Organismic-level acute toxicology profiling of reactive azo dyes

In the present study, organismic-level acute toxicology profile of three reactive azo dyes, viz. Reactive Blue 221, Reactive Red 195, and Reactive Yellow 145, was investigated, by using bacterial (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Listeria monocytogenes, and Bacillus subtilis), fungal (Trichoderma asperellum, Aspergillus flavus, Fusarium fujikuroi, and Rhizoctonia solani), plant (Raphanus sativus, Triticum aestivum, Sorghum bicolor, and Phaseolus mungo), and aquatic (Artemia salina and Daphnia magna) specimens. Microbial test organisms (all the six bacteria and two fungi, i.e., T. asperellum and A. flavus) and D. magna were found to be relatively more sensitive towards the reactive azo dyes and their mixture, as the EC50 values were in the range of 80–330, 135–360, and 108–242 ppm for bacteria, fungi, and D. magna, respectively (but the effect was not acutely toxic). Moreover, the effect of dye mixture was comparable tothe individual dyes in almost all the tested microbial specimens. For plant seeds, the dye mixture was found to be relatively more inhibitory towards T. aestivum and R. sativus than the individual dyes. For S. bicolor and P. mungo seeds, the effect of the dye mixture was almost identical to the individual dyes. However, in all cases, EC50 values were in the range of 950–3500 ppm, which indicates a non-toxic effect on plant seed germination potential. Likewise, the dyes and their mixture were not acutely toxic for Artemia salina larvae (more sensitive to the dye mixture) and Daphnia magna neonates (EC50, 516–950 and 108–242 ppm, respectively).     

Diversity of crude oil-degrading bacteria and alkane hydroxylase (<Emphasis Type="Italic">alkB</Emphasis>) genes from the Qinghai-Tibet Plateau

The aim of this study was to survey the response of the microbial community to crude oil and the diversity of alkane hydroxylase (alkB) genes in soil samples from the Qinghai-Tibet Plateau (QTP). The enrichment cultures and clone libraries were used. Finally, 53 isolates and 94 alkB sequences were obtained from 10 pristine soil samples after enrichment at 10 °C with crude oil as sole carbon source. The isolates fell into the phyla Proteobacteria, Actinobacteria, and Bacteroidetes, with the dominance of Pseudomonas and Acinetobacter. The composition of degraders was different from polar habitats where Acinetobacter sp. is not a predominant responder of alkane degradative microbial communities. Phylogenetic analysis showed that the alkB genes from isolates and enrichment communities formed eight clusters and mainly related with alkB genes of Pseudomonas, Rhodococcus, and Acinetobacter. The alkB gene diversity in the QTP was lower than marine environments and polar soil samples. In particular, a total of 10 isolates exhibiting vigorous growth with crude oil could detect no crude oil degradation-related gene sequences, such as alkB, P450, almA, ndoB, and xylE genes. The Shannon-Wiener index of the alkB clone libraries from the QTP ranged from 1.00 to 2.24 which is similar with polar pristine soil samples but lower than that of contaminated soils. These results indicated that the Pseudomonas, Acinetobacter, and Rhodococcus genera are the candidate for in situ bioremediation, and the environment of QTP may be still relatively uncontaminated by crude oil.     

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.     

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.     

Antibiotic resistance and plasmid profiling of <Emphasis Type="Italic">Vibrio</Emphasis> spp. in tropical waters of Peninsular Malaysia

Vibrio species isolated from four different sampling stations in the west coast of Peninsular Malaysia were screened for their antimicrobial resistance and plasmid profiles. A total of 138 isolates belonging to 15 different species were identified. Vibrio campbellii, V. parahaemolyticus, V. harveyi, and V. tubiashii were found to predominance species at all stations. High incidence of erythromycin, ampicillin, and mecillinam resistance was observed among the Vibrio isolates. In contrast, resistance against aztreonam, cefepime, streptomycin, sulfamethoxazole, and sulfonamides was low. All the Vibrio isolates in this study were found to be susceptible to imipenem, norfloxacin, ofloxacin, chloramphenicol, trimethoprim/sulfamethoxazole, and oxytetracycline. Ninety-five percent of the Vibrio isolates were resistant to one or more different classes of antibiotic, and 20 different resistance antibiograms were identified. Thirty-two distinct plasmid profiles with molecular weight ranging from 2.2 to 24.8 kb were detected among the resistance isolates. This study showed that multidrug-resistant Vibrio spp. were common in the aquatic environments of west coast of Peninsular Malaysia.     

