Underground speciation: Unraveling the systematics and evolution of the highly diverse tuco-tucos (genus Ctenomys) with genomic data.

Subterranean rodents of the genus Ctenomys (tuco-tucos) are endemic to South America and have experienced relatively recent radiation. There are about 67 recognized species that originated in approximately 1-2 MY. They stand out for their species richness, extraordinary chromosomal diversity, and wide range of habitat they occupy in the continent. Phylogenetic relationships among species of tuco-tucos have been challenging to resolve. Groups of closely-related species have been suggested, but their relationships must be resolved. This study estimates the phylogeny of the genus using massive sequencing, generating thousands of independent molecular markers obtained by RADseq, with a taxonomic sampling that includes 66% of the recognized species. The sequences obtained were mapped against the C. sociabilis genome, recovering up to 1,215 widely shared RAD loci with more than 19,000 polymorphic sites. Our new phylogenetic hypothesis corroborated the species groups previously proposed with cytochrome b gene sequences and provided a much greater resolution of the relationships among species groups. The frater group is sister to all other tuco-tucos, whereas some of the earlierliest proposals placed the sociabilis group as sister to all other tuco-tucos. Ctenomys leucodon, previously proposed as an independent lineage, is associated with the frater group with moderate statistical support. The magellanicus and mendocinus are sister groups in a major clade formed by the boliviensis, talarum, tucumanus, torquatus, and opimus groups. Ctenomys viperinus, included in the phylogeny for the first time, belongs to the tucumanus group. This multi-locus phylogenetic hypothesis provides insights into the historical biogeography of understanding this highly diverse genus.

Ultraconserved Elements Improve the Resolution of Difficult Nodes within the Rapid Radiation of Neotropical Sigmodontine Rodents (Cricetidae: Sigmodontinae).

Sigmodontine rodents (Cricetidae, Sigmodontinae) represent the second largest muroid subfamily and the most species-rich group of New World mammals, encompassing above 410 living species and ca. 87 genera. Even with advances on the clarification of sigmodontine phylogenetic relationships that have been made recently, the phylogenetic relationships among the 12 main groups of genera (i.e., tribes) remain poorly resolved, in particular among those forming the large clade Oryzomyalia. This pattern has been interpreted as consequence of a rapid radiation upon the group entrance into South America. Here, we attempted to resolve phylogenetic relationships within Sigmodontinae using target capture and high-throughput sequencing of ultraconserved elements (UCEs). We enriched and sequenced UCEs for 56 individuals and collected data from four already available genomes. Analyses of distinct data sets, based on the capture of 4634 loci, resulted in a highly resolved phylogeny consistent across different methods. Coalescent species-tree-based approaches, concatenated matrices, and Bayesian analyses recovered similar topologies that were congruent at the resolution of difficult nodes. We recovered good support for the intertribal relationships within Oryzomyalia; for instance, the tribe Oryzomyini appears as the sister taxa of the remaining oryzomyalid tribes. The estimates of divergence times agree with the results of previous studies. We inferred the crown age of the sigmodontine rodents at the end of the Middle Miocene, while the main lineages of Oryzomyalia appear to have radiated in a short interval during the Late Miocene. Thus, the collection of a genomic-scale data set with a wide taxonomic sampling provided resolution for the first time of the relationships among the main lineages of Sigmodontinae. We expect the phylogeny presented here will become the backbone for future systematic and evolutionary studies of the group.[Coalescent; Muroidea; Oryzomyalia; phylogenomics; polytomy; Rodentia; Sigmodontalia; species tree; UCEs.].

Genomic and Transcriptomic Basis of Hanseniaspora vineae's Impact on Flavor Diversity and Wine Quality.

