Hair cortisol concentration in finishing pigs on commercial farms: variability between pigs, batches, and farms

Hair cortisol is a stress indicator and could be used to assess the pigs’ exposure to stressors in the weeks/months prior to non-invasive hair sampling. The main aim of this study was to describe the hair cortisol concentration (HCC) variability between individuals within a batch, between farms and between batches within a farm. The secondary aim was to determine how the number of sampled pigs influences the characterization of HCC within a batch. Twenty farrow-to-finish pig farms were recruited considering the diversity of their management practices and health status (data collected). Hair was sampled in two separate batches, 8 months apart. The necks of 24 finishing pigs were clipped per batch the week prior to slaughter. To describe the variability in HCC, an analysis of the variance model was run with three explanatory variables (batch, farm and their interaction). To identify farm clusters, a principal component analysis followed by a hierarchical clustering was carried out with four active variables (means and standard deviations of the two batches per farm) and 17 supplementary variables (management practices, herd health data). We determined how the number of sampled pigs influenced the characterization of HCC within a batch by selecting subsamples of the results. HCC ranged from 0.4 to 121.6 pg/mg, with a mean of 25.9 ± 16.2 pg/mg. The variability in HCC was mainly explained by differences between pigs (57%), then between farms (24%), between batches within the same farm (16%) and between batches (3%). Three clusters of farms were identified: low homogeneous concentrations (n = 3 farms), heterogeneous concentrations with either higher (n = 7) or lower (n = 10) HCC in batch 2 than in batch 1. The diversity of management practices and health statuses allowed to discuss hypotheses explaining the HCC variations observed. We highlighted the need to sample more than 24 pigs to characterize HCC in a pig batch. HCC differences between batches on six farms suggest sampling pigs in more than one batch to describe the HCC at the farm level. HCC variations described here confirm the need to study its links with exposure of pigs to stressors

Cell therapy of Duchenne muscular dystrophy: preclinical trial in GRMD dogs

Duchenne muscular dystrophy (DMD), a genetic progressive X-linked muscular dystrophy, is the most common genetic disease in humans. Cell therapy based on the use of somatic stem cells is a very promising approach. In a dog myopathy model, we isolated a muscle stem cell (MuStem) with the essential requirements for therapeutic use: high amplification capacity, ability to fuse with muscle fibers, renewal of the satellite cell population, dispersion in the whole body after vascular administration, persistence of long-term effect, and dramatic clinical improvement of treated animals. These preclinical results pave the way for a therapeutic trial in children with Duchenne muscular dystrophy.La dystrophie musculaire de Duchenne (DMD) est une maladie génétique progressive du muscle liée au chromosome X. Elle est la maladie génétique la plus fréquente chez l'homme. La thérapie cellulaire basée sur l'utilisation de cellules souches somatiques est une voie thérapeutique riche d'intérêt. Nous avons isolé, chez un modèle de chien myopathe, une cellule souche musculaire (MuStem) qui présente les qualités indispensables à une utilisation thérapeutique: forte capacité d'amplification, capacité à fusionner avec les fibres musculaires, renouvellement du contingent de cellules satellites, dispersion dans l'organisme après administration vasculaire, persistance de l'effet à long terme, spectaculaire amélioration clinique des animaux traités. Ces résultats précliniques ouvrent la voie à un essai thérapeutique chez l'enfant atteint de dystrophie musculaire de Duchenne

Methodologies for Assessing Disease Tolerance in Pigs

Features of intensive farming can seriously threaten pig homeostasis, well-being and productivity. Disease tolerance of an organism is the adaptive ability in preserving homeostasis and at the same time limiting the detrimental impact that infection can inflict on its health and performance without affecting pathogen burden per se. While disease resistance (DRs ) can be assessed measuring appropriately the pathogen burden within the host, the tolerance cannot be quantified easily. Indeed, it requires the assessment of the changes in performance as well as the changes in pathogen burden. In this paper, special attention is given to criteria required to standardize methodologies for assessing disease tolerance (DT) in respect of infectious diseases in pigs. The concept is applied to different areas of expertise and specific examples are given. The basic physiological mechanisms of DT are reviewed. Disease tolerance pathways, genetics of the tolerance-related traits, stress and disease tolerance, and role of metabolic stress in DT are described. In addition, methodologies based on monitoring of growth and reproductive performance, welfare, emotional affective states, sickness behavior for assessment of disease tolerance, and methodologies based on the relationship between environmental challenges and disease tolerance are considered. Automated Precision Livestock Farming technologies available for monitoring performance, health and welfare-related measures in pig farms, and their limitations regarding DT in pigs are also presented. Since defining standardized methodologies for assessing DT is a serious challenge for biologists, animal scientists and veterinarians, this work should contribute to improvement of health, welfare and production in pigs

