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Alternariol (AOH) and alternariol monomethyl ether (AME) are secondary metabolites produced by fungi belonging to the genus Alternaria, which generally contaminate fruits but also cereal crops and vegetables. The objective of this study was to investigate if the co-exposure of the swine epithelial intestinal cell line (IPEC-1) to a mixture of mycotoxins would cause an increase in toxicity as compared with exposure to a single toxin. The effects of individual toxins as well as those of their combination (1:1 ratio), in a range of 1–250 μM, were assessed in vitro for the cell viability of proliferating IPEC-1 cells and then on parameters related to the oxidative stress. Our results indicate that both AOH and AME significantly decreased the IPEC-1 cell viability, but the cytotoxicity induced by the AOH + AME combination was significantly higher than that induced by the exposure to the individual toxins. The main interaction type ranged from slight synergy for the AOH-AME combination affecting 25% of cell viability (CI = 0.88), which evolved into a synergistic effect for a higher level of cytotoxicity IL50 (CI = 0.41) and a strong synergistic effect at IL90 (CI = 0.10). In addition, we investigated the effects of two low concentrations (2.5 μM and 5 μM) of AOH and AME mycotoxins administered individually or in combination on oxidative stress in IPEC-1 cells. Both AOH and AME can induce an increase in reactive oxygen species—ROS (+) cells%—and oxidative damage in porcine IPEC-1 cells. At least an additive effect was observed when the cells were exposed to the combination of AOH-AME, consisting of an increase in the percentage of ROS (+) cells and the oxidation of lipids, proteins, and DNA as compared with the individual toxin effect. A breakdown of the antioxidant defense was observed in IPEC-1 cells after the exposure to individual toxins, related to the decrease in the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), but no additive or synergic effect resulted after the exposure to the mixture of the toxins. In conclusion, our data indicate that both AOH and AME interfere with cell proliferation and oxidative stress. Moreover, the exposure of IPEC-1 cells to the combination of AOH and AME mycotoxins had a dose-dependent synergistic effect on IPEC-1 cell viability. Also, the oxidative damage induced in IPEC-1 cells by the combination of AOH and AME was stronger than the effects of individual toxins. However, the signaling pathways responsible for the toxicity of AOH, AME, and their combinations need further investigations in order to provide important data for risk assessments in swine in the case of the contamination of feed with Alternaria toxins. [ABSTRACT FROM AUTHOR]

Simple Summary: The ban of antibiotics as growth promoters and zinc oxide (ZnO) as medicinal to reduce diarrhea in weaning piglets, and the use of alternative nutritional modulators during weaning stress are under debate and intensively studied. Of these, the dietary inclusion of by-products rich in bioactive compounds could be a promising weaning-induced disturbance in piglets. This review describes nutritional studies which investigated the effects of bioactive compounds derived from fruit (apple) and vegetables (carrot) or their by-products on the intestinal architecture and function, inflammatory processes and oxidative stress at the intestinal level. Data on the associated signaling pathways and on the microbiota modulation by bioactive compounds from these by-products are also presented. At weaning, piglets are exposed to a large variety of stressors, from environmental/behavioral factors to nutritional stress. Weaning transition affects the gastrointestinal tract especially, resulting in specific disturbances at the level of intestinal morphology, barrier function and integrity, mucosal immunity and gut microbiota. All these alterations are associated with intestinal inflammation, oxidative stress and perturbation of intracellular signaling pathways. The nutritional management of the weaning period aims to achieve the reinforcement of intestinal integrity and functioning to positively modulate the intestinal immunity and that of the gut microbiota and to enhance the health status of piglets. That is why the current research is focused on the raw materials rich in phytochemicals which could positively modulate animal health. The composition analysis of fruit, vegetable and their by-products showed that identified phytochemicals could act as bioactive compounds, which can be used as modulators of weaning-induced disturbances in piglets. This review describes nutritional studies which investigated the effects of bioactive compounds derived from fruit (apple) and vegetables (carrot) or their by-products on the intestinal architecture and function, inflammatory processes and oxidative stress at the intestinal level. Data on the associated signaling pathways and on the microbiota modulation by bioactive compounds from these by-products are also presented. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a mycotoxin produced by Fusarium fungi that contaminates food and feed, affecting both human and animal health. Among farm animals, the pig is a great consumer of grains and has a native sensitivity to mycotoxins. As the main route of contamination is oral, the intestine is the first defense barrier that plays an important role in the immune response being able to secrete effector molecules (cytokines). At the European level, there are no regulations regarding the amount of ZEN that can be present in the feed of piglets, only recommendations for piglets 0.100 mg ZEN/kg feed (100 ppb). In this study, the effects of ZEN in concentrations below (75 ppb) and above (290 ppb) EU recommendation on the level of some key markers involved in the oxidative and inflammatory response, as well as the mechanisms and signaling pathways through which ZEN could produce its toxicity, were monitored in the colon of weaned piglets. The exposure of the piglets to the lower concentration of ZEN (75 ppb) did not lead to changes in stress and inflammation markers or in the signaling pathways associated with these processes. [ABSTRACT FROM AUTHOR]

