Mapping the risk of exposure to Crimean-Congo haemorrhagic fever virus in the Iberian Peninsula using Eurasian wild boar (Sus scrofa) as a model.

Crimean-Congo haemorrhagic fever (CCHF) virus (CCHFV) is a tick-borne zoonotic pathogen that can cause a lethal haemorrhagic disease in humans. Although the virus appears to be endemically established in the Iberian Peninsula, CCHF is an emerging disease in Spain. Clinical signs of CCHFV infection are mainly manifested in humans, but the virus replicates in several animal species. Understanding the determinants of CCHFV exposure risk from animal models is essential to predicting high-risk exposure hotspots for public health action. With this objective in mind, we designed a cross-sectional study of Eurasian wild boar (Sus scrofa) in Spain and Portugal. The study analysed 5,291 sera collected between 2006 and 2022 from 90 wild boar populations with a specific double-antigen ELISA to estimate CCHFV serum prevalence and identify the main determinants of exposure probability. To do so, we statistically modelled exposure risk with host- and environment-related predictors and spatially projected it at a 10 × 10 km square resolution at the scale of the Iberian Peninsula to map foci of infection risk. Fifty-seven (63.3 %) of the 90 populations had at least one seropositive animal, with seroprevalence ranging from 0.0 to 88.2 %. Anti-CCHFV antibodies were found in 1,026 of 5,291 wild boar (19.4 %; 95 % confidence interval: 18.3-20.5 %), with highest exposure rates in southwestern Iberia. The most relevant predictors of virus exposure risk were wild boar abundance, local rainfall regime, shrub cover, winter air temperature and soil temperature variation. The spatial projection of the best-fit model identified high-risk foci as occurring in most of western and southwestern Iberia and identified recently confirmed risk foci in eastern Spain. The results of the study demonstrate that serological surveys of CCHFV vector hosts are a powerful, robust and highly informative tool for public health authorities to take action to prevent human cases of CCHF in enzootic and emergency settings.

Occurrence and limited zoonotic potential of Cryptosporidium spp., Giardia duodenalis, and Balantioides coli infections in free-ranging and farmed wild ungulates in Spain.

Little information is currently available on the occurrence and molecular diversity of the enteric protozoan parasites Cryptosporidium spp., Giardia duodenalis, and Balantioides coli in wild ungulates and the role of these host species as potential sources of environmental contamination and consequent human infections. The presence of these three pathogens was investigated in eight wild ungulate species present in Spain (genera Ammotragus, Capra, Capreolus, Cervus, Dama, Ovis, Rupicapra, and Sus) by molecular methods. Faecal samples were retrospectively collected from free-ranging (n = 1058) and farmed (n = 324) wild ungulates from the five Spanish bioregions. Overall infection rates were 3.0% (42/1382; 95% CI: 2.1-3.9%) for Cryptosporidium spp., 5.4% (74/1382; 95% CI: 4.2-6.5%) for G. duodenalis, and 0.7% (9/1382; 95% CI: 0.3-1.2%) for B. coli. Cryptosporidium infection was detected in roe deer (7.5%), wild boar (7.0%) and red deer (1.5%), and G. duodenalis in southern chamois (12.9%), mouflon (10.0%), Iberian wild goat (9.0%), roe deer (7.5%), wild boar (5.6%), fallow deer (5.2%) and red deer (3.8%). Balantioides coli was only detected in wild boar (2.5%, 9/359). Sequence analyses revealed the presence of six distinct Cryptosporidium species: C. ryanae in red deer, roe deer, and wild boar; C. parvum in red deer and wild boar; C. ubiquitum in roe deer; C. scrofarum in wild boar; C. canis in roe deer; and C. suis in red deer. Zoonotic assemblages A and B were detected in wild boar and red deer, respectively. Ungulate-adapted assemblage E was identified in mouflon, red deer, and southern chamois. Attempts to genotype samples positive for B. coli failed. Sporadic infections by canine- or swine-adapted species may be indicative of potential cross-species transmission, although spurious infections cannot be ruled out. Molecular evidence gathered is consistent with parasite mild infections and limited environmental contamination with (oo)cysts. Free-ranging wild ungulate species would not presumably play a significant role as source of human infections by these pathogens. Wild ruminants do not seem to be susceptible hosts for B. coli.

New Earthworm Record from Division Muzaffarabad, Azad Kashmir, Pakistan Supported by Molecular Markers.

