Thiamethoxam degradation by <Emphasis Type="Italic">Pseudomonas</Emphasis> and <Emphasis Type="Italic">Bacillus</Emphasis> strains isolated from agricultural soils

Twelve bacterial species were evaluated to know the degradation pattern of thiamethoxam in liquid medium. All the bacterial species could actively degrade phorate in a mineral salt medium containing phorate (50 μg ml^–1) as sole carbon source. As these species have ability to degrade, we used these for the degradation of thiamethoxam—a neonicoitinoids. Screening of 12 active phorate-metabolizing bacterial species resulted in selection of Bacillus aeromonas strain IMBL 4.1 and Pseudomonas putida strain IMBL 5.2 causing 45.28 and 38.23 % thiamethoxam (50 μg ml^?1) reduction, respectively, in 15 days as potential thiamethoxam degrading species. These two bacterial species grew optimally at 37 °C under shake culture conditions in MSMT medium raised with initial pH of 6.0–6.5 and use of these optimum cultural conditions resulted in improved thiamethoxam degradation by these bacterial species. These species caused maximum thiamethoxam degradation only in the presence of thiamethoxam as sole source of carbon and energy and the same was reduced in the presence of easily metabolize able carbon (C₀ and C₁) and nitrogen ((N₀, N₁ and N₂) sources. This could be attributed to involvement of repressible metabolic pathways, reactions of which are inhibited by the presence of easily available nutrients for growth. Besides above, qualitative analysis of thiamethoxam residues by gas liquid chromatography revealed complete metabolization of thiamethoxam without detectable accumulation of any known thiamethoxam metabolites.     

Assessment of heavy metals in <Emphasis Type="Italic">Averrhoa bilimbi</Emphasis> and <Emphasis Type="Italic">A. carambola</Emphasis> fruit samples at two developmental stages

Though the fruits of Averrhoa bilimbi and A. carambola are economically and medicinally important, they remain underutilized. The present study reports heavy metal quantitation in the fruit samples of A. bilimbi and A. carambola (Oxalidaceae), collected at two stages of maturity. Heavy metals are known to interfere with the functioning of vital cellular components. Although toxic, some elements are considered essential for human health, in trace quantities. Heavy metals such as Cr, Mn, Co, Cu, Zn, As, Se, Pb, and Cd were analyzed by atomic absorption spectroscopy (AAS). The samples under investigation included, A. bilimbi unripe (BU) and ripe (BR), A. carambola sour unripe (CSU) and ripe (CSR), and A. carambola sweet unripe (CTU) and ripe (CTR). Heavy metal analysis showed that relatively higher level of heavy metals was present in BR samples compared to the rest of the samples. The highest amount of As and Se were recorded in BU samples while Mn content was highest in CSU samples and Co in CSR. Least amounts of Cr, Zn, Se, Cd, and Pb were noted in CTU while, Mn, Cu, and As were least in CTR. Thus, the sweet types of A. carambola (CTU, CTR) had comparatively lower heavy metal content. There appears to be no reason for concern since different fruit samples of Averrhoa studied presently showed the presence of various heavy metals in trace quantities.     

Detection of <Emphasis Type="Italic">Bacillus</Emphasis> and <Emphasis Type="Italic">Stenotrophomonas</Emphasis> species growing in an organic acid and endocrine-disrupting chemical-rich environment of distillery spent wash and its phytotoxicity

