[Dietary diversity, anthropometric status and biochemical profile of centenarians of the Nicoya Peninsula, Costa Rica, and their offspring]. / Diversidad dietética, estado antropométrico y perfil bioquímico de centenarios de la Península de Nicoya, Costa Rica, y su descendencia.

BACKGROUND AND OBJECTIVE: Social relationships, physical activity and diet have been proposed as responsible for longevity; however, many of these conditions have changed in modern societies. The objective of this study was to identify if the anthropometric status, biochemical profile and diet of Nicoyan centenarians is different from that of their decency and if the quality and diversity of the diet of these Costa Ricans differs from that of adults from urban areas of the same country. MATERIALS AND METHODS: Anthropometric, biochemical and dietary assessments were carried out on 120 adults residing in the Nicoya Peninsula, including centenarians, their children and grandchildren. The data were analyzed using descriptive and inferential statistics. RESULTS: There is a significantly lower percentage of excess weight, central adiposity and blood lipids in Nicoyan centenarians compared to their children and grandchildren. The consumption of «Dairy¼ and «Other fruits¼ is higher in centenarians than in their descendants. Centenarians showed a higher dietary diversity index (DDI) compared to other age categories, and they also showed a higher DDI and a significantly more diverse diet than the urban population of Costa Rica. CONCLUSION: Nicoyan centenarians presented a better lipid and glycemic profile than their offspring, they have lower weight and a better quality and diversity of diet than their relatives and than adults from the urban area of Costa Rica.

[Review of nutrition and hydration in relation to physical exercise during pregnancy]. / Revisión de la nutrición e hidratación con relación al ejercicio físico en el embarazo.

Studies on nutritional recommendations for pregnant women who exercise are scarce. The objective of this article is not to focus on the diet of pregnant women as a whole, but to review those aspects of it that may be related to physical exercise. To this end, the nutritional and hydration recommendations contained in the main clinical practice guides on physical exercise during pregnancy are collected, including the first Spanish guides. Likewise, the energy requirements required by pregnant women who perform physical exercise during pregnancy for adequate gestational weight gain are addressed, aspects related to macronutrients in the aforementioned population group, and two specific topics, such as nutritional needs in the adolescent who practices physical exercise during pregnancy and eating disorders in pregnant athletes It is concluded by stating that pregnant women who exercise regularly should eat a varied and balanced diet, such as the Mediterranean diet, avoid long periods of fasting to avoid the appearance of hypoglycemia and maintain adequate fluid intake before, during and after physical exercise. Pregnant adolescents who engage in physical exercise require nutritional supervision to achieve adequate gestational weight gain. The pregnant athlete with an eating disorder has a higher risk of complications during pregnancy and childbirth, gynecological, fetal and neonatal, and, therefore, requires close monitoring by specialists in maternal-fetal medicine.

New insights into swine dysentery: faecal shedding, macro and microscopic lesions and biomarkers in early and acute stages of Brachyspira hyodysenteriae infection.

BACKGROUND: Swine dysentery (SD) is a severe mucohaemorrhagic colitis in pigs caused classically by Brachyspira hyodysenteriae. Although several aspects of B. hyodysenteriae infection dynamic are already described, further research in the early stage of this infection is required. In this study, 7-week-old pigs were orally challenged with B. hyodysenteriae to obtain information about faecal shedding, macro and microscopic intestinal lesions and serum acute phase proteins in pigs at the onset of B. hyodysenteriae shedding (early infection group, n = 8), in pigs with mucohaemorrhagic diarrhoea (acute infection group, n = 8) and in non-infected controls (n = 16). RESULTS: First B. hyodysenteriae detection by q-PCR and first loose stools with blood and mucus occurred both at 8 days post-inoculation. The lapse between a positive q-PCR and observation of mucohaemorrhagic diarrhoea ranged from 0 to 3 days, except in a single pig in which this period lasted 5 days. Macroscopic lesions were observed in the large intestine from both infected groups although more frequent and severe in acute infection group. Microscopic observation of the apex mucosa revealed that in early infection only higher ulceration values were observed compared to healthy controls. In contrast, the acute infection group exhibited higher ulceration, neutrophils infiltration and increased mucosal thickness compared to the other two groups. Among the serum biomarkers tested, only haptoglobin, C-reactive protein, and creatine kinase showed a significant increase in pigs in the acute infection period compared to controls, whereas haptoglobin was the only factor with a significant increase at the early infection compared to non-infected animals. CONCLUSIONS: This study provides new insights about SD and remarks the complex and limited options to perform an early detection of infected animals beyond PCR diagnosis.

