A comparison of toxicant-induced succession for five antifouling compounds on marine periphyton in SWIFT microcosms

Five antifouling biocides, chlorothalonile, dichlofluanide, medetomidine, tolylfluanide, and zinc pyrithione, were evaluated regarding their effect on the composition of the periphyton community and the subsequent toxicant-induced succession (TIS). The periphyton communities were exposed in a semi-static setting for 96 h using a SWIFT microcosm. As a measure of community composition, pigment profiles from the exposed communities were used as effect indicators and compared with unexposed parts of the same community using the Bray–Curtis dissimilarity index. Chlorothalonile caused changes in the community starting at 85 μg l^?1 while dichlofluanide had no effect even at the highest concentrations used, 810 μg l^?1. The related substance tolylfluanide only affected the community composition at 2700 μg l^?1. Medetomidine had a different response curve with a small effect on the community composition at 0.8 μg l^?1 which then disappeared only to reappear at 240 μg l^?1. Zinc pyrithione had the largest effect on the periphyton community with changes starting at 10 μg l^?1 and no detectable pigments at 100 μg l^?1. The changes in the community composition for the five substances were also compared using multidimensional scaling. When all substances were analyzed and plotted together, chlorothalonile, dichlofluanide, medetomidine, and tolylfluanide showed surprisingly similar effects compared to zinc pyrithione that gave very different TIS. However, when only chlorothalonile, dichlofluanide, and tolylfluanide were plotted together, clear differences in TIS between the three toxicants were revealed. Dichlofluanide only induced small effects, while concentration-dependent TIS trajectories for chlorothalonile and tolylfluanide took off in opposite directions indicating very different responses of the periphyton communities. This study demonstrates that substances with a similar chemical structure and mechanisms of action can have different effects on the community composition. With the exception of zinc pyrithione, none of the recorded effect levels were at concentrations reported from marine environments so far.     

Recovery of avocado (<Emphasis Type="Italic">Persea americana</Emphasis> Mill.) plants transformed with the antifungal plant defensin gene PDF1.2

Embryogenic avocado cultures derived from ‘Hass’ protoplasts were genetically transformed with the plant defensin gene (pdf1.2) driven by the CaMV 35S promoter in pGPTV with uidA as a reporter gene and bar, the gene for resistance to phosphinothricin, the active ingredient of the herbicide Finale® (Basta) (Bayer Environmental Science, Research Triangle Park, Durham, NC ). Transformation was mediated by Agrobacterium tumefaciens strain EHA105. Transformed cultures were selected in the presence of 3.0 mg l^?1 phosphinothricin in liquid maintenance medium for 3–4 mo. Liquid maintenance medium consisted of modified MS medium containing (per liter) 12 mg NH₄NO₃ and 30.3 mg KNO₃ and supplemented with 0.1 mg l^?1 thiamine HCl, 100 mg l^?1 myo-inositol, 30 g l^?1 sucrose, 3.0 mg l^?1 phosphinothricin, and 0.41 μM picloram. Somatic embryo development from transformed cultures was initiated on MS medium supplemented with 45 g l^?1 sucrose, 4 mg l^?1 thiamine HCl, 100 mg l^?1 myo-inositol, 10% (v/v) filter-sterilized coconut water, 3.0 mg l^?1 phosphinothricin, and 6.0 g l^?1 gellan gum. Limited plant recovery occurred from somatic embryos on semi-solid MS medium supplemented with 3.0 mg l^?1 phosphinothricin, 4.44 μM 6-benzylaminopurine (BA), and 2.89 μM GA₃; transformed shoots were micrografted on in vitro-grown seedling rootstocks. Approximately 1 yr after acclimatization in the greenhouse, transformed shoots were air-layered to recover transformed roots. Genetic transformation of embryogenic cultures, somatic embryos, and regenerated plants was confirmed by polymerase chain reaction (PCR), Southern blot hybridization, the XGLUC reaction for uidA, and application of the herbicide Finale® to regenerated plants.     

