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The purpose of this study was to observe, in a rat animal model, the short and medium term effects of vegan (VEG) or omnivorous (OMNI) diets with different energy partition between nutrients (zone or classic). Six different diets were administered, for 72 days to 120 growing male Sprague-Dawley rats: (i) VEG zone diet; (ii) VEG classic diet; (iii) OMNI zone diet; (iv) OMNI classic diet; (v) OMNI zone diet with added fibre and (vi) OMNI classic diet with added fibre. Zone diets (high protein and low carbohydrates), resulted in better growth , feed efficiency, lower blood glucose and insulin responses. VEG diets have lowered cholesterol blood level. Histopathological analysis evidenced no damage to liver and kidney tissue by the intake of any of the diet types. Further longer animal and human duration studies should be performed to exclude detrimental effect of higher protein diet.

In the present work, Grana Padano (GP) and Trentingrana (TN) cheeses at different ripening time were in vitro digested. To study calcium uptake and utilization, the intact digestates (selected doses that do not alter cell viability and Transepithelial Electrical Resistance) were administered to Caco2/HT-29 70/30 cells, cultured on a semipermeable membrane in transwells, as a model of human intestinal epithelium. Intact digestates as well as the whole basolateral solutions (mimicking the passage of digestates through intestinal cells before reaching the blood flow and bone) in parallel were further administered to human osteoblast-like cells SaOS-2 to study the extracellular bone matrix formation. In vitro digestates deriving from GP and TN promoted calcium uptake and extracellular bone matrix formation independently of both the cheese type and its ripening period (13, 19 or 26months). The present study reports the ability of whole digestates of GP and TN cheeses to improve intestinal calcium absorption and bone matrix formation in vitro. Once fully explored at bone level, this finding could better support the role of cheese in ameliorating calcium deficiencies and associated diseases in vivo., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Consumption of legumes is associated with a number of physiological and health benefits. Legume proteins complement very well those of cereals and are often used to produce gluten-free products. However, legume seeds often contain antinutritional compounds, such as phytate, galactooligosaccharides, phenolic compounds, lectins, enzyme inhibitors, whose presence could affect their nutritional value. Screening natural and induced biodiversity for useful traits, followed by breeding, is a way to remove undesirable components. We used the common bean cv. Lady Joy and the lpa1 mutant line, having different seed composition for absence/presence of lectins,α-amylase inhibitor, (α-AI) and phytic acid, to verify the advantage of their use to make biscuits with improved nutritional properties. We showed that use of unprocessed flour from normal beans (Taylor's Horticulture and Billò) must be avoided, since lectin activity is still present after baking, and demonstrated the advantage of using the cv. Lady Joy, lacking active lectins and having active α-AI. To assess the contribution of bean flour to biscuit quality traits, different formulations of composite flours (B12, B14, B22, B24, B29) were used in combinations with wheat (B14), maize (gluten-free B22 and B29), or with both (B12 and B24). These biscuits were nutritionally better than the control, having a better amino acid score, higher fiber amount, lower predicted glycemic index (pGI) and starch content. Replacement of cv. Lady Joy bean flour with that of lpa1, having a 90% reduction of phytic acid and devoid of α-AI, contributed to about a 50% reduction of phytic acid content. We also showed that baking did not fully inactivate α-AI, further contributing to lowering the pGI of the biscuits. Finally, data from a blind taste test using consumers indicated that the B14 biscuit was accepted by consumers and comparable in terms of liking to the control biscuit, although the acceptability of these products decreased with the increase of bean content. The B22 gluten-free biscuits, although received liking scores that were just above the middle point of the hedonic scale, might represent a good compromise between health benefits (absence of gluten and lower pGI), expectations of celiac consumers and likeness.