Fermented rapeseed meal could be an attractive feed source for piglets after weaning due to its high level of protein with special amino acids and bioactive compounds (polyphenols, PUFA, vitamins B, minerals, fiber) known for their antimicrobial, antioxidative and immunostimulatory effect and lower antinutrients. Lactic acid bacteria were mainly used for rapeseed meal fermentation while yeasts have been less used. The present study evaluated the effect of a diet including 10% rapeseed meal fermented with Saccharomyces cerevisiae on inflammation, oxidative and antioxidative response in spleen of pigs after weaning. The fermentation reduced efficiently glucosinolates level by half in fermented compared to unfermented rapeseed meal and enriched the rapeseed meal in several bioactive compounds such iron, zinc, manganese, n-6 and n-3 unsaturated fatty acids and fiber. Fermented diet reduced the concentration of two important mediators of inflammation, interleukine-1ß (-10.28%) and interleukine-6 (-10.92%) and increased the concentration of IFN-γ (+15.36%), a cytokine involved in the antiviral and antibacterial response. An increase in antioxidant enzymes activity and a reduction in lipid peroxidation was also found in spleen of piglets fed fermented diet. [ABSTRACT FROM AUTHOR]

Inflammatory bowel diseases (IBD) are a major problem for public health, with an increased incidence and impact on life quality. The effect of pre- and probiotic combination has been less studied in IBD. Using genomic and proteomic array technologies, this study examined the efficacy of a new combination of natural alternatives: prebiotics (grape pomace extract, GP) and probiotics (lactobacilli mixture, Lb mix) on inflammation and intracellular signalling routes in a cellular model of inflammation. Caco-2 cells challenged with lipopolysaccharide (LPS) for 4 h were treated with GP extract (50 μg/ml gallic acid equivalent) and Lb combination (3 × 108 colony-forming units/ml total Lb) for 24 h. The profile expressions of forty key inflammatory markers and twenty-six signalling kinases were analysed. Other markers involved in inflammation were also investigated (NF- κ B/RELA, Nrf2, aryl hydrocarbon receptor, Cyp1A1, Cyp1B1); 57·5 and 60 % of investigated genes and proteins, respectively, were down-regulated by the synbiotic combination. Relevant cytokines and chemokines involved in response to microbial infection and inflammation were reduced under the level induced by LPS treatment and toward the unchallenged control. As expected, the reduction effect seems to imply mitogen-activated protein kinase and NF- κ B pathway. Most of the signalling molecules activated by LPS were decreased by GP extract and Lb mix. Our study indicates that the synbiotic combination of GP extract and Lactobacillus sp. mixture exerted anti-inflammatory properties, which are able to decrease the majority of inflammatory genes, their proteins and associated signalling markers. Due to protective role of GP compounds on lactobacilli probiotic, this synbiotic combination might serve as a promising adjunctive therapy in intestinal inflammations. [ABSTRACT FROM AUTHOR]