Induction de tolérance par les cellules dendritiques (prévention du diabète par le transfert des cellules dendritiques chez la souris Non Obese Diabetic)

Les cellules dendritiques jouent un rôle important dans les mécanismes de tolérance centrale et périphérique. Dans le cadre du diabète de type I, la rupture de tolérance, qui mène à l'agression des cellules pancréatiques, pourrait résulter d'un dysfonctionnement du compartiment des cellules dendritiques. Ainsi, l'injection des cellules dendritiques permet de prévenir le développement de la maladie chez la souris NOD (Non obese diabetic mice), modèle du diabète de type I. Cette capacité des cellules dendritiques à induire une tolérance n'est pas entièrement comprise, et notre objectif premier était d'étudier les mécanismes moléculaires qui permettent d'identifier des cellules dendritiques tolérogènes. Nous avons mis en place un modèle de prévention de diabète par le transfert des cellules dendritiques chez la souris NOD. Nos résultats montrent que cette protection s'associé à une une réponse humorale et cellulaire dirigée contre les antigènes bovins, qui est dû à l'injection des cellules dendritiques générées en présence du sérum de veau fœtal. Ces résultats montrent que les observations obtenues dans les protocoles d'immunothérapies utilisant des cellules dendritiques générées en présence du sérum bovin doivent être interprétés avec précautions.In recent years, several investigators have shown that transfer of dendritic cells (DC) prevents development of diabetes in NOD mice. Accumulating evidence that DC cultured in the presence of FCS can induce a dominant unspecific response in tumour models prompted us to investigate the effect of the injection of bone marrow derived DC cultured in the presence of FCS on diabetes development in NOD mice. We showed that two injections, one week apart, with mature bone marrow-derived DC protected NOD mice from developing spontaneous diabetes and induced marked anti-Fetal Calf Serum (FCS) immune responses. Anti-FCS antibodies were detected in serum of DC-injected mice after the first injection, which increased dramatically after the second one and remained higher until 10 months of age. Besides, spleen cells of NOD mice (SC) were isolated one week after DC injection and cultured in RPMI supplemented with FCS, in AIMV-BSA or in RPMI containing autologous mouse serum for autologous mixte lymphocyte reactions. Results showed that SC proliferation increased in the presence of bovine serum proteins suggesting that lymphocytes have been primed against bovine proteins in vivo after DC injection. Also, flow cytometry data using CFSE labelling showed that proliferation was observed in both B and T lymphocytes compartments. All together, our data suggest that injection of DC cultured in FCS-containing medium could switch an aggressive Th1 response to a protective Th2 one. Our results also show that DC grown in the presence of FCS can induce specific anti-FCS immune responses urging cautious interpretation of DC based vaccination protocols.NANTES-BU Sciences (441092104) / SudocSudocFranceF

Type 1 diabetes prone NOD mice have diminished Cxcr1 mRNA expression in polymorphonuclear neutrophils and CD4+ T lymphocytes

In humans, CXCR1 and CXCR2 are two homologous proteins that bind ELR+ chemokines. Both receptors play fundamental roles in neutrophil functions such as migration and reactive oxygen species production. Mouse Cxcr1 and Cxcr2 genes are located in an insulin-dependent diabetes genetic susceptibility locus. The non obese diabetic (NOD) mouse is a spontaneous well-described animal model for insulin-dependent type 1 diabetes. In this disease, insulin deficiency results from the destruction of insulin-producing beta cells by autoreactive T lymphocytes. This slow-progressing disease is dependent on both environmental and genetic factors. Here, we report descriptive data about the Cxcr1 gene in NOD mice. We demonstrate decreased expression of mRNA for Cxcr1 in neutrophils and CD4+ lymphocytes isolated from NOD mice compared to other strains, related to reduced NOD Cxcr1 gene promoter activity. Looking for Cxcr1 protein, we next analyze the membrane proteome of murine neutrophils by mass spectrometry. Although Cxcr2 protein is clearly found in murine neutrophils, we did not find evidence of Cxcr1 peptides using this method. Nevertheless, in view of recently-published experimental data obtained in NOD mice, we argue for possible Cxcr1 involvement in type 1 diabetes pathogenesis

Culture medium modulates the behaviour of human dental pulp-derived cells: technical note.