Hanseniaspora is the main genus of the apiculate yeast group that represents approximately 70% of the grape-associated microflora. Hanseniaspora vineae is emerging as a promising species for quality wine production compared to other non-Saccharomyces species. Wines produced by H. vineae with Saccharomyces cerevisiae consistently exhibit more intense fruity flavors and complexity than wines produced by S. cerevisiae alone. In this work, genome sequencing, assembling, and phylogenetic analysis of two strains of H. vineae showed that it is a member of the Saccharomyces complex and it diverged before the whole-genome duplication (WGD) event from this clade. Specific flavor gene duplications and absences were identified in the H. vineae genome compared to 14 fully sequenced industrial S. cerevisiae genomes. The increased formation of 2-phenylethyl acetate and phenylpropanoids such as 2-phenylethyl and benzyl alcohols might be explained by gene duplications of H. vineae aromatic amino acid aminotransferases (ARO8 and ARO9) and phenylpyruvate decarboxylases (ARO10). Transcriptome and aroma profiles under fermentation conditions confirmed these genes were highly expressed at the beginning of stationary phase coupled to the production of their related compounds. The extremely high level of acetate esters produced by H. vineae compared to that by S. cerevisiae is consistent with the identification of six novel proteins with alcohol acetyltransferase (AATase) domains. The absence of the branched-chain amino acid transaminases (BAT2) and acyl coenzyme A (acyl-CoA)/ethanol O-acyltransferases (EEB1) genes correlates with H. vineae's reduced production of branched-chain higher alcohols, fatty acids, and ethyl esters, respectively. Our study provides sustenance for understanding and potentially utilizing genes that determine fermentation aromas.IMPORTANCE The huge diversity of non-Saccharomyces yeasts in grapes is dominated by the apiculate genus Hanseniaspora Two native strains of Hanseniaspora vineae applied to winemaking because of their high oenological potential in aroma and fermentation performance were selected to obtain high-quality genomes. Here, we present a phylogenetic analysis and the complete transcriptome and aroma metabolome of H. vineae during three fermentation steps. This species produced significantly richer flavor compound diversity than Saccharomyces, including benzenoids, phenylpropanoids, and acetate-derived compounds. The identification of six proteins, different from S. cerevisiae ATF, with diverse acetyltransferase domains in H. vineae offers a relevant source of native genetic variants for this enzymatic activity. The discovery of benzenoid synthesis capacity in H. vineae provides a new eukaryotic model to dilucidate an alternative pathway to that catalyzed by plants' phenylalanine lyases.

[Interaction between antihypertensive therapy and exercise training therapy requires drug regulation in hypertensive patients]. / Interacción entre farmacoterapia hipotensiva y terapia con ejercicio físico requiere regulación farmacológica en pacientes hipertensos.

BACKGROUND: Exercise training could interact with the pharmacological therapy of hypertension, increasing the effects of these medications. AIM: To assess the effects of 12 weeks of physical training on blood pressure of hypertensive or diabetic patients, already receiving pharmacological therapy. MATERIAL AND METHODS: Twelve participants with diabetes and hypertension, 16 participants with hypertension and 18 healthy participants were studied. During six weeks no intervention was carried out, except the treatment for their underlying conditions. During the ensuing six weeks, participants were subjected 12 sessions of physical training. Blood pressure was measured at baseline and after six and twelve weeks of intervention. RESULTS: During the period without physical training, no changes in blood pressure were observed. After the physical training intervention systolic blood pressure decreased by 16, 17 and 20 mm Hg in participants with diabetes and hypertension, participants with hypertension and healthy participants, respectively. Diastolic blood pressure decreased by 9 and 6 mmHg in participants with diabetes and hypertension and hypertension alone, respectively. CONCLUSIONS: There is a decrease in blood pressure among hypertensive participants receiving pharmacological therapy, after a period of physical training.

The influence of ecological and geographical context in the radiation of Neotropical sigmodontine rodents.

BACKGROUND: Much debate has focused on how transitions in life history have influenced the proliferation of some clades. Rodents of the subfamily Sigmodontinae (family Cricetidae) comprise one of the most diverse clades of Neotropical mammals (~400 living species in 86 genera). These rodents occupy a wide range of habitats and lifestyles so that ecological context seems relevant to understand the evolution of this group. Several changes in the landscape of South America through the Neogene might have provided vast resources and opportunity to diversify. The aim of this study was to examine whether transitions between i) lowland and montane habitats, ii) open vegetation and forest, and iii) distinct molar architectures are correlated with shifts in diversification rates and to characterize the general pattern of diversification. RESULTS: Based on a dense taxon sampling of 269 species, we recovered a new phylogeny of Sigmodontinae that is topologically consistent with those of previous studies. It indicates that the subfamily and its major lineages appeared during the Late Miocene. Analyses suggest that vegetation type and elevational range are correlated with diversification rates, but not molar architecture. Tropical lowlands accumulated more lineage diversity than other areas and also supported high speciation rates. Across the radiation the subfamily Sigmodontinae appear to have experienced a decline in diversification rate through time. We detected mixed evidence for lineage-specific diversification rate shifts (e.g., leading to the clades of Akodon, Bibimys, Calomys and Thomasomys). CONCLUSION: We report that the evolution of habitat preference (considering vegetation type and elevational range) was associated with diversification rates among sigmodontine rodents. We propose that the observed diversification slowdown might be the result of ecological or geographical constraints. Our results also highlight the influence of the tropical lowlands -which might have acted as both "a cradle and a museum of species." The tropical lowlands accumulated greater diversity than the remainder of the group's range.

