A Sensitive SERS Method for Determination of Pymetrozine in Apple and Cabbage Based on an Easily Prepared Substrate.

Residual pesticides are one of the major food safety concerns around the world. There is a demand for simple and reliable methods to monitor pesticide residues in foods. In this study, a sensitive method for determination of pymetrozine in apple and cabbage samples using surface-enhanced Raman spectroscopy (SERS) based on decanethiol functionalized silver nanoparticles was established. The proposed method performed satisfactorily with the linear detection range of 0.01-1.00 mg/L and limit of detection (LOD) of 0.01 mg/L in methanol. In addition, it was successfully used to detect pymetrozine in apple and cabbage samples, the LOD was 0.02 and 0.03 mg/L, respectively, and the recoveries of spiked cabbage and apple ranged 70.40-104.00%, with relative standard deviations below 12.18% and 10.33% for intra-day and inter-day tests. Moreover, the results of the correlation test with real cabbage samples of liquid chromatography-tandem mass spectrometry showed that they were highly correlated (slope = 0.9895, R2 = 0.9953). This study provides a sensitive approach for detection of pymetrozine in apple and cabbage, which has great potential for determination of pymetrozine residues in food products.

Impacts on antibiotic-resistant bacteria and their horizontal gene transfer by graphene-based TiO2&Ag composite photocatalysts under solar irradiation.

In recent years, photocatalysis has been considered as a promising method, which provides measures to environmental pollution. Antibiotic resistant bacteria (ARB) and their antibiotic resistance genes (ARGs), as the emerging environmental pollutants, are released into the environment, resulting in antibiotic resistance spread. TiO2-based nanocomposites, as the most common photocatalytic material, may influence ARB and ARGs under photocatalytic conditions. However, the research on this aspect is rare. A novel nanocomposite synthesized from Ag, TiO2 and graphene oxide (GO), was selected as a representative of nanomaterials for investigation. The experimental results indicated that TiO2/Ag/GO nanocomposites significantly affected ARB vitality. 100 mg/L TiO2/Ag/GO will reduce bacterial survival to 12.2% in 10 min under simulated sunlight irradiation. Chloramphenicol as the most representative antibiotic in the water, reduces the effect of ARB inactivation under photocatalytic conditions. The addition of TiO2/Ag/GO could affect tetracycline antibiotic resistance. The level of bacterial tolerance to tetracycline had a significant reduction. The horizontal gene transfer was promoted from 1 to 2 folds with the addition of TiO2/Ag/GO. Even high TiO2/Ag/GO concentration (100 mg/L) sample had a limited promotion, suggesting that TiO2/Ag/GO will not increase the risk of antibiotic resistance spread compared to other nano materials.

Antibiotic resistant bacteria survived from UV disinfection: Safety concerns on genes dissemination.

Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are the emerging contaminants leading to a serious worldwide health problem. Although disinfection like ultraviolet (UV) irradiation could remove part of ARB and ARGs, there still are residual ARB and ARGs in the effluent of wastewater treatment plants. Conjugative transfer is main concern of the risk of ARGs and little is known about the effects of UV disinfection on the transfer ability of the non-inactivated ARB in the effluent which will enter the environment. Hence the influences of UV irradiation and reactivation on ARB conjugative transfer ability were studied under laboratory condition, focusing on the survival bacteria from UV irradiation and the reactivated bacteria, as well as their descendants. The experimental results imply that even 1 mJ/cm2 UV disinfection can significantly decrease the conjugative transfer frequency of the survival bacteria. However, viable but not culturable state cells induced by UV can reactivate through both photoreactivation and dark repair and retain the same level of transfer ability as the untreated strains. This finding is essential for re-considering about the post safety of UV irradiated effluent and microbial safety control strategies were required.

Graphene oxide in the water environment could affect tetracycline-antibiotic resistance.

In recent years, the influence of new materials like nanoparticles in the water environment on biological substances has been widely studied. Antibiotic resistance genes (ARGs) represent a new type of pollutant in the environment. Graphene oxide (GO), as a nano material, because of its unique structure, may have an impact on antibiotic resistance bacteria (ARB) and ARGs; however the research in this area is rarely reported. Therefore, this study mainly investigated the effects of GO on bacterial antibiotic resistance. The results showed that GO had a limited effect on ARB inactivation. A high concentration of GO (>10 mg/L) can damage resistant plasmids to reduce bacterial resistance to antibiotics, but low concentrations of GO (<1 mg/L) led to almost no damage to the plasmid. However, all tested concentrations of GO promoted the conjugative transfer from 1to over 3 folds, with low concentrations and high concentration (1-10 and 100 mg/L) of GO samples the least promoted. The overall effect of GO on antibiotic resistance needs further investigation.