Valorization of Saithe (Pollachius virens) Residuals into Protein Hydrolysates-Silaging as Preservation Technology.

Silaging can be used as preservation technology to valorize currently discarded raw material into protein hydrolysate on board deep-sea vessels. The aim of this study was to investigate the effect of sorting and raw material freshness on the quality and yield of protein hydrolysates obtained through silaging of saithe (Pollachius virens) viscera. Additionally, the effect of using acid-containing antioxidants was tested. Out sorting of the liver prior to silaging resulted in slightly higher hydrolysate yields. The hydrolysates with the highest protein contents were obtained from silages made from fresh raw materials (day 0), and the content decreased significantly after longer storage of the raw material (2-3 days at 4 °C). Storage of the raw material for 1 day did not affect the quality. However, a significantly higher degree of hydrolysis (DH), content of free amino acids (FAA), and total volatile basic nitrogen (TVB-N) were obtained when raw materials were stored for 3 days. The FAA composition was influenced by the raw material's freshness, with increases in free glutamic acid and lysine and a decrease in free glutamine after longer storage. None of the studied parameters were significantly affected by out sorting of liver or the addition of antioxidants. Overall, the results indicate that the whole fraction of the viscera can be utilized without reducing the quality of the hydrolysate and that the raw material should be stored for a maximum of 1 day prior to preservation to optimize the quality.

Exploring the Potential of Atlantic Mesopelagic Species Processed on Board Commercial Fishing Vessels as a Source of Dietary Lipids.

This study investigates the use of untapped mesopelagic species as a source of long-chain polyunsaturated omega-3 fatty acids (LC n-3 PUFAs) to meet the growing demand. The challenges faced by commercial fishing vessels, such as varying catch rates and species distribution affecting lipid levels, are addressed. Marine oils were produced post-catch using thermal separation and enzymatic hydrolysis during four commercial cruises, screening approximately 20,000 kg of mixed mesopelagic species. Maurolicus muelleri and Benthosema glaciale were the dominant species in the catch, while krill was the primary bycatch. The lipid composition varied, with B. glaciale having a higher prevalence of wax esters, while triacylglycerols and phospholipids were more predominant in the other species. LC n-3 PUFAs ranged from 19% to 44% of lipids, with an average EPA + DHA content of 202 mg/g of oil. Both processing methods achieved oil recoveries of over 90%. Estimates indicate that the mesopelagic biomass in the Northeast Atlantic could supply annual recommended levels of EPA + DHA to 1.5 million people, promoting healthy heart and brain functions. These findings offer valuable insights for considering mesopelagic species as a potential source of dietary marine lipids, laying the groundwork for further research and innovation in processing and obtaining valuable compounds from such species.

Salmonella Control in Swine: A Thoughtful Discussion of the Pre- and Post-Harvest Control Approaches in Industrialized Countries.

Pork is among the major sources of human salmonellosis in developed countries. Since the 1990s, different surveys and cross-sectional studies, both national and international (i.e., the baseline studies performed in the European Union), have revealed and confirmed the widespread non-typhoidal Salmonella serotypes in pigs. A number of countries have implemented control programs with different approaches and degrees of success. The efforts could be implemented either at farms, in post-harvest stages, or both. The current review revises the current state of the art in Salmonella in swine, the control programs ongoing or conducted in the past, and their strengths and failures, with particular attention to the weight of pre- and post-harvest control and the implications that both have for the success of interventions or mitigation after outbreaks. This review provides a novel perspective on Salmonella control in swine, a matter that still includes uncertainties and room for improvement as a question of public health and One Health.

Inflammatory bowel disease induces pathological α-synuclein aggregation in the human gut and brain.