Aim of this study was to characterize the effects of an ochratoxin A (181 ± 34 ng/g) contaminated diet on growth performances, blood parameters, systemic cytokine levels, cell stress markers and reactivity of immune system of weaned pigs. Growth performance was not affected by OTA consumption even if OTA levels increased in plasma, kidney and liver. OTA diminished the protein content in the serum and increased levels of TNF-alpha and IL-10 in plasma. HO-1 mRNA, indicative for cells stress, was decreased in the kidney but increased in the liver. Additionally, whole blood of the animals of the OTA-group showed a decreased capacity to respond with cytokine expression (mRNA and protein) to ex vivo challenge with LPS. In conclusion our findings indicate that chronic ingestion with OTA-contaminated feed, even at low level, is hazardous for the animal and virtually for human health, pig being an excellent model for human., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

It was previously reported that injection of anaflatoxin B1 (AnAFB1) conjugated to keyhole limpet hemocyanin (KLH), together with Freund's adjuvant, was effective in inducing in cows a long lasting titer of anti-aflatoxin B1 (AFB1) antibodies (Abs), cross-reacting with other aflatoxins, which were able to hinder, proportionally to their titer, the secretion of aflatoxin M1 (AFM1) into the milk of cows continuously fed with AFB1. According to anti-AFB1 Ab titer, 50% of the vaccinated cows were recognized as high responder animals. In an attempt to prepare a more effective formulation for vaccination of cows, it was compared the immunogenicity, in Holstein Friesian heifers, of AnAFB1 covalently conjugated to KLH or to recombinant diphtheria toxin (CRM197) molecules, and injected together with various adjuvants. This study demonstrated that injection of AnAFB1 conjugated to KLH and mixed with complete (priming) and incomplete Freund's adjuvant (boosters), as in the previous schedule of immunization, was the most effective regimen for inducing Ab responses against AFB1, although pre-calving administration could increase the effectiveness of vaccination, resulting in 100% high responder animals. After one booster dose at the beginning of the milk production cycle, anti-AFB1 Ab titers were comparable to those recorded at the end of the immunization schedule, and proved to be effective in reducing significantly AFB1 carry over, as AFM1, from feed to milk. Pre-calving vaccination of dairy heifers with conjugated AnAFB1, adjuvated with complete and incomplete Freund's adjuvant, may represent the most effective tool for preventing the public health hazard constituted by milk and cheese contaminated with aflatoxins.

The gut microbiota has been shown to be involved in host energy homeostasis and diet-induced metabolic disorders. To gain insight into the relationships among diet, microbiota and the host, we evaluated the effects of a high-fat (HF) diet on the gut bacterial community in weaning mice. C57BL/6 mice were fed either a control diet or a diet enriched with soy oil for 1 and 2 weeks. Administration of the HF diet caused an increase in plasma total cholesterol levels, while no significant differences in body weight gain were observed between the two diets. Denaturing gradient gel electrophoresis (DGGE) profiles indicated considerable variations in the caecal microbial communities of mice on the HF diet, as compared with controls. Two DGGE bands with reduced intensities in HF-fed mice were identified as representing Lactobacillus gasseri and an uncultured Bacteroides species, whereas a band of increased intensity was identified as representing a Clostridium populeti-related species upon sequencing. Quantitative real-time PCR confirmed a statistically significant 1-log decrease in L. gasseri cell numbers after HF feeding, and revealed a significantly lower level of Bifidobacterium spp. in the control groups after 1 and 2 weeks compared with that in the HF groups. These alterations of intestinal microbiota were not associated with caecum inflammation, as assessed by histological analysis. The observed shifts of specific bacterial populations within the gut may represent an early consequence of increased dietary fat.

