Who is where in the Plastisphere, and why does it matter?

To fully understand how plastic is affecting the ocean, we need to understand how marine life interacts directly with it. Besides their ecological relevance, microbes can affect the distribution, degradation and transfer of plastics to the rest of the marine food web. From amplicon sequencing and scanning electron microscopy, we know that a diverse array of microorganisms rapidly associate with plastic marine debris in the form of biofouling and biofilms, also known as the "Plastisphere." However, observation of multiple microbial interactions in situ, at small spatial scales in the Plastisphere, has been a challenge. In this issue of Molecular Ecology Resources, Schlundt et al. apply the combination labelling and spectral imaging - fluorescence in situ hybridization to study microbial communities on plastic marine debris. The images demonstrate the colocalization of abundant bacterial groups on plastic marine debris at a relatively high taxonomic and spatial resolution while also visualizing biofouling of eukaryotes, such as diatoms and bryozoans. This modern imaging technology provides new possibilities to address questions regarding the ecology of marine microbes on plastic marine debris and describe more specific impacts of plastic pollution in the marine food webs.

Synthesis and antiprotozoal activity of nitazoxanide-N-methylbenzimidazole hybrids.

A series of a novel hybrid compounds between nitazoxanide and N-methylbenzimidazole were synthesized starting from the corresponding N-methyl-2-nitroanilines. The new hybrid compounds (1-13) were evaluated in vitro against Giardia intestinalis, Entamoeba histolytica, Trichomonas vaginalis. NTZ, MTZ and ABZ were used as drug standards. Experimental evaluations revealed all of the new compounds (1-13) were active and showed strong activity against the three protozoa, particularly with E. histolytica where the IC50 values ranged between 3 and 69 nM. Overall, compounds 2, 5, 7, 8, 9, 11 and 12 stood out with values lower than 87 nM for all three protozoa, comparatively better than the reference drugs.

New evidences of Roundup (glyphosate formulation) impact on the periphyton community and the water quality of freshwater ecosystems.

Argentina is the second largest world producer of soybeans (after the USA) and along with the increase in planted surface and production in the country, glyphosate consumption has grown in the same way. We investigated the effects of Roundup (glyphosate formulation) on the periphyton colonization. The experiment was carried out over 42 days in ten outdoor mesocosms of different typology: "clear" waters with aquatic macrophytes and/or metaphyton and "turbid" waters with great occurrence of phytoplankton or suspended inorganic matter. The herbicide was added at 8 mg L(-1) of the active ingredient (glyphosate) in five mesocosms while five were left as controls (without Roundup addition). The estimate of the dissipation rate (k) of glyphosate showed a half-life value of 4.2 days. Total phosphorus significantly increased in treated mesocosms due to Roundup degradation what favored eutrophication process. Roundup produced a clear delay in periphytic colonization in treated mesocosms and values of the periphytic mass variables (dry weight, ash-free dry weight and chlorophyll a) were always higher in control mesocosms. Despite the mortality of algae, mainly diatoms, cyanobacteria was favored in treated mesocosms. It was observed that glyphosate produced a long term shift in the typology of mesocosms, "clear" turning to "turbid", which is consistent with the regional trend in shallow lakes in the Pampa plain of Argentina. Based on our findings it is clear that agricultural practices that involve the use of herbicides such as Roundup affect non-target organisms and the water quality, modifying the structure and functionality of freshwater ecosystems.

Metals and linear alkylbenzene sulphonate as inhibitors of the algae Pseudokirchneriella subcapitata acid phosphatase activity.

Sewage sludge applied to soils as a fertilizer often contains metals and linear alkylbenzene sulphonate (LAS) as contaminants. These pollutants can be transported to the aquatic environment where they can alter the phosphatase activity in living organisms. The acid phosphatase of algae plays important roles in metabolism such as decomposing organic phosphate into free phosphate and autophagic digestive processes. The order of in vitro inhibition of Pseudokirchneriella subcapitata acid phosphatase at the highest concentration tested was LAS > Hg2+ = Al3+ > Se4+ = Pb2+ > Cd2+. A non-competitive inhibition mechanism was obtained for Hg2+ (Ki = 0.040 mM) and a competitive inhibition for LAS (Ki = 0.007 mM). In vivo studies with treated algae cultures showed that the inhibition of specific activity was observed in algae exposed during 7 days, in contrast to short term (24 h) treatments with both these chemicals. Our results suggest that the inhibition parameters in vitro did not markedly differ between the two chemicals. On the other hand, in vivo evaluations showed strong differences between both pollutants regarding the concentration values and the degree of response.

