Honeybees (<Emphasis Type="Italic">Apis mellifera</Emphasis>) as a biological barrier for contamination of honey by environmental toxic metals

The aim of the study was to investigate the transfer of toxic metals from honeybee workers (Apis mellifera L.) to bee honey in relation to the ecological state of the environment. The materials of the study consisted of samples of honeybee bodies and varietal honeys taken from the same apiary located in three areas: R1—urbanized (16), R2—ecologically clean (16) and R3—industrialized (15) of south-eastern Poland. The contents of 14 elements in all tested samples, including toxic metals (Cd, Pb, Hg, Al, Ni, Tl) as well as bioelements (K, Mg, Ca, Mn, Fe, Zn, Cu, Se), were analysed by the ICP-OES method with prior microwave mineralization. The concentrations of the majority of the studied elements, excluding aluminum and lead, were significantly higher in bee bodies than in honey samples (P?<?0.05). The pollution of bee bodies by toxic metals was dependent on the environmental cleanliness, and the most pollution was observed in the industrialized area. The bee body was the most effective barrier for Cd and Tl transfer to the honey, while the level of Ni was similar in both tested materials. The Al concentration was significantly higher in honey than bee bodies (14.81?±?24.69 and 6.51?±?5.83 mg kg^?1, respectively), which suggests the possibility of secondary contamination of honey. The greatest sensitivity to heavy metal pollution was observed in honeydew honey compared to nectar honeys (P?<?0.05). It was proved for the first time that bees work as biofilters for toxic metals and prevent honey contamination.     

<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.     

<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.     

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.     

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.     

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.     

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.     

The influence of chemical protection on the content of heavy metals in wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) growing on the soil enriched with granular sludge

The presence of heavy metals in Triticum aestivum L. growing on the soil enriched with granular sludge after chemical protection was observed. The five variants of treatments using herbicide (Chwastox Turbo 340SL) and four fungicides (Topsin M 500SC, Amistar 250SC, Artea 330EC, and Falcon 460EC) were performed. On control and experimental plots, the concentration of Ni, Pb, Cr, and Cu in wheat leaves were in the range 0.32–0.99, 0.92–1.57, 0.89–6.31, and 7.08–12.59 mg/kg and in grains 0.03 to 0.11, 0.14–0.25, 0.11–0.76, and 1.06–1.46 mg/kg, respectively. The concentration of Pb in grain protected by MCPA and 2,4-D with thiophanate-methyl and azoxystrobin was higher than the maximum levels of 0.20 mg/kg D.M. The bioconcentration factor (BCF) differed and depended on chemical protection. The highest value of BCF was achieved for Cd. The statistical analysis showed a significant correlation between concentration of metals and quality parameters of wheat. One observed significant negative correlations between Ni/Zeleny sedimentation value (r = ?0.51) and between Pb/starch content (r = ?0.57). Positive correlations were observed between Cd/yield, the number of grains/ergosterol concentration (respectively, r = 0.41, r = 0.55, r = 0.56), and Zn/thousand grain weight (r = 0.50) at a p ≤ 0.05.     

Metal speciation in sediment and bioaccumulation in <Emphasis Type="Italic">Meretrix lyrata</Emphasis> in the Tien Estuary in Vietnam

The concentrations of seven toxic metals (cadmium (Cd), nickel (Ni), chromium (Cr), arsenic (As), lead (Pb), copper (Cu), and zinc (Zn)) were determined in sediments and the soft tissues of a bivalve species (Meretrix lyrata) collected from the Tien Estuary in Tien Giang Province, South Vietnam. The total metal concentrations in sediments (mg/kg dry weight) increased as Cd (0.06) < Cu (5.0) < Pb (13.9) < As (16.3) < Ni (24) < Cr (50) < Zn (62). Speciation analysis revealed that these metals existed mainly in the residual fraction (43–94%), followed by the Fe-Mn oxide-bound (5–35%) and organic/sulfide-bound (0.6–9.2%) fractions. The metal concentrations in M. lyrata (mg/kg dry weight) were in the ranges of 1.3–1.9 (Cd), 1.5–2.8 (Ni), 1.8–3.4 (Cr), 11–16 (As), 0.3–0.6 (Pb), 6.9–8.7 (Cu), and 95–128 (Zn), which are safe for human consumption. The order of the mean biota-sediment accumulation factor (BSAF) of the metals in the non-residual fractions of the sediment for M. lyrata was Cd > Cu > As > Zn > Cr > Ni > Pb. The Risk Assessment Codes (RACs) suggest that the highest mobility of Cd (with RAC = 37%) poses greater environmental risk to aquatic biota. Correlation analysis results show that M. lyrata can be used as a biomonitor of Cd and Cu pollution in the exchangeable, acid-soluble, and non-residual sediment fractions.     

