Biomarkers at the Individual and Biochemical Level: Effects of Pure and Formulated Lambda-Cyhalothrin in Boana pulchella Tadpoles (Duméril and Bibron, 1841).

We compared the effects of lambda-cyhalothrin as the pure active ingredient and as a formulated product (Zero®), on the larval stage of the autochthonous species Boana pulchella. We evaluated ecotoxicological endpoints, behavioral and developmental alterations, and the biochemical detoxifying, neurotoxic, and oxidative stress responses, covering a wide concentration range from environmental to high application levels. Both pyrethroid preparations displayed similar ecotoxicity (median lethal concentration of ~0.5 mg/L), with the lethal effect of Zero® being more pronounced than that of the active ingredient. Sublethal behavioral alterations in natatory activity were observed at 1000 times lower concentrations, indicating the ecological hazard of tadpole exposure to this pyrethroid at environmentally relevant concentrations. Biochemical endpoints in B. pulchella larvae showed significant responses to lambda-cyhalothrin in the ng/L range; these responses were different for the pure or the formulated product, and they were variable at higher concentrations. Principal components analysis confirmed the prevalence of biochemical responses as early endpoints at the lowest lambda-cyhalothrin concentrations; the Integrated Biomarker Response Index proportionally increased with pyrethroid concentration in a similar way for the pure and the formulated products. We conclude that lambda-cyhalothrin is of concern from an environmental perspective, with particular emphasis on autochthonous anuran development. The battery of biochemical biomarkers included in our study showed a consistent integrated biomarker response, indicating that this is a potent tool for monitoring impacts on amphibians. Environ Toxicol Chem 2024;43:2134-2144. © 2024 SETAC.

Physiological and molecular effects of contaminants of emerging concerns of micro and nano-size in aquatic metazoans: overview and current gaps in Antarctic species.

Although Antarctica is the most isolated continent on Earth, its remote location does not protect it from the impacts of human activities. Antarctic metazoans such as filter-feeding invertebrates are a crucial component of the Antarctic benthos. They play a key role in the benthic-pelagic carbon flux in coastal areas by filtering particles and planktonic organisms from the sediment-water interface. Due to their peculiar ecological niche, these organisms can be considered a wasp-waist in the ecosystem, making them highly sensitive to marine pollution. Recently, anthropogenic particles such as micro-nanoplastics and manufactured nanoparticles (MNP) have been classified as contaminants of emerging concern (CEC) due to their small size range, which also overlaps with the preferred particle size ingested by aquatic metazoans. Indeed, it has been demonstrated that some species such as Antarctic krill can ingest, transform, and release MNPs, making them newly bioavailable for other Antarctic filter-feeding organisms. Similarly, the production and use of anthropogenic MNP are rapidly increasing, leading to a growing presence of materials, such as nano-sized metal-oxides, in the environment. For these reasons, it is important to provide evidence of the adverse effects of such emerging contaminants at sub-lethal concentrations in environmental risk assessments. These contaminants may cause cascade effects with consequences not only on individuals but also at the community and ecosystem levels. In this review, we discuss the state-of-the-art knowledge on the physiological and molecular effects of anthropogenic MNP in Antarctic aquatic metazoans. We further highlight the importance of identifying early biomarkers using sessile metazoans as sentinels of environmental health.

Assessing the environmental pesticides impact of river sediments from a basin in western Mexico: Spatiotemporal distribution, risk assessment of aquatic invertebrates and pesticides prioritization.

The intensive use of pesticides in Mexican agriculture has contributed significantly to the increase in food production, but at the same time represents potential risk to biota. This situation creates a dilemma between the need to increase food production and the preservation of the environment and human health. Aquatic invertebrates play a vital role in the balance of aquatic ecosystems but are sensitive to pesticides contamination. The sensitivity of aquatic invertebrates to pesticides contamination has led them to be used to assess the potential impact of this contamination on aquatic ecosystems. In the present study, conducted in the Ayuquila-Armería basin, the following aims were achieved: 1) quantifying the presence of 20 pesticides in river sediments, 2) assessing the spatiotemporal distribution of pesticides in river sediments, 3) determining the potential risk to aquatic invertebrates, and 4) prioritizing pesticides based on their potential risk. Twelve pesticides were consistently quantified in 192 river sediments samples. The pesticides with the highest concentrations were ametrine, malathion and picloram. The temporal analysis showed seasonality in pesticide concentrations, with higher detection frequencies during the wet season. The risk assessment showed that aquatic invertebrates may be affected by the concentrations of carbofuran, malathion, diazinon and ametrine. Pesticides prioritization identified ametrine, carbofuran, and diazinon as major concerns based on the methodology that considers the Frequency and Extent of Exceedance. This study provides valuable insights into the current pesticides scenario in the Ayuquila-Armería River sediments. The findings underscore the need for sustainable alternatives to mitigate the ecological risks associated with pesticides contamination in this aquatic ecosystem.

Proposal for a tiered regulatory framework for the aquatic risk assessment of pesticides in Brazil.

