.75 cm day^?1. Many of the monitored water quality parameters (biological oxygen demand [BOD], total suspended solids [TSS], total phosphorus [TP], total nitrogen [TN], total ammoniacal nitrogen [TAN], nitrate nitrogen [NO₃], and Escherichia coli) were removed efficiently by the CWs, to the extent that the CW effluent was suitable for use on human food crops grown for raw produce consumption under Victorian state regulations and also suitable for reuse within aquaculture systems.

Results and discussion

The BOD, TSS, TP, TN, TAN, and E. coli removal in the A. donax and P. australis beds was 94%, 67%, 96%, 97%, 99.6%, and effectively 100% and 95%, 87%, 95%, 98%, 99.7%, and effectively 100%, respectively, with no significant difference (p?>?0.007) in performance between the A. donax and P. australis CWs. In this study, as expected, the aboveground yield of A. donax top growth (stems + leaves) (15.0?±?3.4 kg wet weight) was considerably more than the P. australis beds (7.4?±?2.8 kg wet weight). The standing crop produced in this short (14-week) trial equates to an estimated 125 and 77 t ?ha^?1 year^?1 biomass (dry weight) for A. donax and P. australis, respectively (assuming that plant growth is similar across a 250-day (September–April) growing season and a single-cut, annual harvest).

Conclusion

The similarity of the performance of the A. donax- and P. australis-planted beds indicates that either may be used in horizontal subsurface flow wetlands treating aquaculture wastewater, although the planting of A. donax provides additional opportunities for secondary income streams through utilization of the energy-rich biomass produced.

     

Simple technology for recycling phosphate from wastewater to farmland in rural areas

A simple technology for phosphate (P _i ) recovery has been developed using a bifunctional adsorption–aggregation agent. The bifunctional agent was prepared by soaking calcium silicates in hydrochloric acid solution. Importantly, recyclable calcium silicates were available almost free of charge from the cement industry and also from the steel industry. The acid treatment was essential not only for enhancing the ability of calcium silicates to remove P _i from aqueous solution but also for enabling the high settleability of removed P _i . On-site experiments using a mobile plant showed that approximately 80% P _i could be recovered from anaerobic sludge digestion liquor at a wastewater treatment plant. This technology has the potential to offer a simple, compact service for recycling P _i from wastewater to farmland in rural areas.     

Treatment of Arctic wastewater by chemical coagulation,UV and peracetic acid disinfection

Conventional wastewater treatment is challenging in the Arctic region due to the cold climate and scattered population. Thus, no wastewater treatment plant exists in Greenland, and raw wastewater is discharged directly to nearby waterbodies without treatment. We investigated the efficiency of physicochemical wastewater treatment, in Kangerlussuaq, Greenland. Raw wastewater from Kangerlussuaq was treated by chemical coagulation and UV disinfection. By applying 7.5 mg Al/L polyaluminium chloride (PAX XL100), 73% of turbidity and 28% phosphate was removed from raw wastewater. E. coli and Enterococcus were removed by 4 and 2.5 log, respectively, when UV irradiation of 0.70 kWh/m³ was applied to coagulated wastewater. Furthermore, coagulated raw wastewater in Denmark, which has a chemical quality similar to Greenlandic wastewater, was disinfected by peracetic acid or UV irradiation. Removal of heterotrophic bacteria by applying 6 and 12 mg/L peracetic acid was 2.8 and 3.1 log, respectively. Similarly, removal of heterotrophic bacteria by applying 0.21 and 2.10 kWh/m³ for UV irradiation was 2.1 and greater than 4 log, respectively. Physicochemical treatment of raw wastewater followed by UV irradiation and/or peracetic acid disinfection showed the potential for treatment of arctic wastewater.     

Disinfection and removal of human pathogenic bacteria in arctic waste stabilization ponds