Effects of flashing light-emitting diodes on algal biomass productivity

To reduce power consumption and enhance algal biomass productivity in a thin flat-plate bioreactor (called a sliver tank bioreactor), flashing (pulsing) light was used. Biomass productivity and power consumption were monitored in controlled experiments using various photon flux levels, including a constant (non-flashing) flux of 75 μmol photons m^?2 s^?1 and three flashing experiments with photon fluxes of 375, 275, and 175 μmol photons m^?2 s^?1. Flashing experiments were performed at 10 kHz and a duty cycle of 20 %. A sliver tank bioreactor with a chamber width of 6.4 mm was used for its short optical path. Data from the experiments where light was flashed with a photon flux of 375 μmol photons m^?2 s^?1 indicated 9.6 % less power and 2.86 times the biomass productivity compared to the constant photon flux experiments. Similar results were obtained for the other flashing light regimes, which had lower biomass yields but also less input power per unit biomass produced, indicating that a large fraction of the continuously applied photons are shed or wasted, even at levels approximately 1/30th the intensity of full sun.     

Multi-Residue Analysis of Herbicides in Soil with an UPLC-ESI-MS Method

A rapid and sensitive method was developed for the determination of 51 herbicides in soil by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC–ESI–MS). Using acetonitrile effectively extracted 22 kinds of triazine and other basic herbicides, and using 90:10 v/v acetonitrile-phosphate buffer (pH = 7.5) effectively extracted another 29 herbicides. The extract has not cleaned up further. Chromatographic separation was achieved within 10 min using gradient elution with acetonitrile–water as a mobile phase for 22 triazine and phenylurea herbicides, and with 5 mM ammonium acetate containing 0.1% formic acid aqueous solution–acetonitrile as a mobile phase for another 29 herbicides. The response was linear over two orders of magnitude with correlation coefficients (r²) higher than 0.99. The limits of quantification for the herbicides varied from 0.2 to 20 µg kg^?1. The intra- and inter-day precisions (relative standard deviation, RSD) were 2.2–9.3% and 5.7–17.1%, respectively. The average recovery varied from 61.6 to 112% with the RSD of 1.6–11.3%. Analyzing 51 soil samples from 17 counties formed the basis of this method. Three herbicide residues were found in four counties. Atrazine residue in soil for 17 counties was found; its content was 0.4–9.8 μg kg^?1. Nicosulfuron residue in soil for two counties was found, with a high up to 133 or 1317 μg kg^?1. Propazine (0.3 and 1.34 μg kg^?1), atratone (2.14 and 3.93 μg kg^?1), and cynanazine (0.34 μg kg^?1) in soils for some counties were also found. The validated method can ensure the rapid multi-class, multi-residue analysis at low μg kg^?1 level for 47 herbicides in soil. The developed method provides an effective analytical basis for controlling herbicide dosage, investigating their distribution and degradation, and evaluating their hazards on the environment and human health.     

Effect of culture conditions, cytokinins, methyl jasmonate and salicylic acid on the biomass accumulation and production of withanolides in multiple shoot culture of Withania somnifera (L.) Dunal using liquid culture

The influence of cytokinins and culture conditions including medium volume, harvest time and elicitation with abiotic elicitors (SA/MeJ) have been studied for the optimal production of biomass and withanolides in the multiple shoot culture of Withania somnifera. Elicitation of shoot inoculum mass (2 g l^?l FW) with SA at 100 μM in the presence of 0.6 mg l^?l BA and 20 mg l^?l spermidine for 4 h exposure time at the 4th week in 20 ml liquid medium recorded higher withanolides production (withanolides A [8.48 mg g^?l DW], withanolides B [15.47 mg g^?l DW], withaferin A [29.55 mg g^?l DW] and withanone [23.44 mg g^?l DW]), which were 1.14 to 1.18-fold higher than elicitation with MeJ at 100 μM after 5 weeks of culture. SA-elicited cultures did not exhibit much variation in biomass accumulation when compared to control. This cytokinin induces and SA-elicited multiple shoot culture protocol provides a potential alternative for the optimal production of biomass and withanolides utilizing liquid culture.     