Earthworm diversity and ecology in Pakistan is poorly known, especially in the region of Azad Jammu & Kashmir. An earthworm community survey assisted by genetic barcoding detected an unidentified species which could constitute a new record for Pakistan. Morphological study revealed its identity as Perelia kaznakovi. Additionally, Bayesian phylogenetic inference based on five mitochondrial and nuclear molecular markers was performed. Results provided a phylogenetic placement of the genus Perelia within Lumbricidae for the first time, indicating a close relationship with Eophila. This approach should be implemented to Perelia arnoldiana and further representatives of the genus in order to understand their biogeography, diversity and evolutionary history.

Ovary micromorphology in hormogastrid earthworms with a particular emphasis on the organization of the germline cysts.

There is a gap in our knowledge of microorganization and the functioning of ovaries in earthworms (Crassiclitellata) and allied taxa. Recent analyses of ovaries in microdriles and leech-like taxa revealed that they are composed of syncytial germline cysts accompanied by somatic cells. Although the pattern of cyst organization is conserved across Clitellata - each cell is connected via one intercellular bridge (ring canal) to the central and anuclear cytoplasmic mass termed the cytophore - this system shows high evolutionary plasticity. In Crassiclitellata, only the gross morphology of ovaries and their segmental localization is well known, whereas ultrastructural data are limited to lumbricids like Dendrobaena veneta. Here we present the first report about ovarian histology and ultrastructure in Hormogastridae, a small family of earthworms inhabiting the western parts of the Mediterranean sea basin. We analyzed three species from three different genera and showed that the pattern of ovary organization is the same within this taxon. Ovaries are cone-like, with a broad part connected to the septum and a narrow distal end forming an egg string. Ovaries are composed of numerous cysts uniting a small number of cells, eight in Carpetania matritensis. There is a gradient of cysts development along the long ovary axis, and three zones can be distinguished. In zone I, cysts develop in complete synchrony and unite oogonia and early meiotic cells (till diplotene). Then (zone II), the synchrony is lost, and one cell (prospective oocyte) grows faster than the rest (prospective nurse cells). In zone III, oocytes pass the growth phase and gather nutrients; at this time, their contact with the cytophore is lost. Nurse cells grow slightly, eventually die via apoptosis, and are removed by coelomocytes. The most characteristic feature of hormogastrid germ cysts is the inconspicuous cytophore in the form of thread-like thin cytoplasmic strands (reticular cytophore). We found that the ovary organization in studied hormogastrids is very similar to that described for D. veneta and propose the term "Dendrobaena" type of ovaries. We expect the same microorganization of ovaries will be found in other hormogastrids and lumbricids.

Visualizing and evaluating wormholes formation dynamics under flow competition in an intermediate-scale dissolution experiment.

Wormholes are highly conductive channels that develop in high solubility rocks. They are especially important for environmental and industrial sustainability in saline karst aquifers (e.g. Salar de Uyuni, Salar de Atacama). Wormholes dynamics (i.e., the space and time evolution of these preferential flow paths) depends on the hydrodynamic and geochemical conditions during formation, as well as on wormholes competition for flow. Despite the importance of wormholes interaction for their development, experimental efforts have focused on the evolution of a single flow-path. Direct observation and quantification of wormholes dynamics is still lacking. We propose an experimental set-up to visualize and characterize the dynamics of multiple wormholes, which may help to understand the changes in flow and transport behaviour of aquifers. We performed a dissolution experiment in a 2D synthetic evaporitic aquifer, and simultaneous fluorescent tracer tests before and during wormhole growth. We visualized the growth by sequential photographs, with the fluorescent tracer highlighting the evolving structures. We quantified wormholes dynamics by measuring pressure and mass changes of the aquifer, and by image analysis. On the one hand, results show that wormholes tend to form along areas where flux was fastest prior to dissolution. They also show clear evidence of competition for water between wormholes and represent the first quantitative evidence of the amplifying factor that drives the self-organization in wormhole growth. On the other hand, we found that the competition is slower than predicted by current analytical and numerical theories, but consistent with other laboratory results. We conjecture that the discrepancy between theory and experiments can be attributed to the combined effect of non-linear kinetics and particle dragging during dissolution. We then compared experimental tracer breakthrough curves before and during the formation of preferential flow paths. They reflect wormhole growth by an accentuated non-Fickian behaviour, with reduced first arrival and increased tailing.

