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Low genome‐wide homozygosity in 11 Spanish ovine breeds
No full text[EN] The population of Spanish sheep has decreased from 24 to 15 million heads in the last 75 years due to multiple social and economic factors. Such a demographic reduction might have caused an increase in homozygosity and inbreeding, thus limiting the viability of local breeds with excellent adaptations to harsh ecosystems. The main goal of our study was to investigate the homozygosity patterns of 11 Spanish ovine breeds and to elucidate the relationship of these Spanish breeds with reference populations from Europe, Africa and the Near East. By using Ovine SNP50 BeadChip data retrieved from previous publications, we have found that the majority of studied Spanish ovine breeds have close genetic relatedness with other European populations; the one exception is the Canaria de Pelo breed, which is similar to North African breeds. Our analysis has also demonstrated that, with few exceptions, the genomes of Spanish sheep harbor fewer than 50 runs of homozygosity (ROH) with a total length of less than 350 Mb. Moreover, the frequencies of very long ROH (>30 Mb) are very low, and the inbreeding coefficients (FROH) are generally small (FROH < 0.10), ranging from 0.008 (Rasa Aragonesa) to 0.086 (Canaria de Pelo). The low levels of homozygosity observed in the 11 Spanish sheep under analysis might be due to their extensive management and the high number of small to medium farmsSIThanks to Jorge Hugo Calvo from the Centro de Investigación y Tecnología Agroalimentaria de Aragón for providing 50K data from several sheep breeds and criticallyrevising the manuscript. This research was partially fundedby a grant (RZ2011-00015-C03-01) from the Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (Spain). We acknowledge the support of the Spanish Ministerio de Economía y Competitividad for the Centro deExcelencia Severo Ochoa 2016–2019 award SEV-2015-0533 and the CERCA Programme of the Generalitat deCatalunya. Many thanks to the International SheepGenomics Consortium and especially to Dr. Hans Lenstrafor kindly providing ovine 50K data. Taina F Cardoso wasfunded with a fellowship from the CAPES Foundation Coordination of Improvement of Higher Education, Ministry of Education (MEC) of the Federal Government of Brazil. Theauthors are grateful to the Canaria de Pelo, Gallega Ripollesa, Roja Mallorquina and Xisqueta breeding associations for their collaboration in sample collectio
Relación de la fuerza muscular y el rendimiento en ejercicios de halterofilia y gimnásticos en practicantes amateur de CrossFit®
Get PDF[EN] The study focused on replicating and expanding the research conducted by Tibana et al. (2016). It analysed the relationship between strength in Back Squat, Front Squat, and Pull Up exercises and their influence on the performance of specific activities such as Snatch, Clean & Jerk, and Kipping Pull Up. Twenty-two subjects participated in the study, who were divided into two groups based on their performance in the Snatch and Clean & Jerk exercises: high strength group (age, 33.36 ± 6.31 years; body mass, 84.50 ± 10.36 kg; and height, 1.77 ± 0.03 cm) and low strength group (age, 35.35 ± 10.29 years; body mass, 70.97 ± 16.81 kg; and height, 1.73 ± 0.13 cm). All subjects performed a 1 repetition maximum (1RM) test of Back Squat, Front Squat, Pull Up, Snatch, and Clean & Jerk and the maximum repetitions test of Kipping Pull Up. Significant differences (p≤0.05) were observed between the groups in body mass, body mass index, practice time, and training hours. In addition, significant differences (p≤0.05) were detected in the maximum repetitions of all the exercises performed, with the exception of the Kipping Pull Up (p>0.05). Correlations (p≤0.01) were found between most of the values achieved in the different exercises. However, the Kipping Pull Up exercise was only related to the results found in the Pull Up (p0.05). Se encontraron correlaciones (p≤0.01) entre la mayoría de los valores alcanzados en los diferentes ejercicios. Sin embargo, el ejercicio de Kipping Pull Up únicamente se relacionó con los resultados hallados en el Pull Up(p<0,05). Los participantes con mayores valores de fuerza en Back Squat, Front Squaty Pull Uppresentaron un mejor rendimiento en Snatch, Clean & Jerky Kipping Pull UpS
