Carlotta Giromini

Mycotoxin Contamination in the EU Feed Supply Chain: A Focus on Cereal Byproducts

Mycotoxins represent a risk to the feed supply chain with an impact on economies and international trade. A high percentage of feed samples have been reported to be contaminated with more than one mycotoxin. In most cases, the concentrations were low enough to ensure compliance with the European Union (EU) guidance values or maximum admitted levels. However, mycotoxin co-contamination might still exert adverse effects on animals due to additive/synergistic interactions. Studies on the fate of mycotoxins during cereal processing, such as milling, production of ethanol fuels, and beer brewing, have shown that mycotoxins are concentrated into fractions that are commonly used as animal feed. Published data show a high variability in mycotoxin repartitioning, mainly due to the type of mycotoxins, the level and extent of fungal contamination, and a failure to understand the complexity of food processing technologies. Precise knowledge of mycotoxin repartitioning during technological processes is critical and may provide a sound technical basis for feed managers to conform to legislation requirements and reduce the risk of severe adverse market and trade repercussions. Regular, economical and straightforward feed testing is critical to reach a quick and accurate diagnosis of feed quality. The use of rapid methods represents a future challenge.

Whey protein lowers blood pressure and improves endothelial function and lipid biomarkers in adults with prehypertension and mild hypertension: results from the chronic Whey2Go randomized controlled trial

Background: Cardiovascular diseases (CVDs) are the greatest cause of death globally, and their reduction is a key public-health target. High blood pressure (BP) affects 1 in 3 people in the United Kingdom, and previous studies have shown that milk consumption is associated with lower BP. Objective: We investigated whether intact milk proteins lower 24-h ambulatory blood pressure (AMBP) and other risk markers of CVD. Design: The trial was a double-blinded, randomized, 3-way–crossover, controlled intervention study. Forty-two participants were randomly assigned to consume 2 × 28 g whey protein/d, 2 × 28 g Ca caseinate/d, or 2 × 27 g maltodextrin (control)/d for 8 wk separated by a 4-wk washout. The effects of these interventions were examined with the use of a linear mixed-model ANOVA. Results: Thirty-eight participants completed the study. Significant reductions in 24-h BP [for systolic blood pressure (SBP): −3.9 mm Hg; for diastolic blood pressure (DBP): −2.5 mm Hg; P = 0.050 for both)] were observed after whey-protein consumption compared with control intake. After whey-protein supplementation compared with control intake, peripheral and central systolic pressures [−5.7 mm Hg (P = 0.007) and −5.4 mm Hg (P = 0.012), respectively] and mean pressures [−3.7 mm Hg (P = 0.025) and −4.0 mm Hg (P = 0.019), respectively] were also lowered. Flow-mediated dilation (FMD) increased significantly after both whey-protein and calcium-caseinate intakes compared with control intake [1.31% (P < 0.001) and 0.83% (P = 0.003), respectively]. Although both whey protein and calcium caseinate significantly lowered total cholesterol [−0.26 mmol/L (P = 0.013) and −0.20 mmol/L (P = 0.042), respectively], only whey protein decreased triacylglycerol (−0.23 mmol/L; P = 0.025) compared with the effect of the control. Soluble intercellular adhesion molecule 1 and soluble vascular cell adhesion molecule 1 were reduced after whey protein consumption (P = 0.011) and after calcium-caseinate consumption (P = 0.039), respectively, compared with after control intake. Conclusions: The consumption of unhydrolyzed milk proteins (56 g/d) for 8 wk improved vascular reactivity, biomarkers of endothelial function, and lipid risk factors. Whey-protein supplementation also lowered 24-h ambulatory SBP and DBP. These results may have important implications for public health. This trial was registered at clinicaltrials.gov as NCT02090842.

Mycotoxin mechanisms of action and health impact: ‘in vitro’ or ‘in vivo’ tests, that is the question

The aim of this paper is to present examples of in vitro and in vivo tests for mycotoxin mechanisms of action and evaluation of health effects, with a focus on the gut environment and toxicity testing. In vivo investigations may provide information on the net effects of mycotoxins in whole animals, whereas in vitro models represent effective tools to perform simplified experiments under uniform and well-controlled conditions and a suitable alternative to in vivo animal testing providing insights not achievable with animal studies. The main limits of in vitro models are the lack of interactions with other cells and extracellular factors, lack of hormonal or immunological influences, and lack or different levels of in vitro expression of genes involved in the overall response to mycotoxins. The translation of in vitro data into meaningful in vivo effects remains an unsolved problem. The main issues to be considered are the mycotoxin concentration range in accordance with levels encountered in realistic situ...

