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Despite the effectiveness of DNA metabarcoding for gaining insights into biodiversity and environmental species composition, a centralised management and storage option including easy accessibility of already-published data is lacking. Since most data are published as supplementary material or in private repositories, DNA metabarcoding has a huge untapped potential to be used for analyses across multiple taxa, sample locations or multiple research projects. We developed a platform to register, manage and identify amplicon sequence variants (ASVs) or zero-radius OTUs (ZOTUs), respectively, against several barcode reference datasets. Moreover, ASV tables can be uploaded, managed, versioned and published with DOIs thus contributing to the full research Data Life Cycle.

Abstract Affilin proteins, artificial binding proteins based on the ubiquitin scaffold, have been generated by directed protein evolution to yield de-novo variants that bind the extra-domain B (EDB) of oncofetal fibronectin, an established marker of tumor neovasculature. The crystal structures of two EDB-specific Affilin variants reveal a striking structural plasticity of the ubiquitin scaffold, characterised by β-strand slippage, leading to different negative register shifts of the β5 strands. This process recruits amino acid residues from β5 towards the N-terminus to an adjacent loop region and subsequent residues into β5, respectively, remodeling the binding interface and leading to target specificity and affinity. Protein backbone alterations resulting from β-strand register shifts, as seen in the ubiquitin fold, can pose additional challenges to protein engineering as structural evidence of these events is still limited and they are difficult to predict. However, they can surface under the selection pressure of directed evolution and suggest that backbone plasticity allowing β-strand slippages can increase structural diversity, enhancing the evolutionary potential of a protein scaffold.

Background: Per- and polyfluoroalkyl substances (PFAS) are a class of widely used anthropogenic chemicals. Concerns regarding their persistence and potential adverse effects have led to multiple secondary research publications. Here, we aim to assess the resulting evidence base in the systematic secondary literature by examining research gaps, evaluating the quality of reviews, and exploring interdisciplinary connections. Methods: This study employed a systematic evidence-mapping approach to assess the secondary literature on the biological, environmental, and medical aspects of exposure to 35 fluorinated compounds. The inclusion criteria encompassed systematic reviews published in peer-reviewed journals, pre-prints, and theses. Comprehensive searches across electronic databases and grey literature identified relevant reviews. Data extraction and synthesis involved mapping literature content and narrative descriptions. We employed a modified version of the AMSTAR2 checklist to evaluate the methodological rigour of the reviews. A bibliometric data analysis uncovered patterns and trends in the academic literature. A research protocol for this study was previously pre-registered (osf.io/2tpn8) and published (Vendl et al., Environment International 158 (2022) 106973). The database is freely accessible through the interactive and user-friendly web application of this systematic evidence map at https://hi-this-is-lorenzo.shinyapps.io/PFAS_SEM_Shiny_App/. Results: Our map includes a total of 175 systematic reviews. Over the years, there has been a steady increase in the annual number of publications, with a notable surge in 2021. Most reviews focused on human exposure, whereas environmental and animal-related reviews were fewer and often lacked a rigorous systematic approach to literature search and screening. Review outcomes were predominantly associated with human health, particularly with reproductive and children’s developmental health. Animal reviews primarily focused on studies conducted in controlled laboratory settings, and wildlife reviews were characterised by an over-representation of birds and fish species. Recent reviews increasingly incorporated quantitative synthesis methodologies. The methodological strengths of the reviews included detailed descriptions of study selection processes and disclosure of potential conflicts of interest. However, weaknesses were observed in the critical lack of detail in reporting methods. A bibliometric analysis revealed that the most productive authors collaborate within their own country, leading to limited and clustered international collaborations. Conclusions: In this overview of the available systematic secondary literature, we map literature content, assess reviews’ methodological quality, highlight data gaps, and draw research network clusters. We aim to facilitate literature reviews, guide future research initiatives, and enhance opportunities for cross-country collaboration. Furthermore, we discuss how this systematic evidence map and its publicly available database benefit scientists, regulatory agencies, and other stakeholders by providing access to current systematic secondary literature on PFAS exposure.