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.     

Bioaccumulation of metals in common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) from water bodies of Anatolia (Turkey): a review with implications for fisheries and human food consumption

Although fish is widely consumed by humans for its nutritional properties, accumulation of heavy metals can pose serious health hazards. Widespread common carp Cyprinus carpio is cultured worldwide and represents an economically important species for fisheries in several countries. These include Turkey, where C. carpio often makes for a large part of the sales of the locally marketed fish and also for a traditional dish. This study provides a review of bioaccumulation of metals in tissues of C. carpio from water bodies of Anatolia and also includes reference to worldwide studies. From 42 water bodies across the region, 27 metals in total were studied, of which Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn were the most widely analysed, mainly in the muscle, liver and gill tissues. Amongst the potentially toxic metals, Cd, Cr and Pb occurred in several water bodies at concentrations not only above maximum allowed limits but also higher relative to other water bodies worldwide, even though As, Hg and Ni were also sometimes present at potentially hazardous concentrations. The essential metals Cu, Fe, Mn, Se and Zn were detected at various concentrations, with the latter two occasionally above limit. All water bodies flagged as having especially critical (i.e. above limit) concentrations of toxic metals supported C. carpio fisheries from highly populated regions, raising concern about food safety and calling for preventative measures. Given the significantly lower bioaccumulation levels in the muscle relative to the liver and gill tissues, it is suggested that consumption of C. carpio as fillets may be safer than after processing into e.g. meat balls and sausages. The limits of 1.0 μg/g for Cr and 1.15 μg/g for Se, currently lacking from the Turkish food safety legislation, are proposed, and it is suggested that a similar meta-analytical approach as adopted in this study may benefit other countries where C. carpio represents an important fisheries resource.     

Activity of the endophytic fungi <Emphasis Type="Italic">Phlebia</Emphasis> sp. and <Emphasis Type="Italic">Paecilomyces formosus</Emphasis> in decolourisation and the reduction of reactive dyes’ cytotoxicity in fish erythrocytes

The current study investigates the potential for discolouration and degradation of Reactive Blue 19 and Reactive Black 5 textile dyes by endophytic fungi Phlebia sp. and Paecilomyces formosus as well as the potential cytotoxicity of products or by-products generated by the treatments in fish erythrocytes. It was observed at 30 days that both endophytes showed biodegradation activity with 0.1 g mL^?1 of dyes. P. formosus showed highest extracellular and intracellular protein content levels after the 15th day, and Phlebia sp. stands out for production of extracellular laccase, indicating that this enzyme may be associated with the decolouration capacity. The dyes showed toxic effects in fishes at 0.01 g mL^?1 concentration, resulting in the appearance of micronuclei in erythrocyte cells. When degraded dyes treated by endophytes were tested, the frequency of micronuclei reduced approximately 20%, indicating the effectiveness of these endophytic in the treatment of textile dyes with less environmental impact, thus indicating a potential for application of these fungi in bioremediation process.     

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.     

