Metallo-Glycodendrimeric Materials against Enterotoxigenic Escherichia coli.

Conjugation of carbohydrates to nanomaterials has been extensively studied and recognized as an alternative in the biomedical field. Dendrimers synthesized with mannose at the end group and with entrapped zero-valent copper/silver could be a potential candidate against bacterial proliferation. This study is aimed at investigating the bactericidal activity of metal-glycodendrimers. The Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction was used to synthesize a new mannosylated dendrimer containing 12 mannopyranoside residues in the periphery. The enterotoxigenic Escherichia coli fimbriae 4 (ETEC:F4) viability, measured at 600 nm, showed the half-inhibitory concentration (IC50) of metal-free glycodendrimers (D), copper-loaded glycodendrimers (D:Cu) and silver-loaded glycodendrimers (D:Ag) closed to 4.5 × 101, 3.5 × 101 and to 1.0 × 10-2 µg/mL, respectively, and minimum inhibitory concentration (MIC) of D, D:Cu and D:Ag of 2.0, 1.5 and 1.0 × 10-4 µg/mL, respectively. The release of bacteria contents onto broth and the inhibition of ETEC:F4 biofilm formation increased with the number of metallo-glycodendrimer materials, with a special interest in silver-containing nanomaterial, which had the highest activity, suggesting that glycodendrimer-based materials interfered with bacteria-bacteria or bacteria-polystyrene interactions, with bacteria metabolism and can disrupt bacteria cell walls. Our findings identify metal-mannose-dendrimers as potent bactericidal agents and emphasize the effect of entrapped zero-valent metal against ETEC:F4.

The Development of a Risk Assessment Model for Inedible Rendering Plants in Canada: Identifying and Selecting Feed Safety-Related Factors.

The Canadian Food Inspection Agency (CFIA) is developing an establishment-based risk assessment model to categorize rendering plants that produce livestock feed ingredients (ERA-Renderer model) according to animal and human health risks (i.e., feed safety risks) and help in determining the allocation of inspection resources based on risk. The aim of the present study was to identify and select feed-safety-related factors and assessment criteria for inclusion in the ERA-Renderer model. First, a literature review was performed to identify evidence-based factors that impact the feed safety risk of livestock feed during its rendering processes. Secondly, a refinement process was applied to retain only those that met the inclusion conditions, such as data availability, lack of ambiguity, and measurability. Finally, an expert panel helped in selecting factors and assessment criteria based on their knowledge and experience in the rendering industry. A final list of 32 factors was developed, of which 4 pertained to the inherent risk of a rendering plant, 8 were related to risk mitigation strategies, and 20 referred to the regulatory compliance of a rendering plant. A total of 179 criteria were defined to assess factors based on practices in the Canadian rendering industry. The results of this study will be used in the next step of the model development to estimate the relative risks of the assessment criteria considering their impact on feed safety. Once implemented, the CFIA's ERA-Renderer model will provide an evidence-based, standardized, and transparent approach to help manage the feed safety risks in Canada's rendering sector.

Early decrease of ionized calcium and static symmetric dimethylarginine concentration in a model of renal tubular necrosis in corn snakes (Pantherophis guttatus).

OBJECTIVE: To determine if plasma concentrations of symmetric dimethylarginine (SDMA), N-acetyl-beta-d-glucosaminidase (NAG), GGT, ALT, AST, lactate, total calcium, and ionized calcium (iCa) and the calcium:phosphorus ratio are clinically relevant biomarkers to detect early stages of tubular lesions in snakes. ANIMALS: 6 adult corn snakes (Pantherophis guttatus). METHODS: Corn snakes were administered 11 injections of gentamicin at 50 mg/kg, SC, q 24 h in an experimental model of induced tubular necrosis. Plasma biochemistry and blood gas analyses were performed at baseline and after the 3rd and 11th injections. Parameters were compared between time points using a paired Wilcoxon test. In 3 individuals, renal biopsies were collected at baseline before starting injections and at the 3rd and 11th injections, while renal tissue samples were procured after euthanasia in all individuals. RESULTS: Renal proximal and distal tubular necrosis and hepatic steatosis were present in all individuals at necropsy. Compared to baseline, decreased blood concentrations of lactate, ionized calcium, and total calcium and a decreased calcium:phosphorus ratio were noted. A significant decrease of lactate and ionized calcium was observed after 3 days. Conversely, no changes in SDMA, NAG, ALT, AST, GGT, and sodium were detected. CLINICAL RELEVANCE: Ionized calcium and lactate concentrations were the earliest parameters to decrease compared to baseline values in this experimental model. While SDMA is a sensitive indicator of renal disease in mammals, this biomarker did not increase in a model of induced acute tubular necrosis in corn snakes.