The intricate composition, heterogeneity, and hierarchical organization of the human bone marrow hematopoietic microenvironment (HME) present challenges for experimentation, which is primarily due to the scarcity of HME-forming cells, notably bone marrow stromal cells (BMSCs). The limited understanding of non-hematopoietic cell phenotypes complicates the unraveling of the HME’s intricacies and necessitates a precise isolation protocol for systematic studies. The protocol presented herein puts special emphasis on the accuracy and high quality of BMSCs obtained for downstream sequencing analysis. Utilizing CD45 and CD235a as negative markers ensures sufficient enrichment of non-hematopoietic cells within the HME. By adding positive selection based on CD271 expression, this protocol allows for selectively isolating the rare and pivotal bona fide stromal cell population with high precision. The outlined step-by-step protocol provides a robust tool for isolating and characterizing non-hematopoietic cells, including stromal cells, from human bone marrow preparations. This approach thus contributes valuable information to promote research in a field that is marked by a scarcity of studies and helps to conduct important experimentation that will deepen our understanding of the intricate cellular interactions within the bone marrow niche.

Although endangerment towards others is a criterion for an involuntary admission in many countries, research on risk assessment of endangerment among involuntarily admitted individuals is limited. In this retrospective case-control study, we calculated scores for a German-translated version of the Violence Risk Screening-10 (V-RISK-10) and the Brøset Violence Checklist (BVC) in a sample of 111 people undergoing an involuntary admission in Reinickendorf, Berlin. Outcomes were violence, coercive measures, and readmission. In line with our hypotheses, the BVC demonstrated stronger predictive validities for short-term, and V-RISK-10 for long-term events. There was an incremental validity for both instruments for restraint 24 hours after admission and any violence until discharge. These findings support the evidence that structured risk assessment instruments may be useful for individuals undergoing an involuntary admission. Ethical considerations about screening procedures are discussed.

Abstract Per‐ and polyfluoroalkyl substances (PFAS) are a large group of synthetic chemicals that have been in use for over 70 years. Their ubiquitous distribution and harmful effects pose a threat to wildlife worldwide. To provide a comprehensive synopsis and show the gaps and gluts of existing research on PFAS exposure in wildlife, we created a systematic map and bibliographic analysis of the literature. We followed our protocol to conduct a systematic literature search on Scopus, Web of Science and five other databases. In two steps (title/abstract/keywords and full‐text), we screened peer‐reviewed empirical articles, preprints and theses in English that studied the concentration of at least one of 34 PFAS compounds in free‐ranging wildlife or their parts/products. Following the protocol, we extracted data and performed a critical appraisal. We included 581 publications. From the first and only paper in 2001, there was a linear annual increase to 54 papers in 2021. While PFOS (97% of studies), PFOA (91%) and long‐chain PFAS in general were the most measured, few studies investigated new‐generation PFAS (e.g. GenX and ADONA). Across the studied 1042 species from 26 taxonomic classes, the most frequent were the common carp (Cyprinus carpio, 8%), polar bear (Ursus maritimus, 6%) and European perch (Perca fluviatilis, 5%). Most sampling took place in the United States (17%), Norway (13%), Canada (12%) and China (10%), which were also the main publishing countries. Polar regions attracted significant research interest from countries all around the globe. Aquatic habitats (marine: 31%, freshwater: 28%) of temperate zones were the most common locations for sample collection. We encourage researchers to work towards closing the following gaps: investigating new‐generation PFAS, assessing PFAS in mid‐ and low‐income countries and performing more long‐term studies, especially on invertebrates. We note the recent rise in studies on the physiological consequences of PFAS exposure and encourage further work on this crucial topic. Furthermore, we recommend that the statement of potential and actual conflicts of interest, and the provision of raw data and analysis code should be made compulsory by all journals and routinely enforced. This practice will mitigate conflict of interest and ensure reproducibility.