Combining GWAS and RNA-Seq Approaches for Detection of the Causal Mutation for Hereditary Junctional Epidermolysis Bullosa in Sheep
Get PDF[EN] In this study, we demonstrate the use of a genome-wide association mapping together with RNA-seq in a reduced number of samples, as an efficient approach to detect the causal mutation for a Mendelian disease. Junctional epidermolysis bullosa is a recessive genodermatosis that manifests with neonatal mechanical fragility of the skin, blistering confined to the lamina lucida of the basement membrane and severe alteration of the hemidesmosomal junctions. In Spanish Churra sheep, junctional epidermolysis bullosa (JEB) has been detected in two commercial flocks. The JEB locus was mapped to Ovis aries chromosome 11 by GWAS and subsequently fine-mapped to an 868-kb homozygous segment using the identical-by-descent method. The ITGB4, which is located within this region, was identified as the best positional and functional candidate gene. The RNA-seq variant analysis enabled us to discover a 4-bp deletion within exon 33 of the ITGB4 gene (c.4412-4415del). The c.4412-4415del mutation causes a frameshift resulting in a premature stop codon at position 1472 of the integrin β4 protein. A functional analysis of this deletion revealed decreased levels of mRNA in JEB skin samples and the absence of integrin β4 labeling in immunohistochemical assays. Genotyping of c.4412-4415del showed perfect concordance with the recessive mode of the disease phenotype. Selection against this causal mutation will now be used to solve the problem of JEB in flocks of Churra sheep. Furthermore, the identification of the ITGB4 mutation means that affected sheep can be used as a large mammal animal model for the human form of epidermolysis bullosa with aplasia cutis. Our approachevidences that RNA-seq offers cost-effective alternative to identify variants in the species in which high resolution exome-sequencing is not straightforwardSIFunded by Spanish Ministry of Economy and Competitiveness (grant AGL2012-34437 to JJA); Aroa Suarez-Vega is funded by an FPU contract from Spanish Ministry of Education. Beatriz Guterrez-Gil is funded by an Ramon y Cajal contract from the Spanish Ministry of Economy and Competitiveness. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip
Characterization and antimicrobial resistance of Spanish isolates of "Glaesserella parasuis". Evaluation of the efficacy of a vaccine against Glässer's disease based on a modified TbpB (transferrin-binding protein)
No full text120 p.[ES] La industria porcina enfrenta importantes desafíos económicos, siendo la enfermedad de Glässer uno de los más relevantes. Esta enfermedad está causada por Glaesserella (Haemophilus) parasuis, patógeno que coloniza el tracto respiratorio superior de los cerdos y, en condiciones de estrés como el destete o el transporte, puede desencadenar infecciones sistémicas graves. Los lechones en las primeras etapas de lactancia y engorde son especialmente vulnerables, lo que genera altos índices de morbilidad y mortalidad. Esta bacteria adquiere hierro de la transferrina porcina mediante un receptor de superficie compuesto por las proteínas de unión a la transferrina TbpA y TbpB, lo que facilita su supervivencia en entornos con baja disponibilidad de hierro, como las superficies mucosas.
Sin embargo, los mecanismos de virulencia de esta bacteria aún no se comprenden completamente y el desafío se agrava por el preocupante aumento de la resistencia a los antimicrobianos, impulsado por su uso intensivo en la producción porcina. Este doble problema pone de manifiesto la urgencia de profundizar en la investigación sobre los factores de patogenicidad de G. parasuis y de desarrollar estrategias que permitan reducir el uso de estos medicamentos sin comprometer el control de la enfermedad. Este fenómeno compromete la efectividad de los tratamientos y plantea un riesgo adicional de transmisión de genes de resistencia a otras bacterias en el entorno. Sin embargo, ante la ausencia de una vacuna universal comercialmente disponible para prevenir esta enfermedad, el tratamiento con antimicrobianos sigue siendo la principal estrategia de control, a pesar de los desafíos asociados. Esto subraya la necesidad de desarrollar herramientas de diagnóstico precisas, fomentar un uso responsable de los antimicrobianos y promover investigaciones que permitan avances en el diseño de vacunas eficaces y de amplio espectro.