Vitamin D Supplementation Impacts Calcium and Phosphorus Metabolism in Piglets Fed a Diet Contaminated with Deoxynivalenol and Challenged with Lipopolysaccharides.

Using alternative feed ingredients in pig diets can lead to deoxynivalenol (DON) contamination. DON has been shown to induce anorexia, inflammation, and-more recently-alterations in the vitamin D, calcium, and phosphorus metabolisms. Adding vitamin D supplementation in the form of vitamin D3 and 25-OH-D3 to the feed could modify the effects of DON in piglets. In this study, vitamin D3 or 25-OH-D3 supplementation was used in a control or DON-contaminated treatment. A repetitive exposure over 21 days to DON in the piglets led to disruptions in the vitamin D, calcium, and phosphorus metabolisms, resulting in a decreased growth performance, increased bone mineralization, and the downregulation of genes related to calcium and to phosphorus intestinal and renal absorption. The DON challenge also decreased blood concentrations of 25-OH-D3, 1,25-(OH)2-D3, and phosphate. The DON contamination likely decreased the piglets' vitamin D status indirectly by modifying the calcium metabolism response. Vitamin D supplementations did not restore vitamin D status or bone mineralization. After a lipopolysaccharide-induced inflammatory stimulation, feeding a 25-OH-D3 supplementation increased 25-OH-D3 concentration and 1,25-(OH)2-D3 regulations during the DON challenge. DON contamination likely induced a Ca afflux by altering the intestinal barrier, which resulted in hypercalcemia and hypovitaminosis D. The vitamin D supplementation could increase the calcitriol production to face the combined LPS and DON challenge.

Health indicators in surplus calves at the time of arrival at auction markets: Associations with distance from farms of origin in Québec, Canada.

This study evaluated the associations between estimated distance from farms' locations to auction markets, and health indicators of surplus dairy calves sold during summer 2019 and winter 2020 in Québec, Canada. A total of 3,610 animals from 1,331 different farms were used in this cross-sectional cohort study. Geographic coordinates (latitude and longitude) were obtained for each farm and the 2 participating livestock auction markets. Calves' abnormal physical signs (APS) were noted upon arrival at the auction market as they were examined by trained research staff. The haversine distance between the farm and the auction market was evaluated using geographic coordinates and categorized. Generalized linear mixed models were used for statistical analyses. The main APS observed were ocular discharge (34.9%), abnormal hide cleanliness (21.2%), swollen navel (17.2%), dehydration score 1 (at least one of the 2 following clinical signs: persistent skin tent or sunken eye, 12.9%), and dehydration score 2 (both clinical signs mentioned above, 6.5%). Calves from farms located at greater distances from the auction markets (≥110 km) had a higher risk ratio [RR = 1.08; 95% confidence internal (CI) = 1.03, 1.13] for dehydration than those from lesser distances (0-25 km). During the summertime, a RR of 1.18 (95% CI = 1.15, 1.22) was observed for dehydration compared with wintertime. A 2-way interaction between estimated distance and season showed a higher prevalence of ocular discharge for calves from farms at distances greater than or equal to 110 km during the summer (RR = 1.11; 95% CI: 1.04, 1.20) than for calves from farms located at lesser distances (0-25 km). These results demonstrate that calves from farms located at greater distances from the auction markets had more APS, mainly during the summer. A better understanding of the transport conditions and interaction with management at the farm of origin is determinant to mitigate the impact of the journey on surplus calf health.

Inter-Rater Reliability of Scoring Systems for Abomasal Lesions in Quebec Veal Calves.