Aquatic risk assessment is essential to guarantee the sustainable use of pesticides and the conservation of water resources near agricultural fields. This article discusses a proposal for a tiered regulatory framework for the aquatic risk assessment of pesticides in Brazil. The first step is problem formulation, which includes establishing general and specific protection goals. In the exposure assessment, the Estimated Environmental Concentrations in water should be calculated based on realistic worst-case assumptions regarding application rate and frequency, the entry into the edge-of-field water body, and fate in the water body, using scenario-dependent models suggested by the Brazilian Environmental Agency. These calculations can be refined by including Efate studies with variable exposures to reflect realistic environmental conditions accurately and include mitigation measures that impact the modeling. In the hazard assessment, ecotoxicological data for toxicity to fish, aquatic invertebrates, algae, and aquatic plants should be required for all pesticides based on standardized protocols and species. Tier 2 has several refinement options, including incorporating toxicity data from additional test species and effect modeling. In Tier 3, population- and community-level effects are evaluated using semi-field studies. Considering the case study in Brazil, Tier 1 demonstrated that, from the 12 pesticides that were assessed, seven (58%) failed based on the value of the Risk Quotient. In Tier 2, when exposure refinement options and mitigation measures such as buffer zones are considered, all seven pesticides, for which Tier 1 indicated risk, still failed the assessment. The risk for four of these seven pesticides could be refined by considering toxicity information from additional species. Refinement options and mitigation measures that could be applied to the agricultural scenario in Brazil were discussed. In conclusion, the proposed tiered risk assessment is a feasible way to evaluate whether a pesticide will pose an unacceptable risk to aquatic organisms. Integr Environ Assess Manag 2024;20:1514-1528. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Eugenol as anesthetic for Cnesterodon decemmaculatus (Cyprinodontiformes, Poeciliidae).

Anesthetics are commonly used in fish for surgery and to facilitate capture, handling and transport in aquaculture and experimental procedures. In research, the selection of the anesthetic depends on its properties and on the recovery time. Eugenol has been pointed out as an effective anesthetic for fish, alternative to traditional drugs. Although Cnesterodon decemmaculatus is widely used as a model in ecological bioassays, no anesthetic protocol has been reported for this species. The aim of the present study was to evaluate the induction time (i.e. time to reach anesthetic stage VI) and recovery time in individuals of C. decemmaculatus subjected to eugenol at a fixed concentration, according to sex and pregnancy status. Forty-one fish were divided into three groups: males, pregnant females and non-pregnant females. They were measured for total length, standard length and weight, and the condition factor (K) was calculated. No significant differences in induction and recovery times were found for sex, pregnancy status and K between groups. Results are a contribution toward the development protocol of a standard anesthetic protocol for C. decemmaculatus.

Do bio-insecticides affect only insect species? Behavior, regeneration, and sexual reproduction of a non-target freshwater planarian.

Bio-insecticides have been increasingly used worldwide as ecofriendly alternatives to pesticides, but data on their effects in non-target freshwater organisms is still scarce and limited to insects. The aim of this study was to determine the lethal and sub-lethal effects of the bio-insecticides Bac Control (based on Bacillus thuringiensis kurstaki-Btk) and Boveril (based on Beauveria bassiana-Bb) on regeneration, behavioral, and reproductive endpoints of the freshwater planarian Girardia tigrina. The estimated LC50-48h were > 800 mg a.i./L for Btk and 60.74 mg a.i./L for Bb. In addition, exposure to Btk significantly decreased locomotion and feeding activities of planarians (lowest observed effect concentration (LOEC) of 12.5 mg a.i./L Btk) and fecundity rate (LOEC = 3.12 mg a.i./L Btk), whereas exposure to Bb significantly delayed regeneration (LOEC = 0.75 mg a.i./L Bb) and decreased fecundity rate (1.5 mg a.i./L Bb) of planarians. Thus, both bio-insecticides induced deleterious sub-lethal effects on a non-insect freshwater invertebrate species. However, only Bb-based formulation affected the survival, fecundity rate, and regeneration at concentrations below the maximum predicted environmental concentration (PEC = 247 mg/L). Thus, care should be taken when using such formulations as alternatives to chemical insecticides near aquatic ecosystems.

Pesticides detected in surface and groundwater from agroecosystems in the Pampas region of Argentina: occurrence and ecological risk assessment.

The objective of this study was to evaluate the occurrence of pesticides in surface and groundwater of agricultural areas of the Pampas region of Argentina and to develop an ecological risk assessment (ERA) of pesticides in freshwater ecosystems. Eight agricultural sites from south Santa Fe province, in the north of the Pampas region, were sampled seven times between 2016 and 2018. Pesticides were analysed by gas chromatography-mass spectrometry (GC/MS) and liquid chromatography-mass spectrometry (UPLC-LC/MS). Twenty compounds among herbicides, insecticides and fungicides in 84% and 79% of groundwater and surface water samples, respectively, were detected. Atrazine was the most ubiquitous pesticide, following by metolachlor, acetochlor and glyphosate, with maximum concentrations of 28, 24, 77 and 111 µg/L, respectively. An ERA was performed by employing the risk quotient (RQ) method. Atrazine, azoxystrobin, pirimiphos-methyl, acetochlor and epoxiconazole posed a high and very high risk for aquatic organisms (RQ > 1) and glyphosate, metolachlor and 2,4-D exhibited negligible to medium risk. The herbicides were the major contributors to risk. This study is the first contribution on the presence and concentration of pesticides in surface and groundwater from agricultural areas of south Santa Fe province, north Pampas region, Argentina, and a starting point for pesticide ecological risk assessment.