Identification and analysis of polyaromatic hydrocarbons (PAHs)—biodegrading bacterial strains from refinery soil of India

Polyaromatic hydrocarbons (PAHs) utilizing bacteria were isolated from soils of seven sites of Mathura refinery, India. Twenty-six bacterial strains with different morphotypes were isolated. These strains were acclimatized to utilize a mixture of four polycyclic aromatic hydrocarbons, i.e., anthracene, fluorene, phenanthrene, and pyrene, each at 50 mg/L concentration as sole carbon source. Out of total isolates, 15 potent isolates were subjected to 16S rDNA sequencing and identified as a member of diverse genera, i.e., Bacillus, Acinetobacter, Stenotrophomonas, Alcaligenes, Lysinibacillus, Brevibacterium, Serratia, and Streptomyces. Consortium of four promising isolates (Acinetobacter, Brevibacterium, Serratia, and Streptomyces) were also investigated for bioremediation of PAH mixture. This consortium was proved to be efficient PAH degrader resulting in 40–70 % degradation of PAH within 7 days. Results of this study indicated that these genera may play an active role in bioremediation of PAHs.     

Experimental arid land afforestation in Central Anatolia,Turkey

The afforestation of arid lands faces many challenges, and perhaps the most important key for success is choosing one or more species that are adapted well for local environmental conditions. We explored species that would be suitable for the steppe region of Central Anatolia. Intensive site preparation included ripping the subsoil (to 80 cm) and plowing the upper soil before planting seedlings of Elaeagnus angustifolia, Robinia pseudoacacia, Fraxinus angustifolia, and Pinus nigra were used as tree species. We also tested the success of several shrub species: Amygdalus orientalis, Calligonum polygonoides, and Spartium junceum. After five growing seasons, E. angustifolia showed the highest survival, with 80% of planted seedlings remaining. For the shrubs, A. orientalis was the most successful species with a 95% survival rate. Broad-leaved trees grew a cumulative average of 34 cm in height in 5 years, whereas P. nigra seedings grew only 9 cm. The greatest height growth occurred in the shrubs, with A. orientalis gaining 40 cm in height in 5 years. Overall, E. angustifolia and A. orientalis appeared best suited for afforestation in these areas. R. pseodoacacia and F. angustifolia may also be used as alternative species.     

<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 multi-contaminant exposure in a cancer treatment center: a 2-year monitoring of molds,mycotoxins, endotoxins,and glucans in bioaerosols

Indoor air quality in health care facilities is a major public health concern, particularly for immunocompromised patients who may be exposed to microbiological contaminants such as molds, mycotoxins, endotoxins, and (1,3)-ß-D-glucans. Over 2 years, bioaerosols were collected on a monthly basis in a cancer treatment center (Centre F. Baclesse, Normandy, France), characterized from areas where there was no any particular air treatment. Results showed the complexity of mycoflora in bioaerosols with more than 100 fungal species identified. A list of major strains in hospital environments could be put forward due to the frequency, the concentration level, and/or the capacity to produce mycotoxins in vitro: Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus versicolor, Cladosporium herbarum, Purpureocillium lilacinum, and Penicillium brevicompactum. The mean levels of viable airborne fungal particles were less than 30.530 CFU per m³ of air and were correlated to the total number of 0.30 to 20 μm particles. Seasonal variations were observed with fungal particle peaks during the summer and autumn. Statistical analysis showed that airborne fungal particle levels depended on the relative humidity level which could be a useful indicator of fungal contamination. Finally, the exposure to airborne mycotoxins was very low (only 3 positive samples), and no mutagenic activity was found in bioaerosols. Nevertheless, some fungal strains such as Aspergillus versicolor or Penicillium brevicompactum showed toxigenic potential in vitro.     