Genomic databases and the search of protein targets for protozoan parasites.

The development of databases devoted to biological information opened the possibility to integrate, query and analyze biological data obtained from several sources that otherwise would be scattered through the web. Several issues arise in the handling of biological information, mainly due to the diversity of biological subject matter and the complexity of biological approaches towards phenomena of the living world. The integration of genomic data, three-dimensional structures of proteins, biological activity, and drugs availability allows a system approach to the study of the biology. Here we review the current status of these research efforts to develop genomic databases for protozoan parasites, such as the apicomplexan parasites, Trypanosoma cruzi and Leishmania spp. These databases may help in the discovery and development of new drugs against parasite-mediated diseases.

Electron microscopy in antiparasitic chemotherapy: a (close) view to a kill.

Electron microscopy may be useful in chemotherapy studies at distinct levels, such as the identification of subcellular targets in the parasites and the elucidation of the ultimate drug mechanism of action, inferred by the alterations induced by antiparasitic compounds. In this review we present data obtained by electron microscopy approaches of different parasitic protozoa, such as Trypanosoma cruzi, Leishmania spp., Giardia lamblia and trichomonads, under the action of drugs, demonstrating that the cell architecture organization is only determined in detail at the ultrastructural level. The transmission electron microscopy may shed light (i.e. electrons) not only on the affected compartment, but also on the manner it is altered, which may indicate presumable target metabolic pathways as well as the actual toxic or lethal effects of a drug. Cytochemical and analytical techniques can provide valuable information on the composition of the altered cell compartment, permitting the bona fide identification of the drug target and a detailed understanding of the mechanism underneath its effect. Scanning electron microscopy permits the recognition of the drug-induced alterations on parasite surface and topography. Such observations may reveal cytokinetic dysfunctions or membrane lesions not detected by other approaches. In this context, electron microscopy techniques comprise valuable tools in chemotherapy studies.

Natural impacted freshwaters: in situ use of alginate immobilized algae to the assessment of algal response.

The objective of this study was to investigate the feasibility of an in situ phytotoxicity test using alginate-immobilized algae for 60 days, in the assessment of water quality in an impacted small peri-urban stream. After laboratory optimization of algae immobilization/de-immobilization processes, the performance of immobilized/de-immobilized algae was compared to the performance of free algae in terms of specific algal growth and sensitivity. This was done by comparing 72 h EC50 values obtained with zinc and the pesticides clomazone and carbofuran. The results showed a similar performance, which allow us to conclude that immobilization for 60 days do not cause any significant alteration in algae physiology. In the field, immobilized algae were exposed at different times (2, 4 and 7 days) to water samples in both disturbed and undisturbed sites. Both laboratory and field experiments indicated that alginate-immobilized algae for 60 days were sufficiently sensitive for use in the in situ assessment of water quality.

Physico-chemical, microbiological and ecotoxicological evaluation of a septic tank/Fenton reaction combination for the treatment of hospital wastewaters.