Grape wastes, such as grape pomace and grape seeds are industrial wastes from the wine processing which is already known in scientific media, but does not yet have a particular interest in animal nutrition. These wastes are rich in bioactive compounds (polyphenols), with antiinflammatory and antimicrobial properties. The aim of the present study was to investigate the effect of a compound feed containing 8% grape seed meal (GSM) on the humoral, systemic and local (duodenum) inflammatory response, and on oxidative stress parameters in weaning piglets. A total of 12 piglets, 6 piglets per group, were fed with 1) control diet or 2) 8% GSM diet for 30 days. At the end of feeding experiment, piglets were slaughtered. Concentration of immunoglobulin and proinflammatory cytokines and the activities of antioxidant enzymes, the total antioxidant capacity and the lipid peroxidation were assessed. Our results showed that GSM diet did not affect the humoral immune response, plasma concentration of IgA, IgM and IgG remaining unmodified. By contrast, at duodenum level, the GSM diet had anti-inflammatory effect, by reducing the concentrations of proinflammatory cytokines IL-6, IL-1ß and TNF-a. GSM significantly increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) in duodenum and reduced the level of lipid peroxidation. In conclusion, GSM by its content in bioactive compounds (polyphenols) and fibers is a valuable feeding alternative with positive effects in weaning and post-weaning period, with the capacity to reduce the inflammatory response in duodenum and the oxidative stress, by modulation of the antioxidant status at systemic and intestinal level. [ABSTRACT FROM AUTHOR]

Cereals and cereals-based products are frequently contaminated with ochratoxin A (OTA) and aristolochic acid (AA). The aim of this study was to compare the effect of OTA and AA on oxidative stress and inflammation in the swine intestine as major organ involved in the absorption of xenobiotics. Fifteen pigs were randomly distributed in 3 groups (control, OTA and AA) and were fed diets contaminated or not with 250 μg toxin/kg for 28 days. When compared with control group, consumption of a OTA and AA contaminated diet significantly increase the concentration of IL-1 beta and IL-8, as major cytokines involved in the inflammatory response. When compared with OTA intoxicated group, AA group has a higher concentration of IL-8 in the intestine. The exposure of piglets to AA significantly decrease the activity of enzymes involved in the oxidative stress response: CAT and GPx as well as the total antioxidant status. The consumption of the diets contaminated with OTA and AA increase in the lipid peroxidation, as showed by TBARS assay. In conclusion, our results have shown that at the intestine level, both OTA and AA induced inflammation and oxidative stress, AA having probably a more important inflammatory effect in the intestine. [ABSTRACT FROM AUTHOR]

Weaning is a very difficult period for the pig, during which the nature and quality of the feeds is of great influence on the developing systems of digestion and defence. The weaning time is correlated with an increase of local inflammatory response. The supplementation of the weaning diet with ingredients rich in bioactive compounds with antimicrobial properties was lately investigated. This is the case of the polyphenols, which have been studied in depth for their beneficial effect on health and for their antioxidant action, but to a lesser extent, for their antimicrobial action. Grape by-products (grape seeds, grape pomace) could be alternative and cheaper sources with anti-microbial potential which could be used in the weaning diet. In the present study we analysed the effects of 5% grape seed cakes (GS) inclusion in the diet on the blood biochemical parameters and on pro- and anti-inflammatory markers in spleen and lymph nodes. A total of 12 weaned pigs were fed with a control or 5% grape seed cakes (GS) diets for 30 days. Pigs were sacrificed after 30 days, blood and organs were collected and stored at -80°C until analyses. Our results showed that diet included 5% GS did not influence the health status determined by plasma biochemical parameters. Only a tendency for a slight increase of the biochemical parameters associated with energetic profile (glucose, cholesterol, triglycerides) was observed Also, GS diet had no effect on pro- and anti-inflammatory cytokines content in spleen and lymph nodes tissue. Further experiments are needed in order to investigate other rate of dietary inclusion which could provide more evidence about the effect of grape bioactive compounds on general health status and inflammation in weaning piglets. [ABSTRACT FROM AUTHOR]

The objective of this study was to characterize several bioactive xenobiotic phytochemicals from a dried grape pomace (GP) derived from a Romanian winery and further to evaluate their effect on inflammation and oxidative markers in liver of pig. 20 crossbred TOPIG hybrid fattening pigs were randomly assigned (n = 10) to two experimental treatments: a normal diet (control group) and a diet included 5% grape pomace (GP group) for 24 days. The total polyphenols concentration of pomace was 36.2g gallic acid equiv. /100g. The pomace was rich in polyphenols from the flavonoids group, the main class being flavanols (epicatechins, catechin, epigallocatechin, procyanidins) and antocyanins (Malvidin 3-O-glucoside). The highest concentration was recorded for epicatechin (51.96g/100g) and procyanidin dimer (22.79g/100g). A high concentration of total polyunsaturated fatty acids (PUFA) especially ω-6 fatty acids (59.82 g/100g fat) were found also in grape pomace. The GP diet lowered the gene expression and protein concentration of IL-1β, IL-8, TNF-α and IFN-γ cytokines suggesting an anti-inflammatory effect of GP diet. Concentration of hepatic TBARS also decreased significantly. The total antioxidant capacity (liver TEAC) and activity of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) did not differ between the GP and control diet while their gene expression tended to decrease. The results showed that GP diet exerted an anti-inflammatory effect, but 5% dietary inclusion modulated only partially the oxidative response and suggest other rate of inclusion need to be investigated. [ABSTRACT FROM AUTHOR]