Antisecretory factor (AF) is a protein secreted in plasma and other tissue fluids in mammals with proven antisecretory and anti-inflammatory activity; its immunohistological distribution suggests a role in the immune system. The expression level and the distribution of AF protein are altered during an immunological response. Exposure to bacterial toxins induces secretion of AF in plasma, probably reflecting a natural defence mechanism to agents causing diarrhoea, thereby contributing to a favourable clinical outcome and disease termination. An increase of AF levels in plasma by dietary means, such as specially processed cereals (SPC), has been demonstrated in human subjects and animals. Administration of SPC to patients affected by inflammatory bowel disease, gastroenteritis and Ménière's disease relieved symptoms and improved quality of life. A recent study showed the positive effect of SPC diet supplementation on prevention of the effects of exposure to low levels of blast overpressure in rats, reducing the extent of intracranial pressure increase and cognitive function impairment. AF-rich egg yolk powder improved health status in children suffering acute and chronic diarrhoea, reducing the frequency and increasing the consistency of stools. This kind of functional food could be used for prophylaxis in populations exposed to a high risk of morbidity and mortality caused by diarrhoea and as a complementary therapy in patients affected by chronic intestinal inflammatory disease to improve well-being. In pig husbandry AF-inducing diets, owing to their antisecretory activity and anti-inflammatory action, are a suitable option as an alternative to antibiotic growth promoters to counteract post-weaning diarrhoea.

The aim of this research was to determine the fate of aflatoxins (AFs) and fumonisins (FBs) naturally occurring in raw materials (maize grit and malted barley) during four industrial brewing processes. The aflatoxin B(1) (AFB(1)) level in raw materials varied from 0.31 to 14.85 microg kg(-1), while the fumonisin B(1) (FB(1)) level (only in maize grit) varied from 1146 to 3194 microg kg(-1). The concentration in finished beer ranged from 0.0015 to 0.022 microg l(-1) for AFB(1) and from 37 to 89 microg l(-1) for FB(1); the other aflatoxins and fumonisin B(2) were not found in beer samples. The average percentage of toxins recovered in finished beer, referring to the amounts contained in raw materials, were 1.5% +/- 0.8% for AFB(1) and 50.7% +/- 4.7% for FB(1). These results were mainly due to the different solubility of the two mycotoxins during the mashing process. If raw materials comply with the limits fixed by European Commission Regulations, the contribution of a moderate daily consumption of beer to AFB(1) and FB(1) intake does not contribute significantly to the exposure of the consumer.

The occurrence of ochratoxin A (OTA) and aflatoxins (AFs) in liquorice products made in Italy was surveyed. Twenty-eight samples of dried liquorice extract and fifty-four of liquorice confectionery (liquorice content between 2 and 10%) were collected from retail outlets located in northern Italy. After extraction and purification through an immunoaffinity column, OTA and AFs were analysed using both HPLC-FLD and HPLC-MS/MS. OTA occurred in all samples of dried liquorice extract and in 61% of samples of liquorice confectionery, showing very high values for the former (mean 89.6 µg kg(-1), maximum value 990.1 µg kg(-1)), and relatively low levels for the latter (mean 0.96 µg kg (-1), maximum value 8.3 µg kg (-1)). The contribution of dried liquorice extract to OTA intake appears to be non-negligible for children, who are potentially high consumers. AF contamination resulted very low: AFB (1) was detected only in 15.8% of samples (maximum value 7.7 µg kg (-1), mean 0.38 and 0.41 µg kg (-1) for dried liquorice extract and liquorice confectionery, respectively); the other AFs were not detected. To our knowledge, it is the first time that AFB (1) has been detected in liquorice extract samples.

