2b-RAD genotyping for population genomic studies of Chagas disease vectors: Rhodnius ecuadoriensis in Ecuador

Background: Rhodnius ecuadoriensis is the main triatomine vector of Chagas disease, American trypanosomiasis, in Southern Ecuador and Northern Peru. Genomic approaches and next generation sequencing technologies have become powerful tools for investigating population diversity and structure which is a key consideration for vector control. Here we assess the effectiveness of three different 2b restriction site-associated DNA (2b-RAD) genotyping strategies in R. ecuadoriensis to provide sufficient genomic resolution to tease apart microevolutionary processes and undertake some pilot population genomic analyses. Methodology/Principal findings: The 2b-RAD protocol was carried out in-house at a non-specialized laboratory using 20 R. ecuadoriensis adults collected from the central coast and southern Andean region of Ecuador, from June 2006 to July 2013. 2b-RAD sequencing data was performed on an Illumina MiSeq instrument and analyzed with the STACKS de novo pipeline for loci assembly and Single Nucleotide Polymorphism (SNP) discovery. Preliminary population genomic analyses (global AMOVA and Bayesian clustering) were implemented. Our results showed that the 2b-RAD genotyping protocol is effective for R. ecuadoriensis and likely for other triatomine species. However, only BcgI and CspCI restriction enzymes provided a number of markers suitable for population genomic analysis at the read depth we generated. Our preliminary genomic analyses detected a signal of genetic structuring across the study area. Conclusions/Significance: Our findings suggest that 2b-RAD genotyping is both a cost effective and methodologically simple approach for generating high resolution genomic data for Chagas disease vectors with the power to distinguish between different vector populations at epidemiologically relevant scales. As such, 2b-RAD represents a powerful tool in the hands of medical entomologists with limited access to specialized molecular biological equipment. Author summary: Understanding Chagas disease vector (triatomine) population dispersal is key for the design of control measures tailored for the epidemiological situation of a particular region. In Ecuador, Rhodnius ecuadoriensis is a cause of concern for Chagas disease transmission, since it is widely distributed from the central coast to southern Ecuador. Here, a genome-wide sequencing (2b-RAD) approach was performed in 20 specimens from four communities from Manabí (central coast) and Loja (southern) provinces of Ecuador, and the effectiveness of three type IIB restriction enzymes was assessed. The findings of this study show that this genotyping methodology is cost effective in R. ecuadoriensis and likely in other triatomine species. In addition, preliminary population genomic analysis results detected a signal of population structure among geographically distinct communities and genetic variability within communities. As such, 2b-RAD shows significant promise as a relatively low-tech solution for determination of vector population genomics, dynamics, and spread

Identification of Lynch Syndrome Carriers among Patients with Small Bowel Adenocarcinoma

Lynch syndrome; Hereditary cancer; Small bowel adenocarcinomaSíndrome de Lynch; Cáncer hereditario; Adenocarcinoma de intestino delgadoSíndrome de Lynch; Càncer hereditari; Adenocarcinoma d'intestí primBackground: Small bowel adenocarcinoma (SBA) is a rare disease which can be associated with Lynch syndrome (LS). LS tumors are characterized by the presence of microsatellite instability (MSI) and/or the loss of mismatch repair (MMR) protein expression. In SBA, the frequency of MMR deficient (MMRd) tumors varies from 5% to 35%. This study aims to describe the prevalence of LS carriers among patients with MMRd small bowel adenocarcinomas. Methods: A multicenter retrospective study with identification and MMR testing of all consecutive SBA between 2004 and 2020 in a multicenter Spanish study. Demographical data, tumor characteristics, follow-up and survival information were collected. Germline testing was driven by identification of MMRd tumors. Results: A total of 94 individuals diagnosed with SBA were recruited. We observed 20 (21.3%) MMRd tumors. In 9/15 (60%) patients with MMRd tumors, a pathogenic variant was identified (three MLH1, four MSH2, one MSH6 and one PMS2). Accordingly, the prevalence of LS among all SBA cases was 10.1%. Conclusions: More than one-fifth of SBA display MMRd and in more than a half is due to LS. Our data supports the implementation of universal MMR tumor testing among SBA for the identification of LS families.This study was funded by the CERCA Program (Generalitat de Catalunya) and Agència de Gestió d’Ajuts Universitaris i de Recerca (Generalitat de Catalunya, GRPRE 2017SGR21, GRC 2017SGR653). CIBEREHD is funded by the Instituto de Salud Carlos III