Ecotoxicity of losartan potassium in aquatic organisms of different trophic levels.

The intensive use of the antihypertensive losartan potassium (LOS) has culminated in its high occurrence in aquatic environments. However, insufficient studies had investigated its effects in non-target organisms. In this study, ecotoxicity of LOS was assessed in aquatic organisms from distinct trophic levels (Desmodesmus subspicatus, Daphnia magna, and Astyanax altiparanae). Genotoxicity was assessed by the comet assay in D. magna and A. altiparanae, and biochemical biomarkers for the fish. LOS was more toxic to D. subspicatus (EC50(72h) = 27.93 mg L-1) than D. magna (EC50 = 303.69 mg L-1). Subsequently, this drug showed to induce more DNA damage in D. magna than A. altiparanae, when exposed to 2.5 mg L-1. No significant stress responses were observed by the fish biomarkers, suggesting that higher trophic levels organisms are more tolerant to LOS toxicity. LOS showed relatively low toxic potential for a short period of exposure, but with different patterns of toxicity for the organisms from distinct trophic levels, contributing to further risk assessment of LOS.

Toxicological effects of leachate extracts from asphalt mixtures nanomodified under Daphnia magna and Landoltia punctata test organisms.

The incorporation of nanomaterials in binders in the paving area has been studied to improve the mechanical behavior of asphalt mixtures. However, asphalt mixture compounds are susceptible to leaching and deposition in the environment. In this context, this research aimed to investigate the toxic effect of two leachate extracts from asphalt mixtures nanomodified with 2% carbon nanotube and 3% organophilic nanoclay, compared to conventional mixture, using Daphnia magna and Landoltia punctata as test organisms. The nanomaterials were characterized to confirm morphology, stability and effective diameter. Extracts were chemically characterized using the Fourier-Transform Infrared Spectroscopy (FTIR) technique, which indicated presence of functional groups of the asphalt binder in greater intensity in the leachate from conventional mixture. Acute toxicity with D. magna indicated EC50,48h of 83.5 ± 6.2 mL/L for leachate extract from conventional mixture, 306.0 ± 87.6 mL/L for leachate extract from mixture with nanoclay and 464.8 ± 32.1 mL/L for leachate extract from mixture with carbon nanotube. No leachate caused significant chronic toxicity. As for L.punctata, concentrations that caused 50% growth inhibition were 127.5 mL/L for the leachate extract from mixture with carbon nanotube, 196.9 mL/L for the leachate extract from mixture with nanoclay and 205 mL/L for the leachate extract from conventional mixture. For these test organisms, there is no evidence of negative impacts directly associated with the use of the present nanomaterials in asphalt mixtures. The incorporation of these nanos may also reduce the acute toxicity of the mixtures.

A review of antiepileptic drugs: Part 1 occurrence, fate in aquatic environments and removal during different treatment technologies.

Antiepileptic drugs (AEDs) are the main treatment for people with epilepsy. However, in recent years, more and more people are using them for other indications such as: migraine, chronic neuropathic pain, and mood disorders. Consequently, the prescriptions and consumption of these drugs are increasing worldwide. In WWTPs, AEDs can resist degradation processes, such as photodegradation, chemical degradation and/or biodegradation. Until now, only constructed wetlands and photocatalysis have shown good removal rates of AEDs from wastewater. However, their effectiveness depends on the specific conditions used during the treatment. Since the consumption of AEDs has increased in the last decade and their degradation in WWTPs is poor, these drugs have been largely introduced into the environment through the discharge of municipal and/or hospital effluents. Once in the environment, AEDs are distributed in the water phase, as suspended particles or in the sediments, suggesting that these drugs have a high potential for groundwater contamination. In this first part of the AEDs review is designed to fill out the current knowledge gap about the occurrence, fate and removal of these drugs in the aquatic environment. This is a review that emphasizes the characteristics of AEDs as emerging contaminants.

Biofilms of Pseudomonas and Lysinibacillus Marine Strains on High-Density Polyethylene.

Environmental pollution by plastic debris is estimated on a scale of 100 million metric tons, a portion of which is fragmented into micro- and nanoplastics. These fragments are often colonized by bacterial species in marine environments, possibly contributing to the biodegradation of such materials. However, further investigations are necessary to determine the impact of abiotic polymer weathering on biofilm adhesion, as well as the specific biofilm formation strategies employed by marine isolates. Here, we evaluate deep-sea sediment bacterial isolates for biofilm adhesion, extracellular matrix production, and polymer degradation ability. Our study focuses on high-density polyethylene (HDPE) fragments for their high durability and environmental persistence, subjecting fragments to abiotic weathering prior to bacterial colonization. Marine isolates identified as Pseudomonas sp. and Lysinibacillus sp. exhibited decreasing biofilm formation on weathered HDPE, especially over the first 24 h of incubation. This effect was countered by increased extracellular matrix production, likely improving cell adhesion to surfaces roughened by abiotic degradation. These adhesion strategies were contrasted with a reference Pseudomonas aeruginosa strain, which displayed high levels of biofilm formation on non-weathered HDPE and lower extracellular matrix production over the first 24 h of incubation. Furthermore, our results suggest that an increase in biofilm biomass correlated with changes to HDPE structure, indicating that these strains have a potential for biodegradation of plastic fragments. Therefore, this work provides a detailed account of biofilm formation strategies and bacteria-plastic interactions that represent crucial steps in the biodegradation of plastic fragments in marine environments.