Human neutralizing antibodies to cold linear epitopes and to subdomain 1 of SARS-CoV-2

Emergence of SARS-CoV-2 variants diminishes the efficacy of vaccines and antiviral monoclonal antibodies. Continued development of immunotherapies and vaccine immunogens resilient to viral evolution is therefore necessary. Using coldspot-guided antibody discovery, a screening approach that focuses on portions of the virus spike that are both functionally relevant and averse to change, we identified human neutralizing antibodies to highly conserved viral epitopes. Antibody fp.006 binds the fusion peptide and cross-reacts against coronaviruses of the four genera, including the nine human coronaviruses, through recognition of a conserved motif that includes the S2 site of proteolytic cleavage. Antibody hr2.016 targets the stem helix and neutralizes SARS-CoV-2 variants. Antibody sd1.040 binds to subdomain 1, synergizes with antibody rbd.042 for neutralization and, like fp.006 and hr2.016, protects mice when present as bispecific antibody. Thus, coldspot-guided antibody discovery reveals donor-derived neutralizing antibodies that are cross-reactive with Orthocoronavirinae, including SARS-CoV-2 variants. One sentence summaryBroadly cross-reactive antibodies that protect from SARS-CoV-2 variants are revealed by virus coldspot-driven discovery.

Complementary roles of wild boar and red deer to animal tuberculosis maintenance in multi-host communities.

The contribution of wildlife species to pathogen maintenance in multi-host communities has seldom been quantified. To assess the relative contribution of the main wildlife hosts of animal tuberculosis (TB) to its maintenance, we estimated the basic reproduction number (R0) of Mycobacterium tuberculosis complex in wild boar and red deer at 29 sites in the Iberian Peninsula. Host abundance and true TB prevalence were estimated for each species at each site by sampling from distributions incorporating the uncertainty in the proportion of the population harvested each year, sensitivity, and specificity of the diagnostic methods, while excretion of mycobacteria was estimated using site-occupancy models. The distributions of these parameters were then used to estimate, at each site, the R0,wild boar (range 0.1 - 55.9, average 8.7, standard deviation 11.8), and the R0,red deer (0.1 - 18.9, 2.2, 3.9). Animal TB is maintained in epidemiological scenarios ranging from any single species acting as a maintenance host (the wild boar in 18 sites and the red deer in 5), to facultative multi-host disease (6 sites). The prevalence of TB in the red deer is likely an important driver of the epidemiology in multi-host communities. The wild boar was the main maintenance host of TB in most of the study sites and could have an epidemiological role linking the wildlife multi-host community and livestock.

Zoonotic Enterocytozoon bieneusi genotypes in free-ranging and farmed wild ungulates in Spain.

Microsporidia comprises a diverse group of obligate, intracellular, and spore-forming parasites that infect a wide range of animals. Among them, Enterocytozoon bieneusi is the most frequently reported species in humans and other mammals and birds. Data on the epidemiology of E. bieneusi in wildlife are limited. Hence, E. bieneusi was investigated in eight wild ungulate species present in Spain (genera Ammotragus, Capra, Capreolus, Cervus, Dama, Ovis, Rupicapra, and Sus) by molecular methods. Faecal samples were collected from free-ranging (n = 1058) and farmed (n = 324) wild ungulates from five Spanish bioregions. The parasite was detected only in red deer (10.4%, 68/653) and wild boar (0.8%, 3/359). Enterocytozoon bieneusi infections were more common in farmed (19.4%, 63/324) than in wild (1.5%, 5/329) red deer. A total of 11 genotypes were identified in red deer, eight known (BEB6, BEB17, EbCar2, HLJD-V, MWC_d1, S5, Type IV, and Wildboar3) and three novel (DeerSpEb1, DeerSpEb2, and DeerSpEb3) genotypes. Mixed genotype infections were detected in 15.9% of farmed red deer. Two genotypes were identified in wild boar, a known (Wildboar3) and a novel (WildboarSpEb1) genotypes. All genotypes identified belonged to E. bieneusi zoonotic Groups 1 and 2. This study provides the most comprehensive epidemiological study of E. bieneusi in Spanish ungulates to date, representing the first evidence of the parasite in wild red deer populations worldwide. Spanish wild boars and red deer are reservoir of zoonotic genotypes of E. bieneusi and might play an underestimated role in the transmission of this microsporidian species to humans and other animals.