The objective of this study was to determine the inter-rater reliability of current scoring systems used to detect abomasal lesions in veal calves. In addition, macroscopic lesions were compared with corresponding histological lesions. For this, 76 abomasa were retrieved from veal calves in a slaughterhouse in Quebec and scored by four independent raters using current scoring systems. The localisations of the lesions were separated into pyloric, fundic, or torus pyloricus areas. Lesions were classified into three different types, i.e., erosions, ulcers, and scars. To estimate the inter-rater reliability, the coefficient type 1 of Gwet's agreement and Fleiss κ were used for the presence or absence of a lesion, and the intra-class correlation coefficient was used for the number of lesions. All veal calves had at least one abomasal lesion detected. Most lesions were erosions, and most of them were located in the pyloric area. Overall, a poor to very good inter-rater agreement was seen for the pyloric area and the torus pyloricus regarding the presence or absence of a lesion (Fleiss κ: 0.00-0.34; Gwet's AC1: 0.12-0.83), although a higher agreement was observed when combining all lesions in the pyloric area (Fleiss κ: 0.09-0.12; Gwet's AC1: 0.43-0.93). For the fundic area, a poor to very good agreement was also observed (Fleiss κ: 0.17-0.70; Gwet's AC1: 0.90-0.97). Regarding the inter-rater agreement for the number of lesions, a poor to moderate agreement was found (ICC: 0.11-0.73). When using the scoring system developed in the European Welfare Quality Protocol, a poor single random rater agreement (ICC: 0.42; 95% CI: 0.31-0.56) but acceptable average random rater agreement (ICC: 0.75; 95% CI: 0.64-0.83) was determined. Microscopic scar lesions were often mistaken as ulcers macroscopically. These results show that the scoring of abomasal lesions is challenging and highlight the need for a reliable scoring system. A fast, simple, and reliable scoring system would allow for large scale studies which investigate possible risk factors and hopefully help to prevent these lesions, which can compromise veal calves' health and welfare.

Longitudinal Data to Assess Relationships among Plasma Folate, Vitamin B12, Non-esterified Fatty Acid, and ß-Hydroxybutyrate Concentrations of Holstein Cows during the Transition Period.

It is well established that the plasma metabolite profile changes during metabolic dysfunction, such as elevated non-esterified fatty acid (NEFA) release when body reserve mobilization is excessive in early lactation cows. Relationships between changes in plasma concentrations of metabolites caused by a metabolic impairment and the status of vitamins, such as folates and vitamin B12, have barely been studied in cattle. This study was undertaken to assess relationships between peripartum plasma concentrations of folates, vitamin B12, NEFA, and beta-hydroxybutyrate (BHB). Longitudinal data of 48 multiparous Holstein cows from 5 studies were taken from days -14 to 21 relative to calving. Blood samples were taken weekly before calving and either twice or thrice per week postpartum, and plasma was analyzed for folate, vitamin B12, NEFA, and BHB concentrations. Postpartum plasma NEFA and BHB concentrations were negatively related to plasma folate concentration at days -14 and -7 relative to parturition, whereas the opposite relationship was noted for the plasma vitamin B12:folate ratio. The plasma folate and NEFA areas under the curve from the whole studied period were negatively associated, and the opposite was observed with the association between the plasma vitamin B12:folate ratio and NEFA as well as the BHB areas under the curve. The results suggest that there is an increased use of folate for metabolic functions during elevated concentrations of plasma NEFA and BHB. Future research should focus on finding an optimal plasma vitamin B12:folate ratio to favor cow health during the challenging period of parturition.

Assessment of Zearalenone-Induced Cell Survival and of Global Gene Regulation in Mouse TM4 Sertoli Cells.

Zearalenone (ZEA) is a non-steroidal xenoestrogen mycotoxin produced by many Fusarium fungal species, which are common contaminants of cereal crops destined for worldwide human and animal consumption. ZEA has been reported in various male reproduction dysfonctions, including decreased fertility potential. In this report, the direct effect of ZEA on the immature Sertoli TM4 cell line was evaluated. The results show that high concentrations of ZEA increase reactive oxygen species via the activation of MAPK signaling. Transcriptome analysis was performed on the TM4 cell line treated with ZEA, and genes involved in sex differentiation (Fgfr2, Igf1, Notch1, Sox9) and extracellular matrix (ECM) formation (Ctgf, Fam20a, Fbn1, Mmp9, Postn, Sparcl1, Spp1) were identified at the center of the functional protein association network, suggesting that ZEA could be detrimental to the early steps of Sertoli cell differentiation.

