Molecular and physiological characterization of fluoroquinolone resistance in relation to uropathogenicity among Escherichia coli isolates isolated from Wenyu River, China

Increasing antibacterial resistance and pathogenicity in the environment is of growing concern due to its potential human risk. In the present study, 236 Escherichia coli isolates were collected from Wenyu River in China on drugless (48 isolates) and quinolone-containing plates (189 isolates). Their minimum inhibitory concentrations (MICs) were determined ranging from 0.125 μg mL⁻¹ to 128 μg mL⁻¹. Mutation points related to fluoroquinolone resistance were observed at S83 to L and D87 to N or Y in the GyrA subunit and S80 to R or I and E84 to G in the ParC subunit. Generally, MICs of LEV and GAT are dependent on the patterns of these mutation points. The profile with three mutation points was related to LEV-resistant E. coli isolates, and the (S83L, D87N + S80I) mutation profile was most prevalent (65.7%) in LEV-resistant isolates, while a large proportion of isolates, even those with three mutation points, were susceptive to GAT. The incidence of virulence factors in LEV-resistant isolates (44.7%, 59/132) was much higher than in nonresistant isolates (23.1%, 24/104) (χ² = 11.925, 1° of freedom, p < 0.001) indicating that fluoroquinolone-resistant E. coli would pose a potential risk. A similar distribution was also found in isolates resistant to GAT (χ² = 7.843, 1° of freedom, p = 0.0079).     

MSMA Resistance Studies

Abstract

Monosodium methanearsonate (MSMA)-resistant and -susceptible common cocklebur (Xanthium strumarium L.) and cotton (Gossypium hirsutum L.) were treated with MSMA. Plant parameters analyzed were: glutathione synthetase activity, selected amino acid (arginine, glutamic acid, alanine, citrulline, glutamine, and glutathione) content and arsenic content (MSMA, total arsenic, and arsonate). No reduction of arsenic from the parent pentavalent form present in MSMA to the trivalent form was detected. Arginine, glutamic acid, and glutamine content increased in tissue three days after MSMA treatment. Glutathione content decreased during the first three days after treatment; however, five days after treatment the resistant biotype of cocklebur and cotton had elevated glutathione levels (8–20 times greater, respectively). Glutathione Synthetase activity was higher in cotton than in either of the cocklebur biotypes; MSMA did not affect its activity in cotton or either cocklebur biotype. Resistant biotypes have a slightly higher activity than the susceptible biotype. Tolerance of cotton to MSMA may be related to glutathione synthetase activity and possibly to the presence of phytochelatins. Also, increased glutathione levels in the resistant biotype may implicate phytochelatin involvement in the resistance mechanism.     

Estimation of the environmental risk posed by landfills using chemical, microbiological and ecotoxicological testing of leachates

The leachates from 22 municipal solid waste (MSW) landfill sites in Southern Poland were characterized by evaluation of chemical, microbiological and ecotoxicological parameters. Chemical analyses were mainly focused on the identification of the priority hazardous substances according to Directive on Priority Substances, 2008/105/EC (a daughter directive of the WFD) in leachates. As showed, only five substances (Cd, Hg, hexachlorobutadiene, pentachlorobenzene and PAHs) were detected in the leachates. The compounds tested were absent or present at very low concentrations. Among them, only PAHs were found in all samples in the range from 0.057 to 77.2 μg L⁻¹. The leachates were contaminated with bacteria, including aerobic, psychrophilic and mesophilic bacteria, coliform and fecal coliforms, and spore-forming-bacteria, including Clostridium perfringens, and with filamentous fungi. From the analysis of specific microorganism groups (indicators of environmental pollution by pathogenic or opportunistic pathogenic organisms) it can be concluded that the landfill leachates showed sanitary and epidemiological hazard. In the ecotoxicological study, a battery of tests comprised of 5 bioassays, i.e. Microtox®, Spirotox, Rotoxkit F™, Thamnotoxkit F™ and Daphtoxkit F™ magna was applied. The leachate samples were classified as toxic in 13.6%, highly toxic in 54.6% and very highly toxic in 31.8%. The Spirotox test was the most sensitive bioassay used. The percentage of class weight score was very high - above 60%; these samples could definitely be considered seriously hazardous and acutely toxic to the fauna and microflora.No correlations were found between the toxicity values and chemical parameters. The toxicity of leachate samples cannot be explained by low levels of the priority pollutants. It seems that other kinds of xenobiotics present in the samples at subacute levels gave the high aggregate toxic effect.The chemical, ecotoxicological and microbiological parameters of the landfill leachates should be analyzed together to assess the environmental risk posed by landfill emissions.     