Optimization of photosynthesis,growth, and biochemical composition of the microalga <Emphasis Type="Italic">Rhodomonas salina</Emphasis>—an established diet for live feed copepods in aquaculture

The cryptophyte Rhodomonas salina is widely used as feed for copepod cultures. However, culturing conditions to obtain high-quality algae have not yet been efficiently optimized. Therefore, we aimed to develop a cultivation protocol for R. salina to optimize its nutritional value and provide technical recommendations for later large-scale production in algal photobioreactors. We studied photosynthesis, growth, pigments, fatty acid (FA) and free amino acid (FAA) composition of R. salina cultured at different irradiances (10–300 μmol photons m^?2 s^?1) and nutrient availability (deficiency and excess). The optimal range of irradiance for photosynthesis and growth was 60–100 μmol photons m^?2 s^?1. The content of chlorophylls a and c decreased with increasing irradiance while phycoerythrin peaked at irradiances of 40–100 μmol photons m^?2 s^?1. The total FA content was maximal at optimal irradiances for growth, especially under nutrient deficiency. However, highly unsaturated fatty acids, desired components for copepods, were higher under nutrient excess. The total FAA content was highest at limiting irradiances (10–40 μmol photons m^?2 s^?1) but a better composition with a higher fraction of essential amino acids was obtained at saturated irradiances (60–140 μmol photons m^?2 s^?1). These results demonstrate that quality and quantity of FA and FAA of R. salina can be optimized by manipulating the irradiance and nutrient conditions. We suggest that R. salina should be cultivated in a range of irradiance 60–100 μmol photons m^?2 s^?1 and nutrient excess to obtain algae with high production and a balanced biochemical composition as feed for copepods.     

Booster biocides and microfouling

Antifouling (AF) paints are used to prevent the attachment of living organisms to the submerged surfaces of ships, boats and aquatic structures, usually by the release of biocides. Apart from copper, organic booster biocides are the main active components in AF paints, but their use can have a negative impact on the marine environment. The direct effects of biocides on marine bacteria are poorly known. This work investigates the impact of two biocides, viz. diuron and tolylfluanid, on the growth and the viability of marine microorganisms and on their ability to form biofilms. The biocides in solution were found to inhibit growth of two strains of marine bacteria, viz. Pseudoalteromonas and Vibrio vulnificus, at a high concentration (1000 μg ml^?1), but only a small effect on viability was observed. Confocal laser scanning microscopy (CLSM) showed that the booster biocides decreased biofilm formation by both bacteria. At a concentration of 10 μg ml^?1, the biocides inhibited cell attachment and reduced biofilm thickness on glass surfaces. The percentage of live cells in the biofilms was also reduced. The effect of the biocides on two diatoms, Fragilaria pinnata and Cylindrotheca closterium, was also evaluated in terms of growth rate, biomass, chlorophyll a content and attachment to glass. The results demonstrate that diuron and tolylfluanid are more active against diatoms than bacteria.     

Effect of photoperiod,light intensity and carbon sources on biomass and lipid productivities of Isochrysis galbana

Biomass and lipid productivities of Isochrysis galbana were optimized using nutrients of molasses (4, 8, 12 g l^?1), glucose (4, 8, 12 g l^?1), glycerol (4, 8, 12 g l^?1) and yeast extract (2 g l^?1). Combinations of carbon sources at different ratios were evaluated in which the alga was grown at three different light intensities (50, 100 and 150 μmol m^?2 s^?1) under the influence of three different photoperiod cycles (12/12, 18/6 and 24/0 h light/dark). A maximum cell density of 8.35 g l^?1 with 32 % (w/w) lipid was achieved for mixotrophic growth at 100 μmol m^?2 s^?1 and 18/6 h light/dark with molasses/glucose (20:80 w/w). Mixotrophic cultivation using molasses, glucose and glycerol was thus effective for the cultivation of I. galbana.     

