Interindividual variability in energy intake and expenditure during a weight loss intervention.

INTRODUCTION: Behavioral compensations may occur as a response to a negative energy balance. The aim of this study was to explore the associations between changes in energy intake (EI) and changes in physical activity (PA, min/day; kcal/d) as a response to a weight loss (WL) intervention and to understand if interindividual differences occur in EI and energy expenditure (EE). METHODS: Eighty-one participants [mean (SD): age = 42.8 (9.4)y, BMI = 31.2 (4.4)kg/m2, 37% females] divided in intervention (IG, n = 43) and control group (CG, n = 38) were included. The IG underwent a moderate energy restriction (300-500 kcal/d). EI was measured through the intake-balance method. Non-exercise PA (NEPA) and exercise (through logbook) were assessed by accelerometery. The EE in NEPA (NEAT) and in exercise (EiEE) was calculated by applying the Freedson Combination'98 algorithm over the time spent in these activities. Pearson correlations were performed in IG to examine associations between EE components, EI and body composition. To understand if interindividual differences were observed, the SD of individual response (SDIR) and the smallest worthwhile change (SWC, SDbaselineCG×0.2) were calculated. RESULTS: Changes in EI [Δ EI, (kcal/d)] was negatively associated with Δ exercise (min/d:r = -0.413, p = 0.045; %:r = -0.846, p = 0.008) and with Δ EiEE (kcal/d:r = -0.488, p = 0.016; %:r = -0.859, p = 0.006). A negative correlation was found between Δ sedentary time and Δ NEPA (min/d:r = -0.622, p = 0.002; %:r = -0.487, p = 0.018). An interindividual variability was found for EI(SDIR = 151.6, SWC = 72.3) and EE (SDIR = 165, SWC = 134). CONCLUSIONS: Decreases in EI were not associated to compensatory responses such as decreases in PA and/or increases in sedentary time. Interindividual variability was found for EI and EE. Nevertheless, behavioral compensations and the interindividual variability should be considered when implementing WL interventions, to increase the likelihood of achieving sustainable results. (clinicaltrials.gov ID: NCT03031951).

Assessment of milk metabolites as biomarkers for predicting feed efficiency in dairy sheep.

Estimating feed efficiency (FE) in dairy sheep is challenging due to the high cost of systems that measure individual feed intake. Identifying proxies that can serve as effective predictors of FE could make it possible to introduce FE into breeding programs. Here, 39 Assaf ewes in first lactation were evaluated regarding their FE by 2 metrics, residual feed intake (RFI) and feed conversion ratio (FCR). The ewes were classified into high, medium and low groups for each metric. Milk samples of the 39 ewes were subjected to untargeted metabolomics analysis. The complete milk metabolomic signature was used to discriminate the FE groups using partial least squares discriminant analysis. A total of 41 and 26 features were selected as the most relevant features for the discrimination of RFI and FCR groups, respectively. The predictive ability when utilizing the complete milk metabolomic signature and the reduced data sets were investigated using 4 machine learning (ML) algorithms and a multivariate regression method. The orthogonal partial least squares algorithm outperformed other ML algorithms for FCR prediction in the scenarios using the complete milk metabolite signature (R2 = 0.62 ± 0.06) and the 26 selected features (R2 = 0.62 ± 0.15). Regarding RFI predictions, the scenarios using the 41 selected features outperformed the scenario with the complete milk metabolite signature, where the multilayer feedforward artificial neural network (R2 = 0.18 ± 0.14) and extreme gradient boosting (R2 = 0.17 ± 0.15) outperformed other algorithms. The functionality of the selected metabolites implied that the metabolism of glucose, galactose, fructose, sphingolipids, amino acids, insulin, and thyroid hormones was at play. Compared with the use of traditional methods, practical applications of these biomarkers might simplify and reduce costs in selecting feed-efficient ewes.

Epigenetic regulation of functional candidate genes for milk production traits in dairy sheep subjected to protein restriction in the prepubertal stage.