Reductions of bacterial antibiotic resistance through five biological treatment processes treated municipal wastewater.

Wastewater treatment plants are hot spots for antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). However, limited studies have been conducted to compare the reductions of ARB and ARGs by various biological treatment processes. The study explored the reductions of heterotrophic bacteria resistant to six groups of antibiotics (vancomycin, gentamicin, erythromycin, cephalexin, tetracycline, and sulfadiazine) and corresponding resistance genes (vanA, aacC1, ereA, ampC, tetA, and sulI) by five bench-scale biological reactors. Results demonstrated that membrane bioreactor (MBR) and sequencing batch reactor (SBR) significantly reduced ARB abundances in the ranges of 2.80∼3.54 log and 2.70∼3.13 log, respectively, followed by activated sludge (AS). Biological filter (BF) and anaerobic (upflow anaerobic sludge blanket, UASB) techniques led to relatively low reductions. In contrast, ARGs were not equally reduced as ARB. AS and SBR also showed significant potentials on ARGs reduction, whilst MBR and UASB could not reduce ARGs effectively. Redundancy analysis implied that the purification of wastewater quality parameters (COD, NH4 (+)-N, and turbidity) performed a positive correlation to ARB and ARGs reductions.

Insights into the amplification of bacterial resistance to erythromycin in activated sludge.

Wastewater treatment plants are significant reservoirs for antimicrobial resistance. However, little is known about wastewater treatment effects on the variation of antibiotic resistance. The shifts of bacterial resistance to erythromycin, a macrolide widely used in human medicine, on a lab-scale activated sludge system fed with real wastewater was investigated from levels of bacteria, community and genes, in this study. The resistance variation of total heterotrophic bacteria was studied during the biological treatment process, based on culture dependent method. The alterations of bacterial community resistant to erythromycin and nine typical erythromycin resistance genes were explored with molecular approaches, including high-throughput sequencing and quantitative polymerase chain reaction. The results revealed that the total heterotrophs tolerance level to erythromycin concentrations (higher than 32 mg/L) was significantly amplified during the activated sludge treatment, with the prevalence increased from 9.6% to 21.8%. High-throughput sequencing results demonstrated an obvious increase of the total heterotrophic bacterial diversity resistant to erythromycin. Proteobacteria and Bacteroidetes were the two dominant phyla in the influent and effluent of the bioreactor. However, the prevalence of Proteobacteria decreased from 76% to 59% while the total phyla number increased greatly from 18 to 29 through activated sludge treatment. The gene proportions of erm(A), mef(E) and erm(D) were greatly amplified after biological treatment. It is proposed that the transfer of antibiotic resistance genes through the variable mixtures of bacteria in the activated sludge might be the reason for the antibiotic resistance amplification. The amplified risk of antibiotic resistance in wastewater treatment needs to be paid more attention.

Distinguishing effects of ultraviolet exposure and chlorination on the horizontal transfer of antibiotic resistance genes in municipal wastewater.

Growing attention has been paid to the dissemination of antibiotic resistance genes (ARGs) in wastewater microbial communities; however, the disinfection processes, as microbial control technologies, have not been evaluated for their impacts on ARGs transfer. In this study, the effects of ultraviolet (UV) disinfection and chlorination on the frequency of ARGs transfer have been explored based on the conjugative transfer model between Gram-negative strains of E. coli. The results indicated that UV disinfection and chlorination exhibit distinct influences on the conjugative transfer. Low UV doses (up to 8 mJ/cm2) had little influence on the frequency of conjugative transfer, and UV exposure only decreased the bacterial number but did not change the cell permeability. By comparison, low chlorine doses (up to 40 mg Cl min/L) significantly promoted the frequency of conjugative transfer by 2-5-fold. The generated chloramine stimulated the bacteria and improved the cell permeability. More pilus were induced on the surface of conjugative cells, which acted as pathways for ARGs transfer. The frequency of ARG transfers was greatly suppressed by high doses of UV (>10 mJ/cm2) or chlorine (>80 mg Cl min/L).

Fate of antibiotic resistant bacteria and genes during wastewater chlorination: implication for antibiotic resistance control.