Abstract Bovine trypanosomosis, caused by Trypanosoma vivax, is a disease that originated in Africa and currently affects cattle in several South American countries, including almost all Brazilian states. Despite the reports on T. vivax infection in southern Brazil, data on its circulation status is currently unavailable. In this study, we aimed to detect anti-Trypanosoma spp. IgG antibodies in cattle from Rio Grande do Sul and suggest areas with T. vivax transmission risk. A total of 691 serum samples from cattle in the intermediate regions of Rio Grande do Sul were analyzed using indirect immunofluorescence assay (IFA). The overall seroprevalence of anti-Trypanosoma antibodies in cattle was 24.6% (170/691). The detection rate ranged from 0-37.3%, with a high prevalence in the intermediate regions of Ijuí (37.3%), Uruguaiana (30.7%), and Passo Fundo (28.9%). Thus, these regions were suggested as possible bovine trypanosomosis risk areas due to the high seroprevalence. This is the first serological study to determine Trypanosoma spp. infection status in cattle from Rio Grande do Sul, providing data on the epidemiology of trypanosomosis in the state.

Abstract Domestic birds such as Gallus gallus, Meleagris gallopavo, Anser anser and Numida meleagris are widely distributed throughout the world and maintain contact with humans and other animal species considered reservoirs of both Visceral Leishmaniasis (VL) and American Tegumentary Leishmaniasis (ATL), including dogs and cats; wild canids, marsupials; and synanthropic animals such as rodents and chiroptera. Therefore, this study aimed to detect the presence of anti-Leishmania spp. antibodies in birds from a rural area of the municipality of Santa Maria, southern Brazil. From May to December 2022, 262 blood samples were collected from 244 chickens, 8 turkeys, 7 guinea fowl and 3 geese, distributed in 27 rural properties in 6 districts. All the sites visited presented positive birds for the presence of Leishmania spp. Thus, it is inferred that, contact with this protozoan can induce the production of antibodies, suggesting that these animals can be used as sentinels for the circulation of this agent. In addition, the blood of these animals is a preferred food source for insects of the subfamily Phlebotominae, which can be used them as bioindicators of the presence of these phlebotomes.

ABSTRACT: Cattle are considered intermediate hosts of Sarcocystis, which can cause clinical signs and lower performance in the acute phase of infection. Sarcocystis spp. are usually not visible to the naked eye during the post mortem inspection. Moreover, fresh microscopic examination and transmission electron microscopy techniques are difficult to apply to large samples. Therefore, extensive studies on Sarcocystis infection in cattle using molecular and serological methods are required. Here, we investigated Sarcocystis spp. infection in cattle using fresh microscopic examination and polymerase chain reaction of myocardium samples and compared the results with the presence of antibodies against Sarcocystis spp. in corresponding serum samples detected using indirect fluorescent antibody test. Microscopic Sarcocystis were observed in 100% of the myocardial samples, and Sarcocystis DNA was present in 86% (43/50) of these samples. Antibodies against Sarcocystis spp. were detected in 96% (48/50) and 80% (40/50) of the serum samples at 1:25 and 1:200 dilutions, respectively. The three associated methods (fresh microscopic examination, PCR and serology) showed good sensitivity and detection for Sarcocystis spp. compared with fresh microscopic examination (only), and they may facilitate diagnosis in live animals on a large scale as well as monitoring of the herd status.