Vaquitas (Phocoena sinus) continue to die from bycatch not pollutants.

BACKGROUND: The vaquita of Mexico is critically endangered, with a population less than 19 individuals in 2018. The population continues to decline and gillnet use continues in vaquita habitat. METHODS: Nine vaquita carcases were examined from 2016 to 2018 to establish cause of death. Concentrations of polychlorinated biphenyls (PCBs), DDTs and polybrominated diphenyl ethers (PBDEs) in blubber (n=3) were determined using gas chromatography/mass spectrometry and faeces tested for domoic acid and saxitoxin. RESULTS: Carcases were in good nutritional status and had lesions and full stomachs consistent with fisheries bycatch. PCB, DDT and PBDE concentrations ranged between 94 and 180 ng/g, 500 and 1200 ng/g and 97 and 210 ng/g lipid weight, respectively, which are low compared with other marine mammals. No saxitoxin or domoic acid was detected. CONCLUSION: These findings support the conclusion that bycatch is the primary source of vaquita population decline and emphasise the need for stronger measures to eliminate gillnets from the Upper Gulf of California, Mexico.

Fish exposed to water contaminated with eprinomectin show inhibition of the activities of AChE and Na+/K+-ATPase in the brain, and changes in natural behavior.

The objective of this study was to evaluate whether antiparasitic eprinomectin may be an environmental contaminant in water compartment in low concentrations, negatively affecting neurotransmission and, consequently, the natural behavior of the jundiá (Rhamdia quelen). Fish were randomly allocated in tanks and exposed for 24 and 48 h to eprinomectin concentrations in water [0.0 (Control), 1.124 (T1), 1.809 (T2) and 3.976 (T3) µg L-1], followed by 48 h of recovery in eprinomectin-free water, in order to evaluate the behavioral parameters, levels of reactive oxygen species (ROS) in the brain, as well as cerebral enzymatic activities of acetylcholinesterase (AChE) and of the sodium-potassium ATPase pump (Na+/K+-ATPase). Especially at the two highest concentrations of eprinomectin (T2 and T3), the fish showed alterations in natural behavior, particularly hyperlocomotion and longer time on the surface. Furthermore, at these same concentrations, cerebral ROS levels increased and cerebral AChE activity decreased. At the highest concentration (T3) cerebral Na+/K+-ATPase activity was reduced. Increased ROS and impairment of AChE and Na+/K+-ATPase enzymes in the brain may have contributed directly to behavioral changes, due to neuronal damage and synapse impairment. Even after 48 h in water without eprinomectin, behavioral changes and neurotoxic effects were observed in fish, suggesting residual effects of the antiparasitic. In conclusion, eprinomectin even in low concentrations may be a hazardous environmental contaminant for aquatic organisms, as it causes brain damage and affects the natural behavior of fish.

Atividade antimicrobiana e antibiofilme de nanopartículas de prata sobre isolados de Aeromonas spp. obtidos de organismos aquáticos / Antimicrobial and antibiofilm activity of silver nanoparticles against Aeromonas spp. isolated from aquatic organisms

O uso indiscriminado de antimicrobianos tem proporcionado a algumas bactérias patogênicas a seleção de cepas multirresistentes, situação que pode ser agravada pela formação do biofilme. Desta forma, as nanopartículas de prata (AgNPs) vêm se destacando como uma alternativa inovadora, de baixo custo e eficiente contra doenças causadas por bactérias. O objetivo deste estudo foi determinar a atividade antimicrobiana das AgNPs e a interferência na formação do biofilme de Aeromonas spp. obtidas de organismos aquáticos. As AgNPs foram sintetizadas quimicamente utilizando como agente redutor o citrato trissódico e caracterizadas por espectrofotometria ultravioleta-visível (UV-Vis). A atividade antimicrobiana foi realizada contra três isolados pelo método de microdiluição em caldo para determinar a concentração bactericida mínima (CBM) e um cultivo com CCCP, um inibidor da bomba de efluxo, foi realizado para complementar o efeito das AgNPs. A interferência no biofilme foi realizada segundo o protocolo de formação e consolidado, além da caracterização desta estrutura de resistência por microscopia eletrônica de varredura. No teste da CBM, as AgNPs não foram capazes de inativar o crescimento dos isolados, ao passo que o nitrato de prata obteve eficiência em diferentes concentrações. Na presença do inibidor de bomba de efluxo, dos isolados analisados, um passou de resistente a sensível na presença das nanopartículas. As AgNPs foram eficazes em diminuir a formação de biofilme, como também atuaram sobre o biofilme consolidado em todos os isolados testados. Estes resultados indicam o potencial das nanopartículas de prata em interferir com o biofilme de Aeromonas spp. de organismos aquáticos e seres humanos.(AU)