Biomass and terpenoids produced by mutant strains of <Emphasis Type="Italic">Arthrospira</Emphasis> under low temperature and light conditions

The filamentous Cyanobacterium Arthrospira is commercially produced and is a functional, high-value, health food. We identified 5 low temperature and low light intensity tolerant strains of Arthrospira sp. (GMPA1, GMPA7, GMPB1, GMPC1, and GMPC3) using ethyl methanesulfonate mutagenesis and low temperature screening. The 5 Arthrospira strains grew rapidly below 14?°C, 43.75 μmol photons m^?2 s^?1 and performed breed conservation at 2.5?°C, 8.75 μmol photons m^?2 s^?1. We used morphological identification and molecular genetic analysis to identify GMPA1, GMPA7, GMPB1 and GMPC1 as Arthrospira platensis, while GMPC3 was identified as Arthrospira maxima. Growth at different culture temperatures was determined at regular intervals using dry biomass. At 16?°C and 43.75 μmol photons m^?2 s^?1, the maximum dry biomass production and the mean dry biomass productivity of GMPA1, GMPB1, and GMPC1 were 2057?±?80 mg l^?1, 68.7?±?2.5 mg l^?1 day^?1, 1839?±?44 mg l^?1, 60.6?±?1.8 mg l^?1 day^?1, and 2113?±?64 mg l^?1, 77.7?±?2.5 mg l^?1 day^?1 respectively. GMPB1 was chosen for additional low temperature tolerance studies and growth temperature preference. In winter, GMPB1 grew well at mean temperatures <10?°C, achieving 3258 mg dry biomass from a starting 68 mg. In summer, GMPB1 grew rapidly at mean temperatures more than 28?°C, achieving 1140 mg l^?1 dry biomass from a starting 240 mg. Phytonutrient analysis of GMPB1 showed high levels of C-phycocyanin and carotenoids. Arthrospira metabolism relates to terpenoids, and the methyl-d-erythritol 4-phosphate pathway is the only terpenoid biosynthetic pathway in Cyanobacteria. The 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR) gene from GMPB1 was cloned and phylogenetic analysis showed that GMPB1 is closest to the Cyanobacterium Oscillatoria nigro-viridis PCC711. Low temperature tolerant Arthrospira strains could broaden the areas suitable for cultivation, extend the seasonal cultivation time, and lower production costs.     

Differential responses to somatic embryogenesis of different genotypes of Brazilian oil palm (Elaeis guineensis Jacq.)

To assess the potential of different genotypes of Brazilian oil palm (Elaeis guineensis Jacq.) to somatic embryogenesis and somatic embryo proliferation, mature zygotic embryos of nine commercial genotypes of E. guineensis (BRSC2001, BRSC2328, BRSC2301, BRSC3701, BRSCM1115, BRSC7201, BRSC2528, BRSC2501, and BRSCN1637) were used. Explants were incubated on Murashige and Skoog (MS) supplemented with 450 μM picloram, 3.0 % sucrose, 500 mg l^?1 glutamine, and 2.5 g l^?1 activated charcoal, and gelled with 2.5 g l^?1 Phytagel. After induction, for differentiation and maturation, the embryogenic calli (ECs) were transferred into fresh medium supplemented with 0.6 μM naphthaleneacetic acid (NAA) and 12.30 μM 2-isopentenyladenine (2iP) or 40 μM picloram in combination with 0.3 g l^?1 activated charcoal, and 500 mg l^?1 glutamine. Somatic embryos were converted into plants on MS medium with macro- and micro-nutrients at half strength, 2 % sucrose, and 2.5 g l^?1 activated charcoal, and gelled with 2.5 g l^?1 Phytagel. In general, zygotic embryos swelled after 14 days. Primary calli, which were observed in all the genotypes after 45–60 days of culture, eventually progressed to ECs at 90 days. At this time, scanning electron microscopy (SEM) analysis showed cellular differences between compact and friable calli. After 150 days in the induction phase, the ECs with proembryos that were transferred to the medium for differentiation and maturation, differentiated asynchronically into somatic embryos at globular and torpedo stages. The results showed that BRSC2328 and BRSCM1115 had the highest potential for EC formation (90–100 %) and somatic embryo differentiation (40.7 and 52.5 somatic embryos per callus, respectively) when compared to other genotypes. After approximately 90 days of culture on MS basal medium without growth regulators, protrusion of the leaf primordia was observed, characterizing the onset of germination of the somatic embryos into plants.     