AIMS: According to Braak's hypothesis, it is plausible that Parkinson's disease (PD) originates in the enteric nervous system (ENS) and spreads to the brain through the vagus nerve. In this work, we studied whether inflammatory bowel diseases (IBDs) in humans can progress with the emergence of pathogenic α-synuclein (α-syn) in the gastrointestinal tract and midbrain dopaminergic neurons. METHODS: We have analysed the gut and the ventral midbrain from subjects previously diagnosed with IBD and form a DSS-based rat model of gut inflammation in terms of α-syn pathology. RESULTS: Our data support the existence of pathogenic α-syn in both the gut and the brain, thus reinforcing the potential role of the ENS as a contributing factor in PD aetiology. Additionally, we have analysed the effect of a DSS-based rat model of gut inflammation to demonstrate (i) the appearance of P-α-syn inclusions in both Auerbach's and Meissner's plexuses (gut), (ii) an increase in α-syn expression in the ventral mesencephalon (brain) and (iii) the degeneration of nigral dopaminergic neurons, which all are considered classical hallmarks in PD. CONCLUSION: These results strongly support the plausibility of Braak's hypothesis and emphasise the significance of peripheral inflammation and the gut-brain axis in initiating α-syn aggregation and transport to the substantia nigra, resulting in neurodegeneration.

Caecum OX40+CD4 T-cell subset associates with mucosal damage and key markers of disease in treated HIV-infection.

BACKGROUND: Blood OX40-expressing CD4 T-cells from antiretroviral (ART)-treated people living with HIV (PWH) were found to be enriched for clonally-expanded HIV sequences, hence contributing to the HIV reservoir. OX40-OX40L is also a checkpoint regulator of inflammation in multiple diseases. We explored gut mucosal OX40+CD4+ T-cells and their potential significance in HIV disease. METHODS: Biopsies of caecum and terminal-ileum of ART-treated PWH (n = 32) were obtained and mucosal damage and HIV reservoir were assessed. Mucosal OX40+ and Ki67+ CD4 T-cell subsets, as well as several tissue T-cell subsets modulating mucosal integrity and homeostasis (Th17, Th22, Treg, Tc17, Tc22, IL17+TCRγδ, IL22+TCRγδ) were quantified. Inflammatory-related markers, T-cell activation and thymic output were also determined in blood samples. Correlations were explored using Spearman rank test and corrected for multiple comparisons by Benjamini-Hochberg. RESULTS: Compared to healthy controls, a high frequency of mucosal, mainly caecum, CD4 T-cells were OX40+ in PWH. Such frequency strongly correlated with nadir CD4 (r = -0.836; p < 0.0001), CD4/CD8 ratio (r = -0.630; p = 0.002), caecum mucosal damage (r = 0.606; p = 0.008), caecum Th22 (r = -0.635; p = 0.002), caecum Th17 (r = 0.474; p = 0.03) and thymic output (r = -0.686; p < 0.001). It also correlated with Neutrophil-to-Lymphocyte Ratio and blood CD4 T-cell activation and tended to with mucosal HIV reservoir. CONCLUSION: High frequencies of caecum OX40+CD4 T-cells are found in people with HIV (PWH) and successful viral control. Interestingly, this cellular subset reflects key markers of disease and peripheral T-cell activation, as well as HIV-driven mucosal damage. OX40+CD4 T-cells deserve further investigation since they could expand because of T-cell homeostatic proliferation and relate to the Th22/Th17 gut mucosal ratio.

Psychometric Properties of the Death Anxiety Scale for Adult Chronic Patients.

INTRODUCTION: Death anxiety is a predictor of exacerbations in both physical and psychological symptoms of chronic diseases. Therefore, having short and easy-to-apply instruments to assess the presence of death anxiety and adopting a multidisciplinary approach to address it are important. METHOD: This study analyzes the psychometric properties of the Death Anxiety Scale (DAS) developed by Donald Templer in a Colombian population of adult patients diagnosed with a chronic disease. The original instrument was linguistically, conceptually, and culturally adapted to Colombian Spanish to be subsequently applied to 301 adult patients with chronic diseases. RESULTS: The exploratory factor analysis revealed a 3-factor structure, with a variance of 47%. Internal consistency was observed (Cronbach's alpha: 0.71; McDonald's omega: 0.76; Guttman's lambda 6 (G6): 0.74; greatest lower bound: 0.54). A correlation coefficient of 0.64 was found between the total score of the DAS and the Beck Anxiety Inventory. CONCLUSION: When comparing the results with the versions of the DAS in Spanish from Mexico and Spain, variability in the psychometric properties was observed; therefore, language cannot be assumed to be a guarantee of the reliability and validity of the instrument.