Rhipicephalus microplus é uma espécie de carrapato amplamente distribuída que causa perdas diretas e indiretas à produção de gado; portanto, o controle de infestações de carrapatos é essencial. Métodos de tratamento corretos são necessários para evitar sobredosagem ou subdosagem de acaricidas químicos e controlar a resistência aos carrapatos. Além disso, estudos que evidenciem os fatores associados à eficácia dos produtos químicos são extremamente necessários. Portanto, este estudo teve como objetivo avaliar os efeitos da intensidade e do tempo de precipitação sobre a eficácia da aplicação tópica de acaricidas. Fêmeas ingurgitadas foram coletadas de bovinos em fazendas localizadas no Rio Grande do Sul, Brasil e tratadas com Colosso FC30® Ouro Fino Animal Health (15% cipermetrina, 30% clorpirifós, and 15% fention). Após 30 min, 1 h, 2 h, 3 h, 6 h, 9 h, 12 h ou 24 h de tratamento, foram submetidas à simulações de precipitação em intensidade leve ou intensa, para cada tempo. Os resultados mostraram eficácia acaricida de 100% para todos os grupos avaliados e demonstraram que a precipitação de 30 min após o tratamento e em qualquer intensidade, não afetou a eficácia do Colosso FC30. Portanto, este estudo esclareceu a influência da precipitação simulada na eficácia do tratamento tópico Colosso FC30 no controle de R. microplus.

Hematopoiesis is regulated by the bone marrow (BM) stroma. However, cellular identities and functions of the different BM stromal elements in humans remain poorly defined. Based on single-cell RNA sequencing (scRNAseq), we systematically characterized the human non-hematopoietic BM stromal compartment and we investigated stromal cell regulation principles based on the RNA velocity analysis using scVelo and studied the interactions between the human BM stromal cells and hematopoietic cells based on ligand-receptor (LR) expression using CellPhoneDB. scRNAseq led to the identification of six transcriptionally and functionally distinct stromal cell populations. Stromal cell differentiation hierarchy was recapitulated based on RNA velocity analysis and in vitro proliferation capacities and differentiation potentials. Potential key factors that might govern the transition from stem and progenitor cells to fate-committed cells were identified. In situ localization analysis demonstrated that different stromal cells were localized in different niches in the bone marrow. In silico cell-cell communication analysis further predicted that different stromal cell types might regulate hematopoiesis through distinct mechanisms. These findings provide the basis for a comprehensive understanding of the cellular complexity of the human BM microenvironment and the intricate stroma-hematopoiesis crosstalk mechanisms, thus refining our current view on human hematopoietic niche organization.

Estudos sobre doenças de aves silvestres são essenciais no contexto da saúde pública, pois esses animais atuam como sentinelas, permitindo obter informações sobre uma determinada área geográfica. Além disso, as aves são fontes de proteína alimentar para os animais e, portanto, desempenham um papel importante no ciclo de vida do Sarcocystis. Este estudo teve como objetivo identificar Sarcocystis spp. nos músculos do peito de aves de cativeiro naturalmente infectadas. Os músculos do peito de 89 aves foram coletados, e o DNA amplificado pela PCR do gene RNA ribossômico 18S para detecção de Sarcocystis spp. Os produtos da PCR foram sequenciados e 5,61% (5/89) amostras apresentaram 100% de similaridade com o Sarcocystis spp. (um Cyanoliseus patagonus, um Psittacula krameri, dois Pyrrhura frontalis e um Ramphastos dicolorus). O grande número de espécies naturalmente infectadas analisadas por métodos moleculares permitiu a detecção de Sarcocystis spp. em diferentes espécies de aves, corroborando a epidemiologia de Sarcocystis spp. Até onde sabemos, este é o primeiro relato especificando a presença de Sarcocystis spp. no músculo do peito de espécies de aves.

We present pre-stack depth-imaging results for a case study of 3D reflection seismic exploration at the Blötberget iron oxide mining site belonging to the Bergslagen mineral district in central Sweden. The goal of the study is to directly image the ore-bearing horizons and to delineate their possible depth extension below depths known from existing boreholes. For this purpose, we applied a tailored pre-processing workflow and two different seismic imaging approaches, Kirchhoff pre-stack depth migration (KPSDM) and Fresnel volume migration (FVM). Both imaging techniques deliver a well-resolved 3D image of the deposit and its host rock, where the FVM image yields a significantly better image quality compared to the KPSDM image. We were able to unravel distinct horizons, which are linked to known mineralization and provide insights on their possible lateral and depth extent. Comparison of the known mineralization with the final FVM reflection volume suggests a good agreement of the position and the shape of the imaged reflectors caused by the mineralization. Furthermore, the images show additional reflectors below the mineralization and reflectors with opposite dips. One of these reflectors is interpreted to be a fault intersecting the mineralization, which can be traced to the surface and linked to a fault trace in the geological map. The depth-imaging results can serve as the basis for further investigations, drilling, and follow-up mine planning at the Blötberget mining site..