The effects of cyclooxygenase and nitric oxide synthase inhibition on oxidative stress in isolated rat heart

Despite the widespread clinical use of cyclooxygenase (COX) inhibitors, dilemmas still exist about potential impact of these drugs on cardiovascular system. The present study was aimed to estimate the effects of different COX inhibitors (meloxicam, acetylsalicylic acid [ASA], and SC-560) on oxidative stress in isolated rat heart, with special focus on l-arginine/NO system. The hearts of male Wistar albino rats (total number n = 96, each group 12 rats, 8 weeks old, body mass 180–200 g) were retrogradely perfused according to the Langendorff technique at gradually increased perfusion pressure (40–120 cmH₂O). After control experiments the hearts were perfused with the following drugs: 100 μmol/l ASA (Aspirin), alone or in combination with 30 μmol/l l-NAME, 0.3 μmol/l meloxicam (movalis) with or without 30 μmol/l l-NAME, 3 μmol/l meloxicam (alone or in combination with 30 μmol/l l-NAME), 30 μmol/l l-NAME, and administration of 0.25 μmol/l SC-560. In samples of coronary venous effluent the following oxidative stress markers were measured spectrophotometrically: index of lipid peroxidation (measured as thiobarbituric acid reactive substances [TBARS]), superoxide anion radical release (O₂ ^?), and hydrogen peroxide (H₂O₂). While ASA was found to have an adverse influence on redox balance in coronary circulation, and coronary perfusion, meloxicam and SC-560 do not negatively affect the intact model of the heart. Furthermore, all effects were modulated by NOS inhibition. It seems that interaction between COX and l-arginine/NO system truly exists in coronary circulation, and can be one of the possible causes for achieved effects. That means: those effects induced by different inhibitors of COX are modulated by subsequent inhibition of NOS.     

Transgenic <Emphasis Type="Italic">Eustoma grandiflorum</Emphasis> expressing the <Emphasis Type="Italic">bar</Emphasis> gene are resistant to the herbicide Basta<Superscript>®</Superscript>

Lisianthus (Eustoma grandiflorum) is a cut or ornamental flower that is popular all over the world. This ornamental crop, however, lacks an effective weed control method due to its susceptibility to herbicide. In this study, transgenic plants of a lisianthus cultivar were produced using Agrobacterium-mediated delivery of the plasmid pCAMBIA3300, which carried the bialaphos resistance (bar) gene under driven by the CaMV 35S promoter. The transgenic calli were derived from wounded edges of the leaves grown on a shoot regeneration medium containing 100 mg l^?1 cefotaxime and 2 mg l^?1 glufosinate ammonium for 4 weeks. The callus that was detached from the wounded edge of the leaf was transferred to the shoot regeneration medium with 100 mg l^?1 cefotaxime and 5 mg l^?1 glufosinate ammonium for 4 weeks for shoot regeneration. The bar gene integration and expression in the transgenic plants were confirmed by Southern and Northern blot analyses, respectively. Subsequently, the transgenic lines were assessed in vitro and under greenhouse conditions for their resistance to the commercial herbicide Basta^®, which contains glufosinate ammonium as the active component. Six transgenic lines showed high percentages (67–80%) of survival in vitro under the selection condition with glufosinate ammonium (up to 216 mg l^?1). Under greenhouse conditions, the plants from these six lines remained healthy and exhibited a normal phenotype after spraying with glufosinate ammonium (up to 1,350 mg l^?1). This is the first paper to provide a detailed survey of transgenic lisianthus expressing the bar gene and exhibiting herbicide-resistance under greenhouse conditions.     

In vitro propagation of <Emphasis Type="Italic">Psoralea corylifolia</Emphasis> L. by somatic embryogenesis in cell suspension culture