Silage for upcycling oil from saithe (Pollachius virens) viscera - Effect of raw material freshness on the oil quality.

The main objective of this study was to investigate how the freshness of saithe (Pollachius virens) viscera affected the quality, composition and yield of oil obtained by silaging. Minced viscera with and without liver were stored separately for up to 3 days at 4 °C before silaging at pH 3.8 for 6 days at 10 °C. An antioxidant mixture was added to evaluate the effect on the lipid oxidation. Oil was extracted thermally from untreated raw material during storage (day 0-3) and after silaging. For oil obtained after silaging of viscera with liver, the oil yields increased significantly when the raw material was stored for more than one day prior to the treatment. Use of fresh raw material (collected at day 0) led to significantly lower oxidation compared to longer raw material storage. After one day of storage, the oxidation was less dependent on the freshness. Silaging with antioxidants resulted in significantly lower formation of oxidation products compared to acid without antioxidants and the most significant differences were observed after one day of storage. Contents of docosahexaenoic acid (DHA) and total omega-3 fatty acids decreased significantly when the raw material was stored for 1-3 days prior to silaging compared to fresh raw material. Results obtained by high resolution nuclear magnetic resonance (NMR) spectroscopy indicated that oxidation of esterified DHA might explain the DHA decrease. The free fatty acid content was highest when fresh raw material was used and was most likely affected by the formation of cholesteryl esters observed in NMR spectra after longer storage. The study shows that although the oil quality is reduced during silaging, processing shortly after catch and use of antioxidants can optimize the quality resulting in less oxidized oil richer in omega-3 fatty acids.

Detection and genetic characterization of enteric viruses in diarrhoea outbreaks from swine farms in Spain.

BACKGROUND: The aim of this work was to study the prevalence and distribution of Porcine astrovirus (PAstV), Porcine kobuvirus (PKoV), Porcine torovirus (PToV), Mammalian orthoreovirus (MRV) and Porcine mastadenovirus (PAdV) as well as their association with widely recognized virus that cause diarrhoea in swine such as coronavirus (CoVs) and rotavirus (RVs) in diarrhoea outbreaks from Spanish swine farms. Furthermore, a selection of the viral strains was genetically characterized. RESULTS: PAstV, PKoV, PToV, MRV and PAdV were frequently detected. Particularly, PAstV and PKoV were detected in almost 50% and 30% of the investigated farms, respectively, with an age-dependent distribution; PAstV was mainly detected in postweaning and fattening pigs, while PKoV was more frequent in sucking piglets. Viral co-infections were detected in almost half of the outbreaks, combining CoVs, RVs and the viruses studied, with a maximum of 5 different viral species reported in three investigated farms. Using a next generation sequencing approach, we obtained a total of 24 ARN viral genomes (> 90% genome sequence), characterizing for first time the full genome of circulating strains of PAstV2, PAstV4, PAstV5 and PToV on Spanish farms. Phylogenetic analyses showed that PAstV, PKoV and PToV from Spanish swine farms clustered together with isolates of the same viral species from neighboring pig producing countries. CONCLUSIONS: Although further studies to evaluate the role of these enteric viruses in diarrhoea outbreaks are required, their wide distribution and frequent association in co-infections cannot be disregard. Hence, their inclusion into routine diagnostic panels for diarrhoea in swine should be considered.

MicroRNA Dysregulation in Early Breast Cancer Diagnosis: A Systematic Review and Meta-Analysis.