This study investigated fates of nine antibiotic-resistant bacteria as well as two series of antibiotic resistance genes in wastewater treated by various doses of chlorine (0, 15, 30, 60, 150 and 300 mg Cl2 min/L). The results indicated that chlorination was effective in inactivating antibiotic-resistant bacteria. Most bacteria were inactivated completely at the lowest dose (15 mg Cl2 min/L). By comparison, sulfadiazine- and erythromycin-resistant bacteria exhibited tolerance to low chlorine dose (up to 60 mg Cl2 min/L). However, quantitative real-time PCRs revealed that chlorination decreased limited erythromycin or tetracycline resistance genes, with the removal levels of overall erythromycin and tetracycline resistance genes at 0.42 ± 0.12 log and 0.10 ± 0.02 log, respectively. About 40% of erythromycin-resistance genes and 80% of tetracycline resistance genes could not be removed by chlorination. Chlorination was considered not effective in controlling antimicrobial resistance. More concern needs to be paid to the potential risk of antibiotic resistance genes in the wastewater after chlorination.

The sludge loading rate regulates the growth and release of heterotrophic bacteria resistant to six types of antibiotics in wastewater activated sludge.

Wastewater treatment plants are considered as hot reservoirs of antimicrobial resistance. However, the fates of antibiotic-resistant bacteria during biological treatment processes and relevant influencing factors have not been fully understood. This study evaluated the effects of the sludge loading rate on the growth and release of six kinds of antibiotic-resistant bacteria in an activated sludge system. The results indicated that higher sludge loading rates amplified the growth of all six types of antibiotic resistant bacteria. The release of most antibiotic-resistant bacteria through both the effluent and biosolids was amplified with increased sludge loading rate. Biosolids were the main pattern for all antibiotic-resistant bacteria release in an activated sludge system, which was determined primarily by their growth in the activated sludge. A higher sludge loading rate reactor tended to retain more antibiotic resistance. An activated sludge system with lower sludge loading rates was considered more conducive to the control of antibiotic resistance.

Monitoring and assessing the impact of wastewater treatment on release of both antibiotic-resistant bacteria and their typical genes in a Chinese municipal wastewater treatment plant.

Wastewater treatment plants (WWTPs) are important hotspots for the spread of antibiotic resistance. However, the release and impact factors of both antibiotic resistant bacteria and the relevant genes over long periods in WWTPs have rarely been investigated. In this study, the fate of bacteria and genes resistant to six commonly used antibiotics was assessed over a whole year. In WWTP effluent and biosolids, a high prevalence of heterotrophic bacteria resistant to vancomycin, cephalexin, sulfadiazine and erythromycin were detected, each with a proportion of over 30%. The corresponding genes (vanA, ampC, sulI and ereA) were all detected in proportions of (2.2 ± 0.8) × 10(-10), (6.2 ± 3.2) × 10(-9), (1.2 ± 0.8) × 10(-7) and (7.6 ± 4.8) × 10(-8), respectively, in the effluent. The sampling season imposed considerable influence on the release of all ARB. High release loads of most ARB were detected in the spring, while low release loads were generally found in the winter. In comparison, the ARG loads changed only slightly over various seasons. No statistical relevance was found between all ARB abundances and their corresponding genes over the long-term investigation period. This inconsistent behavior indicates that bacteria and genes should both be considered when exploring resistance characteristics in wastewater. A redundancy analysis was adopted to assess the impact of wastewater quality and operational conditions on antibiotic resistance. The results indicated that most ARB and ARG proportions were positively related to the COD and turbidity of the raw sewage, while negatively related to those of the effluent. DO and temperature exhibited strong negative relevance to most ARB prevalence.

Microbial selectivity of UV treatment on antibiotic-resistant heterotrophic bacteria in secondary effluents of a municipal wastewater treatment plant.

Little is known about the microbial selectivity of UV treatment for antibiotic resistant bacteria, and the results of limited studies are conflicting. To understand the effect of UV disinfection on antibiotic resistant bacteria, both total heterotrophic bacteria and antibiotic resistant bacteria (including cephalexin-, ciprofloxacin-, erythromycin-, gentamicin-, vancomycin-, sulfadiazine-, rifampicin-, tetracycline- and chloramphenicol-resistant bacteria) were examined in secondary effluent samples from a municipal wastewater treatment plant. Bacteria resistant to both erythromycin and tetracycline were chosen as the representative of multiple-antibiotic-resistant bacteria and their characteristics after UV treatment were also investigated. UV disinfection results in effective inactivation for total heterotrophic bacteria, as well as all antibiotic resistant bacteria. After UV treatment at a fluence of 5 mJ/cm(2), the log reductions of nine types of antibiotic resistant bacteria varied from 1.0 ± 0.1 to 2.4 ± 0.1. Bacteria resistant to both erythromycin and tetracycline had a similar fluence response as did total heterotrophic bacteria. The findings suggest that UV disinfection could eliminate antibiotic resistance in wastewater treatment effluents and thus ensure public health security. Our experimental results indicated that UV disinfection led to enrichment of bacteria with resistance to sulfadiazine, vancomycin, rifampicin, tetracycline and chloramphenicol, while the proportions of cephalexin-, erythromycin-, gentamicin- and ciprofloxacin-resistant bacteria in the wastewater decreased. This reveals the microbial selectivity of UV disinfection for antibiotic resistant bacteria.