Dating an impressive Neotropical radiation: Molecular time estimates for the Sigmodontinae (Rodentia) provide insights into its historical biogeography.

With about 400 living species and 82 genera, rodents of the subfamily Sigmodontinae comprise one of the most diverse and more broadly distributed Neotropical mammalian clades. There has been much debate on the origin of the lineage or the lineages of sigmodontines that entered South America, the timing of entrance and different aspects of further diversification within South America. The ages of divergence of the main lineages and the crown age of the subfamily were estimated by using sequences of the interphotoreceptor retinoid binding protein and cytochrome b genes for a dense sigmodontine and muroid sampling. Bayesian inference using three fossil calibration points and a relaxed molecular clock estimated a middle Miocene origin for Sigmodontinae (∼12Ma), with most tribes diversifying throughout the Late Miocene (6.9-9.4Ma). These estimates together results of analyses of ancestral area reconstructions suggest a distribution for the most recent common ancestor of Sigmodontinae in Central-South America and a South American distribution for the most recent common ancestor of Oryzomyalia.

Determination of Microcystins in Fish Tissue by ELISA and MALDI-TOF MS Using a Highly Specific Single Domain Antibody.

The development of simple, reliable, and cost-effective methods is critically important to study the spatial and temporal variation of microcystins (MCs) in the food chain. Nanobodies (Nbs), antigen binding fragments from camelid antibodies, present valuable features for analytical applications. Their small antigen binding site offers a focused recognition of small analytes, reducing spurious cross-reactivity and matrix effects. A high affinity and broad cross-reactivity anti-MCs-Nb, from a llama antibody library, was validated in enzyme linked immunosorbent assay (ELISA), and bound to magnetic particles with an internal standard for pre-concentration in quantitative-matrix-assisted laser desorption ionization-time of flight mass spectrometry (Nb-QMALDI MS). Both methods are easy and fast; ELISA provides a global result, while Nb-QMALDI MS allows for the quantification of individual congeners and showed excellent performance in the fish muscle extracts. The ELISA assay range was 1.8-29 ng/g and for Nb-QMALDI, it was 0.29-29 ng/g fish ww. Fifty-five fish from a MC-containing dam were analyzed by both methods. The correlation ELISA/sum of the MC congeners by Nb-QMALDI-MS was very high (r Spearman = 0.9645, p < 0.0001). Using ROC curves, ELISA cut-off limits were defined to accurately predict the sum of MCs by Nb-QMALDI-MS (100% sensitivity; ≥89% specificity). Both methods were shown to be simple and efficient for screening MCs in fish muscle to prioritize samples for confirmatory methods.

Multicompartmental monitoring of legacy and currently used pesticides in a subtropical lake used as a drinking water source (Laguna del Cisne, Uruguay).

A pilot annual monitoring survey (April 2018-March 2019) was conducted to investigate the presence of pesticides in superficial water and fish in Laguna del Cisne, one of the most critical drinking water sources in Uruguay. A total of 25 pesticide residues were detected in superficial water (89.3 % of the samples). Pesticide's temporal distribution was associated with crops and livestock practices, with higher occurrences in spring and summer than in autumn and winter. The most frequent compounds in superficial water were the insecticide chlorantraniliprole, and the herbicides glyphosate (including its metabolite AMPA) and metolachlor. The levels of Organochlorine pesticide, p,p'-DDT, was in some cases two order of magnitude above the international water quality guidelines for Ambient Water Criteria. In fishes, eight different pesticides were detected, at concentrations from 1000 to 453,000 ng·kg-1. The most frequent pesticides found were propiconazole, chlorpyrifos, and p,p'-DDE. The widespread occurrence of pesticides in fish suggests potential exposure effects on fish populations and the aquatic ecosystem. The sampling approach of this work allowed monitoring the continuous concentrations of several pesticides in surface waters and fishes to establish the influence from past and current agriculture practices in Laguna del Cisne basin. For safety measures, continuous monitoring programs must be performed in this system to prevent toxicity impacts on aquatic organisms and human health.

