Search Results (26)

At weaning, piglets deal with numerous changes and stressors, which can lead to reduced feed intake, digestive disturbances, and gut inflammation. In this context, there is a compelling need to develop new and innovative nutritional strategies aimed at restoring intestinal balance in piglets after weaning and controlling the weaning-associated intestinal inflammation. This study investigated the effect of a diet, including a synbiotic additive (a mix of grape seed and camelina meals as the prebiotic and a lactobacilli mixture as the probiotic) on intestinal inflammation in piglets after weaning. An acute inflammation was induced by the intraperitoneally challenge with Escherichia coli lipopolysaccharide (LPS). The experimental groups were as follows: Control group (piglets fed a conventional corn-soybean meal-based diet), LPS group (piglets fed the Control diet and challenged with 80 µg/b.w. of LPS), SYN group (piglets fed a basal diet, including 5% prebiotic mix and 0.1% probiotic mix, SYN diet), and SYN+LPS group (piglets fed the SYN diet and challenged with 80 µg/b.w. of LPS). Using genomic and proteomic techniques, genes and proteins related to intestinal inflammation were measured in both the jejunum and colon. The results showed that the LPS challenge induced an exacerbated inflammatory response in the jejunum and colon of piglets, inducing an overexpression of a large palette of inflammation-related mediators, including lactate dehydrogenase (LDH) activity, cytokines (e.g., interleukin (IL)—IL-1β, IL-6, tumor necrosis factor alpha—TNF-α), and chemokines (e.g., monokine induced by interferon-gamma—MIG, regulated on activation, normal T cell expressed and secreted—RANTES). All these effects are prevented by the SYN diet, which controls the amplitude of intestinal inflammation induced by the LPS challenge in piglets. Overall, these results suggested that piglets, after weaning, fed the synbiotic diet are less susceptible to the LPS challenge. This diet might be used as a nutritional strategy to alleviate intestinal inflammation in piglets after weaning. Full article

Mycotoxins are common natural contaminants of crops and fruits, associated with negative effects on human and animal health. Currently, more than 300 mycotoxins have been identified, but data on their effects and their limits in feed and food are still inconsistent. The European Commission, by directive EC 574/2011, established regulations concerning the maximum limit allowed in farm animals’ feed for aflatoxins, but for all other mycotoxins there are only recommendations (EC 1319/2016) and there are no established limits. Considering their variety and toxic effects, but also the fact that not many details are yet known about the cumulative effects of co-contamination with various mycotoxins, it is necessary to monitor the evolution of their presence in animal feed. The aim of our study was to analyze for a four-year period (2021–2024) the concentrations of six mycotoxins (total aflatoxins-AFT, fumonisins-FB, deoxynivalenol-DON, zearalenone-ZEA, T2/HT2 and ochratoxin (A + B)-OTA), the most frequently encountered in the south area of Romania in poultry, piglets and pig’s complete feed. Our results showed that the maximum highest concentrations were 5.8 ppb for AFT, 4.7 ppm for FB, 1.9 ppm for DON, 62.8 ppb for ZEA, 32.1 ppb for T2/HT2 and 19.7 ppb for OTA irrespective of the type of feed. It should be noted that AFT and ZEA were identified in all samples during the entire monitored period, and the only mycotoxin that exceeded the guidance value was DON, for which the recommendation of 0.9 ppm for pig feed was exceeded. Recent studies demonstrated that sub-chronic and chronic exposure to low concentrations of mycotoxins and specially co-contamination is more common than acute exposure, being able to affect animal health over time by lowering the defense capacity, inducing inflammatory reactions and affecting intestinal health, which in the long term could have important economic consequences. Our survey study can provide important data showing the degree of contamination with mycotoxins in pig and poultry feed including the simultaneous presence of different mycotoxins in this complete feed. Full article