Former food products safety: microbiological quality and computer vision evaluation of packaging remnants contamination

ABSTRACT The use of alternative feed ingredients in farm animal’s diets can be an interesting choice from several standpoints, including safety. In this respect, this study investigated the safety features of selected former food products (FFPs) intended for animal nutrition produced in the framework of the IZS PLV 06/14 RC project by an FFP processing plant. Six FFP samples, both mash and pelleted, were analysed for the enumeration of total viable count (TVC) (ISO 4833), Enterobacteriaceae (ISO 21528-1), Escherichia coli (ISO 16649-1), coagulase-positive Staphylococci (CPS) (ISO 6888), presumptive Bacillus cereus and its spores (ISO 7932), sulphite-reducing Clostridia (ISO 7937), yeasts and moulds (ISO 21527-1), and the presence in 25 g of Salmonella spp. (ISO 6579). On the same samples, the presence of undesired ingredients, which can be identified as remnants of packaging materials, was evaluated by two different methods: stereomicroscopy according to published methods; and stereomicroscopy coupled with a computer vision system (IRIS Visual Analyzer VA400). All FFPs analysed were safe from a microbiological point of view. TVC was limited and Salmonella was always absent. When remnants of packaging materials were considered, the contamination level was below 0.08% (w/w). Of note, packaging remnants were found mainly from the 1-mm sieve mesh fractions. Finally, the innovative computer vision system demonstrated the possibility of rapid detection for the presence of packaging remnants in FFPs when combined with a stereomicroscope. In conclusion, the FFPs analysed in the present study can be considered safe, even though some improvements in FFP processing in the feeding plant can be useful in further reducing their microbial loads and impurity. Graphical Abstract

Short-Communication: A Comparison of the In Vitro Angiotensin-1-Converting Enzyme Inhibitory Capacity of Dairy and Plant Protein Supplements

The consumption of supplements based on dairy or plant proteins may be associated with bioactive potential, including angiotensin-1-converting enzyme inhibitory (ACE-1i) activity, which is linked with blood pressure reduction in vivo. To gain insight into this proposed mechanism, the ACE-1i potential of protein-based supplements, including a selection of dairy (n = 10) and plant (n = 5) proteins were in vitro digested. The total digest was filtered and permeate and retentate were obtained. ACE-1i activity was measured as the ability of proteins (pre-digestion, ‘gastric’, permeate, and retentate) to decrease the hydrolysis of furanacroloyl-Phe-Glu-Glu (FAPGG) substrate for the ACE-1 enzyme. Permeate and retentate of dairy proteins exerted a significantly higher ACE-1i activity (mean of 10 proteins: 27.05 ± 0.2% and 20.7 ± 0.2%, respectively) compared with pre-digestion dairy proteins (16.7 ± 0.3%). Plant protein exhibited high ACE-1i in ‘gastric’ and retentate fractions (mean of five proteins: 54.9 ± 0.6% and 35.7 ± 0.6%, respectively). The comparison of the in vitro ACE-1i activity of dairy and plant proteins could provide valuable knowledge regarding their specific bioactivities, which could inform their use in the formulation of specific functional supplements that would require testing for blood pressure control in human randomly-controlled studies.

Nutritional evaluation of former food products (ex-food) intended for pig nutrition

ABSTRACT Ex-food or former food products (FFPs) have been proposed as one of the categories with great promise as alternative feed ingredients. FFPs’ nutritional potential is not yet fully exploited. Therefore, the aim of this study was to perform a nutritional evaluation of selected FFPs. In particular, six samples of mixed FFPs, all based on bakery products, were analysed for moisture, crude protein, ether extract, crude fibre, neutral detergent fibre, acid detergent fibre, starch and ash. Nitrogen-free extractives and non-structural carbohydrate were also determined. Based on FFPs’ composition data, estimation of digestible energy and metabolisable energy values for pigs were calculated. Further, the in vitro digestibility values of FFPs were investigated using a multi-step enzymatic technique. A wheat sample was included as a control feed ingredient in the study. All data were reported on dry matter basis. FFPs have shown a nutrient composition comparable with cereal grains. In the tested FFPs, the average protein content was 10.0% and the average starch content was 52.4%. Nitrogen-free extractive ranged from 61.2% to 74.7%, whereas non-structural carbohydrate ranged from 58.5% to 79.3%. Compared with wheat, FFPs were characterised by a relative high fat content, averaging about 10.1%. The relatively high nitrogen-free extractive/non-structural carbohydrate/starch and fat concentration designated FFPs as valuable energy sources. Digestible energy and metabolisable energy averages were 17.2 and 16.9 MJ kg–1, respectively. The average in vitro digestibility value of FFPs samples was 88.2% ± 5.8%, comparable with that of wheat (90.6% ± 1.6%). FFPs are a fat-fortified version of common cereals grains. The high energy content and digestibility values elect FFPs as promising non-traditional ingredients for swine.