Inclusion of 1-naphthylacetic acid and 2-(1-naphthyl)acetamide into three typical multiresidue methods for LC/MS/MS analysis of tomatoes and zucchini.

In spite of high plant growth regulator application rates, little has been reported in the literature on determination of their residues in fruits and vegetables. This would be useful in monitoring good manufacturing practices and overall safety through the enforcement of maximum residue levels (MRLs). The present work describes method validation for the determination of 1-naphthylacetic acid (NAA) and 2(1-naphthyl)acetamide (NAAm) in tomato and zucchini using the mini-Luke, ethyl acetate (EtOAc) and acetate-buffered quick, easy, cheap, effective, rugged, and safe (QuEChERS) methods. Samples were spiked at two different levels: 50 and 100 pg/kg for NAA and 20 and 100 pg/kg for NAAm. These compounds were analyzed within the same chromatographic run with LC coupled to triple quadrupole MS (LC/(QqQ)MS/MS) in positive and negative electrospray ionization [ESI(+) and ESI(-)] modes for NAAm and NAA, respectively. For analyte confirmation, LC/ESI(-)QTOF-MS was also investigated given that NAA has only one multiple reaction monitoring transition (185.1-*140.9 m/z). These three common methods were used to determine linearity, recoveries, precision (RSD), matrix effects, repeatability, and reproducibility (n = 5) for the selected matrixes. In terms of the Directorate-General for Health and Consumers (DG-SANCO) guidelines, only insignificant differences were found for the multiresidue methods tested, regardless of the commodity. Matrix-matched calibration was used, and LODs were below 10.1 pg/kg for NAA and 6.0 pg/kg for NAAm, which were lower than the MRLs established in current European Union legislation for these compounds. Obtained recoveries for NAA ranged from 87 to 107% with RSD values below 10% for mini-Luke, 83 to 107% with RSD <11% for EtOAc, and 76 to 85% with RSD <7% for QuEChERS. NAAm recoveries ranged from 74 to 102% with RSD 5 15% for mini-Luke, 76 to 97% with RSD <4% for EtOAc, and 76 to 93% with RSD < 5% for QuEChERS. The linearity of the response over two orders of magnitude was demonstrated (r2 > 0.996) for all methods used. Good repeatability and reproducibility in terms of RSD were obtained (11.3-17.8%) for these analytical methods. The results were acceptable within the studied ranges for both analytes in all methods. As these multiresidue methods are used at the present time in many laboratories, this survey indicates that these analytes should be included in their monitoring scope.

Modeling microplastic with polyethylene (PE) spherical particles: a differential scanning calorimetry approach for quantification.

Detection, identification, and quantification of microplastics have become increasingly relevant for determining their contribution and role in environmental pollution. Thermal analysis is positioned as one of the alternative techniques employed to quantify microplastics. However, a deeper investigation that explores its capabilities is required, since in techniques such as difference scanning calorimetry (DSC), the result of the melting curve is potentially affected by the size of the micro particles. Therefore, to use this technique in the field of quantitative analysis of microplastics, it is necessary to make an evaluation of how the micro particle size affects the signal obtained. We use spherical polyethylene (PE) particles of different sizes (75-710 µm) as a microplastic model to study the effect of particle size and the mixtures of different particle sizes on the melting curve. The effect of possible interferences on the DSC signal was studied and real microplastics isolated from wastewater were tested. It was found that the DSC signal (both melting temperature and peak shape) is affected by the size of the particles, even in the case of mixtures of particles of different sizes. However, through an appropriate sample preparation, it is possible to identify the signals corresponding to microplastics of different sizes and thus quantify their contribution to the mass of the sample. It was evidenced that factors such as the presence of inorganic materials tend to modify the melting temperature. Also, removal of interferences of organic origin is feasible. In addition, the presence of PP, HDPE and LDPE was evidenced in wastewater samples. Our results represent an important advance in the use of the DSC technique in the field of microplastics, since the existence of particles of different sizes can be evidenced in the same sample allowing for an estimation of the number of microplastic particles. Finally, we show the applicability of DSC study on microplastics in environmental matrices.

Determination of glyphosate and AMPA in freshwater and soil from agroecosystems by 9-fluorenylmethoxycarbonyl chloride derivatization and liquid chromatography - fluorescence detection and tandem mass spectrometry.