BACKGROUND: As the prepubertal stage is a crucial point for the proper development of the mammary gland and milk production, this study aims to evaluate how protein restriction at this stage can affect methylation marks in milk somatic cells. Here, 28 Assaf ewes were subjected to 42.3% nutritional protein restriction (14 animals, NPR) or fed standard diets (14 animals, C) during the prepubertal stage. During the second lactation, the milk somatic cells of these ewes were sampled, and the extracted DNA was subjected to whole-genome bisulfite sequencing. RESULTS: A total of 1154 differentially methylated regions (DMRs) were identified between the NPR and C groups. Indeed, the results of functional enrichment analyses of the genes harboring these DMRs suggested their relevant effects on the development of the mammary gland and lipid metabolism in sheep. The additional analysis of the correlations of the mean methylation levels within these DMRs with fat, protein, and dry extract percentages in the milk and milk somatic cell counts suggested associations between several DMRs and milk production traits. However, there were no phenotypic differences in these traits between the NPR and C groups. CONCLUSION: In light of the above, the results obtained in the current study might suggest potential candidate genes for the regulation of milk production traits in the sheep mammary gland. Further studies focusing on elucidating the genetic mechanisms affected by the identified DMRs may help to better understand the biological mechanisms modified in the mammary gland of dairy sheep as a response to nutritional challenges and their potential effects on milk production.

Optically Active Spin Defects in Few-Layer Thick Hexagonal Boron Nitride.

Optically active spin defects in hexagonal boron nitride (hBN) are promising quantum systems for the design of two-dimensional quantum sensing units offering optimal proximity to the sample being probed. In this Letter, we first demonstrate that the electron spin resonance frequencies of boron vacancy centers (V_{B}^{-}) can be detected optically in the limit of few-atomic-layer thick hBN flakes despite the nanoscale proximity of the crystal surface that often leads to a degradation of the stability of solid-state spin defects. We then analyze the variations of the electronic spin properties of V_{B}^{-} centers with the hBN thickness with a focus on (i) the zero-field splitting parameters, (ii) the optically induced spin polarization rate and (iii) the longitudinal spin relaxation time. This Letter provides important insights into the properties of V_{B}^{-} centers embedded in ultrathin hBN flakes, which are valuable for future developments of foil-based quantum sensing technologies.

Isotopic Control of the Boron-Vacancy Spin Defect in Hexagonal Boron Nitride.

We report on electron spin resonance (ESR) spectroscopy of boron-vacancy (V_{B}^{-}) centers hosted in isotopically engineered hexagonal boron nitride (hBN) crystals. We first show that isotopic purification of hBN with ^{15}N yields a simplified and well-resolved hyperfine structure of V_{B}^{-} centers, while purification with ^{10}B leads to narrower ESR linewidths. These results establish isotopically purified h^{10}B^{15}N crystals as the optimal host material for future use of V_{B}^{-} spin defects in quantum technologies. Capitalizing on these findings, we then demonstrate optically induced polarization of ^{15}N nuclei in h^{10}B^{15}N, whose mechanism relies on electron-nuclear spin mixing in the V_{B}^{-} ground state. This work opens up new prospects for future developments of spin-based quantum sensors and simulators on a two-dimensional material platform.

Phase angle, muscle tissue, and resistance training.

The biophysical response of the human body to electric current is widely appreciated as a barometer of fluid distribution and cell function. From distinct raw bioelectrical impedance (BIA) variables assessed in the field of body composition, phase angle (PhA) has been repeatedly indicated as a functional marker of the cell's health and mass. Although resistance training (RT) programs have demonstrated to be effective to improve PhA, with varying degrees of change depending on other raw BIA variables, there is still limited research explaining the biological mechanisms behind these changes. Here, we aim to provide the rationale for the responsiveness of PhA determinants to RT, as well as to summarize all available evidence addressing the effect of varied RT programs on PhA of different age groups. Available data led us to conclude that RT modulates the cell volume by increasing the levels of intracellular glycogen and water, thus triggering structural and functional changes in different cell organelles. These alterations lead, respectively, to shifts in the resistive path of the electric current (resistance, R) and capacitive properties of the human body (reactance, Xc), which ultimately impact PhA, considering that it is the angular transformation of the ratio between Xc and R. Evidence drawn from experimental research suggests that RT is highly effective for enhancing PhA, especially when adopting high-intensity, volume, and duration RT programs combining other types of exercise. Still, additional research exploring the effects of RT on whole-body and regional BIA variables of alternative population groups is recommended for further knowledge development.