Breast cancer continues to be the leading cause of death in women worldwide. Mammography, which is the current gold standard technique used to diagnose it, presents strong limitations in early ages where breast cancer is much more aggressive and fatal. MiRNAs present in numerous body fluids might represent a new line of research in breast cancer biomarkers, especially oncomiRNAs, known to play an important role in the suppression and development of neoplasms. The aim of this systematic review and meta-analysis was to evaluate dysregulated miRNA biomarkers and their diagnostic accuracy in breast cancer. Two independent researchers reviewed the included studies according to the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines. A protocol for this review was registered in PROSPERO with the registration number "CRD42021256338". Observational case-control-based studies analyzing concentrations of microRNAs which have been published within the last 10 years were selected, and the concentrations of miRNAs in women with breast cancer and healthy controls were analyzed. Random-effects meta-analyses of miR-155 were performed on the studies which provided enough data to calculate diagnostic odds ratios. We determined that 34 microRNAs were substantially dysregulated and could be considered biomarkers of breast cancer. Individually, miR-155 provided better diagnostic results than mammography on average. However, when several miRNAs are used to screen, forming a panel, sensitivity and specificity rates improve, and they can be associated with classic biomarkers such us CA-125 or CEA. Based on the results of our meta-analysis, miR-155 might be a promising diagnostic biomarker for this patient population.

Upgrading Marine Oils from Cod (Gadus morhua) On-Board the Deep-Sea Vessels-From Waste to Value.

Significant amounts of marine raw material are lost on-board the deep-sea vessels due to fast quality degradation. Optimal on-board handling and processing strategies can upgrade these resources from waste to food ingredients rich in nutrients such as omega-3 fatty acids. The objective of this study was to investigate the effect of raw material freshness and sorting on the quality, composition and yield of oil produced thermally from cod (Gadus morhua) residuals on-board a commercial trawler. Oil was produced from whole viscera fractions with liver or out-sorted livers right after a catch and after chilled storage for up to 6 days. The results showed that significantly higher oil yields could be obtained if the raw materials were stored for 1 day or longer. However, an undesired emulsion was formed when viscera were stored for 4 days. All oils were rich in health beneficial omega-3 fatty acids, but viscera oils had generally lower quality with higher levels of free fatty acids and oxidation products. However, out-sorting of the liver was not necessary to meet guidelines for high-quality fish oil. Both viscera and liver could be stored for up to 2 days at 4 °C prior to oil production and still meet quality criteria for food applications. These results demonstrate a large potential in upgrading currently wasted marine raw materials into high-quality food ingredients.

Palliative care stay room - designing, testing and evaluating a gamified social intervention to enhance palliative care awareness.

INTRODUCTION: The message of palliative care can be promoted using creative thinking and gamification. It can be an innovative strategy to promote changes in behaviour, promote thinking, and work on skills such as empathy. AIM: Design, test and evaluate a gamified social intervention to enhance palliative care awareness among young university students from non-health background. METHODS: Participatory action research study with mixed methods, Design Thinking and using the Public Engagement strategy. Forty-three undergraduate students participated in a Palliative Care Stay Room and completed the Test of Cognitive and Affective Empathy (TECA) before and after the game. At the end of the game, a ten-minute debriefing was held with the participants, which was concluded with an open conversation. The content analysis was done independently and the sum of the scores of each dimension was compared before and after the activity. FINDINGS: The Stay Room improved the participants' knowledge and new perspectives about palliative care. Before the game, their views focused on the end of life and after the game on their values, highlighting the dedication of the healthcare professionals who do not treat death but the life until death. After de game, participants (N = 43: female = 23; male = 20; x̄ 19.6 years old) presented higher values in perspective adoption (intellectual ability to put oneself in the other's place) p = 0.046 and in emotional understanding (ability to recognize emotional states) p = 0.018, and had high scores on empathic joy (p = 0.08). CONCLUSION: Gamification can be used in teaching and transmitting positive attitudes. Palliative Care and can help young university students to think positively about care issues.

Intralaboratory Validation of a Kinetic Turbidimetric Assay Based on Limulus Amebocyte Lysate (LAL) for Assessing Endotoxin Activity in Cow Milk.

Mastitis, one of the most common diseases in dairy cattle, causes severe losses in the dairy sector worldwide and affects animal welfare. The disease is characterized by an inflammatory reaction of the mammary gland and is mainly caused by bacterial infections, including both Gram-negative and Gram-positive bacteria. The release of endotoxins associated to bacterial lysis is a weighty factor in the clinical course of Gram-negative associated mastitis and should be taken into consideration when using antibiotics in the treatment of these infections. Therefore, endotoxin detection in milk samples would be of help in the management of bovine mastitis. With this aim, we have validated a kinetic turbidimetric assay based on Limulus amebocyte lysate (LAL) for the quantification of endotoxins in milk samples. The assay was adapted to this particular matrix by incorporating filtration and dilution of the milk samples in the procedure. Our results demonstrate the robustness and usefulness of the assay, which allows the identification of coliform mastitis in milk samples from affected cows and the quantification of endotoxin activity in bulk and commercial milk samples. Further studies are required to evaluate the performance of the assay in mastitis milk samples associated to Gram-negative bacteria other than Escherichia coli as well as during the clinical course of these Gram-negative mastitis or after their treatment with antibiotics.