The administration of diets contaminated with low to intermediate doses of deoxynivalenol and supplemented with antioxidants and binding agents slightly affects the growth, antioxidant status, and vaccine response in weanling pigs.

This study aimed to evaluate the impact of grading levels of deoxynivalenol (DON) in the diet of weaned pigs, as well as the effects of a supplementation with antioxidants (AOX), hydrated sodium calcium aluminosilicates (HSCAS), and their combination on the growth, AOX status, and immune and vaccine responses against the porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2). At weaning, 336 piglets were allocated to six dietary treatments according to a randomized complete block design. Treatments were as follows: basal diet (CTRL); basal diet containing DON at 1.2 mg/kg (DON1.2); basal diet containing DON at 2.4 mg/kg (DON2.4); DON2.4 diet + a mix of AOX which included vitamins A and E at 20,000 IU and 200 IU/kg feed respectively, selenized yeast at 0.3 mg/kg, and a grape seed extracts at 100 mg/kg feed (DON2.4 + AOX); DON2.4 diet + the mix of AOX and the modified HSCAS mentioned above (DON2.4 + AOX + HSCAS); DON2.4 + AOX + HSCAS. Pigs were vaccinated against PRRSV and PCV2 at 7 d; on 0, 14, and 35 d, growth performance was recorded, and blood samples were collected in order to evaluate the oxidative status, inflammatory blood markers, lymphocyte blastogenic response, and vaccine antibody response. Increasing intake of DON resulted in a quadratic effect at 35 d in the lymphocyte proliferative response to concanavalin A and PCV2 as well as in the anti-PRRSV antibody response, whereas the catalase activity decreased in DON2.4 pigs compared with the CTRL and DON1.2 groups (P ≤ 0.05). Compared with the DON2.4 diet, the AOX supplementation slightly reduced gain to feed ratio (P = 0.026) and increased the ferric reducing ability of plasma as well as α-tocopherol concentration (P < 0.05), whereas the association of AOX + HSCAS increased the anti-PRRSV IgG (P < 0.05). Furthermore, the HSCAS supplement reduced haptoglobin levels in serum at 14 d compared with the DON2.4 group; however, its concentration decreased in all the experimental treatments from 14 to 35 d and particularly in the DON2.4 + AOX pigs, whereas a different trend was evidenced in the DON2.4 + HSCAS group, where over the same period haptoglobin concentration increased (P < 0.05). Overall, our results show that the addition of AOX and HSCAS in the diet may alleviate the negative effects due to DON contamination on the AOX status and immune response of vaccinated weanling pigs.

Expert Elicitation To Estimate the Feed Safety Impact of Criteria Included in the Canadian Food Inspection Agency Risk Assessment Model for Feed Mills.

ABSTRACT: The Canadian Food Inspection Agency is developing an Establishment-based Risk Assessment (ERA) model for commercial and on-farm mills involved in the manufacture, storage, packaging, labeling, or distribution of livestock feed (ERA-Feed Mill model). This model will help inform the allocation of inspection resources on the basis of feed safety risk, including animal health and food safety risk. In a previous study, 34 risk factors, grouped into inherent, mitigation, and compliance clusters, along with assessment criteria were selected. The objective of this current study was to estimate the relative risk (RR) of the 203 assessment criteria on the basis of the impact on feed safety to design an ERA-Feed Mill model algorithm. Furthermore, the intent of this study was to assess the maximum increase or decrease of risk obtained when multiple criteria belonging to a same cluster were identified in a specific feed mill. To do so, a two-round face-to-face expert elicitation was conducted with 28 Canadian feed experts. Results showed no significant association between respondent profiles (years of experience and work sector) and estimated RR. Uniformity of answers between experts improved between rounds. Criteria having the highest increase in risk (median RR ≥ 4) included the presence of materials prohibited to be fed to ruminants in a facility that produces ruminant feed, the presence of multiple livestock species on-site, and historical noncompliances related to the inspection of the feed mill's process control and end-product control programs. Risk mitigation criteria having the highest impact on decreasing the risk were the implementation of feed safety certifications, the use of dedicated manufacturing lines (prohibited materials or medications), and having a hazard sampling plan in place for finished feed. The median RR assigned to each criterion and cluster will be used to build an algorithm of the Canadian Food Inspection Agency's ERA-Feed Mill model.