International audienc

beta 2-adrenoreceptor stimulation dampens the LPS-induced M1 polarization in pig macrophages

The cross-talk between sympatho-adreno-medullar axis and innate immunity players was mainly studied in rodents. In intensive husbandry, pigs are exposed to multiple stressors inducing repeated releases of catecholamines that bind to adrenoreceptors (AR) on target cells. Among adrenoreceptors, the (beta 2-AR is largely expressed by immune cells including macrophages. We report herein on the effects of catecholamines, through (beta 2-AR stimulation, on pig macrophage functions activated by LPS. beta 2-AR stimulation of porcine macrophages prevented the LPS-induced increase in TINF alpha and IL-8 secretion while increasing IL-10 secretion. In contrast, treatment with a beta 2-agonist had no effect on anti-microbial functions. Lastly, beta 2-AR stimulation of macrophages reduced the expression of genes up regulated by LPS. Altogether, we demonstrated that (beta 2-AR stimulation of porcine macrophages prevented polarization towards a pro-inflammatory phenotype. Since porcine macrophages are a suitable model for human macrophages, our results might be relevant to appreciate catecholamine effects on human macrophages

<i>Cxcr1</i> (A and C) and <i>Cxcr2</i> (B and D) mRNA levels in mouse neutrophils (A and B) and CD4+ lymphocytes (C and D).

<p>Relative <i>Cxcr1</i> and <i>Cxcr2</i> mRNA expression in neutrophils and CD4+ lymphocytes isolated from different mouse strains. Data were analyzed with the relative quantification standard curve method and expression data were normalized to <i>ß-actin</i> and <i>Rpb1</i> housekeeping genes. Each dot represents an independent sample; lines are medians. A, B: pooled data of five independent experiments are shown (<i>Cxcr1</i>- Kruskal-Wallis test: p<0.0001; Dunn’s post test NOD versus NOR, p<0.001; NOD versus C57Bl/6J: p<0.05. <i>Cxcr2</i>- Kruskal-Wallis test, ns). C, D: pooled data of three independent experiments are shown (<i>Cxcr1</i>- Kruskal-Wallis test: p<0.001; Dunn’s post test NOD versus C57Bl/6J, p<0.001; NOD versus Balb/C: p<0.05. <i>Cxcr2</i>- Kruskal-Wallis test: p<0.005; Dunn’s post test NOD versus C57Bl/6J, p<0.01).</p

Dataset of hair cortisol concentration in 950 finishing pigs on 20 commercial farms

This dataset contains the hair cortisol concentrations of 950 finishing pigs. Pig hair was sampled as part of a study funded by the European project HealthyLivestock. Pigs were sampled in two separate batches on 20 farms (24 pigs/batch, two batches/farm. NB: 10 samples could not be analyzed at the laboratory). Farms were located in western France. A reference to the article relating to this dataset will be added when the article will be published. -The first sheet includes the 950 hair cortisol concentrations, distinguishing the batches and farms where pigs were sampled. -The second sheet includes the estimation of the average size of a pig batch on the 20 farms where hair was sampled + the estimation of the percentage of pigs sampled per batch

Dexamethasone stimulates differentiation of odontoblast-like cells in human dental pulp cultures

International audienceRegenerative dental pulp strategies require the identification of precursors able to differentiate into odontoblast-like cells that secrete reparative dentin after injury. Pericytes have the ability to give rise to osteoblasts, chondrocytes, and adipocytes, a feature that has led to the suggestion that odontoblast-like cells could derive from these perivascular cells. In order to gain new insights into this hypothesis, we investigated the effects of dexamethasone (Dex), a synthetic glucocorticoid employed to induce osteogenic differentiation in vitro, in a previously reported model of human dental pulp cultures containing pericytes as identified by their expression of smooth muscle actin (SMA) and their specific ultrastructural morphology. Our data indicated that Dex (10(-8) M) significantly inhibited cell proliferation and markedly reduced the proportion of SMA-positive cells. Conversely, Dex strongly stimulated alkaline phosphatase (ALP) activity and induced the expression of the transcript encoding the major odontoblastic marker, dentin sialophosphoprotein. Nevertheless, parathyroid hormone/parathyroid hormone-related peptide receptor, core-binding factor a1/osf 2, osteonectin, and lipoprotein lipase mRNA levels were not modified by Dex treatment. Dex also increased the proportion of cells expressing STRO-1, a marker of multipotential mesenchymal progenitor cells. These observations indicate that glucocorticoids regulate the commitment of progenitors derived from dental pulp cells to form odontoblast-like cells, while reducing the proportion of SMA-positive cells. These results provide new perspectives in deciphering the cellular and molecular mechanisms leading to reparative dentinogenesis