Sugarcane molasses-based distillery spent wash (DSW) is well known for its toxicity and complex mixture of various recalcitrant organic pollutants with acidic pH, but the chemical nature of these pollutants is unknown. This study revealed the presence of toxic organic acids (butanedioic acid bis(TMS)ester; 2-hydroxysocaproic acid; benzenepropanoic acid, α-[(TMS)oxy], TMS ester; vanillylpropionic acid, bis(TMS)), and other recalcitrant organic pollutants (2-furancarboxylic acid, 5-[[(TMS)oxy] methyl], TMS ester; benzoic acid 3-methoxy-4-[(TMS)oxy], TMS ester; and tricarballylic acid 3TMS), which are listed as endocrine-disrupting chemicals. In addition, several major heavy metals were detected, including Fe (163.947), Mn (4.556), Zn (2.487), and Ni (1.175 mg l^?1). Bacterial community analysis by restriction fragment length polymorphism revealed that Bacillus and Stenotrophomonas were dominant autochthonous bacterial communities belonging to the phylum Firmicutes and γ-Proteobacteria, respectively. The presence of Bacillus and Stenotrophomonas species in highly acidic environments indicated its broad range adaptation. These findings indicated that these autochthonous bacterial communities were pioneer taxa for in situ remediation of this hazardous waste during ecological succession. Further, phytotoxicity assay of DSW with Phaseolus mungo L. and Triticum aestivum revealed that T. aestivum was more sensitive than P. mungo L. in the seed germination test. The results of this study may be useful for monitoring and toxicity assessment of sugarcane molasses-based distillery waste at disposal sites.     

<Emphasis Type="Italic">Mazzaella laminarioides</Emphasis> and <Emphasis Type="Italic">Sarcothalia crispata</Emphasis> as possible bioindicators of heavy metal contamination in the marine coastal zone of Chile

The suitability of Mazzaella laminarioides and Sarcothalia crispata as heavy metal biomonitors of Cd, Cu, Hg, Pb, and Zn was assessed by comparing bioaccumulation of these elements in different life stages and frond sizes in samples from three locations, San Vicente Bay (industrial area), Coliumo, and Quidico (the latter as a reference station), where different degrees of heavy metal pollution are recorded. Bioaccumulation and bioconcentration factors of Cd, Cu, Hg, Pb, and Zn were evaluated. The two macroalgae species showed similar patterns, with higher values of Cu, Hg, Pb, and Zn in polluted areas. M. laminarioides bioaccumulated higher concentrations of all metals assessed than S. crispata, independent of life stage and frond size. The results also showed significantly higher Cu, Hg, Pb, and Zn concentrations (p < 0.05) in water samples from San Vicente Bay than those measured in Coliumo and Quidico. Concentrations of Cd, Hg, Pb, and Zn in San Vicente Bay and Cd, Hg, and Pb in Coliumo and Quidico exceed the mean values considered to represent natural concentrations (Cu = 3.00 μg L^?1; Zn = 5.00 μg L^?1; Pb = 0.03 μg L^?1; Cd = 0.05 μg L^?1; Hg = 0.05 μg L^?1); however, the concentrations recorded do not cause negative effects on the growth and survival of macroalgae. The assessment of heavy metals bioaccumulated in M. laminarioides and S. crispata, particularly Hg, Pb, and Zn, offers a reliable approach for pollution assessment in rocky intertidal environments. Cu and Cd concentrations in seawater samples from San Vicente and Coliumo Bays were significantly higher than in those from Quidico (p value < 0.05); no significant differences in Cd concentrations were observed between San Vicente and Coliumo Bays (p < 0.05). Exceptionally, Cd is bioaccumulated at high levels independent of its availability in the water, thus reaching high concentrations in control areas. High concentrations of metals like Cu and Zn may limit or inhibit Cd uptake in macroalgae, since the transport channels are saturated by some metals, reducing the accumulation of others. These macroalgae species offer good potential for the development of suitable heavy metal pollution survey tools in rocky intertidal environments.     

Performance comparison of three methods for detection of <Emphasis Type="Italic">Giardia</Emphasis> spp. cysts and <Emphasis Type="Italic">Cryptosporidium</Emphasis> spp. oocysts in drinking-water treatment sludge

Detecting pathogenic protozoa in drinking-water treatment sludge is a challenge as existing methods are complex, and unfortunately, there are no specific technical standards to follow. Selecting an efficient analytical method is imperative in developing countries, such as Brazil, in order to evaluate the risk of parasite infection. In this context, three methods to detect Giardia spp. cysts and Cryptosporidium spp. oocysts were tested in sludge generated when water with protozoa and high turbidity was treated. Jar testing was carried out using polyaluminium chloride as a coagulant to generate the residue to be analyzed. The results showed that calcium carbonate flocculation with reduced centrifugation and immunomagnetic separation obtained the highest recoveries in the tested matrix showing 60.2%?±?26.2 for oocysts and 46.1%?±?5 for cysts. The other two methods, the first using the ICN 7× cleaning solution and the second considering the acidification of the sample, both followed by the immunomagnetic separation step, also presented high recoveries showing 41.2%?±?43.3 and 37.9%?±?52.9 for oocysts and 11.5%?±?85.5 and 26%?±?16.3 for cysts, respectively. Evidently, these methods and others should be studied in order to make it possible to detect protozoa in settled residue.     