Detection of hormones in surface and drinking water in Brazil by LC-ESI-MS/MS and ecotoxicological assessment with <Emphasis Type="Italic">Daphnia magna</Emphasis>

The growing use of pharmaceutical drug is mainly due to several diseases in human and in animal husbandry. As these drugs are discharged into waterways via wastewater, they cause a major impact on the environment. Many of these drugs are hormones; in which even at low concentrations can alter metabolic and physiological functions in many organisms. Hormones were found in surface water, groundwater, soil, and sediment at concentrations from nanograms to milligrams per liter of volume—quantities known to cause changes in the endocrine system of aquatic organisms. This study aimed to develop a methodology for hormone detection (estriol, estrone, 17β-estradiol, 17α-ethinylestradiol, progesterone, and testosterone) on surface and treated water samples. Sample toxicity was assessed by ecotoxicology tests using Daphnia magna. A liquid chromatograph coupled to a mass spectrometer with an electrospray ionization source (LC-ESI-MS/MS) was used for the analysis. The results showed that samples were contaminated by the hormones estriol, estrone, progesterone, 17β-estradiol, and 17α-ethinylestradiol during the sampling period, and the highest concentrations measured were 90, 28, 26, 137, and 194 ng·L^?1, respectively. This indicates the inflow of sewage containing these hormones at some points in the Piracicaba River in the State of Sao Paulo—Brazil. Results indicated little toxicity of the hormone estriol in D. magna, indicating that chronic studies with this microcrustacean are necessary.     

The distribution and enrichment characteristics of copper in soil and <Emphasis Type="Italic">Phragmites australis</Emphasis> of Liao River estuary wetland

The aims of the present investigation were to reveal the distribution and enrichment characteristics of copper in soil and Phragmites australis of Liao River estuary wetland. The concentrations of copper in root, stem, leaf, and ear of Phragmites australis as well as in soil were determined to study the absorption capacity of copper by wild Phragmites australis of Liao River estuary wetland. The study was carried out at test pool of the Shenyang Agricultural University, and the experimental materials (soil, irrigating water and Phragmites australis) were derived from Liao River estuary wetland. The concentrations of copper in soil and Phragmites australis were 16.4441 to 49.0209 mg/kg and 0.8621 to 89.5524 mg/kg, respectively. The results indicated that the enrichment coefficients of copper in different tissues of Phragmites australis changed with the growth of Phragmites australis. The results revealed that the enrichment coefficients of copper in the whole Phragmites australis were greater than 1 at each growing stage of the Phragmites australis. The results also showed that the transfer coefficients of Phragmites australis to copper changed with the growth of Phragmites australis. The results revealed that the Phragmites australis had a good removal effect on copper from soil and had some characteristics of copper hyperaccumulator.     

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.     

Waterborne outbreaks in diarrhoea endemic foci of India: a longitudinal exploration and its implications

Diarrhoea remains a global public health enigma raising deep concerns for the health planners since contaminated potable water often spoils the community health structure. We hereby report a 6-year odd continuing outbreak surveillance report based on potable water indices, during which 264 water samples were screened from different districts of West Bengal, India. Samples were analysed for the presence of different enteropathogenic bacterial species by conventional molecular tools and their sensitivity to antibiotics. 78.03% samples were positive for enteropathogenic bacterial organisms and 75% samples harbored Coliform. 45.45, 12.12, and 4.16% samples were positive for E.coli, V. cholerae, and V. mimicus, respectively. Diarrhoeagenic E.coli 7 EPEC, 10 ETEC, and 2 EIEC were isolated along with 2 V. cholerae O1 Ogawa (ctxA and tcpA ElTor positive), one each from tube well and pond. Interestingly, 4 V.cholerae non-O1/non-O139 also harbored hlyA gene. The detection of toxin genes among this bacterial pool of sampled water indicates the fallout of the potable water sources, thus enabling us to establish that it is none other than the contaminated potable water system which often wreaks havoc in the south Bengal diarrhoeal menace. The consequences are further complicated by the presence of drug-resistant pathogenic bacterial pool to fluoroquinolone, beta-lactams, and cephalosporins, in the accessible potable water, with threats of outbreaks exploding into an epidemic, given suitable environment, poor sanitation, and unhygienic practices. Therefore, we strongly recommend re-modelling of ‘point-of-use water disinfection’ measures and adequate personal hygiene for healthier community life.     

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).     