<Emphasis Type="Italic">Noctiluca</Emphasis> and copepods grazing on the phytoplankton community in a nutrient-enriched coastal environment along the southwest coast of India

The relative grazing impact of Noctiluca scintillans (hereafter referred only Noctiluca) and copepods (Acrocalanus gracilis, Paracalanus parvus, Acartia danae and Oithona similis) on the phytoplankton community in an upwelling–mudbank environment along the southwest coast India is presented here. This study was carried out during the Pre-Southwest Monsoon (April–May) to the Late Southwest Monsoon (August) period in 2014. During the sampling period, large hydrographical transformation was evident in the study area (off Alappuzha, Southwest coast of India); warmer Pre-Southwest Monsoon water column condition got transformed into cooler and nitrate-rich hypoxic waters during the Southwest Monsoon (June–August) due to intense coastal upwelling. Copepods were present in the study area throughout the sampling period with a noticeable increase in their abundance during the Southwest Monsoon. On the other hand, the first appearance of Noctiluca in the sampling location was during the Early Southwest Monsoon (mid-June) and thereafter their abundance increased towards the Peak Southwest Monsoon. The grazing experiments carried out as per the food removal method showed noticeable differences in the feeding preferences of Noctiluca and copepods, especially on the different size fractions of phytoplankton. Noctiluca showed the highest positive electivity for the phytoplankton micro-fraction (av. 0.49 ± 0.04), followed by nano-fraction (av. 0.17 ± 0.04) and a negative electivity for the pico-fraction (av. ?0.66 ± 0.06). In total ingestion of Noctiluca, micro-fraction contribution (83.7%) was significantly higher compared to the nano- (15.7%) and pico-fractions (0.58%). On the other hand, copepods showed the highest positive electivity for the phytoplankton nano-fraction (av. 0.38 ± 0.04) followed by micro- (av. -0.17 ± 0.05) and pico-fractions (av. ?0.35 ± 0.05). Similarly, in total ingestion of copepods, nano-fraction (69.7%) was the highest followed by micro- (28.9%) and pico-fractions (1.37%). The grazing pressure of Noctiluca on the total phytoplankton was found to be 27.7% of the standing stock and 45.6% of the production, whereas in the case of copepods, it was 9.95% of the standing stock and 16.6% of the production. The study showed that the grazing pressure of Noctiluca on the total phytoplankton as well as larger phytoplankton fraction was 2.8- and 8-folds higher than that of the copepods. This suggests the leading role of Noctiluca as an effective grazer of larger phytoplankton along the southwest west coast of India, especially during the Peak/Late Southwest Monsoon.     

Seasonal variations and source profile of <Emphasis Type="Italic">n</Emphasis>-alkanes in particulate matter (PM<Subscript>10</Subscript>) at a heavy traffic site,Delhi

Delhi is one of the most polluted cities in the world. The generation of aerosols in the lower atmosphere of the city is mainly due to a large amount of natural dust advection and sizable anthropogenic activities. The compositions of organic compounds in aerosols are highly variable in this region and need to be investigated thoroughly. Twenty-four-hour sampling to assess concentrations of n-alkanes (ng/m³) in PM₁₀ was carried out during January 2015 to June 2015 at Indira Gandhi Delhi Technical University for Women (IGDTUW) Campus, Delhi, India. The total average concentration of n-alkanes, 243.7 ± 5.5 ng/m³, along with the diagnostic tools has been calculated. The values of CPI₁, CPI₂, and CPI₃ for the whole range of n-alkanes series, petrogenic n-alkanes, and biogenic n-alkanes were 1.00, 1.02, and 1.04, respectively, and C _max were at C₂₅ and C₂₇. Diagnostic indices and curves indicated that the dominant inputs of n-alkanes are from petrogenic emissions, with lower contribution from biogenic emissions. Significant seasonal variations were observed in average concentrations of n-alkanes, which is comparatively higher in winter (187.4 ± 4.3 ng/m³) than during the summer season (56.3 ± 1.1 ng/m³).     

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.     