Effect of citric acid on metals mobility in pruning wastes and biosolids compost and metals uptake in <Emphasis Type="Italic">Atriplex halimus</Emphasis> and <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis>

To assess metal mobility in pruning waste and biosolids compost (pH?6.9 and total concentration of metals in milligram per kilogram of Cd 1.9, Cu 132, Fe 8,513, Mn 192, Pb 81, and Zn 313), shrubs species Atriplex halimus and Rosmarinus officinalis were transplanted in this substrate and irrigated with citric acid (4 g?L^?1, pH?2.9) and nutrient solution daily for 60 days. Citric acid significantly increased the concentrations of soluble Mn and Fe in the nutrient substrate solution measured by suction probes, while other metals did not vary in concentration (Cu and Zn) or were not observed at detectable levels (Cd and Pb). In plants, citric acid significantly increased the concentrations of Cu (2.7?±?0.1–3.3?±?0.1 mg?kg^?1), Fe (49.2?±?5.2–76.8?±?6.8 mg?kg^?1), and Mn (7.2?±?1.1–11.4?±?0.7 mg?kg^?1) in leaves of R. officinalis, whereas the concentration of only Mn (25.4?±?0.3–42.2?±?2.9 mg?kg^?1) was increased in A. halimus. Increasing Fe and Mn solubility by citric acid addition indicates the possibility of using it to improve plant nutrition. The mobility of metals in this substrate was influenced for the concentration of the metal, the degree of humification of organic matter and its high Fe content.     

Determination of polychlorinated biphenyls and total mercury in two fish species (<Emphasis Type="Italic">Esox lucius</Emphasis> and <Emphasis Type="Italic">Carassius auratus</Emphasis>) in Anzali Wetland,Iran

The Anzali Wetland is one of the most important ecosystems in the north of Iran, and parts of it were registered as a Ramsar site in 1975. However, even though, due to many problems, including eutrophication produced by inflow of excess nutrients and organic materials, the wetland was also listed on the Montreux Record indicating the need to take urgent remedial action. This study was conducted to study the levels of polychlorinated biphenyls (PCBs) and total mercury (THg) in two fish species (Esox lucius and Carassius auratus) as bio-indicators of the ecosystem condition in eastern part of Anzali Wetland. The sampling was carried out in six different periods between years 2009 and 2010. The results showed that the amounts of PCBs in the muscle of northern pike were below the detection limit of gas chromatography, whereas the average concentration in goldfish was 0.449 mg/kg wet weight. Some possible reasons for the higher levels of PCBs in goldfish in comparison with pike have been discussed. No significant (p < 0.05) correlation was observed between PCBs and biological factors (weight, length, lipid content) for both species. On the other hand, the mean concentration of THg in the muscle of pike and goldfish were 182.22 and 75.27 ng/g dry weight, respectively. Although these concentrations were below US-EPA criterion for human consumption (0.3 mg/kg), it pointed up a significant deterioration of the ecosystem condition during the past years. Finally, statistical analysis revealed a significant correlation between THg with weight and an insignificant correlation with length for pike specimens.     

Bioaccumulation and public health implications of trace metals in edible tissues of the crustaceans <Emphasis Type="Italic">Scylla serrata</Emphasis> and <Emphasis Type="Italic">Penaeus monodon</Emphasis> from the Tanzanian coast