An effective protocol was developed for in vitro propagation of Psoralea corylifolia via somatic embryogenesis in cell suspension culture. Embryogenic callus was obtained on Murashige and Skoog (MS) medium supplemented with 6 μM naphthaleneacetic acid (NAA) and 30 μM glutamine from transverse TCLs from 10-day-old hypocotyl explants with a 96.4% frequency. Embryogenic callus produced a higher number of somatic embryos (123.7 ± 1.24 per gram fresh weight callus) on MS medium containing 30 g l^?1 sucrose, 1 μM NAA, 4 μM benzyladenine (BA), 15 μM glutamine and 2 μM abscisic acid (ABA) after 4 weeks of culture. Somatic embryos successfully germinated (97.6%) on ½ MS medium containing 20 g l^?1 sucrose, 8 g l^?1 agar and supplemented with 2 μM BA, 1 μM ABA and 2 μM gibberellic acid (GA₃) within 2 weeks of culture. Somatic embryos developed into normal plants, which hardened with 100% efficiency in soil in a growth chamber. Plants were successfully transferred to greenhouse and subsequently established in the field. Plant survival percentage in the field differed with seasonal variations. Average psoralen content of 12.9 μg g^?1 DW was measured in different stages of somatic embryo development by high-performance liquid chromatography (HPLC). This protocol will be helpful for efficient propagation of elite clones on a mass scale, conservation efforts of this species and for secondary metabolites production studies.     

midD-encoded ‘rhizomimosinase’ from Rhizobium sp. strain TAL1145 is a C–N lyase that catabolizes L-mimosine into 3-hydroxy-4-pyridone, pyruvate and ammonia

Rhizobium sp. strain TAL1145 catabolizes mimosine, which is a toxic non-protein amino acid present in Leucaena leucocephala (leucaena). The objective of this investigation was to study the biochemical and catalytic properties of the enzyme encoded by midD, one of the TAL1145 genes involved in mimosine degradation. The midD-encoded enzyme, MidD, was expressed in Escherichia coli, purified and used for biochemical and catalytic studies using mimosine as the substrate. The reaction products in the enzyme assay were analyzed by HPLC and mass spectrometry. MidD has a molecular mass of ~45 kDa and its catalytic activity was found to be optimal at 37 °C and pH 8.5. The major product formed in the reaction had the same retention time as that of synthetic 3-hydroxy-4-pyridone (3H4P). It was confirmed to be 3H4P by MS/MS analysis of the HPLC-purified product. The K _m, V _max and K _cat of MidD were 1.27 × 10^?4 mol, 4.96 × 10^?5 mol s^?1 mg^?1, and 2,256.05 s^?1, respectively. Although MidD has sequence similarities with aminotransferases, it is not an aminotransferase because it does not require a keto acid as the co-substrate in the degradation reaction. It is a pyridoxal-5′-phosphate (PLP)-dependent enzyme and the addition of 50 μM hydroxylamine completely inhibited the reaction. However, the supplementation of the reaction with 0.1 μM PLP restored the catalytic activity of MidD in the reaction containing 50 μM hydroxylamine. The catalytic activity of MidD was found to be specific to mimosine, and the presence of its structural analogs including l-tyrosine, l-tryptophan and l-phenylalanine did not show any competitive inhibition. In addition to 3H4P, we also identified pyruvate and ammonia as other degradation products in equimolar quantities of the substrate used. The degradation of mimosine into a ring compound, 3H4P with the release of ammonia indicates that MidD of Rhizobium sp. strain TAL1145 is a C–N lyase.     

Agrobacterium-mediated transformation of Eruca sativa

An Agrobacterium tumefaciens—mediated transformation system was developed for Eruca sativa (eruca). Hypocotyl explants were co-cultivated with bacterial cells carrying a plasmid harboring a uidA:nptII fusion gene along a phosphinothricin acetyl transferase (PAT) gene cassette, for a period of 2 days. These were grown on a high cytokinin/auxin medium containing 5.0 mg l^?1 6-benzyladenine (BA), 1.0 mg l^?1 indole-3-acetic acid (IAA), and 0.1 mg l^?1 α-naphthaleneacetic acid (NAA). Explants were then transferred to a lower cytokinin/auxin medium containing 2.0 mg l^?1 BA and 0.1 mg l^?1 NAA along with 5.0 mg l^?1 silver nitrate and 300 mg l^?1 Timentin®. Upon transfer to a selection medium containing either 20 mg l^?1 kanamycin or 2 mg l^?1 L-phosphinothricin (L-ppt), shoot regenerants were observed. Expression of the transgenes in putative transformants was confirmed using a histochemical GUS assay. Presence of the PAT transgene in GUS-positive T0 plants was confirmed by Southern blot analysis. Moreover, spot tests of T1 seedlings were conducted using the L-ppt herbicide. A transformation frequency of 1.1% was obtained with more than 60% of transgenic lines containing single copies of the transgenes.     