Nanoporous GaN by selective area sublimation through an epitaxial nanomask: AlN versus SixNy.

Nanoporous GaN layers were fabricated using selective area sublimation through a self-organized AlN nanomask in a molecular beam epitaxy reactor. The obtained pore morphology, density and size were measured using plan-view and cross-section scanning electron microscopy experiments. It was found that the porosity of the GaN layers could be adjusted from 0.04 to 0.9 by changing the AlN nanomask thickness and sublimation conditions. The room temperature photoluminescence properties as a function of the porosity were analysed. In particular, a strong improvement (>100) of the room temperature photoluminescence intensity was observed for porous GaN layers with a porosity in the 0.4-0.65 range. The characteristics of these porous layers were compared to those obtained with a SixNynanomask. Furthermore, the regrowth of p-type GaN on light emitting diode structures made porous by using either an AlN or a SixNynanomask were compared.

Interoception and the origin of feelings: A new synthesis.

Feelings are conscious mental events that represent body states as they undergo homeostatic regulation. Feelings depend on the interoceptive nervous system (INS), a collection of peripheral and central pathways, nuclei and cortical regions which continuously sense chemical and anatomical changes in the organism. How such humoral and neural signals come to generate conscious mental states has been a major scientific question. The answer proposed here invokes (1) several distinctive and poorly known physiological features of the INS; and (2) a unique interaction between the body (the 'object' of interoception) and the central nervous system (which generates the 'subject' of interoception). The atypical traits of the INS and the direct interactions between neural and non-neural physiological compartments of the organism, neither of which is present in exteroceptive systems, plausibly explain the qualitative and subjective aspects of feelings, thus accounting for their conscious nature.

Physical fitness trends in a nationally representative sample of Portuguese youth: What has changed from 2008 to 2018?

This investigation aimed to describe the current physical fitness (PF) status of Portuguese youth, compare secular trends from 2008 and 2018, and establish updated age- and sex-specific percentile values for distinct PF tests. In 2008 and 2018, 22 048 and 8960 children and adolescents (10-18 years) were included in two national cross-sectional investigations. PF was evaluated using the FITESCOLA® battery tests and the handgrip strength test. Independent sample t-tests and chi-squared tests were used to model the results. Weight smoothed percentile values were calculated using Cole's Lambda-Mu-Sigma (LMS) method. All analyses were weighted according to age, sex, and geographic region. In 2018, boys surpassed girls in the 20-m shuttle run, curl-ups, push-ups, standing long, and vertical jump tests, while girls performed better in the sit-and-reach (p < 0.05). The percentage of boys and girls meeting the healthy zone in the 20-min shuttle run test did not differ between 2008 and 2018 (p ≥ 0.05). In boys, a higher percentage fell in the healthy zone for the curl-up and push-up tests in 2018 compared to 2008 (85.8% vs. 83.4%, and 57.8% vs. 53.8%; p < 0.05). Girls improved their flexibility component (sit-and-reach test), with a higher percentage meeting the healthy zone in 2018 (32.6% vs. 36.9%; p < 0.05); an opposite trend was seen for boys (65.5% vs. 50.1%; p < 0.05). The present investigation provides new and updated PF percentile curves for Portuguese youth, which can be used as a general overview of the current PF state among the Portuguese young population.

Low-density SNP panel for efficient imputation and genomic selection of milk production and technological traits in dairy sheep.