Bioaccessibility of phenolic compounds in Psidium guajava L. varieties and P. friedrichsthalianum Nied. after gastrointestinal digestion.

Psidium guajava L. and Psidium friedrichsthalianum Nied are part of the Psidium species native to America. Nowadays, it is essential to study the phenolic compound (PC) profile and their changes during digestion and the fractions available for absorption. This study aimed to characterize the PC profile in some Psidium species and their bioaccessibility (BA). Fifty-seven compounds were identified, and forty-six belonged to ten different phenolic classes. PC profiles showed significant differences between the species and the intestinal fraction P. friedrichsthalianum Nied. showed the highest PC content, although it mostly belonged to non-extractable polyphenols. This leads to the lowest BA (37%); P. guajava L. 'Morada' showed the highest (47%). Hydroxycinnamic acids were the most stable PC after gastrointestinal digestion. This study showed relevant differences in the PC content and profile of different Psidium species and changes between the PC in the original matrix and those released in the different stages of gastrointestinal digestion.

Hipospadias: del diagnóstico ecográfico prenatal al manejo posnatal / Hypospadias: from prenatal ultrasound diagnosis to postnatal management

El hipospadias es la localización anormal del meato urinario y es la malformación de genitales externos más frecuentemente diagnosticada. El diagnóstico prenatal es posible mediante ecografía sistemática desde la semana 20 de gestación, siendo más fácil su diagnóstico en el tercer trimestre. Las formas leves suelen ser aisladas, familiares o asociadas a disfunción placentaria o restricción de crecimiento intrauterino, mientras que las formas más graves presentan hasta un 30% de asociación a defectos fetales, anomalías cromosómicas/genéticas o anomalías del desarrollo sexual. La tríada para el diagnóstico ecográfico prenatal consiste en curvatura ventral del pene, anomalía del prepucio dorsal y punta del pene roma. La valoración de la uretra durante la micción y el aspecto del chorro miccional son de gran utilidad para clasificar el defecto. Cuando se diagnostica hipospadias peneano o escrotal es aconsejable realizar una amniocentesis para estudio genético fetal y valorar otros signos de adecuada virilización, como el descenso testicular a partir de la semana 27. El seguimiento tras el parto debe ser multidisciplinario, incluyendo urólogo y endocrinólogo infantil. En hipospadias leves el pronóstico es bueno con reparación quirúrgica en el primer año de vida, pero las formas graves pueden presentar un reto mayor para su corrección funcional y estética.

Hypospadias refers to the abnormal location of the meatus; it is the most common genital malformation detected in the fetus and newborn. Prenatal diagnosis is feasible from 20 weeks onwards with routine ultrasound; however, it is easier to diagnose during the third trimester of pregnancy. Mild defects are usually isolated, familiar o related to placental disfunction or intrauterine growth restriction, while the severe hypospadias are associated to other fetal defects, genetic or chromosomal abnormalities or disorders of sex development. In about 30% of cases. The triad of ultrasound findings prenatally is ventral curvature of the penis, redundant dorsal foreskin and blunt distal penis. The identification of the urethra during the micturition and the direction of the urinary stream help in the classification of the defect. When severe hypospadias is detected, the recommendation is to perform genetic amniocentesis and search for other ultrasound findings related to poor virilization in the fetus, as testicular descent after 27 weeks of gestation. Postnatal follow up should be multidisciplinary including infantile urologist and endocrinologist. The prognosis in distal hypospadias is usually good following surgical repair, however in severe cases surgical interventions may be more challenging in order to obtain satisfactory outcome in terms of function and esthetic.

In vitro evaluation of gentamicin activity against Spanish field isolates of Brachyspira hyodysenteriae.

