Myelin like electrogenic filamentation and Liquid Microbial Fuel Cells Dataset.

Biofilm at water-oil interface of hypoxic water columns of microcosms, prepared from a lacustrine sample, that used diesel as a carbon source was found to show electrogenic properties. These microcosms named, Liquid Microbial Fuel Cells (L-MFCs) were electrically characterized using a custom electronic analyzer; accurate determination of voltage (V), power density (W/m 2), and current density (A/m2) for both charge and discharge phases was carried out. The instrument made it possible to carry out cell characterizations using resistive loads between 0 Ω (Ohm) and 10 kΩ. During the hypoxic and electrogenic phase, the synthesis of a system of "bacterial piping induction", produced filaments of hundreds of micrometers in which the microbial cells are hosted. Ultrastructural microscopy collected by scanning (SEM), transmission (TEM), immunofluorescence, Thunder Imager 3D, confocal laser scanning (CLSM) microscopy revealed a "myelin like" structure during filamentation processes; this "myelin like" structure exhibited cross-reactivity towards different epitopes of the myelin basic protein (MBP) and Claudin 11 (O4) of human oligodendrocytes. The disclosure of these filamentation processes could be helpful to describe further unconventional microbial structures in aquatic ecosystems and of the animal world. The data that support the findings of this study are openly available in at https://data.mendeley.com/datasets/7d35tj3j96/1.

Characterization of microbial response to petroleum hydrocarbon contamination in a lacustrine ecosystem.

Microbiomes of freshwater basins intended for human use remain poorly studied, with very little known about the microbial response to in situ oil spills. Lake Pertusillo is an artificial freshwater reservoir in Basilicata, Italy, and serves as the primary source of drinking water for more than one and a half million people in the region. Notably, it is located in close proximity to one of the largest oil extraction plants in Europe. The lake suffered a major oil spill in 2017, where approximately 400 tons of crude oil spilled into the lake; importantly, the pollution event provided a rare opportunity to study how the lacustrine microbiome responds to petroleum hydrocarbon contamination. Water samples were collected from Lake Pertusillo 10 months prior to and 3 months after the accident. The presence of hydrocarbons was verified and the taxonomic and functional aspects of the lake microbiome were assessed. The analysis revealed specialized successional patterns of lake microbial communities that were potentially capable of degrading complex, recalcitrant hydrocarbons, including aromatic, chloroaromatic, nitroaromatic, and sulfur containing aromatic hydrocarbons. Our findings indicated that changes in the freshwater microbial community were associated with the oil pollution event, where microbial patterns identified in the lacustrine microbiome 3 months after the oil spill were representative of its hydrocarbonoclastic potential and may serve as effective proxies for lacustrine oil pollution.

Electrogenic and hydrocarbonoclastic biofilm at the oil-water interface as microbial responses to oil spill.

The oil-water interface formed during an oil spill represents a challenging environment for pelagic communities living in aquatic ecosystems. At this anoxic barrier, we report the formation of a microbial hydrocarbonoclastic biofilm capable of electron transfer along the water column. This biofilm generated a membrane of surface-active compounds that allowed the spontaneous separation of electrical charges, causing the establishment of an anodic and a cathodic region and, as a result, the spontaneous creation of a liquid microbial fuel cell. Such floating biofilm was connected to the water column underneath by floating filaments that could contribute to oxygen reduction at distance. The filaments revealed an unusual lipid content induced by anoxic conditions, with prominent ultrastructural features similar to myelin found in oligodendrocytes of the vertebrate nervous system. Furthermore, these filaments showed an interesting cross-reactivity towards different epitopes of the myelin basic protein (MBP) and Claudin 11 (O4) of human oligodendrocytes. The presence of a network of filaments similar to myelin suggests the probable existence of evolutionary connections between very distant organisms. Collectively these results suggest a possible mechanism for how lake microbial communities can adapt to oil spills while offering an interesting starting point for technological developments of liquid microbial fuel cells related to the study of hydrocarbon-water interfaces. The data that support the findings of this study are openly available in figshare at https://figshare.com/s/72bc73ae14011dc7920d.

Long-term influence of early human occupations on current forests of the Guiana Shield.