Effects of the mycotoxin metabolite de-epoxy-deoxynivalenol (DOM-1) on embryo development and sperm motility in cattle.

Contamination of animal feed with Fusarium spp results in accumulation of mycotoxins including deoxynivalenol. In animals, deoxynivalenol is metabolized to de-epoxy deoxynivalenol (DOM-1), which is generally considered to be a non-toxic metabolite; however, recent studies demonstrated that DOM-1 can reduce steroid production and induce apoptosis in the bovine ovary. The objectives of this study were to assess the effects of DOM-1 on applied aspects of reproductive function in cattle, specifically sperm function and embryo development in vitro and follicle growth and superovulatory responses in vivo. The effect of naturally contaminated feed on superovulatory responses was assessed; a dose of 6 ppm deoxynivalenol increased blood DOM-1 concentrations to 20 ng/ml, but this did not alter the number of viable embryos recovered on day 7. However, intrafollicular injection of DOM-1 (100 ng/ml) directly into the growing dominant follicle resulted in cessation of follicular growth over the subsequent 3 days. Treatment with DOM-1 reduced motility of bull spermatozoa over a 10-h period in vitro. Addition of DOM-1 to oocytes in vitro during IVM did not alter rates of cumulus expansion and nuclear maturation, but treatment during IVF reduced the rate of blastocyst formation. These data illustrate that DOM-1 is more biologically active than previously thought and negatively impacted reproductive outcomes in cattle.

Interleukins' expression profile changes in granulosa cells of preovulatory follicles during the postpartum period in dairy cows.

The postpartum period in dairy cows is associated with a state of temporary negative energy balance and could induce functional changes into ovarian granulosa cells (GC) resulting in significant impact on the ovarian function and fertility. Yet, the regulation of interleukin receptors (ILRs) in GC as well as ILs expression profile during the postpartum period have not been fully investigated. We hypothesized that the postpartum period is associated with changes in ILs expression profile that could affect follicular development and ovulation rate. First, we aimed to investigate the expression and regulation of different IL and IL receptors in GC at different stages of follicular development and then analyse the changes in target ILs expression profile induced during the postpartum period. In the first objective, normal cycling cows were selected and GC were collected from small follicles (SF), dominant follicles at day 5 of the estrous cycle (DF), and ovulatory follicles, 24 h following hCG injection (OF). In the second objective, dairy cows between 50 and 70 days postpartum were randomly selected, and ß-hydroxybutyrate (BHB) concentrations were measured in blood samples in order to assign cows to the BHB+ group (>1.4 mmol/L) or BHB- group (<1.2 mmol/L). GC were collected from preovulatory follicles by transvaginal aspiration. Total RNA was extracted from GC of all groups for analysis of target ILs and ILRs expression. Steady-state mRNA levels of IL4R was strongest in the DF, while IL15R expression was greatest in the OF, and IL21R showed increased steady-state mRNA levels in the corpus luteum as compared to the different groups of follicles. Overall, expression of IL1A, IL1B, IL8, IL15, IL23 and TNFα was stronger in OF as compared to DF, while IL4 and IL10 expression was stronger in SF than in DF. Similarly, expression of IL1A, IL1B, IL8, IL15, IL23, and TNFα were significantly stronger in GC of BHB+ cows than in the control, while IL4 expression was significantly reduced in BHB+ as compared to control cows. We have established an IL expression profile, which suggest a correlation with BHB levels during the postpartum period. Additionally, we have demonstrated a differential regulation of target ILRs in GC at different stages of follicular development. Overall, these data provide a better understanding of the changes that could affect follicular development and ovulation during the postpartum period and lay the ground for further investigations.

In vivo evaluation of targeted delivery of biological agents using barium sulfate.

