Anti-mycotoxin feed additives: effects on metabolism, mycotoxin excretion, performance, and total tract digestibility of dairy cows fed artificially multi-mycotoxin-contaminated diets.

The aim of this study was to evaluate the effects of different anti-mycotoxin feed additives on the concentration of mycotoxins in milk, urine, and blood plasma of dairy cows fed artificially multi-mycotoxin-contaminated diets. Secondarily, performance, total-tract apparent digestibility of nutrients, and blood parameters were evaluated. Twelve multiparous cows (165 ± 45 d in milk, 557 ± 49 kg body weight, and 32.1 ± 4.57 kg/d milk yield at the start of the experiment) were blocked according to parity, milk yield, and days in milk and used in a 4 × 4 Latin square design experiment with 21-d periods, where the last 7 d were used for sampling and data analysis. Treatments were: 1) Mycotoxin group (MTX), basal diet (BD) without anti-mycotoxin feed additives; 2) Hydrated sodium calcium aluminosilicate (HSCA), HSCA added to the BD at 25g/cow/d; 3) Mycotoxin deactivator 15 (MD15), MD (Mycofix® Plus, dsm-firmenich) added to the BD at 15 g/cow/d; and 4) Mycotoxin deactivator 30 (MD30), MD added to the BD at 30 g/cow/d. Cows from all treatments were challenged with a blend of mycotoxins containing 404 µg aflatoxins B1 (AFB1), 5,025 µg deoxynivalenol (DON), 8,046 µg fumonisins (FUM), 195 µg T2 toxin (T2), and 2,034 µg of zearalenone (ZEN) added daily to the BD during the last 7 d of each period. Neither performance (milk yield and composition) nor nutrient digestibility was affected by treatments. All additives reduced aflatoxin M1 (AFM1) concentration in milk, whereas MD15 and MD30 group had lower excretion of AFM1 in milk than HSCA. DON, FUM, T2, or ZEN were not detected in milk of MD15 and MD30. Concentrations in milk of DON, FUM, T2, and ZEN were similar between MTX and HSCA. Except for AFM1, none of the analyzed mycotoxins were detected in urine of MD30 group. Comparing HSCA to MD treatments, the concentration of AFM1 was greater for HSCA, whereas MD30 was more efficient at reducing AFM1 in urine than MD15. AFM1, DON, FUM, and ZEN were not detected in the plasma of cows fed MD30, and DON was also not detected in MD15 group. Plasma concentration of FUM was lower for MD15, similar plasma FUM concentration was reported for HSCA and MTX. Plasma concentration of ZEN was lower for MD15 than MTX and HSCA. Serum concentrations of haptoglobin and hepatic enzymes were not affected by treatments. Blood concentration of sodium was lower in HSCA compared with MD15 and MD30 groups. In conclusion, the mycotoxin deactivator proved to be effective in reducing the secretion of mycotoxins in milk, urine, and blood plasma, regardless of the dosage. This reduction was achieved without adverse effects on milk production or total-tract digestibility in cows fed multi-mycotoxin-contaminated diets over a short-term period. Greater reductions in mycotoxin secretion were observed with full dose of MD.

Effect of Lactobacillus rhamnosus on growth of Listeria monocytogenes and Staphylococcus aureus in a probiotic Minas Frescal cheese.

This study aimed to evaluate the effect of Lactobacillus rhamnosus GG on growth of Staphylococcus aureus and Listeria monocytogenes, inoculated alone or in combination on surface of Minas Frescal cheeses, during storage for 21 days at 7 °C. Survival percentages of each individual bacterial species after exposure to in vitro simulated gastrointestinal conditions (SGC) were also determined. The addition of L. rhamnosus did not affect (P > 0.05) pH, moisture, fat, protein and texture profile of Minas Frescal cheeses. L. rhamnosus was able to survive in suitable counts (>6 Log CFU/g) in cheeses from the 7th day of storage, with high survival (>74.6-86.4%) after SGC. An inhibitory effect of L. rhamnosus on L. monocytogenes was observed in cheeses (decrease of 1.1-1.6 Log CFU/g) and after SGC (20% reduction in the survival). No inhibitory effect of L. rhamnosus was observed on S. aureus counts (P > 0.05), and this microorganism did not survive the exposure to SGC. In conclusion, the addition of L. rhamnosus in Minas Frescal cheese has a potential for L. monocytogenes inhibition. Further studies are necessary to elucidate the mechanisms involved in the inhibition process and determine the survival ability of the bacterial species evaluated in in vivo experiments.

Food safety systems in a small dairy factory: implementation, major challenges, and assessment of systems' performances.