Phytoplankton community structure in relation to environmental factors from the New Mangalore Port waters along the southwest coast of India

Seasonal and spatial phytoplankton distribution in relation to environmental factors was investigated in New Mangalore Port, a major port along the west coast of India. A well-mixed water column characterized the non-monsoon seasons, whereas it was weakly stratified during monsoon. Water quality index (TRIX) scores indicated good water quality except during pre-monsoon (inner zone surface) and monsoon (near bottom waters). Surface abundance of tychopelagic diatoms (Paralia sulcata, Melosira nummuloides, Cylindrotheca closterium, and Nitzschia sigma) was higher during non-monsoon seasons. Certain centric diatoms, e.g., Leptocylindrus danicus, P. sulcata, and Rhizosolenia imbricata, dominated during pre-monsoon (inner zone) and positively correlated with TRIX. High Skeletonema costatum and dinoflagellate abundance during the monsoon season coincided with high nutrient concentrations. Five potential toxic and fourteen harmful/bloom forming algal species were encountered at abundances below the level that can be considered as harmful to the ecosystem. In addition to a baseline database, this study highlights the potential use of certain diatom species as indicators of hydrography and water quality for monitoring dynamic coastal marine ecosystems.     

Bioindicator responses and performance of plant species along a vehicular pollution gradient in western Himalaya

Loss of green cover, and increasing pollution is a prime global concern. The problem calls for screening of pollution-tolerant tree species that can be integrated into plantation drives. Recognizing this, the study analyzed bio-indicator responses and performance of commonly occurring plant species along a pollution gradient in western Himalaya. Based on distance from the road, three sites viz., highly polluted (HP), moderately polluted (MP), and least polluted (LP), were identified. From these sites, leaves of commonly occurring 26 tree species were collected and analyzed for dust accumulation, total chlorophyll, relative water content (RWC), ascorbic acid, and pH using standard protocols. Later, assessment of Air Pollution Tolerance Index (APTI) and Anticipated Performance Indices (API) was carried out. The results revealed variations in biochemical characteristics. The pH, RWC, and total chlorophyll increased with decreasing pollution while ascorbic acid increased with increasing pollution. Dust capturing potential of Ficus carica (1.191 mg/m²) and Toona ciliata (0.820 mg/m²) was relatively higher. Based on the results of APTI, Grevillea robusta was classified as tolerant. It scored significantly higher values (21.06, 21.19, and 19.61 in LP, MP, and HP sites, respectively). Quercus floribunda, G. robusta (68.75% each), Juglans regia (68.7%), and T. ciliata (62.50%) were good performers in HP sites. Acer caesium, Betula utilis, and Morus alba that had low API scores (43.75%) were predicted as poor performers. Thus, G. robusta, Q. floribunda, J. regia, T. ciliata, and F. carica were evaluated as best performers. They could be integrated into plantations drives for environmental management.     

Impact of mining and forest regeneration on small mammal biodiversity in the Western Region of Ghana

Much of the terrestrial biodiversity in sub-Saharan Africa is supported by tropical rainforest. Natural resource development, particularly surface mining in the rainforest, poses great risks to the region’s rich and endemic biodiversity. Here, we assessed the impact of surface mining and the success of forest rehabilitation on small mammal diversity in the Western Region of Ghana. We surveyed small mammals in the project area and two adjoining forest reserves (control sites) before the mining operation and 10 years after mine closure and forest rehabilitation (topsoil replacement and revegetation). The forest reserves recorded higher species abundance than the mining areas. Majority of the species captured in the forest reserves, including Hylomyscus alleni, Praomys tullbergi, Malacomys cansdalei, and Hybomys trivirgatus, are forest obligate species. Only one individual each of H. alleni and P. tullbergi was captured in the naturally regenerated areas (core areas of mining activities that were allowed to revegetate naturally), while 32 individuals belonging to four species (Lophuromys sikapusi, Mus musculoides, Mastomys erythroleucus, and Crocidura olivieri) were recorded in the rehabilitated areas. Our data suggested negative effects of mining on small mammal diversity and the restoration of species diversity and important ecological processes after rehabilitation of altered habitats. We strongly encourage deliberate conservation efforts, particularly the development of management plans that require the restoration of degraded land resulting from mining activities.     