The fungal-related intracellular parasite Enterocytozoon bieneusi is a worldwide public health and veterinary problem. Here we demonstrated that it was present in wild boar, and wild and farmed red deer in Spain, with genotypes potentially capable of infecting humans, posing a public health risk.

Calidad microbiològica de la carne picada y detección de patógenos en muestras ambientales de carnicerías de la ciudad de Tandil, provincia de Buenos Aires,Argentina / Microbiological quality of fresh ground beef and detection of pathogens in environmental samples taken from butcher shops in the city of Tandil, Buenos Aires Province, Argentina

Resumen Este estudio evaluó las condiciones higiénico-sanitarias de carnicerías de la ciudadde Tandil (provincia de Buenos Aires) mediante una estimación del riesgo basada en encuestasdirigidas a revisar las buenas prácticas de manufactura y de higiene de los establecimientos. Seutilizó una escala de 1 a 100 para clasificar a los establecimientos en las categorías de riesgoalto (0-40), riesgo moderado (41-70) y riesgo bajo (71-100). A su vez, se evaluó la presencia deSalmonella spp., Staphylococcus aureus, Escherichia coli productor de toxina Shiga (STEC) encarne bovina picada y en muestras ambientales como mesada, cuchilla, picadora y manos delcarnicero. Las muestras se tomaron una sola vez e inmediatamente se refrigeraron y transpor-taron al laboratorio para su análisis. En el período de estudio todas las carnicerías (100) fueronclasificadas como de «riesgo bajo¼ y con buenas condiciones higiénico-sanitarias. No obstante,el 75% de las muestras de carne picada no cumplió con al menos uno de los criterios microbiológicos establecidos en el Artículo 255 del Código Alimentario Argentino. Se sugiere estableceruna estrategia tendiente a identificar los desvíos e implementar un plan de mejoras continuasen las carnicerías de la ciudad de Tandil.

Abstract The aim of this work was to evaluate the hygienic-sanitary conditions of butcher shops in Tandil, Buenos Aires Province, by estimating the risk based on good manufacturing and hygiene practices, through surveys of the establishments. The analysis was performed using a scale of 1-100, and classifying them as high risk (0-40), moderate risk (41-70) or low risk (71-100). The presence of Salmonella spp., Staphylococcus aureus and Shiga toxin-producing Escherichia coli (STEC) from both, ground beef and environmental samples such as countertop, cleaver, mincer and butcher's hands, taken at butcher shops was also evaluated. Sampling was performed only once and immediately refrigerated and transported to the laboratory for analysis. All butcher shops evaluated (100) were classified as "low risk'' with good hygienic-sanitary conditions. However, 75% of the ground beef samples analyzed did not meet at least one of the microbiological criteria established in the Código Alimentario Argentino [Argentine Food Code], article 255. We propose to establish a strategy to identify deviations and implement a plan for continuous improvement in butcher shops of Tandil city.

The heat is on: a simple method to increase genome editing efficiency in plants.

BACKGROUND: Precision genome mutagenesis using CRISPR/Cas has become the standard method to generate mutant plant lines. Several improvements have been made to increase mutagenesis efficiency, either through vector optimisation or the application of heat stress. RESULTS: Here, we present a simplified heat stress assay that can be completed in six days using commonly-available laboratory equipment. We show that three heat shocks (3xHS) efficiently increases indel efficiency of LbCas12a and Cas9, irrespective of the target sequence or the promoter used to express the nuclease. The generated indels are primarily somatic, but for three out of five targets we demonstrate that up to 25% more biallelic mutations are transmitted to the progeny when heat is applied compared to non-heat controls. We also applied our heat treatment to lines containing CRISPR base editors and observed a 22-27% increase in the percentage of C-to-T base editing. Furthermore, we test the effect of 3xHS on generating large deletions and a homologous recombination reporter. Interestingly, we observed no positive effect of 3xHS treatment on either approach using our conditions. CONCLUSIONS: Together, our experiments show that heat treatment is consistently effective at increasing the number of somatic mutations using many CRISPR approaches in plants and in some cases can increase the recovery of mutant progeny.

Coordinated maintenance of H3K36/K27 methylation by histone demethylases preserves germ cell identity and immortality.