To decipher the long-term influences of pre-Columbian land occupations on contemporary forest structure, diversity, and functioning in Amazonia, most of the previous research focused on the alluvial plains of the major rivers of the Amazon basin. Terra firme, that is, nonflooded forests, particularly from the Guiana Shield, are yet to be explored. In this study, we aim to give new insights into the subtle traces of pre-Columbian influences on present-day forests given the archaeological context of terra firme forests of the Guiana Shield. Following archaeological prospects on 13 sites in French Guiana, we carried out forest inventories inside and outside archaeological sites and assessed the potential pre-Columbian use of the sampled tree species using an original ethnobotanical database of the Guiana Shield region. Aboveground biomass (320 and 380 T/ha, respectively), basal area (25-30 and 30-35 m2 /ha, respectively), and tree density (550 and 700 stem/ha, respectively) were all significantly lower on anthropized plots (As) than on nonanthropized plots (NAs). Ancient human presence shaped the species composition of the sampled forests with Arecaceae, Burseraceae, and Lauraceae significantly more frequent in As and Annonaceae and Lecythidaceae more frequent in NAs. Although alpha diversity was not different between As and NAs, the presence of pre-Columbian sites enhances significantly the forest beta diversity at the landscape level. Finally, trees with edible fruits are positively associated with pre-Columbian sites, whereas trees used for construction or for their bark are negatively associated with pre-Columbian sites. Half a millennium after their abandonment, former occupied places from the inner Guiana Shield still bear noticeable differences with nonanthropized places. Considering the lack of data concerning archaeology of terra firme Amazonian forests, our results suggest that pre-Columbian influences on the structure (lower current biomass), diversity (higher beta diversity), and composition (linked to the past human tree uses) of current Amazonian forests might be more important than previously thought.

Paralytic Shellfish Toxins and Cyanotoxins in the Mediterranean: New Data from Sardinia and Sicily (Italy).

Harmful algal blooms represent a severe issue worldwide. They affect ecosystem functions and related services and goods, with consequences on human health and socio-economic activities. This study reports new data on paralytic shellfish toxins (PSTs) from Sardinia and Sicily (Italy), the largest Mediterranean islands where toxic events, mainly caused by Alexandrium species (Dinophyceae), have been ascertained in mussel farms since the 2000s. The toxicity of the A. minutum, A. tamarense and A. pacificum strains, established from the isolation of vegetative cells and resting cysts, was determined by high performance liquid chromatography (HPLC). The analyses indicated the highest toxicity for A. pacificum strains (total PSTs up to 17.811 fmol cell-1). The PSTs were also assessed in a strain of A. tamarense. The results encourage further investigation to increase the knowledge of toxic species still debated in the Mediterranean. This study also reports new data on microcystins (MCs) and ß-N-methylamino-L-alanine (BMAA) from a Sardinian artificial lake (Lake Bidighinzu). The presence of MCs and BMAA was assessed in natural samples and in cell cultures by enzyme-linked immunosorbent assay (ELISA). BMAA positives were found in all the analysed samples with a maximum of 17.84 µg L-1. The obtained results added further information on cyanotoxins in Mediterranean reservoirs, particularly BMAA, which have not yet been thoroughly investigated.

Manuseio de emergências cardiológicas em hospitais gerais do estado do Paraná / Management of cardiologic emergencies in general hospitals in the state of Paraná

JUSTIFICATIVA E OBJETIVOS: As diretrizes sobre as condutas em reanimação cardiopulmonar (RCP) foram baseadas na grande evolução dos estudos nessa área, sendo atualizadas em certos períodos de tempo. É por isso que o manuseio correto de uma parada cardíaca depende apenas da atualização do profissional que a atende. O objetivo deste estudo foi avaliar a condutados profissionais responsáveis pelo primeiro atendimento frente a emergências cardiológicas em alguns serviços hospitalares. MÉTODO: Foram avaliados 80 médicos de hospitais gerais, tomando-se como padrão as normas do Advanced Cardiologic Life Support (ACLS) e comparando o índice de acertos em um questionário com alguns quesitos de formação e trabalho do profissional. Os profissionais foram identificados com idade, sexo, estado civil, tempo de formação e local de trabalho, e em seguida responderam a seis questões referentes às condutas utilizadas. RESULTADOS: O índice de acertos foi tabulado e comparado, apresentando uma média de acertos de 3,5 questões. O mínimo de acertos foi zero, e o máximo foram seis questões. Houve correlação inversa significativa entre a idade e o número de acertos, ou seja, médicos mais jovens acertaram mais questões que os médicos de mais idade. Não houve associação significativa entre sexo, estado civil, tempo de formação e local de trabalho em relação ao número de acertos. CONCLUSÃO: Portanto, há a necessidade de atualizações entre os profissionais, buscando o melhor atendimento emergencial.