This study was aimed to monitor the transit through the intestine by X-ray imaging using barium sulfate (BS) as tracer. The in vitro features of monolithic tablets were correlated with their in vivo behavior in order to provide a tool for the development of targeted formulations containing macromolecular bioactive agents. The impact of BS on various matrices (neutral, ionic) was studied in simulated fluids using the disintegration time (DT) as main parameter. Dry tablets were characterized by spectroscopic methods (X-ray diffraction and Infra-Red) and scanning electron microscopy (SEM). The selected formulations were followed in a beagle dog model. The in vivo and in vitro DT of tablets formulated with BS were compared. Results: anionic excipients carboxymethylcellulose (CMC) and carboxymethylstarch (CMS) protected the active ingredient from the gastric acidity, ensuring its targeted delivery in the intestine. The SEM analysis, before and after transit in simulated fluids, showed that BS remained in the tablets allowing their good follow-up in vivo. The incorporation of 30% protein in tablets with 40% BS had no impact on their behavior. In conclusion, BS and X-ray imagery could be a good alternative to scintigraphy for development of targeted formulations containing high molecular weight bioactive agents.

Effects of deoxynivalenol and sodium meta-bisulphite on nutrient digestibility in growing pigs.

Deoxynivalenol (DON), a mycotoxin synthesised by the Fusarium, is known to affect the growth of pigs. This effect can be attenuated with sodium meta-bisulphite (SBS). The aim of this study was to evaluate the effect of SBS with antioxidant blend on nutrient digestibility in pigs fed a diet contaminated naturally with DON. Six crossbred castrated pigs fitted surgically with single-T cannulas in the distal ileum received one of four barley-corn-soybean diets with or without SBS. After 8 d of feeding, faeces and ileal digesta were collected for 2 d. Apparent ileal digestibility (AID) of the dry matter (DM), energy, nutrients and DON, and apparent total tract digestibility (ATTD) of DM, acid detergent fibre (ADF), neutral detergent fibre (NDF), energy and DON were evaluated. The AID of phosphorus, calcium and some amino acids was increased (p < 0.05) in the DON diets whereas the ATTD of DM and energy tended to decrease (p = 0.064 and p = 0.071). SBS reduced the AID of DM, energy, ADF, ether extract, phosphorus and DON (p < 0.05) but had no effect on the ATTD of DM, energy, fibre or DON. These results show that DON improved the AID of some nutrients but tended to reduce the ATTD of energy, which could explain, although anorexia is the main effect of DON on live weight gain, the reported negative effect of DON on pig growth. Finally, SBS with antioxidant blend had reduced AID of some nutrients and intestinal absorption of DON.

Use of antibiotics in broiler production: Global impacts and alternatives.

Antibiotics are used to fight bacterial infections. However, a selective pressure gave rise to bacteria resistant to antibiotics. This leaves scientists worried about the danger to human and animal health. Some strategies can be borrowed to reduce the use of antibiotics in chicken farms. Much research has been carried out to look for natural agents with similar beneficial effects of growth promoters. The aim of these alternatives is to maintain a low mortality rate, a good level of animal yield while preserving environment and consumer health. Among these, the most popular are probiotics, prebiotics, enzymes, organic acids, immunostimulants, bacteriocins, bacteriophages, phytogenic feed additives, phytoncides, nanoparticles and essential oils.

Alternatives to antibiotics in poultry feed: molecular perspectives.

The discovery of the growth promoting property of antibiotics led to their use as antibiotic feed additives (AFAs) in animal feed at sub-therapeutic doses. Although this has been beneficial for animal health and productivity, it has been, essentially, a double-edged sword. The continued and non-judicious use of AFAs has led to the selection and dissemination of antibiotic-resistant strains of poultry pathogens such as Salmonella, Campylobacter and Escherichia coli. The rapid spread of drug-resistant pathogens as well as emergence of antibiotic-related environmental pollutants is of global concern. Hence, the identification and development of new and effective alternatives to antibiotics that do not hinder productivity is imperative. For this, it is essential to understand not only the molecular basis of development of resistance to AFAs but also the mechanisms of action of AFA alternatives and how they differ from AFAs. This review provides a molecular perspective on the alternatives to antibiotics that have been proposed till date and their current trends, as well as novel approaches such as development of improved delivery systems.

The mycotoxin metabolite deepoxy- deoxynivalenol increases apoptosis and decreases steroidogenesis in bovine ovarian theca cells.