Physical and chemical features of a tropical hypertrophic reservoir permanently stratified

Physical and chemical parameters of Pao-Cachinche reservoir (Venezuela) were measured in four stations (S1-S4) during an eighteen months period from September 1997 to February 1999. The tributaries of this reservoir introduce high amounts of nutrients from domestic wastewaters and from poultry and pig farms located in the basin. Thermal stratification was well established throughout the study period. Mountainous topography protects the water surface from mechanical action of wind, preventing water mixing. Hypoxic/anoxic conditions prevailed from 6–7 m depth down to the bottom in the limnetic stations. A strong smell of H₂S was detected below the 10 m level. The low water transparency (max. 1.5 m) may be mainly attributed to a high biogenic turbidity. Water salinity was low, as the maximum conductivity measured (260 μS cm^?1) suggests. The water was alkaline in the upper layers during the day time (pH >8.0), due to the high phytoplanktonic productivity, whereas in deeper layers, where decomposition processes predominate, pH was acidic (close to 6.0). Orthophosphate concentrations were high, reaching maximal concentrations in the deeper part of the reservoir (greater than 800 μg l^?1). Total phosphorus exceeded 1000 μg l^?1 in the hypolimnion during the dry season. Ammonia was the dominant inorganic nitrogen species, and its values were greater than 4000 μg l^?1 in the hypolimnetic layer during the dry season. Nitrates and nitrites were present in relatively low concentrations, except for the Paito stream entrance, where nitrate concentrations remained above 260 μg l^?1. Pao–Cachinche reservoir can be considered hypertrophic, according to Salas & Martinó’s criteria for tropical warm lakes, and could be classified as meromictic and warm monomictic, according to Lewis tropical lake types.     

A novel and highly sensitive acetyl-cholinesterase biosensor modified with hollow gold nanospheres

In this work, a highly sensitive acetylcholinesterase (AChE) inhibition-based amperometric biosensor has been developed. Firstly, a glassy carbon electrode (GCE) was modified with chitosan (Chits). Then, hollow gold nanospheres (HGNs) were absorbed onto the surface of chitosan based on the strong affinity through electrostatic adsorption. After that, l-cysteine (l-cys) was assembled on HGNs through Au–S bond. The hollow gold nanospheres were prepared by using Co nanoparticles as sacrificial templates and characterized by scanning electron microscopy, transmission electron microscopy and ultraviolet spectra, respectively. Finally, AChE was immobilized with covalent binding via –COOH groups of l-cysteine onto the modified GCE. The AChE biosensor fabrication process was characterized by cyclic voltammetry and electrochemical impedance spectroscopy methods with the use of ferricyanide as an electrochemical redox indicator. Under optimum conditions, the inhibition rates of pesticides were proportional to their concentrations in the range of 0.1–150 and 0.1–200 μg L^?1 for chlorpyrifos and carbofuran, respectively, the detection limits were 0.06 μg L^?1 for chlorpyrifos and 0.08 μg L^?1 for carbofuran. Moreover, the biosensor exhibited a good stability and reproducibility and was suitable for trace detection of pesticide residues in vegetables and fruits.     

Effects of Inorganic Turbidity on the Phytoplankton of an Amazonian Lake Impacted by Bauxite Tailings

The dumping of bauxite tailings into Batata Lake, an Amazonian clear-water lake, generated high levels of turbidity and caused a serious decrease in phytoplankton densities, which could possibly be the result of a photosynthetic limitation due to light attenuation together with an increase in algal sinking due to the adhesion of clay particles. This study aimed to investigate the sinking process through the addition of different suspended clay concentrations in columns containing Batata lake water. Since no effect of the suspended clays on Batata Lake phytoplankton sinking was observed, it was then evaluated, under laboratory conditions, whether the low conductivity of the Batata Lake water could interfere with the algae-clay aggregation process. Cultures of two algal species known to be capable to aggregate to Batata Lake suspended clays in algal culture medium: Staurodesmus convergens and Phormidium amoenum, were added to both the low conductivity Batata Lake water (14 μS cm^?1) and the high conductivity algal culture media (WC – 300 μS cm^?1 and Z8 – 560 μS cm^?1) together with Batata lake suspended clays. In both algal culture media and Batata lake water the two species had their sinking accelerated due to clay adhesion. It is thus suggested that the decrease in phytoplankton densities recorded in Batata Lake may not be related to an increase in phytoplankton loss rates due to algal-clay aggregation, but rather are a consequence of decreasing growth rates because of light attenuation.     