Germ cells have evolved unique mechanisms to ensure the transmission of genetically and nongenetically encoded information, whose alteration compromises germ cell immortality. Chromatin factors play fundamental roles in these mechanisms. H3K36 and H3K27 methyltransferases shape and propagate a pattern of histone methylation essential for C. elegans germ cell maintenance, but the role of respective histone demethylases remains unexplored. Here, we show that jmjd-5 regulates H3K36me2 and H3K27me3 levels, preserves germline immortality, and protects germ cell identity by controlling gene expression. The transcriptional and biological effects of jmjd-5 loss can be hindered by the removal of H3K27demethylases, indicating that H3K36/K27 demethylases act in a transcriptional framework and promote the balance between H3K36 and H3K27 methylation required for germ cell immortality. Furthermore, we find that in wild-type, but not in jmjd-5 mutants, alterations of H3K36 methylation and transcription occur at high temperature, suggesting a role for jmjd-5 in adaptation to environmental changes.

Prdm16-mediated H3K9 methylation controls fibro-adipogenic progenitors identity during skeletal muscle repair.

H3K9 methylation maintains cell identity orchestrating stable silencing and anchoring of alternate fate genes within the heterochromatic compartment underneath the nuclear lamina (NL). However, how cell type-specific genomic regions are specifically targeted to the NL is still elusive. Using fibro-adipogenic progenitors (FAPs) as a model, we identified Prdm16 as a nuclear envelope protein that anchors H3K9-methylated chromatin in a cell-specific manner. We show that Prdm16 mediates FAP developmental capacities by orchestrating lamina-associated domain organization and heterochromatin sequestration at the nuclear periphery. We found that Prdm16 localizes at the NL where it cooperates with the H3K9 methyltransferases G9a/GLP to mediate tethering and silencing of myogenic genes, thus repressing an alternative myogenic fate in FAPs. Genetic and pharmacological disruption of this repressive pathway confers to FAP myogenic competence, preventing fibro-adipogenic degeneration of dystrophic muscles. In summary, we reveal a druggable mechanism of heterochromatin perinuclear sequestration exploitable to reprogram FAPs in vivo.

Bead-assisted SARS-CoV-2 multi-antigen serological test allows effective identification of patients

Many new aspects of COVID-19 disease, including different clinical manifestations, have been identified during the pandemic. The wide array of symptoms and variation in disease severity after SARS-CoV-2 infection might be related to heterogeneity in the immune responses of different patients. Here we describe a new method for a simple multi-antigen serological test that generates a full picture of seroconversion in a single reaction. The assay is based on the detection by flow cytometry of multiple immunoglobulin classes (isotypes) specific for four SARS-CoV-2 antigens: the Spike glycoprotein (one of the highly immunogenic proteins), its RBD fragment (the major target for neutralising antibodies), the nucleocapsid protein and the main cysteine-like protease. Until now, most diagnostic serological tests measured antibodies to only one antigen and some patients seemed to not make any antibody response. Our data reveal that while most patients respond against all the viral antigens tested, others show a marked bias to make antibodies against either proteins exposed on the viral particle or those released after cellular infection. Combining all the four antigens and using machine learning techniques, it was possible to clearly discriminate between patients and healthy controls with 100% confidence. Further, combination of antigens and different immunoglobulin isotypes in this multi-antigen assay improved the classification of patients with mild and severe disease. Introduction of this method will facilitate massive screenings of patients to evaluate their immune response. It could also support vaccination campaigns both to select non-immune individuals and to distinguish infected patients from vaccine responders.

COVID-19: Effectiveness of Non-Pharmaceutical Interventions in the United States before Phased Removal of Social Distancing Protections Varies by Region