The extraction efficiency of aflatoxin B1 (AFB1) in cattle feed containing nine adsorbents (ADSs) was investigated using two organic/aqueous solvents composed of methanol/water (80/20 v/v; MeOH) and acetone/water (85/15 v/v; AC). Samples were obtained including a highly AFB1-contaminated (HC) and a low-level AFB(1)-contaminated (LC) feedstuff (15.33 and 7.57 microg kg(-1), respectively), nine ADSs (four clay minerals; one yeast cell wall-based product; one activated carbon and three commercial ADS products) at two different levels of inclusion (10 and 20 g kg(-1)). After solvent extraction and immunoaffinity column clean-up, all samples were analysed for AFB1 by high-performance liquid chromatography (HPLC) with fluorescence detection. For each contamination level (HC and LC), the data obtained were analysed using a factorial arrangement in a completely randomized design. Means were compared with the correspondent controls using the Dunnett's test. No statistical difference was found in AFB1 levels of feedstuffs not containing ADSs when extracted with AC or MeOH, even if numerically higher values were obtained with AC. A dose-dependent effect (p < 0.01) of ADSs inclusion was observed on AFB1 recoveries that were lower when the higher ADS level (20 g kg(-1)) was included in the HC and LC feedstuffs. Higher AFB(1) recoveries were obtained using AC compared with MeOH, both in HC (75.0% versus 12.0%, respectively) and in LC (84.0% versus 22.8%, respectively) ADSs containing feedstuffs. However, when the activated carbon and the sodium bentonite were included in feeds, lower AFB1 concentrations with respect to control values (p < 0.001 and <0.05, respectively) were obtained also using AC. The data obtained in this study indicate that routine use of the MeOH solvent for AFB1 analysis of unknown feedstuffs, can produce misleading results if they contain an ADS.

Background: Organic farming is a production technique that imposes major restrictions on the use of fertilizers, pesticides, feed additives and veterinary drugs and for this reason consumers perceive organic foods to be healthier. The content of health-promoting molecules such as ascorbic acid, beta-carotene, lycopene and salicylic acid are important aspects of the nutritional quality of organic foods., Aim: To evaluate health promoting substances and the heavy metal content of tomato berries grown using conventional, integrated pest management (IPM) and organic farming techniques., Methods: Moisture was determined by drying, crude protein by the Kjeldhal method, and ashes by incineration at 550 degrees C. Ergosterol, ascorbic acid, beta-carotene, lycopene and salicylic acid were determined by HPLC. The levels of heavy metals were measured by atomic absorption spectroscopy., Results: Compared to crops grown using conventional and IPM methods, organic tomatoes contained more salicylic acid but less vitamin C and lycopene. Organic tomatoes had higher Cd and Pb levels but a lower Cu content. Organic fruits had a slightly higher protein content than conventionally cultivated fruits, but the difference was minimal and consequently the nutritive significance was poor., Conclusions: Farming techniques may have an impact on the quality of tomatoes. Their higher salicylate content supports the notion that organic foodstuffs are more wholesome. However, the lower lycopene and ascorbic acid levels of organic tomatoes are not to be regarded as positive. No residues of pesticides and ergosterol were detected.

The aim of the study was to evaluate the effect on broiler performance of transgenic Bacillus thuringiensis (Bt) corn containing the Cry1A(b) protein compared with the corresponding near isogenic corn and to analyze the degradation of the Cry1A(b) gene in the digestive tract. Ross male broilers (432) were fed for 42 consecutive days with diets containing Bt or isogenic corn. Diet, Bt corn, and the isogenic form of the Bt corn were analyzed for composition and aflatoxin B1, fumonisin B1, and deoxynivalenol contents. Broiler body weight and feed intake were recorded at regular intervals (d 0, 21, and 42). The presence of the Cry1A(b) gene and plant-specific genes Zein and Sh-2 in gut contents of crop, gizzard, jejunum, cecum, and samples of blood was determined in 10 animals per treatment at the end of the trial using a PCR technique. Chemical composition was not different between Bt and its isogenic form, whereas the fumonisin B1 content for Bt was lower than for isogenic corn (2,039 vs. 1,1034 ppb; P < 0.05). The results of the growth study showed no difference for average daily weight gain (129.4 vs. 126.0 g/d), feed intake (63.4 vs. 61.8 g/d), and feed conversion ratio (1.95 vs. 2.02) among the groups. No significant relationship was observed between mycotoxins content and growth performances. Feed-derived DNA is progressively degraded along the digestive tract. Detection frequency of short fragments of maize-specific high copy number Zein gene was high but significantly decreased in distal sectors. An 1,800-bp fragment of the Cry1A(b) gene, corresponding to the minimal functional unit, was detected only in crop and gizzard of birds fed Bt corn. Sh-2 showed the same detection frequency of Cry1A(b) and was also found in birds fed isogenic corn. Blood samples were positive with low frequency only for the Zein gene fragment. No significant difference in DNA detection was observed between birds fed Bt and isogenic corn, indicating that DNA derived from transgenic feed undergoes the same fate as isogenic feed.