Wastewater stabilization ponds (WSPs) are commonly used to treat municipal wastewater in Arctic Canada. The biological treatment in the WSPs is strongly influenced by climatic conditions. Currently, there is limited information about the removal of fecal and pathogenic bacteria during the short cool summer treatment season. With relevance to public health, the objectives of this paper were to determine if treatment in arctic WSPs resulted in the disinfection (i.e., removal of fecal indicator bacteria, Escherichia coli) and removal of selected human bacterial pathogens from the treated effluent. The treatment performance, with focus on microbial removal, was assessed for the one-cell WSP in Pond Inlet (Nunavut [NU]) and two-cell WSP in Clyde River (NU) over three consecutive (2012–2014) summer treatment seasons (late June-early September). The WSPs provided a primary disinfection treatment of the wastewater with a 2–3 Log removal of generic indicator E. coli. The bacterial pathogens Salmonella spp., pathogenic E. coli, and Listeria monocytogenes, but not Campylobacter spp. and Helicobacter pylori, were detected in the untreated and treated wastewater, indicating that human pathogens were not reliably removed. Seasonal and annual variations in temperature significantly (p < 0.05) affected the disinfection efficiency. Improved disinfection and pathogen removal was observed for the two-cell system in Clyde River as compared to the one-cell system in Pond Inlet. A quantitative microbial risk assessment should be performed to determine if the release of low levels of human pathogens into the arctic environment poses a human health risk.     

Fate of antimicrobial resistance genes in response to application of poultry and swine manure in simulated manure-soil microcosms and manure-pond microcosms

This study aimed to determine the occurrence, abundance, and fate of nine important antimicrobial resistance genes (ARGs) (sul1, sul2, tetB, tetM, ermB, ermF, fexA, cfr, and Intl1) in the simulated soil and pond microcosms following poultry and swine manure application. Absolute quantitative PCR method was used to determine the gene copies. The results were modeled as a logarithmic regression (N?=?mlnt?+?b) to explore the fate of target genes. Genes sul1, Intl1, sul2, and tetM had the highest abundance following the application of the two manure types. The logarithmic regression model fitted the results well (R ² values up to 0.99). The reduction rate of all genes (except for the genes fexA and cfr) in manure-pond microcosms was faster than those in manure-soil microcosms. Importantly, sul1, intl1, sul2, and tetM had the lowest reduction rates in all the samples and the low reduction rates of tetM was the first time to be reported. These results indicated that ARG management should focus on using technologies for the ARG elimination before the manure applications rather than waiting for subsequent attenuation in soil or water, particularly the ARGs (such as sul1, intl1, sul2, and tetM investigated in this study) that had high abundance and low reduction rate in the soil and water after application of manure.     

Total staphylococci as performance surrogate for greywater treatment

Faecal indicator bacteria (FIB) are commonly used as water quality indicators; implying faecal contamination and therefore the potential presence of pathogenic enteric bacteria, viruses, and protozoa. Hence in wastewater treatment, the most commonly used treatment process measures (surrogates) are total coliforms, faecal coliforms, Escherichia coli (E. coli), and enterococci. However, greywater potentially contains skin pathogens unrelated to faecal load, and E. coli and other FIB may grow within greywater unrelated to pathogens. Overall, FIB occurs at fluctuating and relatively low concentrations compared to other endogenous greywater bacteria affecting their ability as surrogates for pathogen reduction. Therefore, unlike municipal sewage, FIB provides a very limited and unreliable log-reduction surrogate measure for on-site greywater treatment systems. Based on our recent metagenomic study of laundry greywater, skin-associated bacteria such as Staphylococcus, Corynebacterium, and Propionibacterium spp. dominate and may result in more consistent treatment surrogates than traditional FIB. Here, we investigated various Staphylococcus spp. as potential surrogates to reliably assay over 4-log₁₀ reduction by the final-stage UV disinfection step commonly used for on-site greywater reuse, and compare them to various FIB/phage surrogates. A collimated UV beam was used to determine the efficacy of UV inactivation (255, 265 and 285 nm) against E. coli, Enterococcus faecalis, E. faecium, E. casseliflavus, Staphylococcus aureus, and S. epidermidis. Staphylococcus spp. was estimated by combining the bi-linear dose-response curves for S. aureus and S. epidermidis and was shown to be less resistant to UV irradiation than the other surrogates examined. Hence, a relative inactivation credit is suggested; whereas, the doses required to achieve a 4 and 5-log₁₀ reduction of Staphylococcus spp. (13.0 and 20.9 mJ cm^?2, respectively) were used to determine the relative inactivation of the other microorganisms investigated. The doses required to achieve a 4 and 5-log₁₀ reduction of Staphylococcus spp. resulted in a log₁₀ reduction of 1.4 and 4.1 for E. coli, 0.8 and 2.8 for E. faecalis, 0.8 and 3.6 for E. casseliflavus and 0.8 and 1.2 for MS2 coliphage, respectively. Given the concentration difference of Staphylococcus spp. and FIB (3 to 5-log₁₀ higher), we propose the use of Staphylococcus spp. as a novel endogenous performance surrogate to demonstrate greywater treatment performance given its relatively high and consistent concentration and therefore ability to demonstrate over 5-log₁₀ reductions.     