Biotransformation of atrazine and metolachlor within soil profile and changes in microbial communities

Biotransformation studies of atrazine, metolachlor and evolution of their metabolites were carried out in soils and subsoils of Northern Greece. Trace atrazine, its metabolites and metolachlor residues were detected in field soil samples 1 year after their application. The biotransformation rates of atrazine were higher in soils and subsoils of field previously exposed to atrazine (maize field sites) than in respective layers of the field margin. The DT₅₀ values of atrazine ranged from 5 to 18 d in the surface layers of the adapted soils. DT₅₀ values of atrazine increased as the soil depth increased reaching the value of 43 d in the 80-110 cm depth layer of adapted soils. Metolachlor degraded at slower rates than atrazine in surface soils, subsoils of field and field margins with the respective DT₅₀ values ranging from 56 to 72 d in surface soils and from 165 to 186 d in subsoils. Hydroxyatrazine was the most frequently detected metabolite of atrazine. The maximum concentrations of metolachlor-OXA and metolachlor-ESA were detected in the soil layers of 20-40 cm depth after 90 d of incubation. Principal Component Analysis (PCA) of soil Phospholipid Fatty Acids (PLFAs), fungal/bacterial and Gram-negative/Gram-positive ratios of the PLFA profiles revealed that the higher biotransformation rates of atrazine were simultaneously observed with the abundance of Gram-negative bacteria while the respective rates of metolachlor were observed in soil samples with abundance of fungi.     

DOC removal paradigms in highly humic aquatic ecosystems

Background, aim, and scope  Dissolved humic substances (HS) usually comprise 50–80% of the dissolved organic carbon (DOC) in aquatic ecosystems. From a trophic and biogeochemical perspective, HS has been considered to be highly refractory and is supposed to accumulate in the water. The upsurge of the microbial loop paradigm and the studies on HS photo-degradation into labile DOC gave rise to the belief that microbial processing of DOC should sustain aquatic food webs in humic waters. However, this has not been extensively supported by the literature, since most HS and their photo-products are often oxidized by microbes through respiration in most nutrient-poor humic waters. Here, we review basic concepts, classical studies, and recent data on bacterial and photo-degradation of DOC, comparing the rates of these processes in highly humic ecosystems and other aquatic ecosystems. Materials and methods  We based our review on classical and recent findings from the fields of biogeochemistry and microbial ecology, highlighting some odd results from highly humic Brazilian tropical lagoons, which can reach up to 160 mg C L⁻¹. Results and discussion  Highly humic tropical lagoons showed proportionally lower bacterial production rates and higher bacterial respiration rates (i.e., lower bacterial growth efficiency) than other lakes. Zooplankton showed similar δ¹³C to microalgae but not to humic DOC in these highly humic lagoons. Thus, the data reviewed here do not support the microbial loop as an efficient matter transfer pathway in highly humic ecosystems, where it is supposed to play its major role. In addition, we found that some tropical humic ecosystems presented the highest potential DOC photo-chemical mineralization (PM) rates reported in the literature, exceeding up to threefold the rates reported for temperate humic ecosystems. We propose that these atypically high PM rates are the result of a joint effect of the seasonal dynamics of allochthonous humic DOC input to these ecosystems and the high sunlight incidence throughout the year. The sunlight action on DOC is positive to microbial consumption in these highly humic lagoons, but little support is given to the enhancement of bacterial growth efficiency, since the labile photo-chemical products are mostly respired by microbes in the nutrient-poor humic waters. Conclusions  HS may be an important source of energy for aquatic bacteria in humic waters, but it is probably not as important as a substrate to bacterial growth and to aquatic food webs, since HS consumption is mostly channeled through microbial respiration. This especially seems to be the case of humic-rich, nutrient-poor ecosystems, where the microbial loop was supposed to play its major role. Highly humic ecosystems also present the highest PM rates reported in the literature. Finally, light and bacteria can cooperate in order to enhance total carbon degradation in highly humic aquatic ecosystems but with limited effects on aquatic food webs. Recommendations and perspectives  More detailed studies using C- and N-stable isotope techniques and modeling approaches are needed to better understand the actual importance of HS to carbon cycling in highly humic waters.     