Maize samples collected from storage bins and feed mills in Northern Italy between 1995 and 1999 were surveyed for the occurrence of aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON) and fumonisin (FB1); further, ergosterol was analysed as a fungal growth marker. The incidence and mean content of AFB1 were generally low; nevertheless, a remarkable contamination was found in two samples (109 and 158 microg kg(-1)), while five others exceeded 20 microg kg(-1). DON and ZEA mean levels were significantly higher in 1996 (2716 and 453 microg kg(-1)) with respect to the other years, when mean contents ranged from 7 to 30% and from 3 to 17%, respectively, expressed in per cent of 1996 contents. FB1 was present in all samples and was by far the most remarkable mycotoxin in Northern Italian maize, with the exception of samples from 1996. The average level was 3064 microg kg(-1), 69.6% of samples resulted over 1000 microg kg(-1) and 16.9% over 5000 microg kg(-1). Significant correlations were found between ergosterol and the major mycotoxin(s) in each year (FB1 in 1995 and 1997-99; ZEA + DON in 1996). Consequently, ergosterol seems to be a good index of the toxicological quality of maize. Climatic conditions influenced the growth of different fungal species. In 1996, the first 20 days of October were extremely rainy; these weather conditions delayed the harvest until the first week of November and favoured the growth of DON and ZEA producing fungi and the synthesis of mycotoxins. On the contrary, the temperate and dry climate of the other years supported the growth of FB1-producing fungi.

A total of 96 red wines and 15 white dessert wines produced mostly in the years 1995-97 in 19 Italian regions were analysed for ochratoxin A (OTA). The amount of OTA ranged from < 1 to 3856 ng/l the median (mean) was found to be 90 (419) ng/l for the red wines and 8 (736) ng/l for the white dessert wines. Our survey shows that the geographic region of origin has a strong influence on OTA contamination, both for red and for dessert wines: in fact, wines produced in southern Italy were markedly more contaminated. The overall median (mean) OTA concentration in the red wines produced in the four Italian areas (northwest, northeast, centre and south) was 2 (11), 90 (81), 134 (295) and 1264 (1233) ng/l. The same trend was observed for the white dessert wines: OTA concentrations of over 1000 ng/l were found in four out of five samples from southern Italy (1185, 2454, 3477, 3856 ng/l), while central and northern samples showed very low contamination. The contribution of wine to mean daily OTA intake can be considered negligible in the case of people drinking wine manufactured in northern and central Italy; this is not true if a medium drinker constantly consumes red wine produced in southern Italy in this case wine alone could supply the diet with an amount of OTA equal to or even above the tolerable daily intake of 5 ng/kg body weight recommended by the Scientific Committee on Food of the European Commission.