Ultraviolet reduction of erythromycin and tetracycline resistant heterotrophic bacteria and their resistance genes in municipal wastewater.

Antibiotic resistance in wastewater is becoming a major public health concern, but poorly understood about impact of disinfection on antibiotic resistant bacteria and antibiotic resistance genes. The UV disinfection of antibiotic resistant heterotrophic bacteria and their relevant genes in the wastewater of a municipal wastewater treatment plant has been evaluated. Two commonly used antibiotics, erythromycin and tetracycline were selected because of their wide occurrences in regard to the antibiotic resistance problem. After UV treatment at a fluence of 5mJcm(-2), the log reductions of heterotrophic bacteria resistant to erythromycin and tetracycline in the wastewater were found to be 1.4±0.1 and 1.1±0.1, respectively. The proportion of tetracycline-resistant bacteria (5%) was nearly double of that before UV disinfection (3%). Tetracycline-resistant bacteria exhibited more tolerance to UV irradiation compared to the erythromycin-resistant bacteria (p<0.05). Gene copy numbers were quantified via qPCR and normalized to the volume of original sample. The total concentrations of erythromycin- and tetracycline-resistance genes were (3.6±0.2)×10(5) and (2.5±0.1)×10(5) copies L(-1), respectively. UV treatment at a fluence of 5mJcm(-2) removed the total erythromycin- and tetracycline-resistance genes by 3.0±0.1 log and 1.9±0.1 log, respectively. UV treatment was effective in reducing antibiotic resistance in the wastewater.

[Assessment method of the pathogenic microbial exposure caused by aerosolization of reclaimed water].

Inhalation is one of the main exposure routes to pathogenic microorganisms during wastewater reuse. An applicable method is introduced to assess the pathogenic microbial exposure from inhalation. Firstly, emission intensity of pathogenic microorganisms during reclaimed water spraying is determined by the concentration of pathogenic microorganisms in reclaimed water, spraying intensity and aerosolizing efficiency factor. The value of the aerosolizing efficiency factor is usually between 0.003 and 0.01. Then the concentration of pathogenic microorganisms in air is calculated by referring to the model of air pollutant dispersion and considering the microorganisms decay. The value of the microbial viability decay factor is between -0.23 s(-1) and 0 s(-1), and that of the environmental impact factor is between 0.016 and 73, varied by different kinds of microorganisms. And then the exposure from inhalation route can be finally attained by multiplying inhalation rate and exposure time. The daily average inhalation rate is 4.5-17 m3 x d(-1), and the hourly average inhalation rate is 0.3-3.2 m3 x h(-1), differentiated by age, gender, and activity of susceptible population. The method proposed in this study can be directly used to assess the pathogenic microbial exposure caused by reclaimed water used for agriculture irrigation, gardening, or fountain spraying.

[Effect of photoreactivating light intensity on photoreactivation of Escherichia coli and fecal coliform in the tertiary effluent disinfected by UV].

The effect of photoreactivating light intensity on photoreactivation of E. coli and fecal coliform in tertiary effluent after UV disinfection were investigated. The response of the two species to intensity of photoreactivating light varied with UV dose and bacterial species. Photoreactivation of E. coli after UV irradiation of 5 mJ/cm2 achieved the same maximum under three selected intensities of photoreactivating light (0-43 microW/cm2). A threshold existed when UV dose increased to 20 mJ/cm2 and significant photoreactivation was detected only under intensity of light 43 microW/cm2. With different UV doses irradiation, fecal coliform showed little difference under selected intensities of photoreactivating light in this study. The different effects of photoreactivating light intensity on photoreactivation of different bacteria should be considered when proposing the control measurements.

[Dark treatment effects on photoreactivation characteristics of Escherichia coli in a tertiary effluent disinfected by UV].