Effect of Seawater–Sewage Cross-Transplants on Bacterial Metabolism and Diversity

Bioassays experiments were conducted to determine the metabolic and community composition response of bacteria to transplants between relatively pristine coastal seawater and sewage-impacted seawater. There were four treatments: (1) pristine seawater bacteria?+?pristine seawater (Pb?+?Pw), (2) sewage-impacted bacteria?+?sewage-impacted water (Sb?+?Sw), (3) pristine seawater bacteria?+?sewage-impacted water (Pb?+?Sw), and (4) sewage-impacted bacteria?+?pristine seawater (Sb?+?Pw). Sewage-derived DOC was more labile and readily utilized by bacteria, which favored the growth of high nucleic acid (HNA) bacteria, resulting in high bacterial production (BP, 113?±?4.92 to 130?±?15.8 μg C l^?1?day^?1) and low respiration rate (BR, <67?±?11.3 μg C l^?1?day^?1), as well as high bacterial growth efficiency (BGE, 0.68?±?0.09 to 0.71?±?0.05). In contrast, at the relatively pristine site, bacteria utilized natural marine-derived dissolved organic matter (DOM) at the expense of lowering their growth efficiency (BGE, <0.32?±?0.02) with low BP (<62?±?6.3 μg C l^?1?day^?1) and high BR 133?±?14.2 μg C l^?1?day^?1). Sewage DOM input appeared to alter the partitioning of carbon between respiration and production of bacteria, resulting in a shift toward higher BGE, which would not enhance oxygen consumption. Taxonomic classification based on 454 pyrosequencing reads of the 16S rRNA gene amplicons revealed that changes in bacterial community structure occurred when seawater bacteria were transferred to the eutrophic sewage-impacted water. Sewage DOM fueled the growth of Gammma-proteobacteria and Epsilson-proteobacteria and reduced the bacterial richness, but the changes in the community were not apparent when sewage-impacted bacteria were transferred to pristine seawater.     

Salt dynamics in soil profiles during long-term evaporation under different groundwater conditions

To study salt dynamics in soil profiles under different groundwater conditions, a 3-year indoor experiment was carried out under conditions of open-air evaporation. Silt loam soil was treated under three groundwater table depths (0.85, 1.05, and 1.55 m) combined with three groundwater salinities: 0.40 dS m^? 1 (2 g l^? 1), 0.80 dS m^? 1 (4 g l^? 1), and 1.60 dS m^? 1 (8 g l^? 1). A total of nine soil columns (0.14 m internal diameter) were used to simulate different combinations of groundwater depths and salinities. The results obtained showed that salt first accumulated at the bottom of the soil column, and only when soil salinity in this layer had remained relatively stable with time, salt began to accumulate in the adjacent upper soil layers. When all subsoil layers had reached dynamic salinity equilibrium, electrical conductivity (EC) of soils in the surface layer began to increase drastically. With increasing salt accumulation in the surface soil, EC of the subsoil began to rise tardily. The further up the soil layer, the earlier EC started to increase, although the redistribution of salts in the soil profile tended to be homogenous. Groundwater depth did not significantly change subsoil EC values at the same depth; however, it distinctly affected the time needed for the subsoil to reach dynamic salinity equilibrium. Groundwater salinity, on the other hand, did not significantly alter the time point at which soil salinity at the same depth began to increase rapidly or the time period needed to reach dynamic salinity equilibrium. This study explored salt transport processes in the soil profile through a long-term experiment, enabling us to reveal some general laws governing salt dynamics that will be very important to understand the mechanism of soil salinization. The results could be further used to set up strategies to prevent salinization or to improve salt-affected soils.