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ABSTRACT Neospora caninum is the main etiologic agent of neosporosis in domestic animals and its pathogenesis comprises two characteristic phases: acute and chronic. Rodents are used as experimental models to mimic acute and chronic bovine neosporosis. In this study, we inoculated a total of 27 female gerbils, with different doses of N. caninum tachyzoites aiming to induce chronic disease. DNA was extracted from different organs of each animal after spontaneous death or euthanasia. Encephalic tissues were submitted to a highly sensitive real time PCR aiming to detect chronically infected animals. All the other samples were submitted to standard PCR. A total of 11 gerbils died due to acute neosporosis, as confirmed by N. caninum DNA detection in organs. 5x103 tachyzoites/mL of N. caninum was the dosage of antigen that can induce chronic infection in gerbils. In the encephalon sections of some animals that showed clinical signs of persistent infection, we found 70% positive for the anterior encephalon section, suggesting this area as preferential for cyst formation. Therefore, we determined the doses of tachyzoites that cause acute or chronic infection and detection of positive tissues, preferably, systemic organs during acute and encephalon in chronic phases.

ABSTRACT: Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples.

ABSTRACT Free-range chickens may ingest oocysts of T. gondii present in the environment and consequently harbor virulent strains of this parasite in different tissues, without any clinical signs. Isolation of T. gondii through bioassays on mice and cats from naturally infected chicken tissues has been described in several countries, demonstrating the importance of free-range chickens in the transmission of this parasite. The aim of this study was the genotypic characterization of T. gondii isolates obtained from naturally infected free-range chickens in a rural area of the state of Rio Grande do Sul, Brazil. Brain and heart tissue from 12 chickens seropositive for T. gondii were processed using peptic digestion technique for parasite isolation. From 12 samples subjected to mouse bioassay, nine isolates were obtained. RFLP-PCR genotypic characterization was performed using 11 genetic markers: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genetic characterization of the isolates revealed the presence of five atypical genotypes according to ToxoDB (# 11, # 55, # 64, # 140 and # 163). Our results showed a wide genetic diversity of T. gondii in free-range chickens in this region.

ABSTRACT: Serological techniques can detect antibodies against Sarcocystis spp., Neospora caninum and Toxoplasma gondii antigens in single or mixed infections. Immunofluorescent antibody tests (IFAT) is considered the gold standard technique for Sarcocystosis diagnostic in cattle serum and a positive IFAT result reflects Sarcocystis spp. infection. Therefore, the aims of the present study were to compare IFAT and Dot-blot for sarcocystosis diagnostic in experimentally infected mice and to investigate serological cross-reactions with N. caninum and T. gondii in these methods. Mice (Mus musculus) were inoculated intraperitoneally with bradizoites of Sarcocystis spp. or tachyzoites of N. caninum or T. gondii. Serum samples were obtained and analyzed by IFAT and Dot-blot for the three protozoa. Serum from N. caninum and T. gondii experimentally infected mice were tested by IFAT and reacted only to N. caninum or T. gondii antigens, respectively. Specific antibodies against Sarcocystis spp. were present in all animals experimentally infected with this protozoan, with IFAT titers from 10 to 800. Serum samples from mice experimentally infected with Sarcocystis spp., N. caninum and T. gondii and tested by Dot-blot demonstrated no cross reaction between protozoa. A Dot-blot using Sarcocystis spp. antigen appears to be a good alternative to IFAT in the serological diagnosis of Sarcocystosis.