The mycotoxin deoxynivalenol (DON) has been shown to inhibit ovarian granulosa cell function in cattle in vitro, but it is not known whether DON or its metabolite deepoxy-DON (DOM-1) affects theca cell function. The objectives of this study were to determine the effects of DON and of DOM-1 on theca cell steroidogenesis and apoptosis, and to determine the main pathways through which they act. Bovine theca cells were cultured in a nonluteinizing serum-free culture system, and challenged with DON or DOM-1 for 4 days to measure steroidogenesis and apoptosis, for 1-8 h to measure immediate-early genes, and for 5-60 min to measure phosphorylation of intracellular signaling proteins. Addition of DON decreased progesterone secretion at doses as low as 0.5 ng/ml but had no effect on testosterone secretion. Addition of DOM-1 inhibited progesterone and testosterone secretion at 0.5 ng/ml. Treatment of cells with 1 ng/ml DOM-1 increased the proportion of apoptotic cells, whereas DON had no effect. Addition of DON or DOM-1 stimulated phosphorylation of EIF2AK2, MAPK3/1, and AKT. However, DON inhibited and DOM-1 stimulated MAPK14 phosphorylation. DON increased the levels of mRNA encoding early-immediate genes EGR1, EGR3, and FOS, whereas DOM-1 was without effect. DOM-1 but not DON increased abundance of mRNA of the endoplasmic reticulum (ER) stress-related proteins, PRKRA and ATF4. We conclude that DOM-1 has a major impact on theca function in cattle, and possibly induces theca cell apoptosis through ER stress.

The potential effects of antioxidant feed additives in mitigating the adverse effects of corn naturally contaminated with Fusarium mycotoxins on antioxidant systems in the intestinal mucosa, plasma, and liver in weaned pigs.

Seventy-two piglets (6.0 kg BW) were randomly distributed within six different dietary treatments to evaluate the effect of deoxynivalenol (DON) and the potential of four antioxidant feed additives in mitigating the adverse effects of DON on growth performances and oxidative status. Dietary treatments were as follows: control diet 0.8 mg/kg DON; contaminated diet (DON-contaminated diet) 3.1 mg/kg DON; and four contaminated diets, each supplemented with a different antioxidant feed additive, DON + vitamins, DON + organic selenium (Se)/glutathione (GSH), DON + quercetin, and DON + COMB (vitamins + Se/GSH + quercetin from the other treatments). Although DON was the main mycotoxin in the contaminated diet, this diet also contained 1.8 mg/kg of zearalenone (ZEN). The "mycotoxin" effects therefore included the combined effect of these two mycotoxins, DON, and ZEN. The DON-ZEN ingestion did not affect growth performances, average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F ratio), but partially induced oxidative stress in weaned pigs as shown by increased malondialdehyde (MDA) content in the plasma and superoxide dismutase (SOD) activity in liver (P < 0.05). However, no change in the activity of other antioxidant enzymes or GSH concentrations was observed in plasma and liver of piglets fed the DON-contaminated diet (P > 0.05). Supplementation with individual antioxidant feed additive had a limited effect in weaned pigs fed DON-ZEN-contaminated diets. Combination of antioxidants (vitamins A, C, and E, quercetin, and organic Se/GSH) reduced plasma and liver MDA content and SOD activity in liver (P < 0.05) of piglets fed DON-ZEN-contaminated diets. Furthermore, this combination also reduced MDA content in the ileum (P < 0.05), although activity of glutathione peroxidases (GPx), SOD or catalase (CAT) in the ileum was not affected by DON-ZEN contamination or antioxidant supplements. In conclusion, DON-ZEN contamination induced oxidative stress in weaned pigs and combination of antioxidant feed additives restored partially the oxidative status. Further studies will be necessary to assess whether the effects of antioxidant feed additives on oxidative status are specific when feed is contaminated with DON-ZEN.

Efficacy of Fostera PRRS modified live virus vaccine against a Canadian heterologous virulent field strain of porcine reproductive and respiratory syndrome virus.