Hospital wastewater is considered a complex mixture populated with pathogenic microorganisms. The genetic constitution of these microorganisms can be changed through the direct and indirect effects of hospital wastewater constituents, leading to the appearance of antibiotic multi-resistant bacteria. To avoid environmental contamination hospital wastewaters must be treated. The objective of this study was to evaluate the efficiency of hospital wastewater treated by a combined process of biological degradation (septic tank) and the Fenton reaction. Thus, after septic tank biodegradation, batch Fenton reaction experiments were performed in a laboratory-scale reactor and the effectiveness of this sequential treatment was evaluated by a physico-chemical/microbiological time-course analysis of COD, BOD(5), and thermotolerant and total coliforms. The results showed that after 120min of Fenton treatment BOD(5) and COD values decreased by 90.6% and 91.0%, respectively. The BOD(5)/COD ratio changed from 0.46 to 0.48 after 120min of treatment. Bacterial removal efficiency reached 100%, while biotests carried out with Scenedesmus subspicatus and Daphnia magna showed a significant decrease in the ecotoxicity of hospital wastewater after the sequential treatment. The use of this combined system would ensure that neither multi-resistant bacteria nor ecotoxic substances are released to the environment through hospital wastewater discharge.

Mitracarpus frigidus aerial parts exhibited potent antimicrobial, antileishmanial, and antioxidant effects.

The crude extract and the hexane, CH(2)Cl(2), EtOAc, n-BuOH, and hydromethanolic fractions of the aerial parts of Mitracarpus frigidus were evaluated against promastigote forms of two species of Leishmania (L. chagasi and L. amazonensis), 11 strains of bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica sorovar Tythimurium, Shigella sonnei, Klebsiella pneumoniae, Escherichia coli, Micrococcus luteus, Enterococcus faecalis, Enterobacter cloacae, Streptococcus pyogenes and Bacillus cereus) and two yeasts (Candida albicans and Cryptococcus neoformans). The antioxidant activity (DPPH radical scavenging activity and reducing power), cytotoxicity against mammalian cells, and the contents of phenolics and flavonoids were determined. Phytochemical analysis of the major groups of phytoconstituents is also reported. All samples showed antioxidant activity which was positively correlated to the content of phenolic compounds. S. sonnei, B. cereus and C. neoformans were susceptible to all extracts tested, except for the n-BuOH and hydromethanolic fractions, which demonstrated no antimicrobial activity. The lowest MIC was recorded for the CH(2)Cl(2) fraction against C. neoformans (MIC of 10 microg/ml), followed by B. cereus, S. sonnei, and E. cloacae (MIC of 20, 39 and 39 microg/ml, respectively). The CH(2)Cl(2) fraction was the most effective against L. chagasi (IC(50) of 6.7 microg/ml), and the hydromethanolic fraction exhibited the best activity against L. amazonensis (IC(50) of 9 microg/ml). A cytotoxic effect on mammalian cells was observed only for the crude extract and CH(2)Cl(2) fraction at the concentrations of 130 and 31 microg/ml, respectively. These results suggest that M. frigidus has interesting antimicrobial, antileishmanial and antioxidant activities.

In vivo microspectroscopy monitoring of chromium effects on the photosynthetic and photoreceptive apparatus of Eudorina unicocca and Chlorella kessleri.

In microorganisms and plants, chromium (Cr) is not essential for any metabolic process, and can ultimately prove highly deleterious. Due to its widespread industrial use, chromium has become a serious pollutant in diverse environmental settings. The presence of Cr leads to the selection of specific algal populations able to tolerate high levels of Cr compounds. The varying Cr-resistance mechanisms displayed by microorganisms include biosorption, diminished accumulation, precipitation, reduction of Cr(6+) to Cr(3+), and chromate efflux. In this paper we describe the effects of Cr(6+) (the most toxic species) on the photosynthetic and photoreceptive apparatus of two fresh water microalgae, Eudorina unicocca and Chlorella kessleri. We measured the effect of this heavy metal by means of in vivo absorption microspectroscopy of both the thylakoid compartments and the eyespot. The decomposition of the overall absorption spectra in pigment constituents indicates that Cr(6+) effects are very different in the two algae. In E. unicocca the metal induced a complete pheophinitization of the chlorophylls and a modification of the carotenoids present in the eyespot after only 120 h of exposition at a concentration equal or greater than 40 microM, which is the limit for total Cr discharge established by US EPA regulations. In C. kessleri, chromium concentrations a hundred times higher than this limit had no effect on the photosynthetic machinery. The different tolerance level of the two algae is suggested to be due to the different properties of the mucilaginous envelope and cell wall covering, respectively, the colonies of Eudorina and the single cells of Chlorella, which binds chromium cations to a different extent.