Oil contamination in surface sediment of Anzali Wetland in Iran is primarily even carbon number <Emphasis Type="Italic">n</Emphasis>-alkanes

To determine the extent of oil contamination and biodegradation in Anzali Wetland of Iran, we examined aliphatic hydrocarbons in surface sediment of this area (n=20). Petroleum hydrocarbon levels (mean 1585 ± 1117; range 316 to 4358 μg g?¹ dry weight) were similar in value to reports from other highly contaminated areas, such as New York Bight, Saudi and Kuwaiti coasts of the Persian Gulf, and Dubai shorelines. Even carbon homologs dominated distribution of n-alkanes in surface sediment of Anzali, which is rarely reported elsewhere. Multiple factors used in our study point to petrogenic source for n-alkanes in Anzali Wetland. Anzali receives multiple industrial and agricultural runoffs from the surrounding area. Shipping industry and oil industry are responsible for a major portion of pollutants entering Anzali. Municipal wastewater discharges are another source of Anzali pollution. To determine why even carbon number n-alkanes predominate in Anzali, we examined the following indices: existence of unresolved complex mixtures (UCM), ratio of UCM to resolved alkanes (RA), ratio of low-molecular weight to high-molecular weight molecules, presence of degraded oil residue, high-relative biodegradation, and the degree of hydrocarbon weathering in the surface sediment of the area. Our findings corroborate with such predominance.     

Accumulation and health risk assessment of trace elements in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis> from subsidence pools in the Huainan coalfield in China

Microelement (As, Cd, Cr, Cu, Ni, Pb, and Zn) concentrations were determined in the muscle, skin, gill, and liver tissues of Carassius auratus gibelio collected from subsidence pools at three different coal mines in the Huainan coalfield in China. The concentrations of elements in the water were within the allowable levels for raising fish. However, the higher levels of these metals in sediment may pose potential harm on fish. It was found that the concentrations of Cr, Ni, and Zn in all fish tissues were higher, while As, Cd, and Pb levels were relatively low. Microelement accumulation appeared to be more widespread in subsidence pools than that in natural water. Elements accumulated in fish tissues differently: the highest metal concentrations were generally found in the liver tissues of the fish analyzed, whereas the lowest were recorded in the muscles. The mean element concentrations in muscle tissue from C. auratus gibelio collected from subsidence pools (As, 0.16 mg/kg; Cd, 0.06 mg/kg; Cr, 6.21 mg/kg; Cu, 1.61 mg/kg; Ni, 3.88 mg/kg; Pb, 1.76 mg/kg; and Zn, 12.80 mg/kg dry weight) were far below the allowable limit of the hygienic standard in fish proposed by the Ministry of Health in China, suggesting that the fish were safe for human consumption. A health risk assessment also suggested there was no risk from the analyzed elements for inhabitants near the Huainan coalfield that consume fish.     

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.     

Integrating multiple vegetation indices via an artificial neural network model for estimating the leaf chlorophyll content of <Emphasis Type="Italic">Spartina alterniflora</Emphasis> under interspecies competition

The invasive species Spartina alterniflora and native species Phragmites australis display a significant co-occurrence zonation pattern and this co-exist region exerts most competitive situations between these two species, competing for the limited space, directly influencing the co-exist distribution in the future. However, these two species have different growth ratios in this area, which increase the difficulty to detect the distribution situation directly by remote sensing. As chlorophyll content is a key indicator of plant growth and physiological status, the objective of this study was to reduce the effect of interspecies competition when estimating Cab content; we evaluated 79 published representative indices to determine the optimal indices for estimating the chlorophyll a and b (Cab) content. After performing a sensitivity analysis for all 79 spectral indices, five spectral indices were selected and integrated using an artificial neural network (ANN) to estimate the Cab content of different competition ratios: the Gitelson ratio green index, the transformed chlorophyll absorption ratio index/optimized soil-adjusted vegetation index, the modified normalized difference vegetation index, the chlorophyll fluorescence index, and the Vogelmann chlorophyll index. The ANN method yielded better results (R ² = 0.7110 and RMSE = 8.3829 μg cm^?2) on average than the best single spectral index (R ² = 0.6319 and RMSE = 9.3535 μg cm^?2), representing an increase of 10.78% in R ² and a decrease of 10.38% in RMSE. Our results indicated that integrating multiple vegetation indices with an ANN can alleviate the impact of interspecies competition and achieve higher estimation accuracy than the traditional approach using a single index.