Novel therapeutic strategies are urgently needed for advanced metastatic prostate cancer (PCa). Phytochemicals used in Traditional Chinese Medicine seem to exhibit tumor suppressive properties. Therefore, the therapeutic potential of artesunate (ART) on the progressive growth of therapy-sensitive (parental) and docetaxel (DX)-resistant PCa cells was investigated. Parental and DX-resistant PCa cell lines DU145, PC3, and LNCaP were incubated with artesunate (ART) [1-100 µM]. ART-untreated and ‘non-cancerous’ cells served as controls. Cell growth, proliferation, cell cycle progression, cell death and the expression of involved proteins were evaluated. ART, dose- and time-dependently, significantly restricted cell growth and proliferation of parental and DX-resistant PCa cells, but not of ‘normal, non-cancerous’ cells. ART-induced growth and proliferation inhibition was accompanied by G0/G1 phase arrest and down-regulation of cell cycle activating proteins in all DX-resistant PCa cells and parental LNCaP. In the parental and DX-resistant PC3 and LNCaP cell lines, ART also promoted apoptotic cell death. Ferroptosis was exclusively induced by ART in parental and DX-resistant DU145 cells by increasing reactive oxygen species (ROS). The anti-cancer activity displayed by ART took effect in all three PCa cell lines, but through different mechanisms of action. Thus, in advanced PCa, ART may hold promise as a complementary treatment together with conventional therapy.

Background: Per- and polyfluoroalkyl (PFAS) substances are a group of anthropogenic organic chemicals. PFAS have numerous applications in the industrial and commercial fields. They have become abundant in the environment and have been detected in the blood of humans and animals. PFAS exposure likely increases the risk of certain diseases. In recent years, there has been a steep rise in related primary and secondary articles. Objectives: We aim to collate narrative and systematic reviews including both qualitative and quantitative synthesis on biological, environmental, or medical aspects of PFAS exposure and burdens, and to create an evidence review map to fulfil three main objectives. 1) Mappingto reveal what evidence on PFAS has been systematically reviewed and where synthesis may be needed. 2) Bibliometrics: to characterise the interdisciplinarity, independence and connectedness of the reviews. 3) Critical appraisal: to assess the included reviews for the reliability of conclusions and methodological transparency. This overview of reviews will inform future research and policy making. Search and study eligibility: We will perform a systematic search on Scopus, Web of Science, PubMed, and the ‘grey literature’. To be included in the map, a study must be a review from the systematic review family and focus on at least one of predefined 31 PFAS. We will limit inclusions to peer-reviewed publications, pre-prints, and theses without any language restrictions. At least two authors will scan title, abstract and keywords, followed by full-text screening of included reviews. Critical appraisal of reviews and synthesis methods: We will assess each review of the systematic review family referencing the AMSTAR2 checklist. We will collect information on conflict-of-interest statements, funding sources and availability of raw data and analysis code. We will extract relevant data from full texts and bibliographic records, conduct bibliometric analyses, and display findings as tables, figures and a narrative synopsis.

Abstract Per‐ and polyfluoroalkyl substances (PFAS) are a large group of manufactured chemicals. Since the beginning of their commercial production in the 1950s, PFAS have not only found their way into numerous industrial and commercial applications, but also into the bloodstream of much of the human population, the natural environment and wildlife. Exposure to high levels of PFAS poses a health risk for humans and animals, and may exacerbate the effects of other anthropogenic impacts faced by wildlife species. To gain a comprehensive overview of the abundance and distribution of PFAS research on wildlife species, and to better understand the drivers of this research, we will collate the available literature into a systematic evidence map and perform bibliometric analyses. The systematic mapping will present the distribution of research evidence that exists on PFAS in wildlife. The bibliometric analysis will provide an insight into the historical trends, interdisciplinarity, connectedness and the impact of the individual papers. We will conduct a systematic literature search on Scopus, Web of Science and 10 other databases using predefined search strings. We will screen title, abstract and keywords first. We will then screen full‐text papers. Two reviewers will be involved in the screening process. We will only consider publications in English, peer‐reviewed articles, preprints and theses. We will include papers reporting concentrations of any of 34 main PFAS types (based on a previous study) in wild animals. We will assess all publications included in the systematic map for predetermined indicators of quality and potential study‐level biases. In addition, we will use bibliometric records from Scopus to perform impact and network analyses. We will present the results using a narrative summary, tables and colour‐coded maps, bar and network plots. Results and associated database will be available on a dedicated freely accessible website. This study will provide critical insight into the gaps and clusters of the literature with regard to the PFAS concentrations in wildlife. Our study will inform and direct future research efforts to fill the gaps revealed.