The effect of dark treatment on photoreactivation of Escherichia coli in a tertiary effluent disinfected by UV was studied. The effluent of sand filtration of a sewage treatment plant was disinfected by UV, kept in darkness for a period of time, and then put under photoreactivating light to investigate the photoreactivation. The experimental results indicated that the photoreactivation potential could recover immediately after water samples were kept in darkness for less than 6 h. The percentage photoreactivation of the water samples disinfected by UV with doses of 5 mJ/cm2 and 20 mJ/cm2 were 31.1% and 0.4%, respectively, and the photoreactivation rates were 1.67 x 10(4) CFU x (mL x h)(-1) and 125 CFU x (mL x h)(-1). When dark treatment time was prolonged to 24 h, the percentage photoreactivation and photoreactivation rates were lowered to 10.0% and 0.3%, 830 CFU x (mL x h)(-1) and 20 CFU x (mL x h)(-1), respectively. Those suggested that photoreactivation can only be weakened, but can not be controlled completely even after being kept in darkness for 24 h. So the safety concern on photoreactivation after UV disinfection of wastewater should be taken into account.

[Effects of allelochemical EMA isolated from Phragmites communis on algal cell membrane lipid and ultrastructure].

In order to reveal the antialgal mechanisms of allelochemicals, effects of the allelochemical eathyl-2-methyl acetoacetate (EMA) on cell membrane lipid and ultrastructure of Chlorella pyrenoidosa, Microcystis aeruginosa and Chlorella vulagaris were studied in this paper. The lipid fatty acids of the algal membrane were isolated following the Bligh and Dye method and quantified by gas chromatograph/mass spectrometry. The ultrastructure of algal cells was observed with TEM. The results showed that EMA increased the contents of linolenic acid and linolic acid with increment of 14%, while decreased the content of myristic acid and cetylic acid in C. pyrenoidosa, membrane. The content of unsaturated fatty acids C18:1 and C18:2 increased 12% and 10% in M. aeruginosa with the addition of EMA, while the content of saturated fatty acids C18:0 and C16:0 decreased. EMA showed no significant change in the fatty acid composition in C. vulagaris under the experiment condition. EMA broke off cell wall of C. pyrenoidosa and M. aeruginosa. EMA damaged the cell membrane and the inclusion of algal cell leaked out. Nuclear and mitochondrial structure was damaged with the addition of EMA. EMA showed no significant change in the ultrastructure of C. vulgaris.

[Effects of allelochemical isolated from Phragmites communis on algal membrane permeability].

Efflux of K+, Mg2+, Ca2+ ions from algal cells as signals of cell membrane permeability, inductively coupled plasma mass spectrometry (ICP-MS) as detection method of ions, the present research investigated effects of allelochemical eathyl-2-methyl acetoacetate (EMA) isolated from Phragmites communis on cell membrane permeability of Microcystis aeruginosa, Chlorella pyrenoidosa and Chlorella vulagaris. The results showed that, when the cells were boiled for 10 min and the membrane was destroyed absolutely, the K+ efflux of M. aeruginosa and C. pyrenoidosa were 1.45 and 1.59 microg x (10(9) cell) (-1), respectively. When the concentration of EMA was 2 mg x L(-1), the K+ efflux of M. aeruginosa and C. pyrenoidosa were 1.38 and 1.40 microg x (10(9) cell)(-1), respectively. The K+ efflux of M. aeruginosa and C. pyrenoidosa reached 1.44 and 1.58 microg x (10(9) cell)(-1) while the EMA was 4 mg x L(-1). When the concentrations were 2 mg x L(-1) or 4 mg x L(-1) the K+ efflux reached more than 95% of the total ion amount in M. aeruginosa and C. pyrenoidosa cells. But when EMA concentration was 4 mg x L(-1), K+ efflux of C. vulagaris was 0.64 microg x (10(9) cell)(-1), which was only 31.5% of total K+ amount in C. vulagaris. Effects EMA on efflux of Mg2+ and Ca2+ were similar to those of K+. The results indicated that EMA destroyed the cell membrane of M. aeruginosa and C. pyrenoidosa but not C. vulagaris. This is one of the mechanisms of EMA species-selective antialgal.

[Effects of EMA on activities of algal antioxidant enzymes].

Using allelochemicals produced by macrophytes to control harmful algae is a novel antialgal method. The present research investigated effects of the species-specific antialgal allelochemical EMA on activities of antioxidant enzymes of Chlorella pyrenoidosa and Chlorella vulagaris. The results showed that 0.25 mg/L of EMA increased activities of superoxide dismutase (SOD), peroxidase (POD)and catalase (CAT) of C. pyrenoidosa and C. vulagaris. With the increase of EMA concentrations, activities of the 3 enzymes of C. pyrenoidosa decreased sharply. The activity of SOD of C. pyrenoidosa decreased to 0 when the EMA were 4 mg/L. With the increase of EMA concentrations, activities of the 3 enzymes of C. vulagaris increased to as higher as 3-4 folders of that of the control set. The results gave hints to elucidate the species-specific antialgal mechanisms of EMA.