El objetivo principal de esta tesis doctoral es realizar una caracterización detallada de la virulencia y los perfiles de resistencia a antimicrobianos de aislados de G. parasuis presentes en explotaciones porcinas españolas. Paralelamente, se busca analizar la diversidad genética de la proteína TbpB en estos aislados, con el propósito de evaluar la eficacia de una vacuna recombinante basada en TbpB frente a una variedad de serovares clínicos representativos de la enfermedad de Glässer en España.
En primer lugar, se analizaron 68 aislados clínicos de G. parasuis recolectados entre 2018 y 2021 en diversas regiones de España, provenientes de muestras respiratorias y sistémicas de cerdos. Para su caracterización, se utilizó una PCR específica del gen tbpA y una PCR multiplex para identificar los serovares presentes. Los serovares 5, 10, 2, 4 y 1 resultaron ser los más comunes, abarcando aproximadamente el 84 % de los aislados, lo que pone de manifiesto su alta prevalencia en el país.
A continuación, se llevó a cabo un análisis de las secuencias de aminoácidos de la proteína TbpB en 59 de estos aislados, que permitió clasificar las cepas en diez clados distintos. Este hallazgo refleja una notable diversidad genética entre las cepas, que se relaciona tanto con los tipos capsulares como con los diferentes orígenes anatómicos y geográficos de los aislamientos.
Con el fin de caracterizar la virulencia de los aislados, se evaluó la virulencia mediante PCR multiplex de secuencia líder de los genes vtaA, y todos los aislados se identificaron como clínicos (presuntamente virulentos). El patotipado basado en diez genes del pangenoma reveló que el gen virulento HPS_22970 era el más frecuente (83,1%). Se observaron diversos perfiles de patotipo, con 29 combinaciones génicas únicas y dos aislados portadores únicamente de genes del pangenoma potencialmente no virulentos. Posteriormente, se analizaron los perfiles de resistencia a antimicrobianos de los aislados, obteniendo una resistencia generalizada en el fenotipado de resistencias antimicrobianas, con un 63,3% clasificado como multirresistente, y una alta resistencia a la clindamicina (98,3%) y la tilosina (93,3%). Se encontró una asociación estadísticamente significativa entre ciertos genes de patotipos y fenotipos de resistencia a antimicrobianos, en particular entre el gen virulento HPS_22970 y la resistencia a la tetraciclina (p < 0,001).
Por último, en este estudio se evaluó en dos fases la inmunogenicidad y protección de una vacuna oral de dos dosis, basada en una TbpB modificada (TbpBY167A) en lechones privados de calostro. En la primera fase (días 0-45), se analizó la respuesta inmunitaria, y en la segunda (días 45-52), la protección heteróloga tras desafiar a los lechones con cuatro aislados clínicos de G. parasuis de diferentes serovares. La vacuna indujo una respuesta inmune robusta, con un aumento significativo de IgA e IgM. Tras el desafío, se observó una mejora en las tasas de supervivencia y una reducción en los signos clínicos y las lesiones, independientemente del serovar. Además, la vacuna redujo la colonización en el tracto respiratorio y en tejidos sistémicos, como las articulaciones, el hígado y el cerebro. El análisis inmunohistoquímico reveló un menor número de macrófagos en los pulmones de los lechones inmunizados.