The present study describes the implementation of a food safety system in a dairy processing plant located in the State of São Paulo, Brazil, and the challenges found during the process. In addition, microbiological indicators have been used to assess system's implementation performance. The steps involved in the implementation of a food safety system included a diagnosis of the prerequisites, implementation of the good manufacturing practices (GMPs), sanitation standard operating procedures (SSOPs), training of the food handlers, and hazard analysis and critical control point (HACCP). In the initial diagnosis, conformity with 70.7% (n=106) of the items analyzed was observed. A total of 12 critical control points (CCPs) were identified: (1) reception of the raw milk, (2) storage of the raw milk, (3 and 4) reception of the ingredients and packaging, (5) milk pasteurization, (6 and 7) fermentation and cooling, (8) addition of ingredients, (9) filling, (10) storage of the finished product, (11) dispatching of the product, and (12) sanitization of the equipment. After implementation of the food safety system, a significant reduction in the yeast and mold count was observed (p<0.05). The main difficulties encountered for the implementation of food safety system were related to the implementation of actions established in the flow chart and to the need for constant training/adherence of the workers to the system. Despite this, the implementation of the food safety system was shown to be challenging, but feasible to be reached by small-scale food industries.

Mycotoxin occurrence in breast milk and exposure estimation of lactating mothers using urinary biomarkers in São Paulo, Brazil.

In this study, the occurrence of aflatoxins (AFs), fumonisins (FBs), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN) and some of their metabolites were assessed in breast milk and urine of lactating women (N = 74) from Pirassununga, São Paulo, Brazil. Exposure estimations through urinary mycotoxin biomarkers was also performed. Samples were collected in four sampling times (May and August 2018, February and July 2019) and analyzed by liquid chromatography coupled to tandem mass spectrometry. Aflatoxin M1 (AFM1) was not detected in breast milk. However, two samples (3%) presented FB1 at 2200 and 3400 ng/L, while 4 samples (5%) had OTA at the median level of 360 ng/L. In urine, AFM1 and aflatoxin P1 (AFP1) were found in 51 and 11% of samples, respectively (median levels: 0.16 and 0.07 ng/mg creatinine, respectively). Urinary DON (median level: 38.59 ng/mg creatinine), OTA (median level: 2.38 ng/mg creatinine) and ZEN (median level: 0.02 ng/mg of creatinine) were quantified in 18, 8 and 10% of the samples, respectively. Mean probable daily intake (PDI) values based on urinary biomarkers were 1.58, 1.09, 5.07, and 0.05 µg/kg body weight/day for AFM1, DON, OTA, and ZEN, respectively. Although a low mycotoxin occurrence was detected in breast milk, the PDI for the genotoxic AFs was much higher than those reported previously in Brazil, while PDI values obtained for OTA and DON were higher than recommended tolerable daily intakes. These outcomes warrant concern on the exposure of lactating women to these mycotoxins in the studied area.

In vitro and in vivo capacity of yeast-based products to bind to aflatoxins B1 and M1 in media and foodstuffs: A systematic review and meta-analysis.

The aflatoxins are hepatotoxic and carcinogenic metabolites produced by Aspergillus species during growth on crop products. In this regard, a systematic review to collect the quantitative data regarding the in vitro capacity of yeasts-based products to bind to aflatoxin B1 (AFB1) and/or aflatoxin M1 (AFM1) was performed. After screening, 31 articles which met the inclusion criteria was included and then the pooled decontamination of aflatoxins in the defined subgroups (the type of foods, pH, contact time, temperature, yeast species, and aflatoxin type) was calculated by the random effect model (REM). The overall binding capacity (BC) of aflatoxins by yeast was 52.05% (95%CI: 49.01-55.10), while the lowest and highest aflatoxins' BC were associated with Yeast Extract Peptone (2.79%) and ruminal fluid + artificial saliva (96.21%), respectively. Regarding the contact time, temperature, pH and type of aflatoxins subgroups, the binding percentages varied from 50.83% (>300 min) to 52.66% (1-300 min), 50.71% (0-40 °C) to 88.39% (>40 °C), 43.03% (pH: 3.1-6) to 44.56% (pH: 1-3) and 59.35% (pH > 6), and 48.47% (AFB1) to 69.03% AFM1, respectively. The lowest and highest aflatoxins' BC was related to C. fabianii (18.45%) and Z. rouxii (86.40%), respectively. The results of this study showed that variables such as temperature, yeast, pH and aflatoxin type can be considered as the effective factors in aflatoxin decontamination.

Dietary exposure assessment and risk characterization of mycotoxins in lactating women: Case study of São Paulo state, Brazil.