The present study aimed to investigate the ability of an aqueous extract derived from mustard seed meal to counteract the effects of E. coli endotoxin lipopolysaccharide (LPS) on the intestinal epithelium. Caco-2 cells were cultured together with HT29-MTX and used as a cellular model to analyze critical intestinal parameters, such as renewal, integrity, innate immunity, and signaling pathway. Byproducts of mustard seed oil extraction are rich in soluble polysaccharides, proteins, allyl isothiocyanates, and phenolic acids, which are known as powerful antioxidants with antimicrobial and antifungal properties. Cells were seeded at a ratio of nine (Caco-2) to one (HT29-MXT) and treated for 2 h with mustard meal extract (ME, dilution 1/50) and zinc oxide (ZnO, 50 μM) after reaching 80–100% confluence. Then, they were challenged with 5 μg/mL E. coli-LPS and incubated for another 4 h. The results show that LPS did not alter the cell viability but decreased proliferation compared to the control, ME and ZnO treatments. LPS altered the cell membrane integrity and monolayer permeability by decreasing the transepithelial electrical resistance and tight-junction protein expression. In addition, LPS increased the activity of LDH and the expression of Toll-like receptors. The mechanisms by which LPS induces these disturbances involves the overexpression of PKC, p38 MAPK, and NF-κB signaling molecules. The pretreatment with mustard meal and ZnO succeeded in counteracting the impairment of epithelial renewal, the damage of the membrane integrity and permeability as well as in restoring the gene expression of tight-junction proteins. Full article

Mustard seed meal, an oil industry by-product, shows promise as a dietary additive in animal feed due to its rich content of bioactive compounds and antioxidant potential. The present study examined its impact on the gut microbiota of post-weaning piglets exposed to bacterial lipopolysaccharides (LPSs). Twenty-four cross-bred TOPIGS-40 piglets were divided into four groups: C—control group on a basal diet; LPS—group on a basal diet later exposed to lipopolysaccharides; M—group on a basal diet with an 8% w/w mustard seed meal addition; and M+LPS—a combined mustard and LPS group. After 21 days, colonic content analysis using Illumina 16S rRNA sequencing revealed that while LPS challenge led to an increased abundance of Spirochaetota and Desulfobacterota in the LPS group, the presence of mustard seed meal kept these levels low in the M+LPS group. Notably, the Firmicutes to Bacteroidota (F/B) ratio decreased from 2.87 to 1.12 in the M+LPS group. Furthermore, Lactobacillus showed a 4.7-fold increase while Lachnospira decreased by 67.7% in the M+LPS group compared to the LPS group. These findings highlight mustard seed meal’s potential to prevent LPS-induced gut dysbiosis, enhancing gut health in weaning piglets. Full article

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. Full article

Alternariol monomethyl-ether (AME), together with altenuene and alternariol, belongs to the Alternaria mycotoxins group, which can contaminate different substrates, including cereals. The aim of the present study was to obtain a deeper understanding concerning the effects of AME on pig intestinal health using epithelial intestinal cell lines as the data concerning the possible effects of Alternaria toxins on swine are scarce and insufficient for assessing the risk represented by Alternaria toxins for animal health. Our results have shown a dose-related effect on IPEC-1 cell viability, with an IC50 value of 10.5 μM. Exposure to the toxin induced an increase in total apoptotic cells, suggesting that AME induces programmed cell death through apoptosis based on caspase-3/7 activation in IPEC-1 cells. DNA and protein oxidative damage triggered by AME were associated with an alteration of the antioxidant response, as shown by a decrease in the enzymatic activity of catalase and superoxide dismutase. These effects on the oxidative response can be related to an inhibition of the Akt/Nrf2/HO-1 signaling pathway; however, further studies are needed in order to validate these in vitro data using in vivo trials in swine. Full article

Apples are a fruit rich in active biomolecules, and are one of the most important fruits for human food. Apple waste also started to gain importance as potential feedstuff for farm animals after the ban on in-feed antibiotics. Large quantities of apples are consumed as such (~200 g/capita/day), but also as juice, and they are also used as a base for other juices. A large amount of residue containing active nutrients beneficial for health remains available, which could be added as pomace or meals to the feed of farm animals (e.g., pigs). The present study analyzed and compared the composition of three apple cultivars (Granny Smith, Golden, and Red Delicious) and investigated the capacity of the apple meal extract to counteract the membrane damage and pro-inflammatory effect induced by LPS on an in vitro cellular model of pig intestinal epithelial IPEC-1 cells, considering that the epithelium represents the first barrier for nutrient absorption, as well as against toxins and pathogens. The intestine is the first organ affected by inflammation in piglets during the weaning period, in which animals are frequently exposed to infections with pathogens such as E. coli, Salmonella, Rotavirus, etc. Cells were seeded in Transwell inserts in 24 well plates and treated with apple extract for 48 h. After 2 h of incubation, they were challenged with LPS until 48 h. The capacity of apple extract to protect cellular membrane permeability was evaluated by measuring the trans-epithelial electrical resistance (TEER) at 6, 24, and 48 h, and its potential to diminish the pro-inflammatory effect induced by LPS was also assessed by measuring the pro-inflammatory cytokines synthesis (ELISA). Red Delicious apple extract was used for the in vitro studies due to its higher level of micronutrients than the other two. Our results showed that LPS significantly reduced the TEER at all three measured times in a time-dependent manner, suggesting that the endotoxin disrupted the tight junctions’ proteins and as a consequence the epithelial integrity. But apple extract was efficient to defend the cells against the increased membrane permeability caused by LPS. It was also able to prevent the over-production of pro-inflammatory markers triggered by LPS. Full article