Los hallazgos de este estudio destacan la variabilidad genética de G. parasuis y su compleja patogenicidad, evidenciando la necesidad de caracterizar de manera precisa los aislados clínicos para asegurar tratamientos antimicrobianos adecuados y medidas profilácticas efectivas contra las cepas virulentas. A pesar de la diversidad genética observada en los serovares, los hallazgos in silico sugieren que una vacuna basada en la proteína TbpB podría ofrecer una protección amplia contra los brotes de la enfermedad de Glässer en España, destacando su potencial como herramienta de control en la industria porcina. En cuanto al estudio in vivo, la administración oral de la vacuna de subunidad TbpBY167A en lechones ha demostrado ser eficaz, brindando protección heteróloga frente a diferentes aislados virulentos, reduciendo significativamente la colonización bacteriana y la diseminación en los tejidos. Este enfoque supera las limitaciones de las vacunas tradicionales al contrarrestar los efectos de la inmunidad materna, convirtiéndola en una alternativa prometedora para la vacunación universal contra la enfermedad de Glässer.[EN] The swine industry faces significant economic challenges, with Glässer's disease being one of the most relevant. This disease is caused by Glaesserella (Haemophilus) parasuis, a pathogen that colonizes the upper respiratory tract of pigs and, under stressful conditions such as weaning or transportation, can trigger severe systemic infections. In addition, this bacterium is one of the secondary pathogens included in the Porcine Respiratory Disease Complex. Piglets in the early stages of lactation and fattening are particularly vulnerable, leading to high morbidity and mortality rates. This bacterium acquires iron from porcine transferrin through a surface receptor composed of the transferrin-binding proteins TbpA and TbpB, facilitating its survival in iron-limited environments, such as mucosal surfaces.
However, the virulence mechanisms of this bacterium are not yet fully understood, and the challenge is further exacerbated by the concerning rise in antimicrobial resistance, driven by its intensive use in pig production. This dual issue highlights the urgent need to deepen research into the pathogenic factors of G. parasuis and to develop strategies that reduce the use of these drugs without compromising disease control. This phenomenon undermines the effectiveness of treatments and poses an additional risk of resistance gene transmission to other bacteria in the environment. Nonetheless, in the absence of a commercially available universal vaccine to prevent this disease, antimicrobial treatment remains the primary control strategy despite the associated challenges. This underscores the necessity to develop accurate diagnostic tools, promote responsible antimicrobial use, and encourage research to advance the design of effective, broad-spectrum vaccines.
The main objective of this doctoral thesis is to provide a detailed characterization of the virulence and antimicrobial resistance profiles of G. parasuis isolates from Spanish pig farms. In parallel, the genetic diversity of the TbpB protein in these isolates will be analyzed, with the aim of evaluating the effectiveness of a recombinant TbpB-based vaccine against a variety of clinical serovars representative of Glässer's disease in Spain.
First, 68 clinical G. parasuis isolates collected between 2018 and 2021 from various regions of Spain were analyzed. These isolates were obtained from respiratory and systemic samples from pigs. For their characterization, a species-specific PCR targeting the tbpA gene and a multiplex PCR assay were used to identify the present serovars. Serovars 5, 10, 2, 4, and 1 were found to be the most prevalent, accounting for approximately 84% of the isolates, highlighting their high prevalence across the country.
An analysis of the amino acid sequences of the TbpB protein in 59 of these isolates was subsequently conducted, allowing for the classification of the strains into ten distinct clades. This finding reflects significant genetic diversity among the strains, which is associated with both capsular types and the various anatomical and geographical origins of the isolates.
To characterize the virulence of the isolates, virulence was assessed using multiplex PCR with leader sequence targeting the vtaA genes, and all isolates were identified as clinical (presumed virulent). Pangenome-based pathotyping revealed that the virulent gene HPS_22970 was the most frequent (83.1%). Various pathotype profiles were observed, with 29 unique gene combinations and two isolates carrying only potentially non-virulent pangenome genes. Subsequently, the antimicrobial resistance profiles of the isolates were analyzed, revealing widespread resistance in the antimicrobial resistance phenotyping, with 63.3% classified as multi-resistant, and high resistance to clindamycin (98.3%) and tylosin (93.3%). A statistically significant association was found between certain pathotype genes and antimicrobial resistance phenotypes, particularly between the virulent gene HPS_22970 and tetracycline resistance (p < 0.001).
Finally, this study assessed in two phases the immunogenicity and protection of a two-dose oral vaccine based on a modified TbpB (TbpBY167A) in colostrum-deprived piglets. In the first phase (days 0-45), the immune response was analyzed, and in the second phase (days 45-52), heterologous protection was evaluated after challenging the piglets with four clinical isolates of G. parasuis from different serovars. The vaccine induced a robust immune response, with a significant increase in IgA and IgM. Following the challenge, improved survival rates were observed, along with a reduction in clinical signs and lesions, regardless of the serovar. Additionally, the vaccine reduced colonization in the respiratory tract and in systemic tissues such as joints, liver, and brain. Immunohistochemical analysis revealed a lower number of macrophages in the lungs of immunized piglets.