The indiscriminate use of antibiotics has selected some pathogenic bacteria being multidrug-resistant, a situation that can be exacerbated by biofilms formation. Thus, silver nanoparticles (AgNPs) have been highlighted as an innovative alternative, low-cost and effective against bacterial diseases. The aim of this study was to determine the antimicrobial activity of AgNPs and the interference in Aeromonas spp. biofilm formation. The strains were obtained from aquatic organisms. The AgNPs were chemically synthesized using as reducing agent trisodium citrate and characterized by ultraviolet-visible spectroscopy (UV-Vis). The antimicrobial activity was carried out against three isolates by the microdilution broth method for determining minimum bactericidal concentration (CBM) and cultivation of CCCP, an inhibitor of the efflux pump, was carried out to complement the effect of AgNPs. Interference in the biofilm formation was performed according to the protocol and consolidated, within the resistance structure characterization by scanning electron microscopy. In the test of the CBM, the AgNPs were unable to inactivate the growth of the isolates, while the silver nitrate obtained efficiency in different concentrations. In the efflux pump inhibitor presence the isolates were analyzed, one went from resistant to nanoparticles to sensitive. The AgNPs were effective in reducing of biofilm formation and acted on the consolidated biofilm in all tested isolates. These results indicate the silver nanoparticles to interfere with Aeromonas spp. biofilm from aquatic organisms and human bodies.(AU)

Atividade antimicrobiana e antibiofilme de nanopartículas de prata sobre isolados de Aeromonas spp. obtidos de organismos aquáticos / Antimicrobial and antibiofilm activity of silver nanoparticles against Aeromonas spp. isolated from aquatic organisms

O uso indiscriminado de antimicrobianos tem proporcionado a algumas bactérias patogênicas a seleção de cepas multirresistentes, situação que pode ser agravada pela formação do biofilme. Desta forma, as nanopartículas de prata (AgNPs) vêm se destacando como uma alternativa inovadora, de baixo custo e eficiente contra doenças causadas por bactérias. O objetivo deste estudo foi determinar a atividade antimicrobiana das AgNPs e a interferência na formação do biofilme de Aeromonas spp. obtidas de organismos aquáticos. As AgNPs foram sintetizadas quimicamente utilizando como agente redutor o citrato trissódico e caracterizadas por espectrofotometria ultravioleta-visível (UV-Vis). A atividade antimicrobiana foi realizada contra três isolados pelo método de microdiluição em caldo para determinar a concentração bactericida mínima (CBM) e um cultivo com CCCP, um inibidor da bomba de efluxo, foi realizado para complementar o efeito das AgNPs. A interferência no biofilme foi realizada segundo o protocolo de formação e consolidado, além da caracterização desta estrutura de resistência por microscopia eletrônica de varredura. No teste da CBM, as AgNPs não foram capazes de inativar o crescimento dos isolados, ao passo que o nitrato de prata obteve eficiência em diferentes concentrações. Na presença do inibidor de bomba de efluxo, dos isolados analisados, um passou de resistente a sensível na presença das nanopartículas. As AgNPs foram eficazes em diminuir a formação de biofilme, como também atuaram sobre o biofilme consolidado em todos os isolados testados. Estes resultados indicam o potencial das nanopartículas de prata em interferir com o biofilme de Aeromonas spp. de organismos aquáticos e seres humanos.(AU)

The indiscriminate use of antibiotics has selected some pathogenic bacteria being multidrug-resistant, a situation that can be exacerbated by biofilms formation. Thus, silver nanoparticles (AgNPs) have been highlighted as an innovative alternative, low-cost and effective against bacterial diseases. The aim of this study was to determine the antimicrobial activity of AgNPs and the interference in Aeromonas spp. biofilm formation. The strains were obtained from aquatic organisms. The AgNPs were chemically synthesized using as reducing agent trisodium citrate and characterized by ultraviolet-visible spectroscopy (UV-Vis). The antimicrobial activity was carried out against three isolates by the microdilution broth method for determining minimum bactericidal concentration (CBM) and cultivation of CCCP, an inhibitor of the efflux pump, was carried out to complement the effect of AgNPs. Interference in the biofilm formation was performed according to the protocol and consolidated, within the resistance structure characterization by scanning electron microscopy. In the test of the CBM, the AgNPs were unable to inactivate the growth of the isolates, while the silver nitrate obtained efficiency in different concentrations. In the efflux pump inhibitor presence the isolates were analyzed, one went from resistant to nanoparticles to sensitive. The AgNPs were effective in reducing of biofilm formation and acted on the consolidated biofilm in all tested isolates. These results indicate the silver nanoparticles to interfere with Aeromonas spp. biofilm from aquatic organisms and human bodies.(AU)

Bioaccumulation and trophic transfer of metals, As and Se through a freshwater food web affected by antrophic pollution in Córdoba, Argentina.