Biochemical biomarkers in algae and marine pollution: a review.

Environmental pollution by organic compounds and metals became extensive as mining and industrial activities increased in the 19th century and have intensified since then. Environmental pollutants originating from diverse anthropogenic sources have been known to possess adverse values capable of degrading the ecological integrity of marine environment. The consequences of anthropogenic contamination of marine environments have been ignored or poorly characterized with the possible exception of coastal and estuarine waters close to sewage outlets. Monitoring the impact of pollutants on aquatic life forms is challenging due to the differential sensitivities of organisms to a given pollutant, and the inability to assess the long-term effects of persistent pollutants on the ecosystem as they are bio-accumulated at higher trophic levels. Marine microalgae are particularly promising indicator species for organic and inorganic pollutants since they are typically the most abundant life forms in aquatic environments and occupy the base of the food chain. We review the effects of pollutants on the cellular biochemistry of microalgae and the biochemical mechanisms that microalgae use to detoxify or modify pollutants. In addition, we evaluate the potential uses of microalgae as bioindicator species as an early sentinel in polluted sites.

Ultrastructural alterations in organelles of parasitic protozoa induced by different classes of metabolic inhibitors.

Parasitic protozoa such as Leishmania, Trypanosoma, Plasmodium, Toxoplasma gondii, Giardia and Trichomonas are able to cause several diseases affecting millions of people around the world with dramatic consequences to the socio-economic life of the affected countries. Diseases like malaria, leishmaniasis and trypanosomiasis have been classified by the World Health Organization as neglected diseases, because they have been almost completely forgotten by the governments as well as the pharmaceutical companies. The specific chemotherapy currently employed for the treatment of these diseases has serious limitations due to lack of efficacy, toxic side effects, growth of drug-resistance and high costs. Thus, it is urgent to develop new chemotherapeutic agents that are more effective, safe and accessible. In this context, several works have been focused on understanding the effect of different drug-treatments on these parasitic protozoa. Organelles and structures such as mitochondrion, kinetoplast, apicoplast, glycosome, acidocalcisome, hydrogenosome, plasma membrane and the cytoskeleton have been studied using different approaches to identify new targets for the development of new chemotherapeutic agents that are required. Some studies on alterations in the fine structure, as assayed using electron microscopy, have indicated the nature of lesions induced by several drugs, allowing deductions on possible modes of action. Here, we briefly review the available data of the effects of several drugs on the ultrastructure of parasitic protozoa and show how electron microscopy can contribute to elucidate the different mechanisms of these anti-parasitic drugs.

Effect of nitrogen, phosphorous, and their interaction on coral reef algal succession in Glover's Reef, Belize.

Nitrogen and phosphorous fertilizers were used to determine their short-term summer effects on algal colonization, abundance, and species composition in moderate herbivory treatments. Secondary succession of algae on coral skeletons was examined in four treatments: an untreated control, a pure phosphate fertilizer, a pure nitrogen fertilizer, and an equal mix of the two fertilizers. Turf algae cover was the only measure of algae abundance to respond significantly to fertilization. Turf cover was three times higher in treatments with added nitrogen when compared with the pure phosphorus treatment. These turfs were dominated by green and cyanobacteria taxa, namely Enteromorpha prolifera, Lyngbya confervoides, and two species of Cladophora. The phosphate treatment was dominated by encrusting corallines and the cyanobacteria L. confervoides, while the controls had the highest cover of frondose brown algae, namely Padina sanctae-crucis and two species of Dictyota. Results indicate that turf algae were co-limited by nitrogen and phosphorus but enrichment appeared to inhibit brown frondose algae that currently dominate these reefs. Number of species was lowest on the pure phosphorus and nitrogen treatments, highest in the controls and intermediate in the mixed treatments, which suggests that diversity is reduced most by an imbalanced nutrient ratio.

Effects of the herbicide Roundup on freshwater microbial communities: a mesocosm study.

