Occurrence, toxicity, bioaccessibility and mitigation strategies of beauvericin, a minor Fusarium mycotoxin.

Emerging Fusarium mycotoxins include the toxic secondary metabolites fusaproliferin, enniatins, beauvericin (BEA), and moniliform. BEA is produced by some entomo- and phytopathogenic Fusarium species and occurs naturally on corn and corn-based foods and feeds infected by Fusarium spp. BEA has shown various biological activities (antibacterial, antifungal, and insecticidal) and possesses toxic activity, including the induction of apoptosis, increase cytoplasmic calcium concentration and lead to DNA fragmentation in mammalian cell lines. Cereals food processing has an important effect on mycotoxin stability, leading to less-contaminated food compared to the raw materials. Different industrial processes have shown to be effective practices to reduce BEA contents due to thermal food processing applied, such as cooking, boiling, baking, frying, roasting and pasteurization. Some studies demonstrated the capacity of lactic acid bacteria to reduce the presence of the BEA in model solution and in food chain through fermentation processes, modifying this mycotoxin in a less toxic derivate. Prebiotic and probiotic ingredient can modulate the bioaccessibility of BEA reducing the risk of intake of this minor Fusarium mycotoxin. This review summarizes the existing data on occurrence, toxicity and especially on BEA reduction strategies in food and feed such as chemical reduction, biocontrol and food processing.

Effect of electrical stunning frequency and current waveform in poultry welfare and meat quality.

This study aimed to evaluate the effects of different stunning frequencies and electrical current waveforms on chicken welfare and meat quality. Two-hundred-thirty-two Cobb broilers, 48 d of age and 2.76 ± 0.47 live-weight, were randomly assigned into 4 stunning treatments - 2 frequencies (300 Hz and 650 Hz) and 2 current waveforms (direct current [DC] and alternating current [AC]). Broilers were electrically stunned in a water bath in a commercial slaughterhouse (70 V, 100 mA). The electronarcosis and stunning efficiency were confirmed by assessment of visual parameters (absence of rhythmic breathing, ocular reflex, and coordinated wing flapping) and blood parameters (lactate, glucose, creatine kinase, sodium, and potassium), which were measured after bleeding. The incidence of traumas and injuries was assessed after plucking. Meat quality analysis was performed in Pectoralis major (PM), with determinations of pH, breast yield (PMY), water holding capacity (WHC), water absorption capacity (WAC), thawing loss (DL), cooking loss (CL), shear force (SF), and instrumental color (a*, b*, L*, C*, and h). The interaction between waveform and frequency was significant (P < 0.05) only for lactate, sodium, and lightness (L*). Lactate and sodium levels decreased at 300 Hz-DC. Meat lightness increased at 300 Hz-DC. The individual effect of frequency was significant for glucose, creatine kinase, potassium, WHC, PMY, b*, C*, and h. Regarding waveform, AC decreased plasma glucose and DC decreased creatine kinase and WAC. In general, stunning frequency exerts greater influence than waveform on the welfare and meat quality parameters of broilers. The use of frequency at 650 Hz proved to render animals efficiently unconscious and to promote greater meat quality.

Reaction of zearalenone and α-zearalenol with allyl isothiocyanate, characterization of reaction products, their bioaccessibility and bioavailability in vitro.

This study investigates the reduction of zearalenone (ZEA) and α-zearalenol (α-ZOL) on a solution model using allyl isothiocyanate (AITC) and also determines the bioaccessibility and bioavailability of the reaction products isolated and identified by MS-LIT. Mycotoxin reductions were dose-dependent, and ZEA levels decreased more than α-ZOL, ranging from 0.2 to 96.9% and 0 to 89.5% respectively, with no difference (p⩽0.05) between pH 4 and 7. Overall, simulated gastric bioaccessibility was higher than duodenal bioaccessibility for both mycotoxins and mycotoxin-AITC conjugates, with duodenal fractions representing ⩾63.5% of the original concentration. Simulated bioavailability of reaction products (α-ZOL/ZEA-AITC) were lower than 42.13%, but significantly higher than the original mycotoxins. The cytotoxicity of α-ZOL and ZEA in Caco-2/TC7 cells was also evaluated, with toxic effects observed at higher levels than 75µM. Further studies should be performed to evaluate the toxicity and estrogenic effect of α-ZOL/ZEA-AITC.