Predicting bioremediation of hydrocarbons: Laboratory to field scale

There are strong drivers to increasingly adopt bioremediation as an effective technique for risk reduction of hydrocarbon impacted soils. Researchers often rely solely on chemical data to assess bioremediation efficiently, without making use of the numerous biological techniques for assessing microbial performance. Where used, laboratory experiments must be effectively extrapolated to the field scale. The aim of this research was to test laboratory derived data and move to the field scale. In this research, the remediation of over thirty hydrocarbon sites was studied in the laboratory using a range of analytical techniques. At elevated concentrations, the rate of degradation was best described by respiration and the total hydrocarbon concentration in soil. The number of bacterial degraders and heterotrophs as well as quantification of the bioavailable fraction allowed an estimation of how bioremediation would progress. The response of microbial biosensors proved a useful predictor of bioremediation in the absence of other microbial data. Field-scale trials on average took three times as long to reach the same endpoint as the laboratory trial. It is essential that practitioners justify the nature and frequency of sampling when managing remediation projects and estimations can be made using laboratory derived data. The value of bioremediation will be realised when those that practice the technology can offer transparent lines of evidence to explain their decisions.     

Development of a hybrid microbial fuel cell (MFC) and fuel cell (FC) system for improved cathodic efficiency and sustainability: the M2FC reactor

In an effort to improve the efficiency and sustainability of microbial fuel cell (MFC) technology, a novel MFC reactor, the M2FC, was constructed by combining a ferric-based MFC with a ferrous-based fuel cell (FC). In this M2FC reactor, ferric ion, the catholyte in the MFC component, is regenerated by the FC system with the generation of additional electricity. When the MFC component was operated separately, the electricity generation was maintained for only 98 h due to the depletion of ferric ion in the catholyte. In combination with the fuel cell, however, the production of power was sustained because ferric ion was continually replenished from ferrous ion in the FC component. Moreover, the regeneration process of ferric ion by the FC produced additional energy. The M2FC reactor yielded a power density of up to 2 W m⁻² (or time-averaged value of approximately 650 mW m⁻²), density up to 20 times (or approximately six times based on time-averaged value) higher than the corresponding MFC system.     

Do mucilage events influence pico- and nanoplankton size and structure in the Adriatic sea?

Temporal changes in abundance and biomass of picophytoplankton, heterotrophic pico-eukaryotes, and nanoplankton assemblages were investigated along a transect crossing the Adriatic Sea, from the Italian to the Croatian coast. This 15-months (June 1999-August 2000) investigation allowed comparing microbial parameters during summer 1999 (year without mucilage) and summer 2000 when a major mucilage event occurred. Pico- and nanoplankton assemblages displayed significant differences between the 2 summer periods. The main differences can be summarized as: (i) presence of cyanobacterial blooms (up to 10⁸ cells l^-1) in summer 2000, absent in summer 1999; (ii) an increasing fraction of heterotrophic pico-eukaryotes (up to 5.0 × 10⁶ cells l^-1) and heterotrophic nanoplankton (size 2-5 µm) during mucilage event; (iii) a reduced abundance of small-sized (2-3 µm) phototrophic nanoplankton in summer 2000. Changes in community structure were signals of changes in trophic condition of the system, which resulted in a competitive advantage for small sized pico- and nanoheterotrophs. Data presented here indicated that mucilage events are associated with changes in microbial community structure and functioning in ambient water and induced the amplification of 3-step microbial food chain. The potential use of the heterotrophic pico-eukaryotes for describing alterations of the trophic pathways during mucilage events is discussed.     