Although coronavirus disease 2019 (COVID-19) emerged in January 2020, there is no quantified effect size for non-pharmaceutical interventions (NPI) to control the outbreak in the continental US. Objective. To quantify national and sub-national effect sizes of NPIs in the US. Design. This is an observational study for which we obtained daily county level COVID-19 cases and deaths from January 22, 2020 through the phased removal of social distancing protections. A stepped-wedge cluster-randomized trial (SW-CRT) analytical approach is used, leveraging the phased implementation of policies. Data include 3142 counties from all 50 US states and the District of Columbia. Exposures. County-level NPIs were obtained from online county and state policy databases, then classified into four intervention levels: Level 1 (low) - declaration of a State of Emergency; Level 2 (moderate) - school closures, restricting nursing home access, or closing restaurants and bars; Level 3 (high) - non-essential business closures, suspending non-violent arrests, suspending elective medical procedures, suspending evictions, or restricting mass gatherings of at least 10 people; and Level 4 (aggressive) - sheltering in place / stay-at-home, public mask requirements, or travel restrictions. Additional county-level data were obtained to record racial (Black, Hispanic), economic (educational level, poverty), demographic (rural/urban) and climate factors (temperature, specific humidity, solar radiation). Main Outcomes. The primary outcomes are rates of COVID-19 cases, deaths and case doubling times. NPI effects are measured separately for nine US Census Region (Pacific, Mountain, West North Central, East North Central, West South Central, East South Central, South Atlantic, Middle Atlantic, New England). Results. Aggressive NPIs (level 4) significantly reduced COVID-19 case and death rates in all US Census Regions, with effect sizes ranging from 4.1% to 25.7% and 5.5% to 25.5%, respectively, for each day they were active. No other intervention level achieved significance across all US Regions. Intervention levels 3 and 4 both increased COVID-19 doubling times, with effects peaking at 25 and 40 days after initiation of each policy, respectively. The effectiveness of level 3 NPIs varied, reducing case rates in all regions except North Central states, but associated with significantly higher death rates in all regions except Pacific states. Intervention levels 1 and 2 did not indicate any effect on COVID-19 propagation and, in some regions, these interventions were associated with increased COVID-19 cases and deaths. Heterogeneity of NPI effects are associated with racial composition, poverty, urban-rural environment, and climate factors. Conclusion. Aggressive NPIs are effective tools to reduce COVID-19 propagation and mortality. Reducing social and environmental disparities may improve NPI effects in regions where less strict policies are in place.

SARS-Cov-2 cysteine-like protease (Mpro) is immunogenic and can be detected in serum and saliva of COVID-19-seropositive individuals

Currently, there is a need for reliable tests that allow identification of individuals that have been infected with SARS-CoV-2 even if the infection was asymptomatic. To date, the vast majority of the serological tests for SARS-CoV-2 specific antibodies are based on serum detection of antibodies to either the viral spike glycoprotein (the major target for neutralising antibodies) or the viral nucleocapsid protein that are known to be highly immunogenic in other coronaviruses. Conceivably, exposure of antigens released from infected cells could stimulate antibody responses that might correlate with tissue damage and, hence, they may have some value as a prognostic indicator. We addressed whether other non-structural viral proteins, not incorporated into the infectious viral particle, specifically the viral cysteine-like protease, might also be potent immunogens. Using ELISA tests, coating several SARS-CoV-2 proteins produced in vitro, we describe that COVID-19 patients make high titre IgG, IgM and IgA antibody responses to the Cys-like protease from SARS-CoV-2, also known as 3CLpro or Mpro, and it can be used to identify individuals with positive serology against the coronavirus. Higher antibody titres in these assays associated with more severe disease and no cross-reactive antibodies against prior betacoronavirus were found. Remarkably, IgG antibodies specific for Mpro and other SARS-CoV-2 antigens can also be detected in saliva. In conclusion, Mpro is a potent antigen in infected patients that can be used in serological tests and its detection in saliva could be the basis for a rapid, non-invasive test for COVID-19 seropositivity.

Local adaptation fuels cryptic speciation in terrestrial annelids.

Uncovering the genetic and evolutionary basis of cryptic speciation is a major focus of evolutionary biology. Next Generation Sequencing (NGS) allows the identification of genome-wide local adaptation signatures, but has rarely been applied to cryptic complexes - particularly in the soil milieu - as it is the case with integrative taxonomy. The earthworm genus Carpetania, comprising six previously suggested putative cryptic lineages, is a promising model to study the evolutionary phenomena shaping cryptic speciation in soil-dwelling lineages. Genotyping-By-Sequencing (GBS) was used to provide genome-wide information about genetic variability between 17 populations, and geometric morphometrics analyses of genital chaetae were performed to investigate unexplored cryptic morphological evolution. Genomic analyses revealed the existence of three cryptic species, with half of the previously-identified potential cryptic lineages clustering within them. Local adaptation was detected in more than 800 genes putatively involved in a plethora of biological functions (most notably reproduction, metabolism, immunological response and morphogenesis). Several genes with selection signatures showed shared mutations for each of the cryptic species, and genes under selection were enriched in functions related to regulation of transcription, including SNPs located in UTR regions. Finally, geometric morphometrics approaches partially confirmed the phylogenetic signal of relevant morphological characters such as genital chaetae. Our study therefore unveils that local adaptation and regulatory divergence are key evolutionary forces orchestrating genome evolution in soil fauna.