The concentration of metals (Al, Cr, Mn, Fe, Ni, Cu, Zn, Ag, Cd, Hg, Pb, U), As and Se in different ecosystem components (water, sediment, plankton, shrimp, and fish muscle) has been determined in a eutrophic reservoir in the Province of Córdoba (Argentina). Los Molinos Lake (LML) was sampled during the dry (DS) and wet seasons (WS) in order to examine the bioaccumulation and transfer of these inorganic elements through the food web. Stable nitrogen isotope (δ15N) was used to investigate trophic interactions. According to this, samples were divided into three categories: plankton, shrimp (Palaemonetes argentinus) and fish (Silverside, Odontesthes bonariensis). The bioaccumulation factor (BAF) was calculated for the organisms, and it was determined that the elements analyzed undergo bioaccumulation, especially in organisms such as plankton. The invertebrates were characterized by the highest BAF for Cu and Zn in both seasons, As (DS), and Cd and Hg (WS). The fish muscle was characterized by the highest BAF for Se (WS), Ag and Hg (DS). On the other hand, a significant decrease in Al, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Cd and U concentrations through the analyzed trophic web during both seasons was observed. Moreover, a significant increase in Hg levels was observed with increasing trophic levels in the DS, indicating its biomagnification. Despite the increasing impact of metals, As and Se pollution in the studied area due to urban growth and agricultural and livestock activities, no previous study has focused on the behavior and relationships of these pollutants with the biotic and abiotic components of this aquatic reservoir. We expect that these findings may be used for providing directions or guidance for future monitoring and environmental protection policies.

Gene floR e a resistência ao florfenicol em isolados de Aeromonas spp. autóctones de organismos aquáticos / Gene floR and resistance to florfenicol in isolated Aeromonas spp. indigenous aquatic organisms

O gene floR descrito é descrito pela literatura como o responsável pela resistência ao florfenicol, que é um antimicrobiano amplamente utilizado na aquicultura. Esse gene já foi relatado em muitas espécies de bactérias, inclusive no gênero Aeromonas. Essas bactérias causam alta mortalidade na piscicultura trazendo prejuízos econômicos. É importante que haja estudos sobre esse gene e possíveis mutações que possam levar a alterações na estrutura e função da proteína. Os objetivos desse estudo foram caracterizar o gene floR em isolados de Aeromonas spp. obtidas do Vale do São Francisco e verificar se a presença desse gene está associada com a resistência ao florfenicol. Foram realizadas reações em cadeia da polimerase (PCR) para a presença do gene floR em 27 isolados de Aeromonas spp.. Amostras positivas para a presença do gene foram sequenciadas e analisadas quanto à presença de polimorfismos por meio de alinhamentos. Os diferentes haplótipos detectados foram utilizados para análises com os programas SIFT e PolyPhen para predição de alteração de função proteica. A modelagem estrutural da proteina codificada pelo gene floR foi realizada com o programa Modeller e, os modelos foram avaliados pelo Procheck, Verify3D e Whatif. A similaridade da estrutura tridimensional da proteína referência com as estruturas tridimensionais das proteínas codificadas pelos diferentes haplótipos foi comparada através do TM-align. A resistência das bactérias ao florfenicol foi avaliada através do teste de microdiluição em caldo, o qual também foi realizado na presença do carbonil cianeto m-clorofenil hidrazona para verificar o efeito da inibição da bomba de efluxo sobre tal resistência. Dos vinte e sete isolados avaliados quanto a presença do gene floR, 14 isolados foram positivos e 10 foram sequenciados, o que permitiu a identificação de três polimorfismos no gene floR, que levaram a construção de três haplótipos diferentes (TAA, TTA e CTG)...(AU)

The floR gene is described in related literature as responsible for resistance to florfenicol, which is a widely used antimicrobial agent in aquaculture. This gene has been reported in many species of bacteria, including the genus Aeromonas. These bacteria cause high mortality in fish farming bringing economic losses. It is important that studies of this gene and possible mutations that can lead to changes in the structure and function of the protein. The aim of this study was to characterize the floR gene in isolates of Aeromonas spp. and check if the presence of this gene is associated with resistance to florfenicol in Aeromonas spp. obtained from the San Francisco Valley. PCR (Polymerase Chain Reaction) were also performed to verify the presence of the floR gene in 27 isolates of Aeromonas spp. Positive samples for the presence of the gene were sequenced and analyzed for the presence of polymorphisms using alignments. Different haplotypes detected were used for analysis with the SIFT and PolyPhen programs for prediction of changes in protein function. The structural modeling of protein encoded by the floR gene was performed using the Modeller software, and the models were evaluated by Procheck, Verify3D and Whatif. The similarity of the dimensional structure of reference protein with the dimensional structures of the proteins encoded by the different haplotypes was compared by TM-align. Bacterial resistance to florfenicol was assessed by the microdilution test, which was also performed in the presence of carbonyl cyanide m-chlorophenyl hydrazone to verify the effect of inhibiting the efflux pump. 14 isolates were positive for the presence of floR gene and 10 were sequenced and allowed the identification of three polymorphisms in the floR gene, which led to construction of three different haplotypes (TAA TTA and CTG). The analyzes carried out with the SIFT and PolyPhen...(AU)