StringENT test suite: ENT battery revisited for efficient P value computation

Random numbers play a key role in a wide variety of applications, ranging from mathematical simulation to cryptography. Generating random or pseudo-random numbers is not an easy task, especially when hardware, time and energy constraints are considered. In order to assess whether generators behave in a random fashion, there are several statistical test batteries. ENT is one of the simplest and most popular, at least in part due to its efficacy and speed. Nonetheless, only one of the tests of this suite provides a p value, which is the most useful and standard way to determine whether the randomness hypothesis holds, for a certain significance level. As a consequence of this, rather arbitrary and at times misleading bounds are set in order to decide which intervals are acceptable for its results. This paper introduces an extension of the battery, named StringENT, which, while sticking to the fast speed that makes ENT popular and useful, still succeeds in providing p values with which sound decisions can be made about the randomness of a sequence. It also highlights a flagrant randomness flaw that the classical ENT battery is not capable of detecting but the new StringENT notices, and introduces two additional tests.Depto. de Estadística e Investigación OperativaFac. de Ciencias MatemáticasTRUEpu

Ileal apparent and standardized amino acid digestibility of soybean and colza meal in diets for finishing pigs

Agri-food chains generate by-products such as soybean and colza meal for animal feed. Their nutritional value is variable and should be analysed for better quality control before their inclusion in balanced diets. The objective was to determine the content and ileal apparent and standardized digestibility of protein and amino acids (AA) of soybean (PS) and colza meal (PC) as a source of protein in diets for finishing pigs. The experimental units were arranged in a 3×3 Latin square design repeated. The treatments (T) were T1: PS+corn starch, T2: PC+corn starch, and T3: corn starch (Control), which were randomly assigned to six pigs cannulated in distal ileum (PV 75±1.2 kg). The variables were contents and apparent ileal and standardized ileal digestibility of amino acids in soybean and colza meal. To determine digestibility, chromic oxide was added to the diets. The protein and fat contents were higher in the soybean meal. Ash, crude fibre, neutral detergent fibre, acid detergent fibre, phosphorus and calcium contents were higher in the colza meal. Essential amino acid content was higher in soybean meal, but methionine was higher in colza meal. Apparent ileal digestibility (DIA) of total amino acids was similar (p > 0.05) among protein ingredients; but for lysine and threonine it was higher (p ≤ 0.05) in soybean meal, except methionine. Methionine DIA was 89.78 %, 3.88 % higher (p ≤ 0.05) in colza meal. Standardized ileal digestibility (DIE) of total amino acids was similar (p > 0.05) among ingredients, but lysine and threonine were higher (p ≤ 0.05) in soybean meal. DIE was different (p ≤ 0.05) among ingredients for all amino acids except methionine. Ileal and standardized digestibility of all amino acids were higher in soybean meal, except methionine, which was higher in colza meal. The DIA of total amino acids was similar in both protein ingredients, except lysine and threonine in soybean meal and methionine in colza meal

Diagnostic challenges of brucellosis in humans and livestock in Tanzania: A thematic review

This research article was published by CABI One Health, in 2023Brucellosis is an endemic bacterial zoonosis in Tanzania, and is among the most prioritized zoonotic diseases in the country. Brucellosis affects public health and livestock production in developing countries. Most human and livestock cases are not detected by the existing surveillance systems resulting in significant underestimation of the disease burden, and poor management of human cases by using nonspecific antibiotics may potentially contribute to antimicrobial resistance. To quantify the factors related to underreporting and those associated with the challenges in the diagnosis of brucellosis in Tanzania, search terms including “Brucella” “diagnosis” and “challenges” were used to query in Google search engine and publisher databases such as MEDLINE, PUBMED, NCBI, Springer, Hindawi, and Elsevier. The search parameters were limited to publications between 1995 and 2020. The searches returned 319 publications and grey articles which were screened and 57 were eligible for inclusion in this study. Four main areas were identified that cause underreporting of brucellosis and hinder brucellosis diagnosis: (1) inadequate knowledge of brucellosis among stakeholders in the livestock value chain, (2) limited diagnostic capacity for brucellosis due to unawareness of diagnostic tests and lack of epidemiological background of brucellosis among human and livestock health service workers (3) challenges associated with diagnostic tests, which include unreliable availability of diagnostic tests and unskilled workers, and (4) the uneven distribution of brucellosis surveillance studies in the country. This study suggests that there is a need for (1) training on public health education and brucellosis awareness among stakeholders in the livestock value chain; (2) providing scheduled continuing professional education with regard to brucellosis and other zoonotic diseases to health and livestock workers; (3) future brucellosis surveillance studies must focus on unrepresented regions; and (4) lastly, we recommend that the rose Bengal plate test (RBPT) and competitive enzyme-linked immunosorbent assay (cELISA) should be considered in brucellosis diagnostic schemes as a complementary tool to hasten the implementation of an ongoing national strategy for the prevention and control of brucellosis in humans and livestock in Tanzania. We suggest that these recommendations be considered for inclusion in the national strategy for brucellosis control in Tanzania. One Health Impact Statement Brucellosis is a disease caused by bacteria of genus Brucella and is transmitted from animals to humans. Humans get infected through ingestion of infected animal products but also through contact with bacteria via broken skin or inhalation of aerosolized bacteria particles during culture in the laboratory. Humans working with animals or animal products and laboratory personnel are at higher risk of infection. Tanzania has recently established a One-Health desk in the Prime Minister’s Office to address all matters related to One Health. However, the desk is in the infancy stage, more studies must be done to fill knowledge gaps so that working instruments became realistic. This review provides a synthesis of information that could be used by the One Health desk and other One Health stakeholders in the country on how to improve the existing brucellosis surveillance structures for improvement of One-Health service delivery in the country