BACKGROUND The effect of a diet containing 15% grape pomace ( GP) on the general health status and milk quality of dairy cows was assessed by plasma biochemistry and total polyphenol ( TP) content, milk polyphenols, milk composition and milk protein fractions. RESULTS Among the polyphenols measured by liquid chromatography-mass spectroscopy in GP, in feed containing GP ( GP+) or not containing GP ( GP-), gallic acid and epicatechin were present in the highest concentrations (67.58 and 19.23 µg mL−1, respectively). Higher amounts of TP were also detected in the blood plasma of GP+ cows (114.06 and 83.93 mg GAE L−1, respectively) but not in their milk (233.17 and 245.75 mg GAE L−1, respectively). Also a significant increase was found for lactose and β-lactoglobulin, although there was no effect on α-lactalbumin, albumin, secretory components and caseins. CONCLUSION Inclusion of 15% GP in the diets of dairy cows is beneficial for overall normal blood constituent metabolism and helps to maintain cow health. The milk of cows fed with a GP diet preserves the normal levels of fat, protein and caseins, and has increased levels of components that make this milk a versatile ingredient material for the food industry (e.g. model whey powders, stability of lactose-rich powders). © 2016 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Weaning is a very difficult period for the pig, during which the nature and quality of the feeds is of great influence on the developing systems of digestion and defence. The weaning time is correlated with an increase of local inflammatory response. The supplementation of the weaning diet with ingredients rich in bioactive compounds with antimicrobial properties was lately investigated. This is the case of the polyphenols, which have been studied in depth for their beneficial effect on health and for their antioxidant action, but to a lesser extent, for their antimicrobial action. Grape by-products (grape seeds, grape pomace) could be alternative and cheaper sources with anti-microbial potential which could be used in the weaning diet. In the present study we analysed the effects of 5% grape seed cakes (GS) inclusion in the diet on the blood biochemical parameters and on pro- and anti-inflammatory markers in spleen and lymph nodes. A total of 12 weaned pigs were fed with a control or 5% grape seed cakes (GS) diets for 30 days. Pigs were sacrificed after 30 days, blood and organs were collected and stored at -80°C until analyses. Our results showed that diet included 5% GS did not influence the health status determined by plasma biochemical parameters. Only a tendency for a slight increase of the biochemical parameters associated with energetic profile (glucose, cholesterol, triglycerides) was observed Also, GS diet had no effect on pro- and anti-inflammatory cytokines content in spleen and lymph nodes tissue. Further experiments are needed in order to investigate other rate of dietary inclusion which could provide more evidence about the effect of grape bioactive compounds on general health status and inflammation in weaning piglets. [ABSTRACT FROM AUTHOR]

The rapidly increase of the incidence of severe chronic intestinal inflammation diseases have a huge impact on animal and human life quality. The counteracting of intestinal inflammation using prebiotics was lately investigated. Grape pomace (GP) is an industrial waste from the wine processing rich in polyphenols which could be an alternative, less expensive source of antiinflammatory bioactive compounds (polyphenols). Using an in vitro model of intestinal inflammation, we evaluated the effect of the grape pomace extract enriched in polyphenols on several key markers of inflammation. Thus, expression of cytokines, TLRs and signalling molecules were analysed in epithelial CaCO2 cell culture. After cell culture treatments, tRNA was extracted and gene expressions were evaluated by qPCR. Our results showed that grape pomace extract is able to counteract the LPS-induced intestinal inflammation by decreasing the gene expression of pro-inflammatory cytokines: TNF-a, IL-1ß, IL-6 and Il-8 and by slightly increase of the IL-10, antiinflammatory cytokine mRNA. Also, our results showed that grape pomace extract could modulate the expressions of TLRs genes, as modulators of the innate immune response. In conclusion, our study contributes to a better understanding of the grape pomace extract enriched in bioactive compounds, in modulation the intestinal inflammation, as well as the consequence in terms of susceptibility to enteric disease. [ABSTRACT FROM AUTHOR]