Vaccination is a useful option to control infection with porcine reproductive and respiratory syndrome virus (PRRSV), and several modified live-PRRSV vaccines have been developed. These vaccines have shown some efficacy in reducing the incidence and severity of clinical disease as well as the duration of viremia and virus shedding but have failed to provide sterilizing immunity. The efficacy of modified live-virus (MLV) vaccines is greater against a homologous strain compared with heterologous PRRSV strains. The objective of this study was to evaluate the efficacy of Fostera PRRS MLV vaccine in protecting against challenge with a heterologous field strain widely circulating in the swine herds of eastern Canada. Forty-six piglets were divided into 4 groups: nonvaccinated-nonchallenged; nonvaccinated-challenged; vaccinated-challenged; and vaccinated-nonchallenged. The animals were vaccinated at 23 d of age with Fostera PRRS and challenged 23 d later with a heterologous field strain of PRRSV (FMV12-1425619). Overall, the vaccine showed some beneficial effects in the challenged animals by reducing the severity of clinical signs and the viral load. A significant difference between nonvaccinated and vaccinated animals was detected for some parameters starting 11 to 13 d after challenge, which suggested that the cell-mediated immune response or other delayed responses could be more important than pre-existing PRRSV antibodies in vaccinated animals within the context of protection against heterologous strains.

La vaccination est une option utile pour limiter l'infection par le virus du syndrome reproducteur et respiratoire porcin (VSRRP), et plusieurs vaccins VSRRP vivants modifiés ont été développés. Ces vaccins ont démontré une certaine efficacité à réduire l'incidence et la sévérité de la maladie clinique ainsi que la durée de la virémie et de l'excrétion virale mais ont failli à produire une immunité stérilisante. L'efficacité des vaccins vivants modifiés (VVM) est supérieure contre une souche homologue comparativement à des souches hétérologues de VSRRP. L'objectif de la présente étude était d'évaluer l'efficacité du vaccin Fostera, un VVM contre le VSSRP, à protéger contre une infection défi avec une souche de terrain hétérologue circulant librement dans les troupeaux porcins de l'est du Canada. Quarante-six porcelet ont été répartis en quatre groupes : non vaccinés-non infectés; non vaccinés-infectés; vaccinés-infectés; et vaccinés-non infectés. Les animaux ont été vaccinés à 23 jours d'âge avec Fostera SRRP et infectés 23 jours plus tard avec une souche de terrain hétérologue du VSRRP (FMV12-1425619). De manière générale, le vaccin a démontré quelques effets bénéfiques chez les animaux infectés en réduisant la sévérité des signes cliniques et la charge virale. Une différence significative entre les animaux non vaccinés et ceux vaccinés a été détectée pour quelques paramètres et débutant 11 à 13 j suite à l'infection, ce qui suggère que la réponse de l'immunité à médiation cellulaire ou d'autres réponses retardées pourraient être plus importantes que la présence d'anticorps anti-SRRP existants chez des animaux vaccinés dans le contexte d'une protection contre des souches hétérologues.(Traduit par Docteur Serge Messier).

Prevention of deoxynivalenol- and zearalenone-associated oxidative stress does not restore MA-10 Leydig cell functions.

The worldwide contamination of grains designated to human and animal feeding with Fusarium mycotoxins is a significant problem. Among Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZEA) are the most prevalent mycotoxins found in cereals. Co-occurrence of DON and ZEA is also very frequent and indicates that these mycotoxins might be involved in a wide range of synergistic or additive interactions. Both mycotoxins have been linked to various male reproduction problems including downregulation of steroidogenesis. In this study, the impact of DON and ZEA alone or in combination on the viability and steroid production of Leydig cell line MA-10 was determined. The ability of vitamin E, sesamin and their combination to prevent oxidative stress and restore progesterone secretion in DON- and ZEA-exposed cells was also determined. Results showed that MA-10 cells were more sensitive to the effect of DON compared to ZEA. DON and ZEA also significantly reduced MA-10 progesterone secretion after forskolin activation but no significant interactions between DON and ZEA were detected. Preventive treatment with the combination of vitamin E and sesamin significantly reduced ROS production and increased cell survival after exposition to DON and ZEA. However this treatment failed to restore normal progesterone secretion. In conclusion, both DON and ZEA are deleterious to steroidogenesis in Leydig cells. Prevention of oxidative stress caused by DON and ZEA was effective to restore cell viability but failed to restore other functions of Leydig cells suggesting that ROS production is not the main cause of steroidogenic failure in DON and ZEA treated MA-10 cells.