Use of halophytes in pilot-scale horizontal flow constructed wetland treating domestic wastewater

Recent findings encourage the use of halophytes in constructed wetlands for domestic wastewater treatment due to their special physiological characteristics as the ability to accumulate heavy metals and salts in their tissues makes them ideal candidates for constructed wetland vegetation. In this particular study, we investigated the application of halophytic plants in a horizontal flow constructed wetland for domestic wastewater treatment purposes. The pilot plant which was situated in Crete (Greece) was planted with a polyculture of halophytes (Tamarix parviflora, Juncus acutus, Sarcocornia perrenis, and Limoniastrum monopetalum). The system’s performance was monitored for a period of 11 months during which it received primary treated wastewater from the local wastewater treatment plant. Results show that halophytes developed successfully in the constructed wetland and achieved organic matter and pathogen removal efficiencies comparable to those reported for reeds in previous works (63% and 1.6 log units, respectively). In addition, boron concentration in the effluent was reduced by 40% in comparison with the influent. Salinity as expressed by electrical conductivity did not change during the treatment, indicating that the accumulation of salts in the leaves is not able to overcome electrical conductivity increasing due to evapotranspiration. The results indicate an improvement in the treatment of domestic wastewater via the use of halophyte-planted CWs.     

Molecular characterization of microbial communities and quantification of <Emphasis Type="Italic">Mycobacterium immunogenum</Emphasis> in metal removal fluids and their associated biofilms

A number of human health effects have been associated with exposure to metal removal fluids (MRFs). Multiple lines of research suggest that a newly identified organism, Mycobacterium immunogenum (MI), appears to have an etiologic role in hypersensitivity pneumonitis (HP) in case of MRFs exposed workers. However, our knowledge of this organism, other possible causative agents (e.g., Pseudomonads), and the microbial ecology of MRFs in general, is limited. In this study, culture-based methods and small subunit ribosomal RNA gene clone library approach were used to characterize microbial communities in MRF bulk fluid and associated biofilm samples collected from fluid systems in an automobile engine plant. PCR amplification data using universal primers indicate that all samples had bacterial and fungal contaminated. Five among 15 samples formed colonies on the Mycobacteria agar 7H9 suggesting the likely presence of Mycobacteria in these five samples. This observation was confirmed with PCR amplification of 16S rRNA gene fragment using Mycobacteria specific primers. Two additional samples, Biofilm-1 and Biofilm-3, were positive in PCR amplification for Mycobacteria, yet no colonies formed on the 7H9 cultivation agar plates. Real-time PCR was used to quantify the abundance of M. immunogenum in these samples, and the data showed that the copies of M. immunogenum 16S rRNA gene in the samples ranges from 4.33?×?10⁴ copy/ml to 4.61?×?10⁷ copy/ml. Clone library analysis revealed that Paecilomyces sp. and Acremonium sp. and Acremonium-like were dominant fungi in MRF samples. Various bacterial species from the major phylum of proteobacteria were found and Pseudomonas is the dominant bacterial genus in these samples. Mycobacteria (more specifically MI) were found in all biofilm samples, including biofilms collected from inside the MRF systems and from adjacent environmental surfaces, suggesting that biofilms may play an important role in microbial ecology in MRFs. Biofilms may provide a shield or sheltered microenvironment for the growth and/or colonization of Mycobacteria in MRFs.     

Phenological development stages variation versus mercury tolerance,accumulation, and allocation in salt marsh macrophytes <Emphasis Type="Italic">Triglochin maritima</Emphasis> and <Emphasis Type="Italic">Scirpus maritimus</Emphasis> prevalent in Ria de Aveiro coastal lagoon (Portugal)