The findings of this study highlight the genetic variability of G. parasuis and its complex pathogenicity, underscoring the need for precise characterization of clinical isolates to ensure appropriate antimicrobial treatments and effective prophylactic measures against virulent strains. Despite the genetic diversity observed in the serovars, in silico findings suggest that a vaccine based on the TbpB protein could provide broad protection against outbreaks of Glässer’s disease in Spain, emphasizing its potential as a control tool in the swine industry. Regarding the in vivo study, the oral administration of the subunit vaccine TbpBY167A in piglets has proven effective, offering heterologous protection against different virulent isolates, significantly reducing bacterial colonization and dissemination in tissues. This approach overcomes the limitations of traditional vaccines by counteracting the effects of maternal immunity, making it a promising alternative for universal vaccination against Glässer’s disease
Effect of several packaging conditions on the microbiological, physicochemical and sensory properties of ostrich steaks during refrigerated storage
Get PDF1).- MATERIAS (SUBJETS): 1a).- Tecnología de los Alimentos; 1b).- Medicina. Salud[EN]. A total of 365 ostrich steaks were packaged in air (AIR), vacuum (VAC), MAP1 (70% O-2 + 30% CO2), MAP2 (30% O-2 + 30% N-2 + 40% CO2), MAP3 (20% O-2 + 30% N-2 + 50% CO2), MAP4 (50% N-2 + 50% CO2), MAP5 (20% N-2 + 80% CO2) or MAP6 (100% CO2). Microbial counts (10 groups), pH, Aw and sensory properties (nine-point hedonic scale) were determined on days 0, 1, 3, 7 and 15 of storage (4 degrees C). On day 0, microbial counts (log(10) cfu/g) ranged from undetectable levels (Brochothrix thermosphacta, enterococci) to 3.21 +/- 0.63 (total aerobic counts -TAC-). The highest and the lowest microbial loads throughout storage were observed in AIR and MAP6, respectively. On day 15 TAC as high as 9.96 +/- 0.20 log(10) cfu/g were found in AIR. The shelf-life (time until overall acceptability score fell below 5) was 3 days (MAP1, MAP2), 7 days (MAP3, AIR) or 15 days (MAP4, MAP5, MAP6). Only for VAC the shelf-life limit extended beyond 15 daysSIThe authors wish to thank the Universidad de León (BB178) for the financial support
Safety of frozen and dried formulations from migratory locust (Locusta migratoria) as a Novel food pursuant to Regulation (EU) 2015/2283
Get PDF[EN] Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Food and Food Allergens (NDA) was asked to deliver an opinion on the safety of frozen and dried formulations from migratory locust (Locusta migratoria) as a novel food pursuant to Regulation (EU) 2015/2283. The term migratory locust refers to the adult of the insect species Locusta migratoria. The NF is proposed in three formulations i) frozen without legs and wings; ii) dried without legs and wings; iii) ground with legs and wings. The main components of the NF are protein, fat and fibre (chitin) in the dried form of the NF, and water, protein, fat and fibre (chitin) in the frozen form of the NF. The Panel notes that the concentration of contaminants in the NF depends on the occurrence levels of these substances in the insect feed. The Panel notes that there are no safety concerns regarding the stability of the NF if the NF complies with the proposed specification limits during its entire shelf-life. The NF has a high protein content, although the true protein levels in the NF are overestimated when using the nitrogen-to-protein conversion factor of 6.25, due to the presence of non-protein nitrogen from chitin. The applicant proposed to use the NF as frozen, dried and ground in the form of snack, and as a food ingredient in a number of food products. The target population proposed by the applicant is the general population. The Panel notes that considering the composition of the NF and the proposed conditions of use, the consumption of the NF is not nutritionally disadvantageous. The submitted history of use and toxicity studies from literature did not raise safety concerns. The Panel considers that the consumption of the NF might trigger primary sensitisation to L. migratoria proteins and may cause allergic reactions in subjects with allergy to crustaceans, mites and molluscs. Additionally, allergens from the feed may end up in the NF. The Panel concludes that the NF is safe under the proposed uses and use levelsSIThe Panel wishes to thank Petra Gergelova for the support provided to this scientific opinio