The herbicide glyphosate (GLY) and its metabolite aminophosphonic acid (AMPA) are troublesome compounds for analysis in the environment. Here we report a reliable technique for GLY and AMPA determination in freshwater and soils by means of derivatization with 9-fluorenylmethoxycarbonyl chloride (FMOC-Cl) and further liquid chromatography with fluorescence detection (FLD) and tandem mass spectrometry (MS/MS) analysis. Selected experiments were carried out to evaluate selectivity, sensitivity, repeatability, linearity and quantification performance in both matrices.

Human microbiota drives hospital-associated antimicrobial resistance dissemination in the urban environment and mirrors patient case rates.

BACKGROUND: The microbial community composition of urban environments is primarily determined by human activity. The use of metagenomics to explore how microbial communities are shaped in a city provides a novel input that can improve decisions on public health measures, architectural design, and urban resilience. Of note, the sewage system in a city acts as a complex reservoir of bacteria, pharmaceuticals, and antimicrobial resistant (AMR) genes that can be an important source of epidemiological information. Hospital effluents are rich in patient-derived bacteria and can thus readily become a birthplace and hotspot reservoir for antibiotic resistant pathogens which are eventually incorporated into the environment. Yet, the scope to which nosocomial outbreaks impact the urban environment is still poorly understood. RESULTS: In this work, we extensively show that different urban waters from creeks, beaches, sewage spillways and collector pipes enclose discrete microbial communities that are characterized by a differential degree of contamination and admixture with human-derived bacteria. The abundance of human bacteria correlates with the abundance of AMR genes in the environment, with beta-lactamases being the top-contributing class to distinguish low vs. highly-impacted urban environments. Indeed, the abundance of beta-lactamase resistance and carbapenem resistance determinants in the urban environment significantly increased in a 1-year period. This was in line with a pronounced increase of nosocomial carbapenem-resistant infections reported during the same period that was mainly driven by an outbreak-causing, carbapenemase-producing Klebsiella pneumoniae (KPC) ST-11 strain. Genome-resolved metagenomics of urban waters before and after this outbreak, coupled with high-resolution whole-genome sequencing, confirmed the dissemination of the ST-11 strain and a novel KPC megaplasmid from the hospital to the urban environment. City-wide analysis showed that geospatial dissemination of the KPC megaplasmid in the urban environment inversely depended on the sewage system infrastructure. CONCLUSIONS: We show how urban metagenomics and outbreak genomic surveillance can be coupled to generate relevant information for infection control, antibiotic stewardship, and pathogen epidemiology. Our results highlight the need to better characterize and understand how human-derived bacteria and antimicrobial resistance disseminate in the urban environment to incorporate this information in the development of effluent treatment infrastructure and public health policies. Video Abstract.

Salt marshes as the final watershed fate for meso- and microplastic contamination: A case study from Southern Brazil.

Plastics pose a major threat to aquatic ecosystems especially in smaller size fractions. Salt marshes play a crucial role in maintaining the coastal zone and aquatic food web, yet their contamination, including by plastic materials, is still poorly investigated. This work investigated meso- (MEP, 5-25 mm) and microplastic (MIP, 1 µm-5 mm) contamination of a salt marsh, which reached average levels of 279.63 ± 410.12 items kg-1, 366.92 ± 975.18 items kg-1, and 8.89 ± 8.75 items L-1 in surface sediment, sediment cores and water, respectively. Photomicrographs revealed a complex fouling community on plastics surface for both different salt marsh zones and plastic formats. Abundance of plastics in sediment was higher in the dryer, vegetated zones compared to flooded, unvegetated zones. This is consistent with the role of vegetation as a trap for solid litter and final fate of plastic deposition, but also with local hydrodynamics influencing deposition pattern. Plastics were detected up to 66 cm-depth, presenting higher levels at surface sediments. It was also possible to identify the main groups of microorganisms (1638 bacterial cells, 318 microalgae cells, and 20049.93 µm2 of filamentous fungi) composing the Plastisphere communities on all plastic items recorded in the different zones. These results are a pioneer contribution, highlighting that regional salt marshes participate in sequestration and longstanding accumulation of plastic particles in estuarine environments, before exportation to the ocean.

Behavior of amoxicillin in wastewater and river water: identification of its main transformation products by liquid chromatography/electrospray quadrupole time-of-flight mass spectrometry.