Efficient and sustainable management of rapidly mounting environmental issues has been the focus of current intensive research. The present study aimed to investigate the impact of plant phenological development stage variation on mercury (Hg) tolerance, accumulation, and allocation in two salt marsh macrophytes Triglochin maritima and Scirpus maritimus prevalent in historically Hg-contaminated Ria de Aveiro coastal lagoon (Portugal). Both plant samples and the sediments vegetated by monospecific stands of T. maritima and S. maritimus were collected from reference (R) and sites with moderate (M) and high (H) Hg contamination in Laranjo bay within Ria de Aveiro lagoon. Hg tolerance, uptake, and allocation in T. maritima and S. maritimus, physico-chemical traits (pH, redox potential, and organic matter content) and Hg concentrations in sediments vegetated by these species were impacted differentially by phenological development stages variation irrespective of the Hg contamination level. In T. maritima, Hg concentration increased with increase in Hg contamination gradient where root displayed significantly higher Hg followed by rhizome and leaf maximally at H. However, in S. maritimus, the highest Hg concentration was perceptible in rhizome followed by root maximally at M. Between the two studied plant species, S. maritimus displayed higher Hg tolerance index (depicted by higher plant dry mass allocated to reproductive stage) and higher available Hg at M (during all growth stages) and H (during senescent stage) when compared to T. maritimus. Both plant species proved to be Hg excluder (low root/rhizome–leaf Hg translocation). Additionally, T. maritima also acted as Hg stabilizer while, S. maritimus as Hg accumulator. It can be inferred from the study that (a) the plant phenological development stage variations significantly influenced plant Hg sensitivity by impacting sediment chemistry, plant growth (in terms of plant dry mass), Hg accumulation, and its subsequent allocation capacity, contingent to Hg contamination gradient; (b) S. maritimus accumulated higher Hg but restricted its translocation to above-ground part using exclusion process at both M and H due to its accelerated growth during Hg-tolerant reproductive/metabolically active phenological development stage greater than its counterpart T. maritima; and (c) the studied salt marsh plants although hailed from the same C3 and monocot group did not necessarily display similar phenotypic plasticity and behavior towards Hg-contaminated scenario during their life cycle.     

Expression of copper-resistance genes in microbial communities under copper stress and oxic/anoxic conditions

Microorganisms have developed copper-resistance mechanisms in order to survive in contaminated environments. The abundance and expression of the copper-resistance genes cusA and copA, encoding respectively for a Resistance Cell Nodulation protein and for a P-type ATP-ase pump, was assessed along a gradient of copper concentration in microcosms prepared from Seine estuary mudflat sediment. We demonstrated that the abundance of copA and cusA genes decreased with the increase of copper concentration and that cusA gene was up to ten times higher than the copA gene. Only the copA gene was expressed in both oxic and anoxic conditions. The abundance and activity of the microbial community remained constant whatever the concentrations of copper along the gradient. The molecular phylogeny of the two copper-resistance genes was studied and revealed that the increase of copper increased the diversity of copA and cusA gene sequences.     

Microbial communities in pesticide-contaminated soils in Kyrgyzstan and bioremediation possibilities

In Kyrgyzstan, many former storehouses and dump sites for obsolete pesticides exist. In 2009/2010, an inventory and assessment of these sites including risks of environmental hazard has been conducted by FAO and the World Bank. Monitoring revealed high concentration of pesticides listed as persistent organic pollutants (POPs). The purpose of this research was to study the microbial structural complexes of the pesticide-contaminated soils in these dumping zones, and to search for and select microorganism’s destructors with cytochrome P450 genes for pesticide degradation. Culture-dependent and culture-independent approaches were used to determine the taxonomic composition of these bacterial communities. The universal primer set for the 16S ribosomal RNA (rRNA) gene and the specific primer set P450R were used to amplify the cytochrome P450 hydroxylase gene. In soils from Suzak A and B and soils from Balykchy dumping sites, the bacteria from the Actinobacteria phylum (Micrococcus genus) were dominant. These bacteria made up 32–47% of the indigenous local microflora; bacteria species from the Pseudomonas genus (Gammaproteobacteria phylum) made up 23% in Suzak, 12% in Balykchy soils. Bacillus species from the Firmicutes phylum were found only in Suzak soils. The 16S rRNA analyses and the specific primer set P450R have revealed bacteria with cytochrome genes which are directly involved in the degradation process of organic carbon compounds. Experiments were carried out to help select active degraders from the bacterial populations isolated and used to degrade Aldrin in laboratory. Active bacterial strains from the Pseudomonas fluorescens and Bacillus polymyxa population were selected which demonstrated high rates of degradation activity on Aldrin.     