The present study aimed to ascertain how different strategies for leveraging genomic information enhance the accuracy of estimated breeding values for milk and cheese-making traits and to evaluate the implementation of a low-density (LowD) SNP chip designed explicitly for that aim. Thus, milk samples from a total of 2,020 dairy ewes from 2 breeds (1,039 Spanish Assaf and 981 Churra) were collected and analyzed to determine 3 milk production and composition traits and 2 traits related to milk coagulation properties and cheese yield. The 2 studied populations were genotyped with a customized 50K Affymetrix SNP chip (Affymetrix Inc.) containing 55,627 SNP markers. The prediction accuracies were obtained using different multitrait methodologies, such as the BLUP model based on pedigree information, the genomic BLUP (GBLUP), and the BLUP at the SNP level (SNP-BLUP), which are based on genotypic data, and the single-step GBLUP (ssGBLUP), which combines both sources of information. All of these methods were analyzed by cross-validation, comparing predictions of the whole population with the test population sets. Additionally, we describe the design of a LowD SNP chip (3K) and its prediction accuracies through the different methods mentioned previously. Furthermore, the results obtained using the LowD SNP chip were compared with those based on the 50K SNP chip data sets. Finally, we conclude that implementing genomic selection through the ssGBLUP model in the current breeding programs would increase the accuracy of the estimated breeding values compared with the BLUP methodology in the Assaf (from 0.19 to 0.39) and Churra (from 0.27 to 0.44) dairy sheep populations. The LowD SNP chip is cost-effective and has proven to be an accurate tool for estimating genomic breeding values for milk and cheese-making traits, microsatellite imputation, and parentage verification. The results presented here suggest that the routine use of this LowD SNP chip could potentially increase the genetic gains of the breeding selection programs of the 2 Spanish dairy sheep breeds considered here.

Flat Bands and Giant Light-Matter Interaction in Hexagonal Boron Nitride.

Dispersionless energy bands in k space are a peculiar property gathering increasing attention for the emergence of novel electronic, magnetic, and photonic properties. Here, we explore the impact of electronic flat bands on the light-matter interaction. The van der Waals interaction between the atomic layers of hexagonal boron nitride induces flat bands along specific lines of the Brillouin zone. The macroscopic degeneracy along these lines leads to van Hove singularities with divergent joint density of states, resulting in outstanding optical properties of the excitonic states. For the direct exciton, we report a giant oscillator strength with a longitudinal-transverse splitting of 420 meV, a record value, confirmed by our ab initio calculations. For the fundamental indirect exciton, flat bands result in phonon-assisted processes of exceptional efficiency, that compete with direct absorption in reflectivity, and that make the internal quantum efficiency close to values typical of direct band gap semiconductors.

Study on the concordance between different SNP-genotyping platforms in sheep.

Different SNP genotyping technologies are commonly used in multiple studies to perform QTL detection, genotype imputation, and genomic predictions. Therefore, genotyping errors cannot be ignored, as they can reduce the accuracy of different procedures applied in genomic selection, such as genomic imputation, genomic predictions, and false-positive results in genome-wide association studies. Currently, whole-genome resequencing (WGR) also offers the potential for variant calling analysis and high-throughput genotyping. WGR might overshadow array-based genotyping technologies due to the larger amount and precision of the genomic information provided; however, its comparatively higher price per individual still limits its use in larger populations. Thus, the objective of this work was to evaluate the accuracy of the two most popular SNP-chip technologies, namely, Affymetrix and Illumina, for high-throughput genotyping in sheep considering high-coverage WGR datasets as references. Analyses were performed using two reference sheep genome assemblies, the popular Oar_v3.1 reference genome and the latest available version Oar_rambouillet_v1.0. Our results demonstrate that the genotypes from both platforms are suggested to have high concordance rates with the genotypes determined from reference WGR datasets (96.59% and 99.51% for Affymetrix and Illumina technologies, respectively). The concordance results provided in the current study can pinpoint low reproducible markers across multiple platforms used for sheep genotyping data. Comparing results using two reference genome assemblies also informs how genome assembly quality can influence genotype concordance rates among different genotyping platforms. Moreover, we describe an efficient pipeline to test the reliability of markers included in sheep SNP-chip panels against WGR datasets available on public databases. This pipeline may be helpful for discarding low-reliability markers before exploiting genomic information for gene mapping analyses or genomic prediction.

Criterion validity of a single-item question for assessment of daily breaks in sedentary time in adults.