The identification of transformation products (TPs) of pharmaceuticals in the environment is essentially a challenging task due to the lack of standards and the instrumental capabilities required to detect compounds (sometimes unknowns) that are produced under environmental conditions. In this work, we report the use of liquid chromatography/electrospray quadrupole time-of-flight mass spectrometry (LC/QTOF-MS/MS) as a tool for the identification of amoxicillin (AMX) and its main TPs in wastewater and river water samples. Laboratory degradation experiments of AMX were performed in both alkaline and acidic media in order to confirm that the expected transformation pathway in the aquatic media is through the ß-lactam ring cleavage. A thorough study was carried out with both standards and real samples (wastewater and river water samples). Four compounds were identified as main TPs: both amoxicillin diketopiperacine-2',5' and amoxilloic acid diastereomers. Amoxilloic acid stereoisomers are reported for the first time in environmental matrices. The transformation product (5R)-amoxicillin diketopiperacine-2',5' was frequently detected in river waters. Besides, another AMX transformation product formed during analysis was also structurally elucidated for the first time (amoxicilloic acid methyl ester) via accurate mass measurements. Collected data show that although AMX is not present as such in environmental samples, different TPs occur. This study represent a valuable indicator of the potential of LC/QTOF-MS/MS for the identification and structural elucidation of TPs in the environment using accurate MS/MS experiments, enabling thus the recognition of the environmental transformation pathway.

Sulfur precursor and citric acid effect on SnS2 nanoparticles and their influence on the photodegradation activity of selected organic compounds.

Semiconductor nanoparticle-mediated photocatalysis is an attractive option for water decontamination, being the semiconductors as SnS2 with a bandgap in the visible region, the most promising materials. In the present work, we evaluated the influence of important parameters in the photocatalytic application of SnS2 nanoparticles. Our results show that the presence of citric acid (used as a capping agent) restricts the formation of hexagonal nanoparticles. We also demonstrated that using thioacetamide as a sulfur source results in smaller nanoparticles than thiourea, 24.0 nm and 616 nm respectively. Moreover, small hexagonal nanoparticles play a key role in the photocatalytic activity of SnS2 nanoparticles. Compared with TiO2 performance, SnS2 nanoparticles exhibited faster kinetics for methyl orange (MO) degradation, Kapp = 0.0102 min-1, and 0.029 min-1, respectively. We proved that SnS2 is capable of breaking the azo bond of methyl orange by direct reduction. Furthermore, our analyses indicate that SnS2 nanoparticles do not degrade atrazine and imazapic, but the photocatalytic route of metribuzin competed with photolysis, resulting in a particular transformation product that was not obtained with light irradiation only. We demonstrated that SnS2 nanoparticles have high bond selectivity for azo breaking. Furthermore, they represent an advance for the development of designed materials (such as heterostructures), where the properties of SnS2 can be tuned.

Transfer of pesticides to the brew during mate drinking process and their relationship with physicochemical properties.

In order to evaluate the extraction of pesticide residues that are transferred to the brew during mate drinking process of P.U.1 yerba mate leaves (Ilex paraguariensis), a special device to simulate the way in which mate is drunk in Uruguay was developed. The transfer to the brew of 12 organophosphates, 5 synthethic pyrethroids and one organochlorine pesticide from spiked samples was studied. The relationship between the transfer data thus obtained and physicochemical properties like water solubility (Ws), octanol-water coefficient (Kow) and Henry's constant (H) was evaluated. The extractability of the pesticide residues from yerba mate can be correlated with log Ws and log Kow. These transfer values allowed the calculation of ARLs (acceptable residue level) for the pesticides following Food and Agriculture Organization (FAO), World Health Organizaion (WHO) guidelines. These results can help the future establishment of maximum residue levels (MRLs).

Identification of microplastics in wastewater samples by means of polarized light optical microscopy.