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. Full article

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. Full article

Alternariol is a metabolite produced by Alternaria fungus that can contaminate a variety of food and feed materials. The objective of the present paper was to provide a prediction of Phase I and II metabolites of alternariol and a detailed ADME/Tox profile for alternariol and its metabolites using an in silico working model based on the MetaTox, SwissADME, pKCMS, and PASS online computational programs. A number of 12 metabolites were identified as corresponding to the metabolomic profile of alternariol. ADME profile for AOH and predicted metabolites indicated a moderate or high intestinal absorption probability but a low probability to penetrate the blood–brain barrier. In addition to cytotoxic, mutagenic, carcinogenic, and endocrine disruptor effects, the computational model has predicted other toxicological endpoints for the analyzed compounds, such as vascular toxicity, haemato-toxicity, diarrhea, and nephrotoxicity. AOH and its metabolites have been predicted to act as a substrate for different isoforms of phase I and II drug-metabolizing enzymes and to interact with the response to oxidative stress. In conclusion, in silico methods can represent a viable alternative to in vitro and in vivo tests for the prediction of mycotoxins metabolism and toxicity. Full article

Pigs are the most sensitive animal to zearalenone (ZEN) contamination, especially after weaning, with acute deleterious effects on different health parameters. Although recommendations not to exceed 100 µg/kg in piglets feed exists (2006/576/EC), there are no clear regulations concerning the maximum limit in feed for piglets, which means that more investigations are necessary to establish a guidance value. Due to these reasons, the present study aims to investigate if ZEN, at a concentration lower than the EC recommendation for piglets, might affect the microbiota or induce changes in SCFA synthesis and can trigger modifications of nutritional, physiological, and immunological markers in the colon (intestinal integrity through junction protein analysis and local immunity through IgA production). Consequently, the effect of two concentrations of zearalenone were tested, one below the limit recommended by the EC (75 µg/kg) and a higher one (290 µg/kg) for comparison reasons. Although exposure to contaminated feed with 75 µg ZEN/kg feed did not significantly affect the observed parameters, the 290 µg/kg feed altered several microbiota population abundances and the secretory IgA levels. The obtained results contribute to a better understanding of the adverse effects that ZEN can have in the colon of young pigs in a dose-dependent manner. Full article

Food and feed safety is an issue of great concern for both animal and human health, due to the frequent contamination of food and feed with pathogens, such as bacteria, viruses, yeasts, molds, and parasites. The present paper assumes the possibility that a mycotoxin contamination could occur at the same time as a bacterial infection and investigates the effects of such co-contamination, in comparison with the individual effects of bacterial challenge. For this purpose, we have investigated the effects of simultaneous contamination of swine peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS) (as a model for bacterial contamination) and mycotoxins (ochratoxin or zearalenone) on cell viability, cell cycles, oxydative stress, and inflammation. Our results show important additive/synergistic effects of co-exposure to fungal and bacterial toxins, and that these effects are more important when the cells are co-exposed to LPS and ochratoxin (OTA). The exposure of PBMCs to both OTA and LPS induced an exacerbation of the increase in the inflammation of concentrations of pro-inflamatory cytokines, compared with that of LPS-challenged cells: IL-1β (4.1 times increase), TNF-α (3.2 times increase), IL-6 (3.1 times increase). There was also a decrease in antioxidant defence (i.e., a significant decrease in the total antioxidant capacity and catalase activity) and a significant increase in the percentage of cells undergoing necrosis (24.3% vs. 15.3% in LPS-treated cells). The exposure to zearalenone (ZEA) and LPS led to less important effects and concerned mainly the parameters related to oxidative stress (i.e., a decrease in total antioxidant capacity). The present study provides important data for risk assessment, as the concomitant contamination with bacteria and mycotoxins can lead to a higher toxicity than that which results after an individual infection with Gram-negative bacteria. Full article