BACKGROUND: As the implementation of sensor-based assessment for sedentary time (ST) and physical activity (PA) has practical limitations when applied on a large-scale, most studies rely on subjective data. We aimed to examine the criterion validity of a single-item question to assess daily breaks in ST and other PA-related outcomes for the first time using sensor-based data as the criterion. METHODS: In a sample of 858 adults, breaks in ST and other PA-related parameters were assessed through sensor-based accelerometry and subjective data, which included a comprehensive questionnaire with a specific question ('During the day, do you usually sit for a long time in a row or interrupt frequently?') with a three-level closed answer. The Spearman's rank correlation coefficient was used to determine the agreement between the single-item question and sensor-based data. RESULTS: Positive correlations were found for self-reported breaks in ST with sensor-based breaks in ST in both women (ρ=0.37; 95% CI=0.29-0.44) and men (ρ=0.15; 95% CI=0.04-0.26). Self-reported breaks in ST were inversely correlated with ST in women (ρ =-0.33; 95% CI=-0.40 to 0.25). For both sexes, self-reported breaks in ST showed a positive correlation with light-intensity PA (ρ=0.39; 95% CI=0.31-0.46 women; ρ=0.13; 95% CI=0.02-0.24 men), however, positive correlations between self-reported breaks in ST and moderate-to-vigorous PA (ρ=0.13; 95% CI=0.02-0.24) were found only in men. CONCLUSIONS: Our single-item question can be used as an indication for ranking people's breaks in ST during the waking day, although acknowledging that some misclassification will occur, especially in men. There must be an effort to include this question in future national and international surveys to replicate these findings.

Genome-wide association studies (GWAS) and post-GWAS analyses for technological traits in Assaf and Churra dairy breeds.

This study aimed to perform a GWAS to identify genomic regions associated with milk and cheese-making traits in Assaf and Churra dairy sheep breeds; second, it aimed to identify possible positional and functional candidate genes and their interactions through post-GWAS studies. For 2,020 dairy ewes from 2 breeds (1,039 Spanish Assaf and 981 Churra), milk samples were collected and analyzed to determine 6 milk production and composition traits and 6 traits related to milk coagulation properties and cheese yield. The genetic profiles of the ewes were obtained using a genotyping chip array that included 50,934 SNP markers. For both milk and cheese-making traits, separate single-breed GWAS were performed using GCTA software. The set of positional candidate genes identified via GWAS was subjected to guilt-by-association-based prioritization analysis with ToppGene software. Totals of 84 and 139 chromosome-wise significant associations for the 6 milk traits and the 6 cheese-making traits were identified in this study. No significant SNPs were found in common between the 2 studied breeds, possibly due to their genetic heterogeneity of the phenotypes under study. Additionally, 63 and 176 positional candidate genes were located in the genomic intervals defined as confidence regions in relation to the significant SNPs identified for the analyzed traits for Assaf and Churra breeds. After the functional prioritization analysis, 71 genes were identified as promising positional and functional candidate genes and proposed as targets of future research to identify putative causative variants in relation to the traits under examination. In addition, this multitrait study allowed us to identify variants that have a pleiotropic effect on both milk production and cheese-related traits. The incorporation of variants among the proposed functional and positional candidate genes into genomic selection strategies represent an interesting approach for achieving rapid genetic gains, specifically for those traits difficult to measure, such as cheese-making traits.

Estimation of genetic parameters for cheese-making traits in Spanish Churra sheep.

The global production of sheep milk is growing, and the main industrial use of sheep milk is cheese making. The Spanish Churra sheep breed is one of the most important native dairy breeds in Spain. The present study aimed to estimate genetic parameters for a wide range of traits influencing the cheese-making ability of Churra sheep milk. Using a total of 1,049 Churra ewes, we studied the following cheese-making traits: 4 traits related to milk coagulation properties (rennet coagulation time, curd-firming time, and curd firmness at 30 and 60 min after addition of rennet), 2 traits related to cheese yield (individual laboratory cheese yield and individual laboratory dried curd yield), and 3 traits measuring curd firmness over time (maximum curd firmness, time to attain maximum curd firmness, and syneresis). In addition, a list of milk traits, including the native pH of the milk and several milk production and composition traits (milk yield; the fat, protein, and dried extract percentages; and the somatic cell count), were also analyzed for the studied animals. After discarding the noncoagulating samples (only 3.7%), data of 1,010 ewes were analyzed with multiple-trait animal models by using the restricted maximum likelihood method to estimate (co)variance components, heritabilities, and genetic correlations. In general, the heritability estimates were low to moderate, ranging from 0.08 (for the individual laboratory dried curd yield trait) to 0.42 (for the fat percentage trait). High genetic correlations were found within pairs of related traits (i.e., 0.93 between fat and dried extract percentages, -0.93 between the log of the curd-firming time and curd firmness at 30 min, 0.70 between individual laboratory cheese yield and individual laboratory dried curd yield, and -0.94 between time to attain maximum curd firmness and syneresis). Considering all the information provided here, we suggest that in addition to the current consideration of the protein percentage trait for improving cheese yield traits, the inclusion of the pH of milk as a measured trait in the Churra dairy breeding program would represent an efficient strategy for improving the cheese-making ability of milk from this breed.