The coastal population in East Africa is growing rapidly but sewage treatment and recycling facilities in major cities and towns are poorly developed. Since estuarine mangroves are the main hotspots for pollutants, there is a potential for contaminants to accumulate in edible fauna and threaten public health. This study analysed trace metals in muscle tissues of the giant mud crabs (Scylla serrata) and the giant tiger prawns (Penaeus monodon) from the Tanzanian coast, in order to determine the extent of bioaccumulation and public health risks. A total of 180 samples of muscle tissues of S. serrata and 80 of P. monodon were collected from nine sites along the coast. Both species showed high levels of trace metals in the wet season and significant bioaccumulation of As, Cu and Zn. Due to their burrowing and feeding habits, mud crabs were more contaminated compared to tiger prawns sampled from the same sites. Apart from that, the measured levels of Cd, Cr and Pb did not exceed maximum limits for human consumption. Based on the current trend of fish consumption in Tanzania (7.7 kg/person/year), the measured elements (As, Cd, Co, Cu, Mn, Pb and Zn) are not likely to present health risks to shellfish consumers. Nevertheless, potential risks of As and Cu cannot be ruled out if the average per capita consumption is exceeded. This calls for strengthened waste management systems and pollution control measures.     

A new <Emphasis Type="Italic">P. putida</Emphasis> instrumental toxicity bioassay

Here, we present a new toxicity bioassay (CO₂-TOX), able to detect toxic or inhibitory compounds in water samples, based on the quantification of Pseudomonas putida KT2440 CO₂ production. The metabolically produced CO₂ was measured continuously and directly in the liquid assay media, with a potentiometric gas electrode. The optimization studies were performed using as a model toxicant 3,5-DCP (3,5-dichlorophenol); later, heavy metals (Pb²⁺, Cu²⁺, or Zn²⁺) and a metalloid (As⁵⁺) were assayed. The response to toxics was evident after 15 min of incubation and at relatively low concentrations (e.g., 1.1 mg/L of 3,5-DCP), showing that the CO₂-TOX bioassay is fast and sensitive. The EC₅₀ values obtained were 4.93, 0.12, 6.05, 32.17, and 37.81 mg/L for 3,5-DCP, Cu²⁺, Zn²⁺, As⁵⁺, and Pb²⁺, respectively, at neutral pH. Additionally, the effect of the pH of the sample and the use of lyophilized bacteria were also analyzed showing that the bioassay can be implemented in different conditions. Moreover, highly turbid samples and samples with very low oxygen levels were measured successfully with the new instrumental bioassay described here. Finally, simulated samples containing 3,5-DCP or a heavy metal mixture were tested using the proposed bioassay and a standard ISO bioassay, showing that our test is more sensible to the phenol but less sensible to the metal mixtures. Therefore, we propose CO₂-TOX as a rapid, sensitive, low-cost, and robust instrumental bioassay that could perform as an industrial wastewater-process monitor among other applications.     

Impact of collection on bark regeneration from <Emphasis Type="Italic">Stryphnodendron rotundifolium</Emphasis> Mart. in northeastern Brazil

The growing commercial demand for products with medicinal use has caused overexploitation of several plant species worldwide. To prevent the decline of these populations, the collection of these resources should be done in a sustainable way considering the time of its replacement in natural stocks. This study was designed to identify the relationship between different intensities of extraction of bark from the trunk of Stryphnodendron rotundifolium Mart. and its regeneration speed. For this, we selected two areas of Cerrado in the Northeast of Brazil, where a monitoring experiment with duration of 24 months was performed. This experiment consisted in simulating different extractive damage to assess the regeneration of bark. In each area, we selected 20 individuals, among which four treatments with five repetitions were implemented. The data showed that in both study areas, the trees regenerated their shells faster when subjected to higher collection intensities. However, this regeneration was not related to variations in rainfall in the environment.     

Phytoremediation assessment of <Emphasis Type="Italic">Gomphrena globosa</Emphasis> and <Emphasis Type="Italic">Zinnia elegans</Emphasis> grown in arsenic-contaminated hydroponic conditions as a safe and feasible alternative to be applied in arsenic-contaminated soils of the Bengal Delta