Concepciones acerca de la Evaluacion Sumativa en Estudiantes de Tercero Primaria en las Instituciones Leon XIII y Normal Superior

This article investigates the conceptions of summative evaluation in the teaching-learning process from the voices of students in third grade of primary school in two institutions; Addressing exploratory-descriptive methodology, applying instruments of: participant observation, interview and focus groups, the imaginaries and conceptions of summative evaluation were analyzed in a sample of 63 students. In the institutions, students recognized summative evaluation, a perception that helps to value learning at the end of the two-month period as a tool for improvement. It is manifested to involve families with a previous preparation at the time of carrying out the evaluations, the students perceive the bimonthly evaluations as an evaluation of everything learned during the school year implying that they review the topics seen and put into operation the thinking skills; to process information based on their knowledge. The perceptions of the students were known to take them into account by the teachers, in the design and development of summative evaluations, because knowing these appreciations achieves improvements in the results and opens the possibility of renewing the teaching and learning process so that it adjusts to the institutional needs from the integrality of each individual.El presente artículo indaga sobre las concepciones de la evaluación sumativa en el proceso de enseñanza-aprendizaje desde las voces de estudiantes en tercer grado de básica primaria en dos Instituciones; abordando metodología exploratoria-descriptiva, aplicando instrumentos de: observación participante, la entrevista y grupos focales, se analizaron los imaginarios y concepciones de la evaluación sumativa en una muestra de 63 estudiantes. En las instituciones se evidencio por parte de los estudiantes, reconocimiento de la evaluación sumativa, una percepción que ayuda a valorar los aprendizajes al final del bimestre como herramienta de mejoramiento. Se manifiesta involucrar a las familias con una preparación previa al momento de realizar las evaluaciones, los estudiantes perciben las evaluaciones bimestrales como una evaluación de todo lo aprendido durante el año escolar implicando que repasen los temas vistos y poner en funcionamiento las habilidades de pensamiento; para procesar la información a partir de sus conocimientos.  Se conoció las percepciones de los estudiantes para tenerlas en cuenta por parte de los docentes, en el diseño y desarrollo de evaluaciones sumativas, pues conociendo estas apreciaciones se logra mejoras en los resultados y abre la posibilidad de renovar el proceso de enseñanza y aprendizaje de manera que se ajuste a las necesidades institucionales desde la integralidad de cada individuo

Effect of Boriding on the Mechanical Properties of AISI 1045 Steel

Efecto en las propiedades mecánicas de un acero borurado 1045.Some mechanical properties of AISI 1045 borided steels were estimated in the present work. The boriding process was carried out by the powder pack method at 950°C with 8 h of treatment. The fatigue strength on borided notched specimens was evaluated with rotating bending tests (R=1) considering a stress concentration factor (Kt) of 2.53. Likewise, the presence of residual stresses in boride layers was established by the XRD technique. The Daimler-Benz Rockwell C test was used, also, to estimate the strength adhesion of the coated system. The results show a decrease in the fatigue strength of AISI borided steels due to the presence of high porosity in the layers. Finally, the Rockwell-C adhesion test showed no coating failure for the boride layer