ABSTRACT: Giardiasis is an important and prevalent zoonosis in dogs and humans caused by Giardia spp. The close relationship between pets and humans has physical, emotional and social benefits. The dogs have an important role in Giardia duodenalis cycle and transmission. This study aimed to verify the occurrence of the parasite in dogs from Central Region, in Santa Maria, Rio Grande do Sul State, Brazil, from April to October 2018. Dog feces (230) were submitted to Faust coproparasitological and molecular analyses. The positive samples in the nested-PCR (β-giardin gene) were sent for DNA sequencing and phylogenetic analyses (Neighbor-Joining). The occurrence of G. duodenalis, was 5.6% (13/230) and 4.3% (10/230) detected by coproparasitological technique and nested-PCR, respectively. There was no difference in the sensitivity of the tests used. From the faecal samples analyzed, there were no differences among the variables: diagnostic techniques, local, sex, and age of the animals (p>0.05). Only in the stool examination methodology a difference was observed between the ages (p

Per- and polyfluoroalkyl substances (PFAS) are found ubiquitously in wastewater treatment plants (WWTPs) due to their multiple sources in industry and consumer products. In Australia, limited spatial data are available on PFAS levels in WWTPs influent, while no temporal data have been reported. The aim of this study was to investigate the occurrence and temporal trend of PFAS in the influent of two large WWTPs in Australia (WWTP A and B) over a four-year period. Daily influent samples were collected over one week at different seasons from 2014 to 2017. Eleven perfluoroalkyl acids (PFAA) (i.e. seven perfluoroalkyl carboxylic acids (PFCAs) and four perfluoroalkyl sulfonic acids (PFSA)) were detected with mean Σ11PFAA concentrations of 57 ± 3.3–94 ± 17 ng/L at WWTP A, and 31 ± 6.1–142 ± 73 ng/L at WWTP B. The highest mean concentrations were observed for perfluorohexanoate (PFHxA) (20 ± 2 ng/L) in WWTP A, and perfluorooctane sulfonate (PFOS) (17 ± 13 ng/L) in WWTP B. The precursor 6:2 fluorotelomer sulfonate was detected over five sampling periods from Aug 2016 to Oct 2017, with mean concentrations of 37 ± 18–138 ± 51 ng/L for WWTP A and 8.8 ± 4.5–29 ± 5.1 ng/L for WWTP B. Higher concentration of 6:2 FTS (1.8–11 folds) than those of PFOA and PFOS in WWTP A indicate a likely substitution of C8 PFAA by fluorotelomer-based PFAS in this catchment. Temporal trends (annual and seasonal) in per-capita mass load were observed for some PFAA, increasing for PFPeA, PFHxA, PFHpA, PFNA, and PFHxS, while decreasing for PFBS and PFOS in either WWTPs. Notably, elevated levels of PFOS in October 2017 were observed at both WWTPs with the highest per capita mass load of up to 67 μg/day/inhabitant. For some PFAS release trends, longer sampling periods would be required to achieve acceptable statistical power. Keywords: Per- and polyfluoroalkyl substances (PFAS), 6:2 fluorotelomer sulfonate (6:2 FTS), Temporal trends, Wastewater treatment plant (WWTP), Influent, Power, Effect size

