The toxicity of zearalenone (ZEA) was evaluated in swine spleen, a key organ for the innate and adaptative immune response. Weaned pigs were fed for 18 days with a control or a ZEA contaminated diet. The effect of ZEA was assessed on wide genome expression, pro- (TNF-α, IL-8, IL-6, IL-1β, IFN-γ) and anti-inflammatory (IL-10, IL-4) cytokines, other molecules involved in inflammatory processes (MMPs/TIMPs), as well as signaling molecules, (p38/JNK1/JNK2-MAPKs) and nuclear receptors (PPARγ/NFkB/AP-1/STAT3/c-JUN). Microarray analysis showed that 46% of total number of differentially expressed genes was involved in cellular signaling pathway, 13% in cytokine network and 10% in the inflammatory response. ZEA increased expression and synthesis of pro- inflammatory (TNF-α, IL-8, IL-6, IL-1β) and had no effect on IFN-γ, IL-4 and IL-10 cytokines in spleen. The inflammatory stimulation might be a consequence of JNK pathway activation rather than of p-38MAPK and NF-kB involvement whose gene and protein expression were suppressed by ZEA action. In summary, our findings indicated the role of ZEA as an immune disruptor at spleen level.
Zearalenone (ZEA) is an oestrogenic mycotoxin produced by Fusarium species, considered to be a risk factor from both public health and agricultural perspectives. In the present in vivo study, a feeding trial was conducted to evaluate the in vivo effect of a ZEA-contaminated diet on immune response in young pigs. The effect of ZEA on pro-inflammatory (TNF-a, IL-8, IL-6, IL-1b and interferon-g) and antiinflammatory (IL-10 and IL-4) cytokines and other molecules involved in inflammatory processes (matrix metalloproteinases (MMP)/ tissue inhibitors of matrix metalloproteinases (TIMP), nuclear receptors: PPARg and NF-kB1, mitogen-activated protein kinases (MAPK): mitogen-activated protein kinase kinase kinase 7 (TAK1)/mitogen-activated protein kinase 14 (p38a)/mitogen-activated protein kinase 8 ( JNK1)/ mitogen-activated protein kinase 9 ( JNK2)) in the liver of piglets was investigated. The present results showed that a concentration of 316 parts per billion ZEA leads to a significant decrease in the levels of pro-and anti-inflammatory cytokines at both gene expression and protein levels, correlated with a decrease in the levels of other inflammatory mediators, MMP and TIMP. The results also showed that dietary ZEA induces a dramatic reduction in the expressions of NF-kB1 and TAK1/p38a MAPK genes in the liver of the experimentally intoxicated piglets, and has no effect on the expression of PPARg mRNA. The present results suggest that the toxic action of ZEA begins in the upstream of the MAPK signalling pathway by the inhibition of TAK1, a MAPK/NF-kB activator. In conclusion, the present study shows that ZEA alters several important parameters of the hepatic cellular immune response. From an economic point of view, these data suggest that, in pigs, ZEA is not only a powerful oestrogenic mycotoxin but also a potential hepatotoxin when administered through the oral route. Therefore, the present results represent additional data from cellular and molecular levels that could be taken into account in the determination of the regulation limit of the tolerance to ZEA.
The bioavailability of pollen bioactive compounds for humans is limited. In this study, our aim was to enhance the health-related benefits of pollen by fermentation with a Kombucha/SCOBY (symbiotic culture of bacteria and yeasts) consortium. We performed the fermentation of pollen suspended from the beginning with SCOBY on sweetened green tea or on Kombucha vinegar, by adding pollen after 20 days of Kombucha fermentation. We analyzed: formation of bioactive compounds (anti-oxidant polyphenols, soluble silicon, hydroxy-acids, short chain fatty acids—SCFA); parameters related to Kombucha fermentation (dynamics of lactic acid bacteria—LAB, formation of organic acids, soluble sugar evolution on Kombucha vinegar); the influence of Kombucha fermentation on pollen morphology and ultrastructure; in vitro cytotoxic and antitumoral effects of the Kombucha fermented pollen. The pollen addition increases LAB proportion in the total number of SCOBY microbial strains. SEM images highlight the adhesion of the SCOBY bacteria to pollen. Ultrastructural analysis reveals the release of the pollen content. The content of bioactive compounds (polyphenols, soluble silicon species and SCFA) is higher in the fermented pollen and the product shows a moderate antitumoral effect on Caco-2 cells. The health benefits of pollen are enhanced by fermentation with a Kombucha consortium.
Ochratoxins, are toxic fungal metabolites produced by certain moulds of the genera Aspergillus and Penicillium that grow on a wide range of raw food commodities. The most relevant toxin is ochratoxin A (OTA) and the European Commission has established guidance values for OTA concerning complementary and complete feeding stuff recommending that for pigs a maximum concentration of 0.05 mg/kg. These guidance values represent only a recommendation of the Commission and the establishment of a legal regulation needs additional toxicological data generated from farm animal experiments. The aim of this paper was to investigate the effect of OTA – at the recommended EU guidance value of 0.05 mg/kg – on liver health. For this purpose, twelve crossbred, weaned piglets were fed for 33 days a maize-soybean-meal-based diet contaminated or not with 0.05 mg/kg OTA. Blood plasma samples were collected at the end of this period and subjected to biochemical analyses, whereas liver samples were analysed for cytokine concentration (ELISA), enzyme activity and expression of selected genes (qRT-PCR) involved in liver metabolism. Exposure to OTA resulted in a significant decrease in the concentrations of total protein, albumin and nitric oxide in plasma, and interleukin-6 in the liver. OTA exposure also resulted in a significant increase of alanine aminotransferase and triglycerides in plasma and of superoxide dismutase in the liver. In conclusion, the administration of 0.05 mg/kg of OTA, to weaned piglets for a period of 33 days caused measurable hepatocellular injury in the toxin-exposed. Additional in vivo studies should be performed with larger numbers of animals in order to confirm our results and to provide robust data for the establishment of safe concentrations of OTA in swine feeds.
Smoking is an excellent food preservation method but also a source of contamination of foodstuffs with carcinogenic polycyclic aromatic hydrocarbons (PAHs). Herein we investigated the influence of smoking temperature, smoking time, and type of wood sawdust used as smoke source on PAH levels attained through controlled smoking of pork sausages. Four PAHs (benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[a]pyrene) were monitored, as required by European Commission Regulation 835/2011. PAH concentrations increased continuously both with higher temperatures (55−95 °C) and with longer smoking periods (2−9 h), although the level of benzo[a]pyrene exhibited a tendency to plateau after 6 h. Among seven types of hardwoods tested, plum, alder, and birch yielded PAH concentrations considerably higher than that of commonly used beech, and oak showed similar levels to beech while apple and, to a lesser extent, walnut caused lower levels of sausage contamination. These findings could guide the establishment of good practices in the smoked meat industry.
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