Assessment of the application for renewal of authorisation of 6‐phytase produced by Trichoderma reeseiCBS 122001 as a feed additive for pigs and poultry, from Roal Oy
Get PDF[EN] Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the assessment of the application for renewal of authorisation of 6-phytase produced by Trichoderma reeseiCBS 122001 (brand name: Finase®EC). The applicant has provided evidence that the additive currently in the market complies with the existing conditions of authorisation. The Panel concludes that the additive remains safe for poultry for fattening, breeding and laying, and all pigs, the consumer and the environment under the authorised conditions of use. Regarding user safety, the Panel reiterates that the additive is not a skin or eye irritant or sensitiser but should be considered a potential respiratory sensitiser. There is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation. These conclusions also apply to the new proposed liquid formulation Finase®EC 5 LSIThe Panel wishes to acknowledge the contribution of Yolanda Garcia Cazorla to this opinio
Herramientas de evaluación como medio para conocer las interacciones familiares ante tareas de competencia escrita
No full text[ES] Estudio de las herramientas de evaluación como medio para conocer las interacciones familiares ante tareas de competencia escrit
Older Men's Participation in Cognitive Training: A Review
Get PDF[EN] Background: Cognitive training for healthy older adults living in a community is an essential resource that allows them to live at home for as long as possible. Objective: The purpose of the review is to examine the degree of participation of males and females in longitudinal studies of cognitive training. Moreover, we want to identify if these studies include the gender variable in their analyzes or reflect on its importance. Method: This review considered longitudinal cognitive training studies were published in English and Spanish and conducted with healthy older adults living in a community. Results: The Advanced Cognitive Training for Independent and Vital Elderly (ACTIVE) study (in English) and the Memoria Mejor (MM) Longitudinal Study (in Spanish), both illustrate the trend of sex/gender treatment of the studies reviewed: a) high participation of older people seventy and more years b) recruiting stratified by age and sex; c) males are disproportionately underrepresented in cognitive training studies [24% - 14%]; d) the evaluation measures (baseline, follow-up, and final) and dropout data are provided but not stratified by age and sex/gender. Conclusions: Researchers demonstrate awareness about the impact of sex/gender differences but do not focus on it. Understanding sex/gender differences are necessary for understanding not only that these differences occur, but also why they occur; this will allow policies or intervention programs with approaches that are more equitable for both sexes/genders to be formulated.S
Identificación de un nuevo haplogrupo del cromosoma Y en el ganado autóctono español
No full text[EN] The aim of this work was to perform a thorough analysis of the diversity of Y-haplotypes in
Spanish cattle. A total of 207 Bos taurus males were sampled across 25 European breeds,
with a special focus on rare, local Spanish populations. Animals were genotyped with five
Y-specific microsatellites (INRA189, UMN0103, UMN0307, BM861 and BYM1), two
indels (ZFY10 and USP9Y) and one SNP (UTY19). A new haplogroup, distinct from those
described by Gotherstr € om€ et al. (2005), was identified and named Y1.2. Samples
representing the three B. taurus Y-haplogroups were genotyped for four additional
Y chromosome SNPs (rs121919254, rs121919281, rs121919323 and rs137049553).
Among these SNPs, only rs121919281 was informative in B. taurus and helped to confirm
the new Y1.2 haplogroup. Analysis of a larger dataset of standardized haplotypes for 1507
individuals from 57 populations from Spain, other European countries and Africa showed
the new Y1.2 haplogroup to be found exclusively in Spanish breeds. This finding reinforces
the importance of local Spanish cattle as reservoirs of genetic diversity as well as the
importance of the Iberian Peninsula in the history of cattle.SIR. Pelayo was partially founded by grant MHE2011-00117. C. Ginja was supported by a contract grant from the Fundação para a Ciência e a Tecnologia (Ref. IF/00866/2014). The work was partially supported by the Veterinary Genetics Laboratory, University of California, Davis, USA. We acknowledge the collaboration of the Lidia cattle breed association (Agrupación Española de Ganaderos de Reses Bravas)