Ochratoxins are fungal secondary metabolites produced during storage by fungus of the genera Aspergillus, mainly in tropical and warmer regions and by Penicillium verrucosum, in temperate and colder areas. The most commonly occurring and most toxic member is ochratoxin A (OTA). OTA has multiple toxicity, being nephrotoxic, genotoxic and immunotoxic. According to the Commission Recommendation no. 576/2006 concerning the presence of OTA in products intended for animal feeding, the maximum level admitted for OTA in complementary and complete feeding stuffs for pigs is 0.05ppm. However, these guidance values are only a recommendation, and the establishment of a legal regulation needs additional toxicological data issued from farm animal experiments. The aim of the present paper was to investigate the effect of the EU recommended OTA maximal tolerable feed concentration (0.05 mg/kg) on the spleen of weaned pigs. [ABSTRACT FROM AUTHOR]

In the present study an in vivo feeding experiment was carried out to evaluate the immunotoxic and genotoxic effect of a diet contaminated with 100 ppb zearalenone (the maximal tolerance level recommended by CE/576/2006) in young pigs. Six weaned pigs received the diet contaminated with 100 ppb of zearalenone (ZEA), while other six pigs receiving a non-contaminated diet were used as control for 33 days. The effect of ZEA on pro-inflammatory cytokines and several molecules involved in oxidative stress (CAT, SOD, GPx, iNOS, eNOS) and related pathways (NF-kB1, p38-MAPK, Nrf2) was investigated in internal organs (liver, spleen, kidney and intestine) collected at the end of the experiment. Real-time PCR and ELISA analyses were used to perform gene expression and protein concentration of the above mentioned markers. Our qPCR results showed that the level of 100 ppb of ZEA in the pig diet decreased the gene expression of markers involved in inflammatory process in intestine (duodenum) and liver, but not in spleen, kidney and colon in which an increase in the expression of these genes was found. The modulation of gene expression was not statistically different for the majority of pro-inflammatory cytokines compared to control. Also, throughout ELISA analyses no differences were detected at the protein level for these markers. However, a significant difference was found in gene expression for IL-8 in spleen (+320%) and duodenum (-89%), IL-6 in spleen (+127%) and kidney (+120%) and for COX2 in liver (-88%) and colon (+279%) respectively, when compared to control. Gene encoding for enzymes involved in the oxidative stress were affected (up-regulation) especially in liver and kidney and less in the other organs. The corroboration of these results with other markers of metabolic and physiological processes might contribute to the establishment of the dose accepted as maximal limits in the diet designated to young pigs. [ABSTRACT FROM AUTHOR]

Grape pomace (GP), resulting from wine making, is rich in antioxidant polyphenols originating from the input material, the grapes. Because of the high production volumes of grape pomace, environmental impact and nutritional content, new ways for its valorization are experimented. In order to incorporate it in animal feed (cow and pig), we determined its content in total polyphenols by the Folin Ciocalteu method, the antioxidant activity by the DPPH assay and its stability using the UV-Vis spectroscopy for different extraction media. In terms of total polyphenols the acetone extraction was the best (4667.1 mg GAE/100g sample) comparing with ethanol and water (2140.4 mg GAE/100g sample respectively 2083.9 mg GAE/100g sample), and the ruminal fluid (732.9 mg GAE/100g sample) and pepsin (712.2 mg GAE/100g sample). The highest antioxidant activity expressed as an antiradical activity against the DPPH radical, was registered for the acetone extract (32.8 μM Trolox Equivalents(TE)), followed by the ethanol (6.5 μM TE) and water extraction equal with pepsin extraction (4 μM TE), and the mixture of ruminal fluid (0.5 μM TE). The results show that even though the extraction of polyphenols in organic solvents is the best, when applied to the digestive media the situation changes. In both ruminal fluid and pepsin were measured almost the same amount of total polyphenols but the antioxidant activity was much lower in the ruminal fluid-8 times lower. The UV- Vis spectroscopy shows that the acetone extract is stable over time when kept at--20 C. [ABSTRACT FROM AUTHOR]