Genotoxic effects and gene expression in <Emphasis Type="Italic">Danio rerio</Emphasis> (Hamilton 1822) (Cypriniformes: Cyprinidae) exposed to mining-impacted tributaries in Manizales,Colombia

The zebrafish (Danio rerio) is one of the most studied aquatic organisms for water biomonitoring, due to its sensitivity to environmental degradation and resistance to toxic substances. This study determined the presence of micronuclei and nuclear abnormalities in peripheral blood erythrocytes, and assessed the gene expression of caspase-3 (CASP-3) and metallothionein 1 (MT-1) in the gills and liver of D. rerio. The study fish (n = 45) were exposed to water collected from two stations with mining impact (E2 and E3) and a reference station without evident mining contamination (E1), all located in La Elvira stream (Manizales-Colombia). In addition, a positive control (PC) with HgCl₂ (50 μg/L) and negative control (NC) with tap water were included. The fish from the PC and E2 and E3 treatments displayed genotoxic effects and changes in gene expression, with significant differences in micronuclei formation and the presence of blebbed nuclei. The cytochrome oxidase subunit I (COI) gene was used as reference and proved to be stable compared to the β-actin and 28S ribosomal RNA (28S) genes. In gills, CASP-3 expression was higher in the PC, and MT-1 expression was higher in the PC and E3 treatment. In liver, CASP-3 was expressed in the E2 treatment, and MT-1 expression was low. These results show that the genotoxic effects and differential gene expression observed in fish exposed to water from La Elvira stream could also be affecting the organisms present in this habitat.     

The first detection of potentially toxic <Emphasis Type="Italic">Microcystis</Emphasis> strains in two Middle Atlas Mountains natural lakes (Morocco)

Aguelmam Azizgza (LAZ) and Dayet Afourgah (DAF) are two Moroccan natural lakes located in a humid hydrographic basin of the Middle Atlas Mountains. Both are considered important reservoirs of plant and animal biodiversity. In addition, they are extensively used for recreational and fishing activities and as a water source for irrigation of agricultural crops. Recurrent cyanobacteria scum episodes in the two water bodies have been reported, Microcystis being the main genus in the scums. Here, we report on the toxic potential of three Microcystis aeruginosa strains isolated from those lakes: Mic LAZ and Mic B7 from LAZ and Mic DAF isolated from DAF. The toxic potential was checked by their microcystin (MC) content and the presence of mcy genes involved in MC synthesis. The identification and quantification of MC variants were performed by high-performance liquid chromatography-photo-diode array. The detection of mcy genes was achieved by whole-cell multiplex PCR that allowed the simultaneous amplification of DNA sequences corresponding to specific mcy regions. MC content of cultured cells, as MC-LR equivalents per gram cell biomass, was slightly higher in Mic LAZ (ca. 860) than in Mic B7 (ca. 700) and Mic DAF (ca. 690). Four MC variants were identified in the three isolates: MC-WR, MC-RR, MC-DM-WR, and MC-YR. The presence of toxic Microcystis strains in the two studied lakes may be regarded as an environmental and health hazard, especially during periods of bloom proliferation. It would be recommended the use of two complementary techniques, as those utilized herein (HPLC and mcy detection) to alert on highly probable toxicity of such lakes.     

Epilithic diatom assemblages and their relationships with environmental variables in the Nilüfer Stream Basin,Bursa, Turkey

Patterns of epilithic diatom species distribution in relation to environmental variables from 12 sampling sites on the main stream and some of its tributaries in the Nilüfer Stream Basin were determined using multivariate statistical techniques. The stream basin has been heavily influenced by anthropogenic effects. The upper part of the basin that is distant from pollution sources mostly has a spring water quality, while the lower part where the stream flows through the urban area and receives domestic and industrial wastewater has a quite low quality. Ordination techniques using both diatom taxa and 21 environmental variables revealed non- to slightly polluted upper basin sites and highly polluted lower basin sites along the stream. The results showed that the stream catchment is polluted gradually from upstream to downstream and that most of the downstream sites have very low water quality especially in summer months. A total of 134 epilithic diatom taxa belonging to 50 genera were recorded for 12 sample sites. Partial CCA results indicated that water temperature (T), discharge (Q), and total phosphorus (TP) were the most important variables affecting the distribution of diatom species. Unpolluted or slightly polluted upper basin sites were dominated by Achnanthidium minutissimum, Cocconeis placentula var. euglypta, Gomphonema olivaceum, and Navicula tripunctata. Highly polluted lower basin sites were characterized by high levels of organic and inorganic matters and low dissolved oxygen (DO) values. Species widespread in the highly polluted lower basin sites such as Nitzschia umbonata, Nitzschia amphibia, Nitzschia capitellata, Nitzschia palea, Nitzschia paleacea, Luticola mutica, and Stephanodiscus niagarae were mostly related to pollution indicator variables such as ammonium nitrogen (NH₄ ⁺–N), sodium (Na⁺), total phosphorus (TP), and total organic matter (TOM).