A hierarchy of correlates impacting adults' sensor-based physical activity and sedentary time.

Investigation into correlates across all levels of the socio-ecological model predictive of objectively measured physical activity has rarely been assessed in adults. While considering a diversity of correlates, we determined which correlates best predict sensor-based moderate-to-vigorous physical activity (MVPA) and sedentary-time (ST) in adults. A Chi-squared Automatic Interaction Detection algorithm was used to hierarchize the correlates associated with high ST (≥66.6thpercentile) and sufficient MVPA (≥150 min/week) in 865 adults. The main correlate predictive of being active was currently partaking in sport/exercise. The following relevant correlates were being male for the exercisers and having trees in the neighbourhood for the non-exercisers. The final correlate to boost male exercisers' MVPA was having lots of shops in the neighbourhood and not having television in the bedroom for women. The primary correlate for high ST was job activity level, with individuals having highly active jobs being less likely to exhibit high levels of ST; being single, male, and a former athlete also increased the chances of being highly sedentary. To increase adults' MVPA, promotion of sport participation, neighbourhood landscape planning, shop availability, as well as limiting television in the bedroom must be prioritized. For counteracting ST, increasing workplace activity level is warranted.

Strongly Confined Excitons in GaN/AlN Nanostructures with Atomically Thin GaN Layers for Efficient Light Emission in Deep-Ultraviolet.

Fascinating optical properties governed by extremely confined excitons have been so far observed in 2D crystals like monolayers of transition metal dichalcogenides. These materials, however, are limited for production by epitaxial methods. Besides, they are not suitable for the development of optoelectronics for the challenging deep-ultraviolet spectral range. Here, we present a single monolayer of GaN in AlN as a heterostructure fabricated by molecular beam epitaxy, which provides extreme 2D confinement of excitons, being ideally suited for light generation in the deep-ultraviolet. Optical studies in the samples, supplemented by a group-theory analysis and first-principle calculations, make evident a giant enhancement of the splitting between the dark and bright excitons due to short-range electron-hole exchange interaction that is a fingerprint of the strongly confined excitons. The practical significance of our results is in the observation of the internal quantum yield of the room-temperature excitonic emission as high as ∼75% at 235 nm.

Use of bacteriophage vB_Pd_PDCC-1 as biological control agent of Photobacterium damselae subsp. damselae during hatching of longfin yellowtail (Seriola rivoliana) eggs.

AIMS: This study describes the effect of phage therapy on hatching of longfin yellowtail (Seriola rivoliana) eggs challenged with Photobacterium damselae subsp. damselae. METHODS AND RESULTS: A lytic phage (vB_Pd_PDCC-1) against P. damselae subsp. damselae was isolated and characterized. The use of phage vB_Pd_PDCC-1 increased the hatching rate of eggs, and reduced presumptive Vibrio species to non-detectable numbers, even in non-disinfected eggs. High-throughput 16S rRNA gene sequencing analysis revealed that phage vB_Pd_PDCC-1 caused significant changes in the composition and structure of the associated microbiota, allowing that members (e.g. those belonging to the family Vibrionaceae) of the class Gammaproteobacteria to be displaced by members of the class Alphaproteobacteria. CONCLUSIONS: To the best of our knowledge, this represents the first study evaluating phage therapy to control potential negative effects of P. damselae subsp. damselae during hatching of longfin yellowtail eggs. SIGNIFICANCE AND IMPACT OF THE STUDY: The Seriola genus includes several important commercial fish species due to its rapid growth and easy adaptability to confinement conditions. However, bacterial infections (especially those caused by Vibrio and Photobacterium species) are among the main limiting factors for the intensification of marine fish aquaculture, particularly during early development stages. Therefore, the use of phages, which are natural killers of bacteria, represents a promising strategy to reduce the mortality of farmed organisms caused by pathogenic bacteria.