Phytoplankton dynamics with a special emphasis on harmful algal blooms in the Mar Piccolo of Taranto (Ionian Sea,Italy)

The response of phytoplankton assemblages to the closure of urban sewage outfalls (USOs) was examined for the Mar Piccolo of Taranto (Mediterranean Sea), a productive semi-enclosed coastal marine ecosystem devoted to shellfish farming. Phytoplankton dynamics were investigated in relation to environmental variables, with a particular emphasis on harmful algal blooms (HABs). Recent analyses evidenced a general reduction of the inorganic nutrient loads, except for nitrates and silicates. Also phytoplankton biomass (chlorophyll a) and abundances were characterized by a decrease of the values, except for the inner area of the basin (second inlet). The phytoplankton composition changed, with nano-sized species, indicators of oligotrophic conditions, becoming dominant over micro-sized species. If the closure of the USOs affected phytoplankton dynamics, however, it did not preserve the Mar Piccolo from HABs and anoxia crises. About 25 harmful species have been detected throughout the years, such as the potentially domoic acid producers Pseudo-nitzschia cf. galaxiae and P seudo-nitzschia cf. multistriata, identified for the first time in these waters. The presence of HABs represents a threat for human health and aquaculture. Urgent initiatives are needed to improve the communication with authorities responsible for environmental protection, economic development, and public health for a sustainable mussel culture in the Mar Piccolo.     

<Emphasis Type="Italic">Ailanthus Altissima</Emphasis> and <Emphasis Type="Italic">Phragmites Australis</Emphasis> for chromium removal from a contaminated soil

The comparative effectiveness for hexavalent chromium removal from irrigation water, using two selected plant species (Phragmites australis and Ailanthus altissima) planted in soil contaminated with hexavalent chromium, has been studied in the present work. Total chromium removal from water was ranging from 55 % (Phragmites) to 61 % (Ailanthus). After 360 days, the contaminated soil dropped from 70 (initial) to 36 and 41 mg Cr/kg (dry soil), for Phragmites and Ailanthus, respectively. Phragmites accumulated the highest amount of chromium in the roots (1910 mg Cr/kg_{(dry tissue)}), compared with 358 mg Cr/kg_{(dry tissue)} for Ailanthus roots. Most of chromium was found in trivalent form in all plant tissues. Ailanthus had the lowest affinity for Cr^VI reduction in the root tissues. Phragmites indicated the highest chromium translocation potential, from roots to stems. Both plant species showed good potentialities to be used in phytoremediation installations for chromium removal.     

Heavy metal and trace elements in riparian vegetation and macrophytes associated with lacustrine systems in Northern Patagonia Andean Range

Vegetation associated with lacustrine systems in Northern Patagonia was studied for heavy metal and trace element contents, regarding their elemental contribution to these aquatic ecosystems. The research focused on native species and exotic vascular plant Salix spp. potential for absorbing heavy metals and trace elements. The native species studied were riparian Amomyrtus luma, Austrocedrus chilensis, Chusquea culeou, Desfontainia fulgens, Escallonia rubra, Gaultheria mucronata, Lomatia hirsuta, Luma apiculata, Maytenus boaria, Myrceugenia exsucca, Nothofagus antarctica, Nothofagus dombeyi, Schinus patagonicus, and Weinmannia trichosperma, and macrophytes Hydrocotyle chamaemorus, Isöetes chubutiana, Galium sp., Myriophyllum quitense, Nitella sp. (algae), Potamogeton linguatus, Ranunculus sp., and Schoenoplectus californicus. Fresh leaves were analyzed as well as leaves decomposing within the aquatic bodies, collected from lakes Futalaufquen and Rivadavia (Los Alerces National Park), and lakes Moreno and Nahuel Huapi (Nahuel Huapi National Park). The elements studied were heavy metals Ag, As, Cd, Hg, and U, major elements Ca, K, and Fe, and trace elements Ba, Br, Co, Cr, Cs, Hf, Na, Rb, Se, Sr, and Zn. Geochemical tracers La and Sm were also determined to evaluate contamination of the biological tissues by geological particulate (sediment, soil, dust) and to implement concentration corrections.     

Endogenous <Emphasis Type="Italic">trans</Emphasis>-zeatin content in plants with different metal-accumulating ability: a field survey