Atividade antimicrobiana e antibiofilme de nanopartículas de prata sobre isolados de Aeromonas spp. obtidos de organismos aquáticos

ABSTRACT: The indiscriminate use of antibiotics has selected some pathogenic bacteria being multidrug-resistant, a situation that can be exacerbated by biofilms formation. Thus, silver nanoparticles (AgNPs) have been highlighted as an innovative alternative, low-cost and effective against bacterial diseases. The aim of this study was to determine the antimicrobial activity of AgNPs and the interference in Aeromonas spp. biofilm formation. The strains were obtained from aquatic organisms. The AgNPs were chemically synthesized using as reducing agent trisodium citrate and characterized by ultraviolet-visible spectroscopy (UV-Vis). The antimicrobial activity was carried out against three isolates by the microdilution broth method for determining minimum bactericidal concentration (CBM) and cultivation of CCCP, an inhibitor of the efflux pump, was carried out to complement the effect of AgNPs. Interference in the biofilm formation was performed according to the protocol and consolidated, within the resistance structure characterization by scanning electron microscopy. In the test of the CBM, the AgNPs were unable to inactivate the growth of the isolates, while the silver nitrate obtained efficiency in different concentrations. In the efflux pump inhibitor presence the isolates were analyzed, one went from resistant to nanoparticles to sensitive. The AgNPs were effective in reducing of biofilm formation and acted on the consolidated biofilm in all tested isolates. These results indicate the silver nanoparticles to interfere with Aeromonas spp. biofilm from aquatic organisms and human bodies.

RESUMO: O uso indiscriminado de antimicrobianos tem proporcionado a algumas bactérias patogênicas a seleção de cepas multirresistentes, situação que pode ser agravada pela formação do biofilme. Desta forma, as nanopartículas de prata (AgNPs) vêm se destacando como uma alternativa inovadora, de baixo custo e eficiente contra doenças causadas por bactérias. O objetivo deste estudo foi determinar a atividade antimicrobiana das AgNPs e a interferência na formação do biofilme de Aeromonas spp. obtidas de organismos aquáticos. As AgNPs foram sintetizadas quimicamente utilizando como agente redutor o citrato trissódico e caracterizadas por espectrofotometria ultravioleta-visível (UV-Vis). A atividade antimicrobiana foi realizada contra três isolados pelo método de microdiluição em caldo para determinar a concentração bactericida mínima (CBM) e um cultivo com CCCP, um inibidor da bomba de efluxo, foi realizado para complementar o efeito das AgNPs. A interferência no biofilme foi realizada segundo o protocolo de formação e consolidado, além da caracterização desta estrutura de resistência por microscopia eletrônica de varredura. No teste da CBM, as AgNPs não foram capazes de inativar o crescimento dos isolados, ao passo que o nitrato de prata obteve eficiência em diferentes concentrações. Na presença do inibidor de bomba de efluxo, dos isolados analisados, um passou de resistente a sensível na presença das nanopartículas. As AgNPs foram eficazes em diminuir a formação de biofilme, como também atuaram sobre o biofilme consolidado em todos os isolados testados. Estes resultados indicam o potencial das nanopartículas de prata em interferir com o biofilme de Aeromonas spp. de organismos aquáticos e seres humanos.

Gene floR e a resistência ao florfenicol em isolados de Aeromonas spp. autóctones de organismos aquáticos / Gene floR and resistance to florfenicol in isolated Aeromonas spp. indigenous aquatic organisms

O gene floR descrito é descrito pela literatura como o responsável pela resistência ao florfenicol, que é um antimicrobiano amplamente utilizado na aquicultura. Esse gene já foi relatado em muitas espécies de bactérias, inclusive no gênero Aeromonas. Essas bactérias causam alta mortalidade na piscicultura trazendo prejuízos econômicos. É importante que haja estudos sobre esse gene e possíveis mutações que possam levar a alterações na estrutura e função da proteína. Os objetivos desse estudo foram caracterizar o gene floR em isolados de Aeromonas spp. obtidas do Vale do São Francisco e verificar se a presença desse gene está associada com a resistência ao florfenicol. Foram realizadas reações em cadeia da polimerase (PCR) para a presença do gene floR em 27 isolados de Aeromonas spp.. Amostras positivas para a presença do gene foram sequenciadas e analisadas quanto à presença de polimorfismos por meio de alinhamentos. Os diferentes haplótipos detectados foram utilizados para análises com os programas SIFT e PolyPhen para predição de alteração de função proteica. A modelagem estrutural da proteina codificada pelo gene floR foi realizada com o programa Modeller e, os modelos foram avaliados pelo Procheck, Verify3D e Whatif. A similaridade da estrutura tridimensional da proteína referência com as estruturas tridimensionais das proteínas codificadas pelos diferentes haplótipos foi comparada através do TM-align. A resistência das bactérias ao florfenicol foi avaliada através do teste de microdiluição em caldo, o qual também foi realizado na presença do carbonil cianeto m-clorofenil hidrazona para verificar o efeito da inibição da bomba de efluxo sobre tal resistência. Dos vinte e sete isolados avaliados quanto a presença do gene floR, 14 isolados foram positivos e 10 foram sequenciados, o que permitiu a identificação de três polimorfismos no gene floR, que levaram a construção de três haplótipos diferentes (TAA, TTA e CTG). As análises realizadas com os programas SIFT e PolyPhen apontaram que os haplótipos TTA e TAA provavelmente poderiam alterar a estrutura e função da proteína. As proteínas modeladas para os três haplótipos demonstraram possuir praticamente a mesma conformação estrutural entre si. Todos os isolados que apresentaram o gene foram resistentes ao florfenicol e aqueles que não apresentavam foram sensíveis. O teste na presença do Carbonil Cianeto m-Clorofenil Hidrazona foi realizado para três isolados, cada isolado representando um haplótipo, sendo possível observar a inibição do crescimento bacteriano em todas as concentrações independente do haplótipo. Os resultados obtidos nesse estudo mostram que a resistência ao flofenicol em Aeromonas spp. pode ser explicada pela presença do gene floR, e que esse gene está relacionado com uma bomba de efluxo. As mutações verificadas no gene floR, parecem não estar envolvidas com alteração de estrutura e função da proteína codificada por esse gene.(AU)