(1) The present study tested in vitro the capacity of a fermented rapeseed meal extract to reduce medicinal ZnO, which will be banned at the EU level from 2023 onwards because of its potential to cause environmental pollution and the development of Zn resistance in gut bacteria. Rapeseed meal could be an important ZnO substitute as it has antioxidant/radical scavenging properties due to its content of bioactive compounds (e.g., polyphenols). (2) Protein array and flow cytometry were used to detect apoptosis, oxidative stress production, and inflammatory and signaling-related molecules in Caco-2 and goblet HT29-MTX co-culture cells challenged with Escherichia coli lipopolysaccharides and treated with ZnO and FRSM. (3) LPS induced cell death (21.1% vs. 12.7% in control, p < 0.005); apoptosis (16.6%); ROS production; and overexpression of biomarkers related to inflammation (63.15% cytokines and 66.67% chemokines), oxidative stress, and signaling proteins when compared to untreated cells. ZnO was effective in counteracting the effect of LPS, and 73.68% cytokines and 91.67% of chemokines were recovered. FRSM was better at restoring normal protein expression for 78.94% of cytokines, 91.67% of chemokines, and 61.11% of signaling molecules. FRSM was able to mitigate negative effects of LPS and might be an alternative to ZnO in pig diets. Full article

(1) Background: The weaning period is a very important stage in the pig life, as during weaning, the animals are very susceptible to pathogens and develop postweaning diarrhoea. The aim of our study was to counteract weaning stress and to improve piglets’ gut health by using a nutritional intervention consisting of a mix of agro-industrial wastes (grapeseed, flaxseed and sea blackthorn meals) rich in omega-3 PUFA. (2) Methods: Twelve cross-bred TOPIG hybrid piglets with an average body weight of 11.25 kg were randomly distributed to one of the two experimental groups: a control group fed basic corn soybean diet (control diet) and an experimental group fed a diet with a 10% mixture of grapeseed, flaxseed and sea buckthorn meals in a ratio of 3:4:1 (GFS diet). (3) Results: the GFS diet had no effect on the performance, biochemical parameters or the total antibody synthesis. GFS diet was able to significantly reduce the concentration of proinflammatory cytokines IL-1 beta and TNF-alpha and to significantly increases the expression of junction proteins (occludin, claudin 4, claudin 7 and extracellular protein matrix) at the gene or protein level as compared with control. The presence of GFS in the diet increased the abundance of Bifidobacterium and Lactobacillus species in the colonic content as well as the concentration of propionic and butyric acids. (4) Conclusions: Taken together, our results showed that agro-industrial wastes rich in omega-3 PUFA can be used as an ecological, environmentally friendly nutritional intervention for improving the negative effects associated with the weaning stress. Full article

Zearalenone (ZEA) is an estrogenic fusariotoxin, being classified as a phytoestrogen, or as a mycoestrogen. ZEA and its metabolites are able to bind to estrogen receptors, 17β-estradiol specific receptors, leading to reproductive disorders which include low fertility, abnormal fetal development, reduced litter size and modification at the level of reproductive hormones especially in female pigs. ZEA has also significant effects on immune response with immunostimulatory or immunosuppressive results. This review presents the effects of ZEA and its derivatives on all levels of the immune response such as innate immunity with its principal component inflammatory response as well as the acquired immunity with two components, humoral and cellular immune response. The mechanisms involved by ZEA in triggering its effects are addressed. The review cited more than 150 publications and discuss the results obtained from in vitro and in vivo experiments exploring the immunotoxicity produced by ZEA on different type of immune cells (phagocytes related to innate immunity and lymphocytes related to acquired immunity) as well as on immune organs. The review indicates that despite the increasing number of studies analyzing the mechanisms used by ZEA to modulate the immune response the available data are unsubstantial and needs further works. Full article