Many reports state the potential hazards of microplastics (MPs) and their implications to wildlife and human health. The presence of MP in the aquatic environment is related to several origins but particularly associated to their occurrence in wastewater effluents. The determination of MP in these complex samples is a challenge. Current analytical procedures for MP monitoring are based on separation and counting by visual observation or mediated with some type of microscopy with further identification by techniques such as Raman or Fourier-transform infrared (FTIR) spectroscopy. In this work, a simple alternative for the separation, counting and identification of MP in wastewater samples is reported. The presented sample preparation technique with further polarized light optical microscopy (PLOM) observation positively identified the vast majority of MP particles occurring in wastewater samples of Montevideo, Uruguay, in the 70-600 µm range. MPs with different shapes and chemical composition were identified by PLOM and confirmed by confocal Raman microscopy. Rapid identification of polyethylene (PE), polypropylene (PP) and polyethylene terephthalate (PET) were evidenced. A major limitation was found in the identification of MP from non-birefringent polymers such as PVC (polyvinylchloride). The proposed procedure for MP analysis in wastewater is easy to be implemented at any analytical laboratory. A pilot monitoring of Montevideo WWTP effluents was carried out over 3-month period identifying MP from different chemical identities in the range 5.3-8.2 × 103 MP items/m3.

Determination of multiclass, semi-polar pesticide residues in fatty fish muscle tissue by gas and liquid chromatography mass spectrometry.

With the aim of monitoring multiclass semi-polar pesticide residues in freshwater fatty fish, two QuEChERS approaches (so-called acetate buffered and unbuffered versions) were evaluated for the determination of 77 pesticide residues. Compounds were selected according to the dominant rainfed agriculture activities in South America. Unbuffered QuEChERS was finally chosen for validation purposes owing that it provided the best results in terms of recovery yields. Method performance was evaluated in two instrumental systems, liquid chromatography - tandem mass spectrometry (LC-MS/MS) in Scheduled MRM™ algorithm available on hybrid quadrupole - linear ion trap (QLIT) instrument, and gas chromatography - mass spectrometry (GC-MS) under selected ion monitoring (SIM) mode. Spiking experiments were carried out to determine the trueness, precision, linearity, limit of quantification of the method as well as matrix effect. The Unbuffered QuECHERS method described here: •Was validated for the analysis of 67 pesticide residues in fish muscle tissue.•Presented quantification limits in the range 1-15 µg kg-1 for the vast majority of the studied compounds.•Enable environmental monitoring of pesticide residues in fish due to their low LOQs.

Basin-scale monitoring and risk assessment of emerging contaminants in South American Atlantic coastal lagoons.

Emerging contaminants (ECs) such as pharmaceuticals, personal care products, drugs of abuse and polar pesticides are under particular attention due to their high consumption, frequent detection in the environment and reported ecotoxicological risk. This study investigates the occurrence and distribution of multiclass of ECs in surface waters at basin scale of two Atlantic coastal lagoons of Uruguay, South America. For this purpose, a target screening approach covering up to 362 compounds was employed using nanoflow liquid chromatography - high resolution mass spectrometry (nanoLC/HRMS). 56 compounds were identified including five banned pesticides in the European Union: atrazine, carbendazim, chlorpyrifos ethyl, diazinon, and ethion. Pharmaceuticals, hormones and drugs of abuse showed maximum detection frequencies and concentrations downstream cities. The highest occurrence of pesticides was found in lagoons and streams with neighboring agricultural activity. ECs were also found in coastal sea. Environmental risk assessment revealed that the hormones 17α-ethinylestradiol and 17-ß-estradiol showed the highest risk to aquatic organisms in these basins. This study represents the first basin- scale monitoring of ECs in superficial waters encompassing streams, lagoons, and coastal seas in Uruguay, South America.

Identification and quantitation of semi-crystalline microplastics using image analysis and differential scanning calorimetry.

There are several techniques used to analyze microplastics. These are often based on a combination of visual and spectroscopic techniques. Here we introduce an alternative workflow for identification and mass quantitation through a combination of optical microscopy with image analysis (IA) and differential scanning calorimetry (DSC). We studied four synthetic polymers with environmental concern: low and high density polyethylene (LDPE and HDPE, respectively), polypropylene (PP), and polyethylene terephthalate (PET). Selected experiments were conducted to investigate (i) particle characterization and counting procedures based on image analysis with open-source software, (ii) chemical identification of microplastics based on DSC signal processing, (iii) dependence of particle size on DSC signal, and (iv) quantitation of microplastics mass based on DSC signal. We describe the potential and limitations of these techniques to increase reliability for microplastic analysis. Particle size demonstrated to have particular incidence in the qualitative and quantitative performance of DSC signals. Both, identification (based on characteristic onset temperature) and mass quantitation (based on heat flow) showed to be affected by particle size. As a result, a proper sample treatment which includes sieving of suspended particles is particularly required for this analytical approach.