Intestinal Alkaline Phosphatase: Potential Roles in Promoting Gut Health in Weanling Piglets and Its Modulation by Feed Additives - A Review.

The intestinal environment plays a critical role in maintaining swine health. Many factors such as diet, microbiota, and host intestinal immune response influence the intestinal environment. Intestinal alkaline phosphatase (IAP) is an important apical brush border enzyme that is influenced by these factors. IAP dephosphorylates bacterial lipopolysaccharides (LPS), unmethylated cytosine-guanosine dinucleotides, and flagellin, reducing bacterial toxicity and consequently regulating toll-like receptors (TLRs) activation and inflammation. It also desphosphorylates extracellular nucleotides such as uridine diphosphate and adenosine triphosphate, consequently reducing inflammation, modulating, and preserving the homeostasis of the intestinal microbiota. The apical localization of IAP on the epithelial surface reveals its role on LPS (from luminal bacteria) detoxification. As the expression of IAP is reported to be downregulated in piglets at weaning, LPS from commensal and pathogenic gram-negative bacteria could increase inflammatory processes by TLR-4 activation, increasing diarrhea events during this phase. Although some studies had reported potential IAP roles to promote gut health, investigations about exogenous IAP effects or feed additives modulating IAP expression and activity yet are necessary. However, we discussed in this paper that the critical assessment reported can suggest that exogenous IAP or feed additives that could increase its expression could show beneficial effects to reduce diarrhea events during the post weaning phase. Therefore, the main goals of this review are to discuss IAP's role in intestinal inflammatory processes and present feed additives used as growth promoters that may modulate IAP expression and activity to promote gut health in piglets.

In vitro antifungal activity of allyl isothiocyanate (AITC) against Aspergillus parasiticus and Penicillium expansum and evaluation of the AITC estimated daily intake.

Isothiocyanates (ITCs) are natural compounds derived from cruciferous vegetables produced by enzymatic conversion of metabolites called glucosinolates. They are potentially useful antimicrobial compounds for food applications have been shown to be promising agents against cancer in human cell culture, animal models, and in epidemiological studies. In this study, the antifungal activity of the allyl isothiocyanate (AITC) was evaluated on two mycotoxigenic fungi as Aspergillus parasiticus and Penicillium expnsum, aflatoxins (AFs) and patulin (PAT) producers, employing an assay on solid medium. Also an approximation of the risk evaluation associated to the intake of food treated with the AITC to reduce the risk of fungi spoilage has been evaluated. On solid medium and after 20 days incubation the strain of Penicillium expansum was inhibited with AITC quantities highest than 50 mg, whereas the strain of A. parasiticus was sensible to AITC doses highest than 5 mg. The analysis of the risk assessment associated to the intake of several food classes treated with the bioactive compound AITC to prevent fungi spoilage evidenced that this product can be considered as safe due that the estimated daily intakes (EDIs) are always lower than the AITC Admissible Daily intake (ADI).

Evaluation of antimicrobial resistance of Campylobacter spp. isolated from broiler carcasses.

1. The aim of the present study was to evaluate the antimicrobial resistance of Campylobacter strains (C. jejuni, C. coli and C. lari) isolated from broiler carcasses processed in the State of Paraná, Brazil. 2. Rates of microbial resistance and susceptibility were assessed by both Disk Diffusion (DD) and Etest (Minimum Inhibitory Concentration) techniques. Antibiotics were tested using DD (12 antibiotics) and/or MIC (7 antibiotics) methods. 3. A total of 95.8% of the strains were resistant to at least two agents. In terms of multidrug resistance, 75% of strains were resistant to three or more groups of antibiotics. The highest rates of resistance were detected for cefalotin, ciprofloxacin, tetracycline and nalidixic acid. A high rate of susceptibility of the strains to erythromycin (95.8%) was found confirming that this is considered the agent of choice for treating campylobacteriosis. Comparison of the microbial resistance and susceptibility, as determined simultaneously by the two methods, found the techniques to be statistically equivalent for 5 out of the 6 antibiotics tested. 4. The results of this study suggest the need for adopting measures to control the use of antibiotics in broiler production to prevent multidrug resistance of Campylobacter strains and reduce the risk of serious human diseases caused by the consumption of contaminated chicken meat.

A chemical approach for the reduction of beauvericin in a solution model and in food systems.

Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains with a strong antibacterial, antifungal, and insecticidal activities. This study evaluated the reduction of BEA added at 25 mg/kg in phosphate buffer saline (PBS) solutions at pH of 4, 7 and 10, or to different cereal products (kernels and flours) by the bioactive compounds phenyl isothiocyanate (PITC) and benzyl isothiocyanate (BITC). The concentration of the mycotoxin was evaluated using liquid chromatography coupled to the diode array detector (LC-DAD). In solution, BEA reduction ranged from 9% to 94% on a time-dependent fashion and lower pH levels resulted in higher BEA reduction. Cereal kernels and flours treated with gaseous PITC and BITC (50, 100 and 500 µL/L) presented BEA reduction from 9% to 97% and was dose-dependent. Among the crops, corn was the vehicle where BEA was mostly affected by the action of the ITCs, followed by wheat and rice, and lastly barley. Overall, PITC caused higher reduction of BEA and should be chosen over BITC as a fumigant to decrease the levels of this mycotoxin in grains and flours.

Study of the chemical reduction of the fumonisins toxicity using allyl, benzyl and phenyl isothiocyanate in model solution and in food products.

Fumonisins (FBs) are bioactive compounds produced by several strains of Fusarium spp. which contain a polyketide structure similar to sphinganine. These mycotoxins contain a free amino group that could work as an electron donor and react with the electrophile carbon present within the isothiocyanate (ITC) group. The objective of this study was to determine the effect of ITCs (allyl, benzyl and phenyl) on the stability of FB(1), FB(2) and FB(3). Firstly, PBS solutions at three pH levels (4, 7 and 9) were prepared and added with pairs of one FB (1 mg/L) plus one ITC (1 mg/L). Then, gaseous ITC was used to fumigate corn kernels and corn flour contaminated with FBs produced by Gibberella moniliformis CECT 2987 in situ. Mycotoxin levels were evaluated using liquid chromatography coupled to mass spectrometry in tandem (LC-MS/MS), while products formed from the reaction of FBs and ITCs were examined by liquid chromatography coupled to mass spectrometry-linear ion trap (LC-MS-LIT). The reduction of FB(1) and FB(2) in solution ranged from 42 to 100% on a time-dependent manner. This variance was greatly influenced by pH. In general, lower pH levels eased the reaction between ITCs and FBs. ITC fumigation treatment (50, 100 and 500 µL/L) was able to reduce 53-96% of FB(1) levels, 29-91% of FB(2) and 29-96% of FB(3). Four reaction products between the bioactive compounds employed in this study were identified, corresponding to FB + ITC conjugates.

Chemical reduction of the mycotoxin beauvericin using allyl isothiocyanate.

Beauvericin (BEA) is a bioactive compound produced by the secondary metabolism of several Fusarium strains and known to have various biological activities. This study investigated the reduction of BEA present in the concentration of 25mg/kg on a solution model (phosphate buffer saline at pH 4 and 7) and in wheat flour using allyl isothiocyanate (AITC) as a reactant. The concentration of the mycotoxin studied was evaluated using liquid chromatography coupled to the diode array detector (LC-DAD), whereas adducts formed between the BEA and AITC were examined by liquid chromatography coupled to mass spectrometry-linear ion trap (LC-MS-LIT). In solution, BEA reduction ranged from 20% to 100% on a time-dependent fashion and no significant differences were evidenced between the two pH levels employed. In the food system composed by wheat flour treated with gaseous AITC (50, 100 and 500 µL/L), the BEA reduction varied from 10% to 65% and was dose-dependent. Two reaction products between the bioactive compounds employed in this study were identified, corresponding to BEA conjugates containing one or two AITC molecules.

Effect of free-SH containing compounds on allyl isothiocyanate antimicrobial activity against Escherichia coli O157:H7.

Escherichia coli O157:H7 contamination is a significant meat safety issue in many countries. Allyl isothiocyanate (AIT) is a natural compound found to limit the survival of E. coli O157:H7 and other pathogens in meat and meat products. In the present study, it was found that glutathione and cysteine naturally present in meat can interfere with AIT antimicrobial activity. Spectroscopy, HPLC, and LC-MS were used to confirm that glutathione was able to react with AIT and formed a conjugate with no or low bactericidal activity against the tested organisms. The same reaction also occurred at pH values of 4.9 and 5.8 at 25 and 4 degrees C, respectively, which broadly represent storage conditions in raw beef (pH 5.8) and during fermented sausage (pH 4.9) manufacture. Reactions observed help to explain reduction in antimicrobial potency of AIT in food (meat) systems.