Identification of potential functional variants underlying ovine resistance to gastrointestinal nematode infection by using RNA-Seq.

In dairy sheep flocks from Mediterranean countries, replacement and adult ewes are the animals most affected by gastrointestinal nematode (GIN) infections. In this study, we have exploited the information derived from an RNA-Seq experiment with the aim of identifying potential causal mutations related to GIN resistance in sheep. Considering the RNA-Seq samples from 12 ewes previously classified as six resistant and six susceptible animals to experimental infection by Teladorsagia circumcincta, we performed a variant calling analysis pipeline using two different types of software, gatk version 3.7 and Samtools version 1.4. The variants commonly identified by the two packages (high-quality variants) within two types of target regions - (i) QTL regions previously reported in sheep for parasite resistance based on SNP-chip or sequencing technology studies and (ii) functional candidate genes selected from gene expression studies related to GIN resistance in sheep - were further characterised to identify mutations with a potential functional impact. Among the genes harbouring these potential functional variants (930 and 553 respectively for the two types of regions), we identified 111 immune-related genes in the QTL regions and 132 immune-related genes from the initially selected candidate genes. For these immune-related genes harbouring potential functional variants, the enrichment analyses performed highlighted significant GO terms related to apoptosis, adhesion and inflammatory response, in relation to the QTL related variants, and significant disease-related terms such as inflammation, adhesion and necrosis, in relation to the initial candidate gene list. Overall, the study provides a valuable list of potential causal mutations that could be considered as candidate causal mutations in relation to GIN resistance in sheep. Future studies should assess the role of these suggested mutations with the aim of identifying genetic markers that could be directly implemented in sheep breeding programmes considering not only production traits, but also functional traits such as resistance to GIN infections.

Silver nanoparticles are lethal to the ciliate model Tetrahymena and safe to the pike silverside Chirostoma estor.

Ciliate ectoparasites are one of the most important groups of pathogens in fish culture, and the traditional treatments are sometimes harmful to the fish and the environment. Thus, the search for novel compounds that are effective at low concentrations and safe for fish are necessary to optimise treatments in aquaculture. The antiprotozoal capacity of silver nanoparticles (AgNPs) against the ciliate Tetrahymena has been documented; however, their toxicity may vary with the synthesis methodology and nanoparticle size. The objectives of this study were a) to evaluate the acute toxicity in vitro of two AgNPs (Argovit™ and UTSA) on Tetrahymena sp., a biological model for ciliated ectoparasites of fish and b) to test the safety of lethal and higher doses of UTSA AgNPs for ciliates on the fish C. estor. Light microscopy and scanning electron microscopy (SEM) were used to determine whether AgNPs affected the structure of the cell surface of Tetrahymena. The mortality, histopathological alterations and metagenomics of the fish were used to determine the major effects of UTSA AgNPs. In Tetrahymena, the median lethal concentration (LC50) for Argovit™ was 2501 ± 1717 ng/L at 15 min and 796 ± 510 ng/L at 60 min, while the LC50 for UTSA AgNPs was 4 ± 2 and 1 ± 0.6 ng/L at 15 min and 60 min, respectively. A concentration of 3300 ng/L Argovit™ and 10.6 ng/L UTSA AgNPs for 15 and 60 min, respectively, was 100% effective against Tetrahymena. After 60 min of exposure to 0.25 and 0.50 ng/L UTSA AgNPs, the number of cilia significantly reduced, there were small holes on the cell surface, and the cellular membrane was ruptured. In fish exposed to lethal (10.6 ng/L) and higher (31.8 and 95.4 ng/L) doses of UTSA, the AgNPs did not affect fish survival after 96 h, and there were no signs of histopathological damage or gut microbial changes. This study is the first report on microscopic and ultrastructural changes in Tetrahymena after exposure to significantly low concentrations of UTSA AgNPs with antiprotozoal efficacy without evidence of harmful effects on fish. These results provide the basis for further studies of both pet aquarium and commercial fish that may validate these findings at a larger experimental scale, taking into account AgNPs bioaccumulation, safety for human consumption and environmental impact.