A field survey was conducted to evaluate soil metal pollution and endogenous trans-zeatin content in the leaves of plants growing at six sites in a metal-polluted area located in Gejiu, Yunnan, China. Five plant species were collected, and the physicochemical properties and concentrations of five metals in the soil were analyzed. The trans-zeatin content in plant leaves was measured by high-performance liquid chromatography. Based on the Nemerow pollution index, the six sites were classified into four levels of pollution (i.e., low, medium, high, and severely high). The degree of soil metal pollution was cadmium (Cd) > arsenic (As) > lead (Pb) > zinc (Zn) > copper (Cu). The leaf trans-zeatin content in Pteris vittata (an arsenic hyperaccumulator) increased significantly by 98.6 % in soil with a severely high level of pollution compared with soil at a low level of pollution. However, in non-hyperaccumulators Bidens pilosa var. radiata and Ageratina adenophora, a significant decrease in leaf trans-zeatin content of 35.6 and 87.6 %, respectively, was observed. The leaf trans-zeatin content in Artemisia argyi also decreased significantly by 73.6 % in high metal-polluted soil compared with that in medium metal-polluted soil. Furthermore, significant correlations were observed between leaf trans-zeatin content in Pteris vittata and As, Pb, and Cd concentrations in the soil; however, either no correlation or a negative one was observed in the other plant species. Therefore, a high content of trans-zeatin in the leaves of Pteris vittata may play an important role in its normal growth and tolerance to metals.     

Biodegradation of toluene vapor in coir based upflow packed bed reactor by <Emphasis Type="Italic">Trichoderma asperellum</Emphasis> isolate

In the present study, a new biofiltration system involving a selective microbial strain isolated from aerated municipal sewage water attached with coir as packing material was developed for toluene degradation. The selected fungal isolate was identified as Trichoderma asperellum by 16S ribosomal RNA (16S rRNA) sequencing method, and pylogenetic tree was constructed using BLASTn search. Effect of various factors on growth and toluene degradation by newly isolated T. asperellum was studied in batch studies, and the optimum conditions were found to be pH 7.0, temperature 30 °C, and initial toluene concentration 1.5 (v/v)%. Continuous removal of gaseous toluene was monitored in upflow packed bed reactor (UFPBR) using T. asperellum. Effect of various parameters like column height, flow rate, and the inlet toluene concentration were studied to evaluate the performance of the biofilter. The maximum elimination capacity (257 g m^?3 h^?1) was obtained with the packing height of 100 cm with the empty bed residence time of 5 min. Under these optimum conditions, the T. asperellum showed better toluene removal efficiency. Kinetic models have been developed for toluene degradation by T. asperellum using macrokinetic approach of the plug flow model incorporated with Monod model.     

Amphibian recovery after a decrease in acidic precipitation

We here report the first sign of amphibian recovery after a strong decline due to acidic precipitation over many decades and peaking around 1980–90. In 2010, the pH level of ponds and small lakes in two heavily acidified areas in southwestern Scandinavia (Aust-Agder and Østfold in Norway) had risen significantly at an (arithmetic) average of 0.14 since 1988–89. Parallel with the general rise in pH, amphibians (Rana temporaria, R. arvalis, Bufo bufo, Lissotriton vulgaris, and Triturus cristatus) had become significantly more common: the frequency of amphibian localities rose from 33% to 49% (n = 115), and the average number of amphibian species per locality had risen from 0.51 to 0.88. In two other (reference) areas, one with better buffering capacity (Telemark, n = 21) and the other with much less input of acidic precipitation (Nord-Trøndelag, n = 106), there were no significant changes in pH or amphibians.     

PCB dechlorination hotspots and reductive dehalogenase genes in sediments from a contaminated wastewater lagoon

Polychlorinated biphenyls (PCBs) are a class of persistent organic pollutants that are distributed worldwide. Although industrial PCB production has stopped, legacy contamination can be traced to several different commercial mixtures (e.g., Aroclors in the USA). Despite their persistence, PCBs are subject to naturally occurring biodegradation processes, although the microbes and enzymes involved are poorly understood. The biodegradation potential of PCB-contaminated sediments in a wastewater lagoon located in Virginia (USA) was studied. Total PCB concentrations in sediments ranged from 6.34 to 12,700 mg/kg. PCB congener profiles in sediment sample were similar to Aroclor 1248; however, PCB congener profiles at several locations showed evidence of dechlorination. The sediment microbial community structure varied among samples but was dominated by Proteobacteria and Firmicutes. The relative abundance of putative dechlorinating Chloroflexi (including Dehalococcoides sp.) was 0.01–0.19% among the sediment samples, with Dehalococcoides sp. representing 0.6–14.8% of this group. Other possible PCB dechlorinators present included the Clostridia and the Geobacteraceae. A PCR survey for potential PCB reductive dehalogenase genes (RDases) yielded 11 sequences related to RDase genes in PCB-respiring Dehalococcoides mccartyi strain CG5 and PCB-dechlorinating D. mccartyi strain CBDB1. This is the first study to retrieve potential PCB RDase genes from unenriched PCB-contaminated sediments.