The impact of the widely used herbicide glyphosate has been mainly studied in terrestrial weed control, laboratory bioassays, and field studies focusing on invertebrates, amphibians, and fishes. Despite the importance of phytoplankton and periphyton communities at the base of the aquatic food webs, fewer studies have investigated the effects of glyphosate on freshwater microbial assemblages. We assessed the effect of the commercial formulation Roundup using artificial earthen mesocosms. The herbicide was added at three doses: a control (without Roundup) and two treatments of 6 and 12 mg/L of the active ingredient (glyphosate). Estimates of the dissipation rate (k) were similar in the two treatments (half-lives of 5.77 and 7.37 d, respectively). The only two physicochemical parameters showing statistically significant differences between treatments and controls were the downward vertical spectral attenuation coefficient kd(lambda), where lambda is wavelength, and total phosphorus concentration (TP). At the end of the experiment, the treated mesocosms showed a significant increase in the ratio kd(490 nm)/k(d)(550 nm) and an eightfold increase in TP. Roundup affected the structure of phytoplankton and periphyton assemblages. Total micro- and nano-phytoplankton decreased in abundance in treated mesocosms. In contrast, the abundance of picocyanobacteria increased by a factor of about 40. Primary production also increased in treated mesocosms (roughly by a factor of two). Similar patterns were observed in the periphytic assemblages, which showed an increased proportion of dead: live individuals and increased abundances of cyanobacteria (about 4.5-fold). Interestingly, the observed changes in the microbial assemblages were captured by the analysis of the pigment composition of the phytoplankton, the phytoplankton absorption spectra, and the analysis of the optical properties of the water. The observed changes in the structure of the microbial assemblages are more consistent with a direct toxicological effect of glyphosate rather than an indirect effect mediated by phosphorus enrichment.

Secondary metabolites help biocontrol strain Pseudomonas fluorescens CHA0 to escape protozoan grazing.

In soil ecosystems, bacteria must cope with predation activity, which is attributed mainly to protists. The development of antipredation strategies may help bacteria maintain higher populations and persist longer in the soil. We analyzed the interaction between the root-colonizing and biocontrol strain Pseudomonas fluorescens CHA0 and three different protist isolates (an amoeba, a flagellate, and a ciliate). CHA0 produces a set of antibiotics, HCN, and an exoprotease. We observed that protists cannot grow on CHA0 but can multiply on isogenic regulatory mutants that do not produce the extracellular metabolites. The in vitro responses to CHA0 cells and its exoproducts included growth inhibition, encystation, paralysis, and cell lysis. By analyzing the responses of protists to bacterial supernatants obtained from different isogenic mutants whose production of one or more exometabolites was affected and also to culture extracts with antibiotic enrichment, we observed different contributions of the phenolic antifungal compound 2,4-diacetylphloroglucinol (DAPG) and the extracellular protease AprA to CHA0 toxicity for protists and to the encystation-reactivation cycle. The grazing pressure artificially produced by a mixture of the three protists in a microcosm system resulted in reduced colonization of cucumber roots by a regulatory isogenic CHA0 mutant unable to produce toxins. These results suggest that exometabolite production in biocontrol strain CHA0 may contribute to avoidance of protist grazing and help sustain higher populations in the rhizosphere, which may be a desirable and advantageous trait for competition with other bacteria for available resources.

A sediment quality triad assessment of the impact of copper mine tailings disposal on the littoral sedimentary environment in the Atacama region of northern Chile.

A sediment quality triad (SQT) assessment was made of the impact of copper mine tailings disposal on littoral meiofaunal assemblages in the Atacama region of northern Chile. This situation is unusual in that the disposal is direct into the high-energy coastal system and not via a river estuary or other low-energy environment. This situation also allows for the examination of the impact of copper mine tailings in the absence of confounding effects from other pollutants. The three components of the SQT were: 1. an analysis of the bioavailable metals in both the sedimentary porewater and the adjacent seawater, 2. a microcosm bioassay of both sediments and seawaters using meiofaunal assemblages, and 3. quantitative field samples of the meiofaunal assemblages. Twelve study sites with varying degrees of impact were used, including three reference sites. The study identified that both the meiofaunal assemblage densities and taxa diversities decrease with increasing levels of bioavailable copper, that the Foraminifera and Harpacticoida are sensitive to copper, and that otoplanid Turbellaria are often characteristic of impacted sites; tailings also have both chemical and physical impacts on the environment. In some cases the physical impact of tailings is more important in excluding some organisms e.g. the interstitial polychaete, Saccocirrus sonomacus, from a site than is their chemical impact.