Primary malignant melanoma of the biliary tract: A case report and literature review.

BACKGROUND: Primary malignant melanoma of the biliary tract (MBT) is a rare condition whose diagnosis requires excluding a primary origin in another location. This paper reviews the most important characteristics of MBT cases published in the literature and reports a new case. The patient reported here is the first case of primary malignant melanoma of the biliary tract with pulmonary metastasis treated with immunotherapy. This patient remains disease-free 36 mo after the treatment of metastatic lung lesions. CASE SUMMARY: A 51-year-old man was admitted to the gastrointestinal department to study obstructive jaundice of a 1 wk clinical course. Magnetic resonance cholangiopancreatography revealed dilatation of the intrahepatic biliary tract and stenosis of the common hepatic duct. Given the suspicion of biliary tract neoplasia, cholecystectomy and resection of the common hepatic duct were performed with hepatic jejunostomy free of complications. Anatomo-pathological diagnosis was melanoma. After intervention, the patient was referred to the Department of Medical Oncology, where a primary origin was excluded in the skin, mucosa, and eyes. This confirmed diagnosis of primary biliary tract melanoma. Computed tomography was performed 12 mo after the procedure revealed several subcentimetric lung nodules. Wedge resection was performed. After confirming the diagnosis of pulmonary metastasis of primary melanoma of the biliary tract, the patient was started on immunotherapy with nivolumab. Tolerance to treatment was excellent. The patient remains disease-free 36 mo after the treatment of metastatic lung lesions. CONCLUSION: The patient reported here is the first case of primary malignant melanoma of the biliary tract with lung metastases successfully treated with immunotherapy.

Análisis histórico tendencial del desarrollo de la enseñanza aprendizaje del inglés con fines médicos en Cuba / Historical-trend analysis of the development of English teaching learning process for medical purposes in Cuba

RESUMEN La enseñanza-aprendizaje del inglés con fines médicos ha experimentado cambios metodológicos ante la necesidad de formar profesionales de la salud altamente competitivos ante las exigencias de la colaboración médica internacional. El presente trabajo tiene como objetivo realizar un análisis histórico-tendencial del desarrollo de la enseñanza-aprendizaje del inglés con fines médicos en Cuba. En la investigación se asumió la dialéctica materialista como método general de la ciencia. Se utilizaron métodos teóricos, empíricos y matemático-estadísticos que posibilitaron el conocimiento de la evolución y desarrollo del objeto de investigación para revelar su historia, las etapas principales en su desarrollo y las conexiones históricas fundamentales, así como el análisis crítico de la información contentiva en documentos normativos y materiales sobre la temática. Se estableció la existencia de tres etapas fundamentales en el desarrollo de su enseñanza y aprendizaje determinadas por las exigencias didácticas y los sustentos teóricos asumidos.

ABSTRACT The teaching-learning process of English for medical purposes has undergone methodological changes in view of the need to train highly competitive health professionals in the face of the demands of international medical collaboration. The objective of this work is to perform a historical-trend analysis of the development of teaching-learning process of English for medical purposes in Cuba. In the investigation, the materialist dialectic was assumed as the general method of science. Theoretical, empirical and mathematical-statistical methods were used that allowed the knowledge of the evolution and development of the research object to reveal its history, the main stages in its development and the fundamental historical connections, as well as the critical analysis of the content information in normative documents and materials on the subject. It was established the existence of three fundamental stages in the development of their teaching and learning determined by the didactic demands and the assumed theoretical foundations.

Mineral profile exploratory analysis for rice grains traceability.

Rice geographical traceability requires analytical procedures and data evaluation capable of linking its composition to the producing area. In this work, major and trace elements in soil and rice grains and husk from 9 cities and 17 producers were evaluated. Arsenic species were measured solely in rice grains. The rice mineral profile evaluated by principal component analysis allowed the identification of controlling variables and origin fingerprints. Vectors controlling data variability were linked to the geographical area, to crop management, producers and in a lower extent to soil composition. Elemental discrimination through 3D models was proposed. Arsenic species in the grains and elemental husk composition were decisive to achieve the required discrimination. Rice discrimination was obtained by cities, producers and varieties. The present work model was compared with others from similar studies.