BACKGROUND AND OBJECTIVES: The cardiopulmonary resuscitation (CPR) guidelines were based on the large evolution of the studies in this area, being updated in certain periods of time. That is why the correct management of a cardiac arrest only depends on the professionals updating. The goal of the study was to evaluate the behavior of professionals responsible for primary care when they are faced with cardiologic emergencies in some hospitals. METHOD: Eighty physicians of general hospitals were evaluated, taking as base the Advance Cardiologic Life Support (ACLS) guidelines and comparing the rates of correct answers in a questionnaire with questions about professional training and work. The professionals were identified by age, gender, civil state, time of training and work place, and then they answered six questions about the procedures undertaken. RESULTS: The correct answers were tabulated and compared, demonstrating an average number of right answers of 3.5. The minimum was zero right answers and the maximum, six. There was a significant inverse correlation between the age and the number of correct answers, i.e., younger physicians had more correct answers than older ones. There were not significant relations among gender, civil state, time of training, and work place and number of correct answers. CONCLUSION: Therefore, professionals should keep themselves updated in order to deliver better emergency care.

A peptidomic approach for monitoring and characterising peptide cyanotoxins produced in Italian lakes by matrix-assisted laser desorption/ionisation and quadrupole time-of-flight mass spectrometry.

In recent years, the occurrence of cyanobacterial blooms in eutrophic freshwaters has been described all over the world, including most European countries. Blooms of cyanobacteria may produce mixtures of toxic secondary metabolites, called cyanotoxins. Among these, the most studied are microcystins, a group of cyclic heptapeptides, because of their potent hepatotoxicity and activity as tumour promoters. Other peptide cyanotoxins have been described whose structure and toxicity have not been thoroughly studied. Herein we present a peptidomic approach aimed to characterise and quantify the peptide cyanotoxins produced in two Italian lakes, Averno and Albano. The procedure was based on matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry mass spectrometry (MALDI-TOF-MS) analysis for rapid detection and profiling of the peptide mixture complexity, combined with liquid chromatography/electrospray ionisation quadrupole time-of- flight tandem mass spectrometry (LC/ESI-Q-TOF-MS/MS) which provided unambiguous structural identification of the main compounds, as well as accurate quantitative analysis of microcystins. In the case of Lake Averno, a novel variant of microcystin-RR and two novel anabaenopeptin variants (Anabaenopeptins B(1) and Anabaenopeptin F(1)), presenting homoarginine in place of the commonly found arginine, were detected and characterised. In Lake Albano, the peculiar peptide patterns in different years were compared, as an example of the potentiality of the peptidomic approach for fast screening analysis, prior to fine structural analysis and determination of cyanotoxins, which included six novel aeruginosin variants. This approach allows for wide range monitoring of cyanobacteria blooms, and to collect data for evaluating possible health risks to consumers, through the panel of the compounds produced along different years.

Seasonal succession of Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum blooms with cylindrospermopsin occurrence in the volcanic Lake Albano, Central Italy.

The cyanobacterial toxin cylindrospermopsin is rapidly spreading in the European temperate Countries. Cylindrospermopsin was detected for the first time in Italy in 2004; in this study, the presence of this toxin in Albano Lake (Central Italy) has been correlated to the cyanobacterial species Cylindrospermopsis raciborskii and Aphanizomenon ovalisporum and their population dynamics. In 2004, these two species succeeded in the lake during spring, summer, and early autumn without overlapping, causing superficial blooms. Cylindrospermopsin was detected in lake samples by LC-MS/MS and ELISA immunoassay, showing extracellular superficial values ranging from 2.6 to 126 microg/L, and water column values ranging from 0.41 to 18.4 microg/L. Twenty-six of 30 positive water samples (86%) exceeded the recommended limit of 1 microg/L. Intracellular values up to 42.3 microg/g were measured. Moreover, cylindrospermopsin was detected in tissues from two Salmo trutta trouts (up to 2.7 ng/g) and in a well for drinking water supply (1.6 microg/L). For the first time, two cyanobacterial species producing cylindrospermopsin were detected in the same lake in Italy.

Determination of cylindrospermopsin in freshwaters and fish tissue by liquid chromatography coupled to electrospray ion trap mass spectrometry.