Several agricultural fields show high contents of arsenic because of irrigation with arsenic-contaminated groundwater. Vegetables accumulate arsenic in their edible parts when grown in contaminated soils. Polluted vegetables are one of the main sources of arsenic in the food chain, especially for people living in rural arsenic endemic villages of India and Bangladesh. The aim of this study was to assess the feasibility of floriculture in the crop rotation system of arsenic endemic areas of the Bengal Delta. The effects of different arsenic concentrations (0, 0.5, 1.0, and 2.0 mg As L^?1) and types of flowering plant (Gomphrena globosa and Zinnia elegans) on plant growth and arsenic accumulation were studied under hydroponic conditions. Total arsenic was quantified using atomic absorption spectrometer with hydride generation (HG-AAS). Arsenic was mainly accumulated in the roots (72 %), followed by leaves (12 %), stems (10 %), and flowers (<1 %). The flowering plants studied did not show as high phytoremediation capacities as other wild species, such as ferns. However, they behaved as arsenic tolerant plants and grew and bloomed well, without showing any phytotoxic signs. This study proves that floriculture could be included within the crop rotation system in arsenic-contaminated agricultural soils, in order to improve food safety and also food security by increasing farmer’s revenue.     

HPLC-FLD determination of 4-nonylphenol and 4-<Emphasis Type="Italic">tert</Emphasis>-octylphenol in surface water samples

A simple, sensitive and reliable HPLC-FLD method for the routine determination of 4-nonylphenol, 4-NP and 4-tert-octylphenol, 4-t-OP content in water samples was developed. The method consists in a liquid–liquid extraction of the target analytes with dichloromethane at pH  3.0–3.5 followed by the HPLC-FLD analysis of the organic extract using a Zorbax Eclipse XDB C8 column, isocratic elution with a mixed solvent acetonitrile/water 65:35, at a flow rate of 1.0 mL/min and applying a column temperature of 40°C. The method was validated and then applied with good results for the determination of 4-NP and 4-t-OP in Ialomi?a River water samples collected each month during 2006. The concentration levels of 4-NP and 4-t-OP vary between 0.08–0.17 μg/L with higher values of 0.24–0.37 μg/L in the summer months for 4-NP, and frequently <0.05 μg/L but also between 0.06–0.09 μg/L with higher values of 0.12–0.16 μg/L in July and August for 4-t-OP and were strongly influenced by sesonial and anthropic factors. The method was also applied on samples collected over 2 years 2007 and 2008 from urban wastewaters discharged into sewage or directly into the rivers by economic agents located in 30 Romanian towns. Good results were obtained when the method was used for analysis of effluents discharged into surface waters by 16 municipal wastewater treatment plants, during the year 2008.     

Identification and detection sensitivity of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> from mixed and field samples using MALDI-TOF MS

To verify the applicability of identifying Microcystis aeruginosa by matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry (MALDI-TOF MS), mixed and field samples were employed to study the sensitivity and the analysis power, respectively. Series diluted samples and artificially mixed samples by the M. aeruginosa NIES-843 strain were designed to verify the sensitivity. The lowest detection limit was 1.955?×?10⁶ cells in pure samples, while for mixed samples, the lowest detection limit and ratio of NIES-843 strain were 2.88?×?10⁶ cells and 33.7%, respectively. The results provided a reference for the reasonable volume of the water sample in which the M. aeruginosa could be detected. Ribosomal protein biomarkers for identifying M. aeruginosa which were successfully detected from the field samples in Taihu Lake, indicated that the identification of M. aeruginosa by MALDI-TOF MS could be applied in field samples. Furthermore, different genetic types of M. aeruginosa strains were also detected at different locations in Taihu Lake, which revealed the diversity of M. aeruginosa and the detection power of MALDI-TOF MS at the strain level for the field samples. The sensitivity and detection power in the analysis of M. aeruginosa by the MALDI-TOF MS demonstrated the applicability of this method in routine environmental monitoring.     

<Emphasis Type="Italic">Synechococcus</Emphasis> production and grazing loss rates in nearshore tropical waters