Combining E-Nose and Lateral Flow Immunoassays (LFIAs) for Rapid Occurrence/Co-Occurrence Aflatoxin and Fumonisin Detection in Maize

The aim of this study was to evaluate the potential use of an e-nose in combination with lateral flow immunoassays for rapid aflatoxin and fumonisin occurrence/co-occurrence detection in maize samples. For this purpose, 161 samples of corn have been used. Below the regulatory limits, single-contaminated, and co-contaminated samples were classified according to the detection ranges established for commercial lateral flow immunoassays (LFIAs) for mycotoxin determination. Correspondence between methods was evaluated by discriminant function analysis (DFA) procedures using IBM SPSS Statistics 22. Stepwise variable selection was done to select the e-nose sensors for classifying samples by DFA. The overall leave-out-one cross-validated percentage of samples correctly classified by the eight-variate DFA model for aflatoxin was 81%. The overall leave-out-one cross-validated percentage of samples correctly classified by the seven-variate DFA model for fumonisin was 85%. The overall leave-out-one cross-validated percentage of samples correctly classified by the nine-variate DFA model for the three classes of contamination (below the regulatory limits, single-contaminated, co-contaminated) was 65%. Therefore, even though an exhaustive evaluation will require a larger dataset to perform a validation procedure, an electronic nose (e-nose) seems to be a promising rapid/screening method to detect contamination by aflatoxin, fumonisin, or both in maize kernel stocks.

Former Food Products Safety Evaluation: Computer Vision as an Innovative Approach for the Packaging Remnants Detection

Former food products (FFPs) represent a way by which leftovers from the food industry (e.g., biscuits, bread, breakfast cereals, chocolate bars, pasta, savoury snacks, and sweets) are converted into ingredients for the feed industry, thereby keeping food losses in the food chain. FFPs represent an alternative source of nutrients for animal feeding. However, beyond their nutritional value, the use of FFPs in animal feeding implies also safety issues, such as those related to the presence of packaging remnants. These contaminants might reside in FFP during food processing (e.g., collection, unpacking, mixing, grinding, and drying). Nowadays, artificial senses are widely used for the detection of foreign material in food and all of them involve computer vision. Computer vision technique provides detailed pixel-based characterizations of colours spectrum of food products, suitable for quality evaluation. The application of computer vision for a rapid qualitative screening of FFP’s safety features, in particular for the detection of packaging remnants, has been recently tested. This paper presents the basic principles, the advantages, and disadvantages of the computer vision method with an evaluation of its potential in the detection of packaging remnants in FFP.

Polyphenolic content and antioxidant activity in agri-food wastes and by-products using different extraction methods

Agri-food wastes (AFW) and by-products chain still have the potential to be reprocessed into other production systems. AFW and by-products may contain components that could be valorised for their bioactivity, such as polyphenols and antioxidant molecules that can be used as a source of functional ingredients for feed industries. However the bioaccessibility of these products are higly variable and dependent on a range of factors, one of the most important being food matrix characteristic. The aim of this study was to determine the total phenolic content and the antioxidant capacity of several AFW (fruit and vegetable waste (FVW), citrus pulp, strawberry and orange dried) and by-products (grape marc, Camilina sativa cake, olive pomace and whey) using different extraction protocols. A total of 24 samples were processed using two different extraction methods: chemical extraction and in vitro physiological extraction. Afterwards, the polyphenolic content was assessed by Folin–Ciocalteu assay while antioxidant capacity was determined by 2, 2-Azino-bis-3 ethylbenzothiazoline-6-sulfonic Acid (ABTS) assay. Soy and wheat samples were included as controls in all the experiments. Results obtained showed that the chemical extracts of by-products and AFW contain different amount of polyphenols; in particular, as expected, the grape marc showed the highest polyphenolic content with a value of 4.5% w/w, followed by Camilina sativa cake, olive pomace, FVW, orange and strawberry dried showed a polyphenolic content of 1.3, 0.7, 1.3, 1.6 and 1.3 %w/w, respectively. Considering the antioxidant capacity, grape marc exhibited a significant (P<0.05) value of 573.6 μmol Trolox equivalent/g after chemical extraction compared to the other samples considered. The physiological extraction yielded high polyphenolic content and antioxidant capacity, suggesting that during the digestion the bioaccessibility of phenolic and antioxidant compounds was improved. The results obtained in this study indicate that AFW and by-products could be considered a promising source of antioxidants and phenolic compounds.

Effect of Zinc Oxide and Zinc Chloride on Human and Swine Intestinal Epithelial Cell Lines

Zinc (Zn) salts are often used as nutritional additives in order to promote gut health. The aim of the present study was to assess the effect of two widely used additives in feedstuff, on the intestinal epithelium. In particular, the effect of zinc oxide (ZnO) and zinc chloride (ZnCl 2 ) was investigated in human (INT-407) and porcine (IPI-2I) cell line models. The effect of Zn sources on IPI-21 and INT-407 cell lines was evaluated by a colorimetric viability test using an incubation period of 3 and 24 hours under serum-free conditions. INT407 and IPI-2I showed to be a suitable model of the intestine and a simple tool to investigate the role of Zn supplements. INT407 showed to be the most sensible model to Zn supplements considered, whereas IPI-2I were more resistant. The results of this study contribute to determine the role of zinc in human and swine intestinal epithelium. However, further in vivo experiments may be done to clarify the contribution of Zn supplements in gut health and to improve Zn supplementation in animal feed and in human formulations.