In vitro affinity tests were conducted to assess the effectiveness of 19 activated carbons (ACs), hydrated sodium calcium aluminosilicate (HSCAS), and sepiolite (S) in binding fumonisin B 1 (FB 1 ) from solution. Relationships between adsorption ability and physicochemical parameters of ACs (specific surface area, iodine value, and methylene blue index) were tested. When 5 ml of a 4-μg/ml aqueous solution of FB 1 was treated with 10 mg of AC, ACs adsorbed 0.46 to 100% of the FB 1 . HSCAS and S were not effective in binding FB 1 . In two saturation tests carried out with decreased amounts of sorbent (5 and 2 mg, respectively), three ACs also showed high adsorption ability (adsorbing 96.48 to 99.20% of the FB 1 ) A general relationship between adsorption ability and the physicochemical parameters of the ACs was observed, supporting the inference of a close relationship between molecule trapping and surface physicochemical adsorption processes. The methylene blue index was more reliable than iodine number and surface area for predicting ability of ACs to adsorb FB 1. In tests of simultaneous adsorption ability carried out using 5 ml of a solution containing 10 μg/ml FB 1 plus 50 μg/ml aflatoxin B 1 (AFB 1 )and 2 or 5 mg of sorbent, ACs showed a higher affinity for AFB 1 than for FB 1 . However, two ACs bound ca. 100% of the two mycotoxins. When 5 ml of an aqueous extract solution obtained from naturally contaminated corn containing 1.84 μg/ml FB 1 and 0.042 μg/ml AFB 1 was treated with 10 mg of sorbent, one AC adsorbed ca. 95% and 99% of FB 1 and AFB 1 , respectively. It is concluded that certain ACs have high in vitro affinity for FB 1 and AFB 1 singly or in combination, and may hold promise as multi-mycotoxin sequestering agents. However, further in vivo investigations are neededto confirm the abilities of ACs to sequester the most important mycotoxins singly or in combinations, establish the amounts to be added to feeds, and determine any long-term effects they may have on gastrointestinal absorption of essential nutrients.

A total of 223 samples of Grana Padano cheese manufactured in 4 years (1991-94) by dairies in 11 provinces of the Po valley were checked for aflatoxin M1. Grated cheese was extracted with chloroform and the defatted extract was purified by an immunoaffinity column; aflatoxin M1 was determined by HPLC using a fluorescence detector. From the analysis of the data it has emerged that only one sample exceeded the maximum tolerated level in cheese in some European countries (250 ng/kg). Most samples (91%) were in the range 5-100 ng/kg and only 15 (6.7%) in the range 100-250 ng/kg. Notwithstanding a diffuse microcontamination, the situation regarding the AFM1 levels can be considered fairly satisfactory. Mean contamination levels of 1992 and 1994 were significantly higher (P < 0.05) than those of 1993 and 1991. No significant difference was observed among provinces or dairies of origin.

According to a double-reversal experimental design on 12 late-lactation Friesian cows the effect of two activated carbons (ACs) (CAC1 and CAC2) and a hydrated sodium calcium aluminosilicate (HSCAS) on carryover of aflatoxin B 1 (AFB 1 ) from feed to aflatoxin M 1 (AFM 1 ) in milk was determined. Cows were fed a basal diet containing AFB 1 naturally contaminated corn meal and copra, During week 1 cows were fed diets containing AFB 1 alone (11.28 μg of AFB 1 /kg of feed); in week 2 the diets contained AFB 1 plus 2.0% sorbent; and in week 3 the diets again contained AFB 1 alone (13.43 μg of AFB 1 /kg of feed). ACs reduced the analytical content of AFB 1 in the pelleted feed by from 40.6% to 73.6%, whereas reduction by HSCAS was 59.2%, The AFM 1 concentrations in milk in weeks 1 and 3 were higher than that in week 2, Decreases in the AFM 1 excreted in the milk by addition to feed of 2% of the sorbents ranged from 22% to 45%. CAC1 and HSCAS were significantly different from each other in reducing the AFM 1 concentration in milk (45.3% versus 32.5%); these reductions were significantly higher than that of CAC2 (22.0%). Carryover reduction by addition of CAC1 (50%) was significantly higher than that of HSCAS (36%). Addition of 2% CAC2 did not allow pelleting of feed because of the caking action of this carbon, The lower performance of CAC2 could be related to the unsuccessful pelleting. The addition of ACs did not influence feed intake, milk production, milk composition, or body weight. Our results suggest that ACs, high-affinity sorbents for AFB 1 in vitro, are efficacious in reducing AFB 1 carryover from cow feed to milk. Further in vivo investigations should establish lower amounts of ACs which can be efficacious.