Foraminiferal responses to polluted sediments in the Montevideo coastal zone, Uruguay.

A study of foraminiferal assemblages was carried out in 24 sediment samples collected from the Montevideo coastal zone (south-eastern coastal region of South America) to assess the response of the benthic foraminifera to the polluted sediments. The area is affected by different pollutants such as sewage, hydrocarbons and heavy metals derived from different sources. Biological data were analyzed with multivariate techniques of cluster analysis and a principal component analysis (PCA) was performed for abiotic factors. The results allowed the recognition of different species assemblages corresponding to different sub-environments, which reflected the prevalent ecological conditions. The Montevideo Bay, particularly, its inner part showed an extremely poor foraminiferal fauna-including a totally azoic station-and high percentages of abnormal tests, when compared with the adjacent Punta Carretas and Punta Yeguas zones. Mean faunal density showed a strong relationship with organic matter, oxygen and heavy metal concentrations, as well as redox potential and pH values of each sub-environment. Although the adjacent zones presented a moderate pollution degree, it was noticed that a positive effect on the foraminiferal density specially on Ammonia tepida, caused by the sewage pipe located in Punta Carretas, a pure organic contamination. Differences among foraminiferal assemblages seemed to be related to the combined action of the several kinds of pollutants and the natural abiotic variables, like the rapid salinity changes that occurred in this area.

Comparative toxicity of effluents processed by different treatments in V79 fibroblasts and the algae Selenastrum capricornutum.

The efficacy of ozonation and of photocatalysis processing in the treatment of pulp mill ECF (elementary chlorine free) bleaching and textile effluents was evaluated by determining total organic carbon reduction (TOC) and the toxicity. The chronic toxicity of the effluents was evaluated by the ability to inhibit the growth of algae Selenastrum capricornutum. Cultured hamster V79 fibroblasts were used to assess the cytotoxicity of effluents submitted to different detoxification processes. Two endpoints were measured in V79 cells: 3-(4,5-dimethylthiazole-2-yl)-2,5-biphenyl tetrazolium bromide (MTT) reduction and neutral red uptake (NRU). Both treatment processes were able to reduce the TOC, although ozonization was less effective for pulp mill ECF bleaching. The pulp mill ECF bleaching and textile effluents reduced the growth of S. capricornutum by 39% and 27%, respectively. However, at the highest concentration tested, the textile effluents treated by photochemical process for 60 min showed increased cytotoxicity in V79 cells compared to the untreated effluent when assessed by the NRU and MTT reduction assays (increases of 30% and 40%, respectively). Pulp mill ECF bleaching effluent treated by ozonization had a similar cytotoxicity to that of untreated effluent in the NRU assay. In contrast, the MTT reduction assay indicated that effluents treated with ozone were around 20% more cytotoxic than untreated effluents. These results show that cultured fibroblasts may be useful for studying cellular responses to pollutants and may be included in tests to monitor the efficiency of effluent detoxification processes.

Cissampeloflavone, a chalcone-flavone dimer from Cissampelos pareira.

From the aerial parts of Cissampelos pareira L. (Menispermaceae), a chalcone-flavone dimer has been isolated which, mainly from NMR spectroscopic and MS data, was proved to be 2-(4-hydroxy-3-methoxyphenyl)-7-(4-methoxyphenyl)-6-(2-hydroxy-4,6-dimethoxybenzoyl)-furano[3,2-g]benzopyran-4-one. This has been assigned the trivial name cissampeloflavone. The compound has good activity against Trypanosoma cruzi and T. brucei rhodesiense and has a low toxicity to the human KB cell line.