Temporal variation of Synechococcus, its production (μ) and grazing loss (g) rates were studied for 2 years at nearshore stations, i.e. Port Dickson and Port Klang along the Straits of Malacca. Synechococcus abundance at Port Dickson (0.3–2.3 × 10⁵ cell ml^?1) was always higher than at Port Klang (0.3–7.1 × 10⁴ cell ml^?1) (p < 0.001). μ ranged up to 0.98 day^?1 (0.51 ± 0.29 day^?1), while g ranged from 0.02 to 0.31 day^?1 (0.15 ± 0.07 day^?1) at Port Klang. At Port Dickson, μ and g averaged 0.47 ± 0.13 day^?1 (0.29–0.82 day^?1) and 0.31 ± 0.14 day^?1 (0.13–0.63 day^?1), respectively. Synechococcus abundance did not correlate with temperature (p > 0.25), but nutrient and light availability were important factors for their distribution. The relationship was modelled as log Synechococcus = 0.37Secchi ? 0.01DIN + 4.52 where light availability (as Secchi disc depth) was a more important determinant. From a two-factorial experiment, nutrients were not significant for Synechococcus growth as in situ nutrient concentrations exceeded the threshold for saturated growth. However, light availability was important and elevated Synechococcus growth rates especially at Port Dickson (F = 5.94, p < 0.05). As for grazing loss rates, they were independent of either nutrients or light intensity (p > 0.30). In nearshore tropical waters, an estimated 69 % of Synechococcus production could be grazed.     

Cytotoxic and genotoxic effects of silver nanoparticle/carboxymethyl cellulose on <Emphasis Type="Italic">Allium cepa</Emphasis>

Several mutagenic agents may be present in substances released in the environment, which may cause serious environmental impacts. Among these substances, there is a special concern regarding the widespread use of silver nanoparticles (AgNP) in several products due to their widely known bactericidal properties, including in the medical field and the food industry (e.g., active packaging). The assessment of the effects of AgNP released in the environment, having different concentrations, sizes, and being associated or not to other types of materials, including polymers, is therefore essential. In this research, the objective was to evaluate the genotoxic and cytotoxic effects of AgNP (size range between 2 and 8 nm) on root meristematic cells of Allium cepa (A. cepa). Tests were carried out in the presence of colloidal solution of AgNP and AgNP mixed with carboxymethylcellulose (CMC), using distinct concentrations of AgNP. As a result, when compared to control samples, AgNP induced a mitotic index decrease and an increase of chromosomal aberration number for two studied concentrations. When AgNP was in the presence of CMC, no cytotoxic potential was verified, but only the genotoxic potential for AgNP dispersion having concentration of 12.4 ppm.     

Cultivable microbiome and its resistance to antimicrobials isolated from <Emphasis Type="Italic">Zaprionus indianus</Emphasis>

The objective of this study was to identify Z. indianus in PEJC and PESCAN. Listed are the following methods: to isolate bacteria from the integument of the Z. indianus species collected and to check the resistance of microorganisms to antibiotics. Collections of Z. indianus were performed in four seasons of the two parks. The results obtained suggest that the low amount of Z. indianus collected can be justified by environmental factors such as high average temperature and low average humidity. It is noted that there is a predominance of bacteria of the Enterobacteriaceae family found in both the PEJC and the PESCAN. The antibiogram performed for the isolated PEJC bacteria shows statistical significance when comparing the edge and inside values of the park. Studies with fungi were also carried out, and it was evidenced that Trichophyton spp. was the genus that most inhabited the two environments studied. The ability of fluconazole and ketoconazole to inhibit fungal growth was also investigated, and considering the concentration tested may suggest that they have good action spectra. Plasmid profile data show that 60% of antibiotic-resistant bacteria have plasmids. The values found show that Z. indianus can act as vectors of microorganisms that affect the healthy animals and humans and that these organisms may be influenced by seasons.     

Assessment of physicochemical parameters and prevalence of virulent and multiple-antibiotic-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> in treated effluent of two wastewater treatment plants and receiving aquatic <Emphasis Type="Italic">milieu</Emphasis> in Durban,South Africa