The floR gene is described in related literature as responsible for resistance to florfenicol, which is a widely used antimicrobial agent in aquaculture. This gene has been reported in many species of bacteria, including the genus Aeromonas. These bacteria cause high mortality in fish farming bringing economic losses. It is important that studies of this gene and possible mutations that can lead to changes in the structure and function of the protein. The aim of this study was to characterize the floR gene in isolates of Aeromonas spp. and check if the presence of this gene is associated with resistance to florfenicol in Aeromonas spp. obtained from the San Francisco Valley. PCR (Polymerase Chain Reaction) were also performed to verify the presence of the floR gene in 27 isolates of Aeromonas spp. Positive samples for the presence of the gene were sequenced and analyzed for the presence of polymorphisms using alignments. Different haplotypes detected were used for analysis with the SIFT and PolyPhen programs for prediction of changes in protein function. The structural modeling of protein encoded by the floR gene was performed using the Modeller software, and the models were evaluated by Procheck, Verify3D and Whatif. The similarity of the dimensional structure of reference protein with the dimensional structures of the proteins encoded by the different haplotypes was compared by TM-align. Bacterial resistance to florfenicol was assessed by the microdilution test, which was also performed in the presence of carbonyl cyanide m-chlorophenyl hydrazone to verify the effect of inhibiting the efflux pump. 14 isolates were positive for the presence of floR gene and 10 were sequenced and allowed the identification of three polymorphisms in the floR gene, which led to construction of three different haplotypes (TAA TTA and CTG). The analyzes carried out with the SIFT and PolyPhen programs showed that the TTA and TAA haplotypes could probably change the protein structure-function. Proteins modeled for the three haplotypes were found to have substantially the same structural conformation with each other. All isolates presenting the gene were resistant to florfenicol and those who did not have were sensitive. The test in the presence of carbonyl cyanide m-chlorophenylhydrazone was conducted for three isolates, representing each single haplotype and was observed inhibition of bacterial growth at all concentrations independent of the haplotype. The results of this study show that resistance to flofenicol in Aeromonas spp. may be explained by the presence of floR gene and that this gene is associated with an efflux pump. Mutations observed in floR gene do not appear to be involved with chenges in structure and function of the protein encoded by gene.(AU)

Aquatic ecotoxicity of ashes from Brazilian savanna wildfires.

In a global scenario of climate change, several studies have predicted an increase in fires in different parts of the world. With the occurrence of rains following the fires in the Brazilian savanna (Cerrado biome), the compounds present in ashes may enter aquatic environments and cause adverse effects to these ecosystems. In this context, this study evaluated the potential toxicity of ashes from two areas of Cerrado and an area of pasture, through ecotoxicological bioassays and using three aquatic species from distinct trophic levels, which were exposed to different dilutions of ashes: the microcrustacean Ceriodaphnia dubia, the fish Danio rerio and the mollusc Biomphalaria glabrata. The ashes from the three sampled areas showed higher concentrations of some elements in relation to the soil samples (B, Ca, K, Mg, Mn, P, S, Si, Sr, Zn), but only a small quantity of these compounds was solubilised. Our data showed that all ash samples caused acute toxicity to C. dubia (48hs-LC50 = 13.4 g L-1; 48hs-LC50 = 6.33 g L-1; 48hs-LC50 = 9.73 g L-1 respectively for transition area, pasture, typical cerrado areas), while in relation to D. rerio and B. glabrata, no acute toxicity was observed when they were exposed to ashes from native Cerrado vegetation and pasture areas. Ashes from a transition area showed toxicity for D. rerio (48hs-LC50 = 25.0 g L-1); possibly, this was due to the combination of multiple preponderant inorganic elements of ashes with other organic compounds not analysed, such as polycyclic aromatic hydrocarbons (PAHs). In summary, these results suggest that wildfires may pose risks to zooplankton communities and emphasize the need for more studies to better understand the complexity of the ecological effects of fire on aquatic ecosystems.