The exposure and risk characterization of lactating women to aflatoxins (AFs), fumonisins (FBs), ochratoxin A (OTA) and zearalenone (ZEN) due to consumption of different types of food products in Pirassununga, São Paulo, Brazil, was assessed. Lactating women (N = 74) provided samples of foods stored and available at their households between April-August/2018, totaling 184 samples. Mycotoxins were determined in food samples by liquid chromatography-tandem mass spectrometry. According to findings, 20% (n = 36) of all food samples were contaminated with AFs at median concentrations ranging from 9.2 to 18.5 µg/kg, while OTA was detected only in three samples (rice, bread and pasta) at concentrations of 22.3, 23.8 and 48.7 µg/kg, respectively. ZEN was detected in 34 samples (18%) at median levels of 62-195 µg/kg, and FBs at median levels of 58-1546 µg/kg was observed in 22 samples (12%). Moreover, the concentration of AFs, OTA, ZEN and FBs exceeded their respective maximum permitted levels in 11 (6%), 3 (2%), 8 (4%) and 5 (3%) from total samples, respectively. Twenty-eight samples (15%) were contaminated with two or three types of mycotoxins. Corn products contributed for the highest mean probable daily intakes (PDI) of AFs (0.119 ± 0.193 µg/kg body weight (bw)/day), ZEN (0.325 ± 0.097 µg/kg bw/day) and FBs (2.936 ± 1.541 µg/kg bw/day), while wheat-based products contributed for the highest PDI of OTA (0.035 ± 0.028 µg/kg bw/day). The Margin of Exposure (MoE) value for AFs (3.72) demonstrated a high cancer risk (MoE < 10,000), and the Hazard Quotient (HQ) obtained for OTA (24.66), ZEN (4.24) and total FBs (5.01) also resulted in a non-tolerable risk (HQ > 1) via consumption of the investigated food products. Results of this trial indicate high exposure levels of lactating women to dietary mycotoxins in the studied area, which warrant concern about the possible transfer of residual mycotoxins into breast milk.

Aflatoxin in foodstuffs: Occurrence and recent advances in decontamination.

Aflatoxins are highly toxic compounds produced as secondary metabolites by some Aspergillus species, whose occurrence have been reported predominantly in several types of foods of low moisture content, while aflatoxin biotransformation products have been reported mainly in milk and milk products. This review deals with the occurrence of aflatoxins in some of the major food products in the last 5 years including regulatory aspects, and recent advances in detoxification strategies for contaminated foods. Aflatoxin contamination in cereals including corn and peanut is still a public health problem for some populations, especially in African countries. Despite that most of physical and chemical methods for aflatoxin detoxification may affect the nutritional properties of food, or are not safe for human consumption, gamma-radiation and ozone applications have demonstrated great potential for detoxification of aflatoxins in some food matrices. Biological methods based on removal or degradation of aflatoxins by bacterial and yeast have good perspectives, although further studies are needed to clarify the detoxification mechanisms by microorganisms and determine practical aspects of the use of these methods in food products, especially their potential effects on sensory characteristics of foods.

Staphylococcus aureus in Some Brazilian Dairy Industries: Changes of Contamination and Diversity.

Staphylococcus aureus, a major food-poisoning pathogen, is a common contaminant in dairy industries worldwide, including in Brazil. We determined the occurrence of S. aureus in five dairies in Brazil over 8 months. Of 421 samples, 31 (7.4%) were positive for S. aureus and prevalence varied from 0 to 63.3% between dairies. Sixty-six isolates from the 31 samples were typed by Multi-Locus Sequence Typing to determine if these isolates were persistent or continuously reintroduced. Seven known sequence types (STs), ST1, ST5, ST30, ST97, ST126, ST188 and ST398, and four new ST were identified, ST3531, ST3540, ST3562 and ST3534. Clonal complex (CC) 1 (including the four new ST), known as an epidemic clone, was the dominant CC. However, there were no indications of persistence of particular ST. The resistance toward 11 antibiotic compounds was assessed. Twelve profiles were generated with 75.8% of strains being sensitive to all antibiotic classes and no Methicillin-resistant S. aureus (MRSA) strains were found. The enterotoxin-encoding genes involved in food-poisoning, e.g., sea, sed, see, and seg were targeted by PCR. The two toxin-encoding genes, sed and see, were not detected. Only three strains (4.5%) harbored seg and two of these also harbored sea. Despite the isolates being Methicillin-sensitive S. aureus (MSSA), the presence of CC1 clones in the processing environment, including some harboring enterotoxin encoding genes, is of concern and hygiene must have high priority to reduce contamination.