Effect of land management on soil microbial N supply to crop N uptake in a dry tropical cropland in Tanzania

In Sub-Saharan Africa, conservation of available soil N during early crop growth, when N loss by leaching generally occurs, is important to improve crop productivity. In a dry tropical cropland in Tanzania, we assessed the potential role of soil microbes as a temporal N sink-source to conserve the available soil N until later crop growth, which generally requires substantial crop N uptake. We evaluated the effect of land management [i.e., no input, plant residue application before planting (P plot) with or without fertilizer application, fertilizer application alone, and non-cultivated plots] on the relationship between soil N pool [microbial biomass N (MBN) and inorganic N] and crop N uptake throughout the ∼120-d crop growth period in two consecutive years. In the P plot, MBN clearly increased (∼14.6-29.6 kg N ha⁻¹) early in the crop growth period in both years because of immobilization of potentially leachable N, and it conserved a larger soil N pool (∼10.5-21.2 kg N ha⁻¹) than in the control plot. Especially in one year in which N leaching was critical, increased MBN maintained a larger soil N pool in the P plot throughout the experimental period, and a delay of increased MB C:N ratio and a substantial decrease in MBN was observed, indicating better soil microbial N supply for crop N uptake during later crop growth. Therefore, plant residue application before planting should enhance the role of soil microbes as a temporal N sink-source, leading to the conservation of potentially leachable N until later phase of crop growth, especially in years in which N leaching is relatively severe. Although further studies are necessary, our results suggest that plant residue application before planting is a promising option to achieve better N synchronization.     

Differences in nitrous oxide fluxes from red soil under different land uses in mid-subtropical China

Red soil may play an important role in nitrous oxide (N₂O) emissions due to its recent land use change pattern. To predict the land use change effect on N₂O emissions, we examined the relationship between soil N₂O flux and environmental determinants in four different types of land uses in subtropical red soil. During two years of study (January 2005-January 2007), biweekly N₂O fluxes were measured from 09:00 to 11:00 a.m. using static closed chamber method. Objectives were to estimate the seasonal and annual N₂O flux differences from land use change and, reveal the controlling factors of soil N₂O emission by studying the relationship of dissolved organic carbon (DOC), microbial biomass carbon (MBC), water filled pore space (WFPS) and soil temperature with soil N₂O flux. Nitrous oxide fluxes were significantly higher in hot-humid season than in the cool-dry season. Significant differences in soil N₂O fluxes were observed among four land uses; 2.9, 1.9 and 1.7 times increased N₂O emissions were observed after conventional land use conversion from woodland to paddy, orchard and upland, respectively. The mean annual budgets of N₂O emission were 0.71-2.21 kg N₂O-N ha⁻¹ year⁻¹ from four land use types. The differences were partly attributed to increased fertilizer use in agriculture land uses. In all land uses, N₂O fluxes were positively related to soil temperature and DOC accounting for 22-48% and 30-46% of the seasonal N₂O flux variability, respectively. Nitrous oxide fluxes did significantly correlate with WFPS in orchard and upland only. Nitrous oxide fluxes responded positively to MBC in all land use types except orchard which had the lowest WFPS. We conclude that (1) land use conversion from woodland to agriculture land uses leads to increased soil N₂O fluxes, partly due increased fertilizer use, and (2) irrespective of land use, soil N₂O fluxes are under environmental controls, the main variables being soil temperature and DOC, both of which control the supply of nitrification and denitrification substrates.