A criação de equinos geralmente está articulada como atividade secundária nas propriedades, nas quais os animais são utilizados para lazer ou trabalho. Dessa forma é importante considerar a sanidade desses animais, buscando entender quais os principais agentes que circulam em determinada propriedade ou região. Com isso, o presente trabalho teve por objetivo avaliar a presença de anticorpos IgG anti- Sarcocystis spp., anti-Toxoplasma gondii e anti-Neospora spp. em equinos na região Oeste de Santa Catarina. Foram analisadas 100 amostras de soro de equinos de 10 propriedades, localizadas nos municípios de Faxinal dos Guedes, Xaxim e Xanxerê, no período de junho a dezembro de 2019. As amostras foram avaliadas por Reação de Imunofluorescência Indireta (RIFI) para detecção de anticorpos da classe IgG contra os três agentes. Os resultados obtidos demonstram que os equinos da região apresentam soropositividade contra os três agentes estudados, sendo 11%, para Sarcocystis spp. 21%, para T. gondii e 20 %, para Neospora spp. Na análise da presença destes protozoários em diferentes ambientes, observou-se que tanto equinos estabulados ou criados de forma extensiva ambos apresentaram já terem entrado em contato com os agentes pesquisados. Animais que vivem apenas estabulados, apresentaram maior ocorrência de anticorpos IgG anti – Sarcocystis (7/32; 21,88%) e anti-T. gondii (34.38%-11/32). Quando avaliados os animais criados a pasto, de forma semi-extensiva e extensiva, houve um percentual maior de soropositividade para N. caninum (26.47% -9/34). A alta ocorrência de animais soropositivos aos protozoários estudados reforça a importância com o controle e prevenção desses agentes na região, uma vez que estes têm potencial para causar grande impacto na equideocultura, envolvendo principalmente as perdas reprodutivas, além da mieloencefalite protozoária equina (EPM).

Human bone marrow stromal cells (BMSC) are key elements of the hematopoietic environment and they play a central role in bone and bone marrow physiology. However, how key stromal cell functions are regulated is largely unknown. We analyzed the role of the immediate early response transcription factor EGR1 as key stromal cell regulator and found that EGR1 was highly expressed in prospectively-isolated primary BMSC, down-regulated upon culture, and low in non-colony-forming CD45neg stromal cells. Furthermore, EGR1 expression was lower in proliferative regenerating adult and fetal primary cells compared to adult steady-state BMSC. Overexpression of EGR1 in stromal cells induced potent hematopoietic stroma support as indicated by an increased production of transplantable CD34+CD90+ hematopoietic stem cells in expansion co-cultures. The improvement in bone marrow stroma support function was mediated by increased expression of hematopoietic supporting genes, such as VCAM1 and CCL28. Furthermore, EGR1 overexpression markedly decreased stromal cell proliferation whereas EGR1 knockdown caused the opposite effects. These findings thus show that EGR1 is a key stromal transcription factor with a dual role in regulating proliferation and hematopoietic stroma support function that is controlling a genetic program to co-ordinate the specific functions of BMSC in their different biological contexts.

Toxoplasma gondii is a protozoan that has great genetic diversity and is prevalent worldwide. In 2018, an outbreak of toxoplasmosis occurred in Santa Maria, Brazil, which was considered the largest outbreak ever described in the world. This paper describes the isolation and molecular characterization of Toxoplasma gondii from the placenta of two pregnant women with acute toxoplasmosis who had live births and were receiving treatment for toxoplasmosis during the outbreak. For this, placental tissue samples from two patients underwent isolation by mice bioassay, conventional PCR and genotyping using PCR-RFLP with twelve markers. Both samples were positive in isolation in mice. The isolate was lethal to mice, suggesting high virulence. In addition, the samples were positive in conventional PCR and isolates submitted to PCR-RFLP genotyping presented an atypical genotype, which had never been described before. This research contributes to the elucidation of this great outbreak in Brazil.