Cylindrospermopsin (CYN) is a toxic alkaloid-like compound produced by some strains of cyanobacteria, procariotic organisms occurring in water blooms, observed worldwide in eutrophic lakes and drinking water reservoirs. Methods for determination of CYN in freshwater and fish muscle by liquid chromatography coupled to electrospray ion trap mass spectrometry are herein described. The performances of both methods are reported; ion trap LC/ESI-MS/MS resulted highly selective and reliable in unambiguous identification of CYN, based on monitoring the precursor ion and three product ions. The methods developed showed satisfactory mean recoveries (higher than 63.6%) and relative standard deviations, ranging from 5.8 to 9.8%. The limits of quantification at 0.10 ng/mL in freshwaters and 1.0 ng/g in fish muscle, respectively, allow for determination of CYN also in early contamination stages. Ion trap LC/ESI-MS/MS was successfully applied to the identification and quantification of CYN in water and cyanobacteria extracts from Lake Averno, near Naples, representing the first case of contamination described in southern Italy.

Liquid chromatography coupled to quadruple time-of-flight tandem mass spectrometry for microcystin analysis in freshwaters: method performances and characterisation of a novel variant of microcystin-RR.

Cyanobacteria, also called blue-green algae, occur worldwide within water blooms in eutrophic lakes and drinking water reservoirs, producing several biotoxins (cyanotoxins). Among these, microcystins (MCs) are a group of cyclic heptapeptides showing potent hepatotoxicity and activity as tumour promoters. So far, at least 89 MCs from different cyanobacteria genera have been characterised. Herein, ion trap, matrix-assisted laser desorption/ionisation time-of-flight (MALDI-ToF) and quadruple time-of-flight (Q-ToF) mass spectrometry (MS)-based methods were tested and compared for analysing MCs in freshwaters. Method performances in terms of limit of detection, limit of quantification, mean recoveries, repeatability, and specificity were evaluated. In particular, a liquid chromatography/electrospray ionisation (LC/ESI)-Q-ToF-MS/MS method was firstly described to analyse MCs in freshwaters; this technique is highly selective and sensitive, and allowed us to characterise the molecular structure of an unknown compound. Indeed, the full structural characterisation of a novel microcystin variant from a bloom of Planktothrix rubescens in the Lake Averno, near Naples, was attained by the study of the fragmentation pattern. The new cyanotoxin was identified as the 9-acetyl-Adda variant of microcystin-RR.

Characterisation of biotoxins produced by a cyanobacteria bloom in Lake Averno using two LC-MS-based techniques.

Cyanobacteria (blue-green algae) cause blooms in eutrophic lakes and drinking water reservoirs. They also produce biotoxins, including microcystins (MCs), highly toxic cyclic heptapeptides that cause poisoning in animals and human. In this paper, we present a method for the analysis of four MCs by ion trap LC-MS and MALDI-TOF/MS. The data are compared to evaluate the performance and reliability of the different MS detection systems. The method was applied to the analysis of water and algae samples from Lake Averno, near Naples, as a consequence of a cyanobacteria bloom. The analysis of algae cell extracts showed no contamination by known microcystins, but the three main substances were detected. MALDI-TOF/MS was successful for screening of the biotoxins in the samples, identifying anabaenopeptin B and anabaenopeptin F as the major contaminants on the basis of literature mass spectrometry data. The structure of the third compound was not identified and is under further investigation. The method could characterise the biotoxins produced in Lake Averno for evaluating health risks related to their presence.

Simple and rapid determination of anatoxin-a in lake water and fish muscle tissue by liquid-chromatography-tandem mass spectrometry.

Anatoxin-a (AN) is a powerful neurotoxin that can be produced by cyanobacteria in eutrophic freshwaters. Consequently, AN can contaminate lakes, rivers and basins destined for drinking water and aquaculture. Two simple, specific and sensitive procedures for determining AN in lake water and fish muscle tissue are presented. Both analytical protocols are based on liquid-chromatography (LC)-tandem mass spectrometry (MS) with electrospray ionization. MS data were acquired in the multi reaction monitoring mode by selecting four precursor to product ion transitions. After filtration, AN in lake water was analyzed by directly injecting 0.5 ml of the aqueous sample in the LC column. Analysis of AN in fish muscle tissue involved the matrix solid-phase dispersion technique. The analyte was extracted from tissue by 4 ml of water acidified to pH 2 and heated at 80 degrees C. After acidification and filtration, 0.2 ml of the aqueous extract was injected in the LC column. Analyte recovery ranged between 71 and 79% and was not substantially affected by both the analyte concentration and the type of fish. Phenylalanine is an essential amino acid invariably present in any animal tissue. Like AN, this amino acid produces a pseudo molecular ion at m/z 166, it has a very similar fragmentation pattern and LC retention time. This method is able to prevent identifying phenylalanine for AN as the latter compound is eluted more than 1 min before the former one and the two compounds have remarkably different relative ion signal intensities. On the basis of a signal-to-noise ratio of 10, limits of quantification of AN in water and fish fillet were estimated to be 13 ng/l and 0.5 ng/g, respectively.