The poor operational status of some wastewater treatment plants often result in the discharge of inadequately treated effluent into receiving surface waters. This is of significant public health concern as there are many informal settlement dwellers (ISDs) that rely on these surface waters for their domestic use. This study investigated the treatment efficiency of two independent wastewater treatment plants (WWTPs) in Durban, South Africa and determined the impact of treated effluent discharge on the physicochemical and microbial quality of the receiving water bodies over a 6-month period. Presumptive Escherichia coli isolates were identified using biochemical tests and detection of the mdh gene via PCR. Six major virulence genes namely eae, hly, fliC, stx1, stx2, and rfbE were also detected via PCR while antibiotic resistance profiles of the isolates were determined using Kirby-Bauer disc diffusion assay. The physicochemical parameters of the wastewater samples ranged variously between 9 and 313.33 mg/L, 1.52 and 76.43 NTUs, and 6.30 and 7.87 for COD, turbidity, and pH respectively, while the E. coli counts ranged between 0 and 31.2?×?10³ CFU/ml. Of the 200 selected E. coli isolates, the hly gene was found in 28 %, fliC in 20 %, stx2 in 17 %, eae in 14 %, with stx1 and rfbE in only 4 % of the isolates. Notable resistance was observed toward trimethoprim (97 %), tetracycline (56 %), and ampicillin (52.5 %). These results further highlight the poor operational status of these WWTPs and outline the need for improved water quality monitoring and enforcement of stringent guidelines.     

Effects of field storage method on <Emphasis Type="Italic">E. coli</Emphasis> concentrations measured in storm water runoff

Storm water runoff is increasingly assessed for fecal indicator organisms (e.g., Escherichia coli, E. coli) and its impact on contact recreation. Concurrently, use of autosamplers along with logistic, economic, technical, and personnel barriers is challenging conventional protocols for sample holding times and storage conditions in the field. A common holding time limit for E. coli is 8 h with a 10 °C storage temperature, but several research studies support longer hold time thresholds. The use of autosamplers to collect E. coli water samples has received little field research attention; thus, this study was implemented to compare refrigerated and unrefrigerated autosamplers and evaluate potential E. coli concentration differences due to field storage temperature (storms with holding times ≤24 h) and due to field storage time and temperature (storms >24 h). Data from 85 runoff events on four diverse watersheds showed that field storage times and temperatures had minor effects on mean and median E. coli concentrations. Graphs and error values did, however, indicate a weak tendency for higher concentrations in the refrigerated samplers, but it is unknown to what extent differing die-off and/or regrowth rates, heterogeneity in concentrations within samples, and laboratory analysis uncertainty contributed to the results. The minimal differences in measured E. coli concentrations cast doubt on the need for utilizing the rigid conventional protocols for field holding time and storage temperature. This is not to say that proper quality assurance and quality control is not important but to emphasize the need to consider the balance between data quality and practical constraints related to logistics, funding, travel time, and autosampler use in storm water studies.     

Relative proportions of <Emphasis Type="Italic">E. coli</Emphasis> and <Emphasis Type="Italic">Enterococcus spp.</Emphasis> may be a good indicator of potential health risks associated with the use of roof harvested rainwater stored in tanks

A total of 285 water samples were collected from 71 roof harvested rainwater tanks from four villages in different provinces over a two-year (2013–2014) period during the early (October to December) and late (January to March) rainy season. Water quality was evaluated based on Escherichia coli, faecal coliforms and Enterococcus spp. prevalence using the IDEXX Quanti-Tray quantification system. Real-Time PCR was used to analyse a subset of 168 samples for the presence of Shigella spp., Salmonella spp. and E. coli virulence genes (stx1, stx2 and eaeA). Escherichia coli were detected in 44.1% of the samples, Enterococcus spp. in 57.9% and faecal coliforms in 95.7%. The most prevalent E. coli concentrations in harvested rainwater were observed in 29.1% of samples and 22.5% for Enterococcus spp. and, were within 1–10 cfu/100 ml and 10–100 cfu/100 ml, respectively, whereas those for faecal coliforms (36.6%) were within 100–1000 cfu/100 ml. On average 16.8% of the samples had neither E. coli nor Enterococcus spp. detected, while 33.9% had only Enterococcus spp. and 23.7% had only E. coli. E. coli and Enterococcus spp. were detected together in 25.5% of the samples. Evaluation of samples for potential pathogenic bacteria showed all tested samples to be negative for the Shigella spp. ipaH gene, while five tested positive for Salmonella ipaB gene. None of the samples tested positive for the stx1 and stx2 genes, and only two tested positive for the eaeA gene. These findings are potentially useful in the development of a simplified risk assessment strategy based on the concentrations of indicator bacteria.