BACKGROUND: The treatment of swine dysentery (SD) has become constrained in recent years due to the limited availability of effective drugs combined with a rise in antimicrobial resistance. Gentamicin, an aminoglycoside antibiotic, is authorised for the control of this disease in several European countries but has not been extensively used so far. In this study, the in vitro susceptibility of 56 Brachyspira hyodysenteriae field isolates was evaluated against gentamicin using a broth microdilution test. The molecular basis of decreased susceptibility to gentamicin was also investigated by sequencing the 16S rRNA gene and phylogenetic relatedness by multiple-locus variable number tandem-repeat analysis (MLVA). RESULTS: Most B. hyodysenteriae isolates presented low minimum inhibitory concentration (MIC) values to gentamicin, with a mode of 2 µg/mL, a median or MIC50 of 4 µg/mL and percentile 90 or MIC90 of 16 µg/mL. The distribution of these values over the period studied (2011-2019) did not show a tendency towards the development of resistance to gentamicin. Differences in susceptibility among isolates could be explained by two point-mutations in the 16S rRNA gene, C990T and A1185G, which were only present in isolates with high MICs. These isolates were typed in three different MLVA clusters. Analyses of co-resistance between gentamicin and antimicrobials commonly used for the treatment of SD revealed that resistance to tiamulin and valnemulin was associated with low MICs for gentamicin. CONCLUSIONS: The results provide an accurate characterisation of antimicrobial sensitivity to gentamicin and possible mechanisms of resistance in Spanish B. hyodysenteriae isolates. These findings allow us to propose gentamicin as an alternative in the antibiotic management of SD, particularly in outbreaks caused by pleuromutilin resistant isolates.

Genomic Insights into the Mobilome and Resistome of Sentinel Microorganisms Originating from Farms of Two Different Swine Production Systems.

Antimicrobial resistance (AMR) is a threat to public health due to long-term antimicrobial use (AMU), which promotes the bacterial acquisition of antimicrobial resistance determinants (ARDs). Within food-producing animals, organic and extensive Iberian swine production is based on sustainable and eco-friendly management systems, providing an excellent opportunity to evaluate how sustained differences in AMU impact the development and spread of AMR. Here, through a whole-genome sequencing approach, we provide an in-depth characterization of the resistome and mobilome and their interaction in 466 sentinel bacteria, namely, Escherichia coli, Enterococcus spp., Campylobacter coli, and Staphylococcus spp., recovered from 37 intensive and organic-extensive pig farms. Both ARDs and mobile genetic elements (MGEs) were primarily taxon-associated, with higher similarities among bacteria which were closely phylogenetically related. E. coli exhibited the most diverse resistome and mobilome, with 85.4% mobilizable ARDs, 50.3% of which were plasmid-associated. Staphylococcus spp. exhibited a broad repertoire of ARDs and MGEs, with 52.3% of its resistome being mobilizable. Although Enterococcus spp. carried the highest number of ARDs per isolate and its plasmidome was similar in size to that of E. coli, 43.7% of its resistome was mobilizable. A narrow spectrum of ARDs constituted the C. coli resistome, with point mutations as its main AMR driver. A constrained AMU, as observed in organic-extensive herds, determined a reduction in the quantitative composition of the resistome and the complexity of the resistome-mobilome interaction. These results demonstrate taxon-associated AMR-MGE interactions and evidence that responsible AMU can contribute to reducing AMR pressure in the food chain. IMPORTANCE This study provides the first integral genomic characterization of the resistome and mobilome of sentinel microorganisms for antimicrobial resistance (AMR) surveillance from two different swine production systems. Relevant differences were observed among taxa in the resistomes and mobilomes they harbored, revealing their distinctive risk in AMR dissemination and spread. Thus, Escherichia coli and, to a lesser extent, Staphylococcus spp. constituted the main reservoirs of mobilizable antimicrobial resistance genes, which were predominantly plasmid-associated; in contrast to Campylobacter coli, whose resistome was mainly determined by point mutations. The reduced complexity of mobilome-resistome interaction in Enterococcus spp. suggested its limited role in AMR dissemination from swine farms. The significant differences in antimicrobial use among the studied farms allowed us to assess the suitability of whole-genome sequencing as a rapid and efficient technique for the assessment of mid- to long-term on-farm interventions for the reduction of antimicrobial use and the evaluation of AMR status.