Monitoring algal toxins in lake water by liquid chromatography tandem mass spectrometry.

Microcystins (MCs) and cylindrospermopsin (CYL) are potent natural toxins produced by cyanobacteria (blue-green algae) that grow worldwide in eutrophic freshwaters and cause animal and human water-based toxicoses. The main purpose of this work has been assessing the contamination levels of some MCs and CYL in eutrophic Italian lake (Albano) water. To do this, we have developed an original analytical method involving MC extraction with a sorbent (Carbograph 4) cartridge. CYL is a highly polar compound that is scarcely retained by any sorbent material. To analyze this toxin, we directly injected 0.5 mL of filtered lake water into the liquid chromatography (LC) column. Analytes were quantified by LC coupled to tandem mass spectrometry in the multireaction monitoring mode. The recovery of five selected MCs added to an analyte free lake water sample at three different concentrations (50, 150, and 500 ng/L) ranged between 93 and 103% with RSD values no larger than 8%. Limits of quantification (LOQ) of the five MCs were within the 2-9 ng/L range, whereas the LOQ of CYL was 300 ng/L. The occurrence and abundance of cyanotoxins in Lake Albano was monitored over four months (Sept-Dec 2004) by analyzing water samples collected monthly at the center of the lake and at different depths (from 0 to -30 m). During survey and with the MS/MS system operating in the parent ion scan mode, we individuated two demethylated forms of MC-RR and one demethylated variety of MC-LR. Demethylated MC-RRs are known to be even more toxic than MC-RR toward zooplanktic grazers. CYL was the most-abundant toxin during the first three monitoring months. To the best of our knowledge, this is the first work reporting concentration levels of CYL in lake water.

Simple assay for analyzing five microcystins and nodularin in fish muscle tissue: hot water extraction followed by liquid chromatography-tandem mass spectrometry.

A simple, specific, and sensitive procedure for determining six cyanotoxins, that is, microcystins RR, LR, YR, LA, and LW and nodularin, in fish muscle tissue is presented. This method is based on the matrix solid-phase dispersion technique with heated water as extractant followed by liquid chromatography (LC)-tandem mass spectrometry (MS) equipped with an electrospray ion source. Target compounds were extracted from tissue by 4 mL of water acidified to pH 2 and heated at 80 degrees C. After acidification and filtration, 0.2 mL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the multireaction monitoring mode, with at least two precursor ion > product ion transitions selected for each target compound. Analyte recovery ranged between 61 and 82% and was not substantially affected by either the analyte concentrations or the type of fish. The nonexcellent recovery of some of the microcystins was traced to binding of these compounds to protein phosphatases in fish tissue occurring during sample treatment. The existence of covalently bound microcystins in fish has been evidenced by several studies. Compared to an older sample preparation procedure, this one extracted larger amounts of the analytes in a simpler and much more rapid way. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to range between 1.6 and 4.0 ng/g. The effects of temperature and volume of the extractant on the analyte recovery were studied.

Anatoxin-a toxin in the cyanobacterium Planktothrix rubescens from a fishing pond in northern Italy.

A heavy algal bloom occurring in a fishing pond in northern Italy full of Salmo trutta was examined for algae taxonomy and toxic production. The dominant algal species (98%) was identified as the cyanobacterium Planktothrix rubescens (D.C. ex GOMONT) Komarek Anagnostidis, based on morphological examination, and it was revealed to be toxic in mouse and Vibrio fischeri bioassays. The toxin was identified as anatoxin-a using high-performance liquid chromatography and confirmed using liquid chromatography-mass spectrometry (LC-MS). The mouse bioassay gave signs of poisoning, as previously reported for anatoxin-a. The LC-MS confirmed the presence of an anatoxin-a peak at m/z 166 (M+H+). The content of toxin in the field population was estimated at 12.13 microg/g of fresh cells. The bloom was sustained by the very high N/P ratio in the water. This is the first report in Italy of an anatoxin-a-producing Planktothrix rubescens population.