On Ochratoxin A Research Articles

Spirobenzofuran Mitigates Ochratoxin A-Mediated Intestinal Adverse Effects in Pigs through Regulation of Beta Defensin 1.

Antimicrobial peptides (AMPs) function to extensively suppress various problematic factors and are considered a new alternative for improving livestock health and enhancing immunomodulation. In this study, we explored whether AMP regulation has positive influences on Ochratoxin A (OTA) exposure using a porcine intestinal epithelial cell line (IPEC-J2 cells). We constructed a beta-defensin 1 (DEFB1) expression vector and used it to transfection IPEC-J2 cells to construct AMP overexpression cell lines. The results showed that OTA induced cytotoxicity, decreased cell migration, and increased inflammatory markers mRNA in IPEC-J2 cells. In DEFB1 overexpressing cell lines, OTA-induced reduced cell migration and increased inflammatory markers mRNA were alleviated. Additionally, a natural product capable of inducing DEFB1 expression, which was selected through high-throughput screening, showed significant alleviation of cytotoxicity, cell migration, and inflammatory markers compared to OTA-treated IPEC-J2 cells. Our finding provides novel insights and clues for the porcine industry, which is affected by OTA exposure.

Removal of ochratoxin A in wine by Cryptococcus albidus and safety assessment of degradation products.

Ochratoxin A (OTA) is a mycotoxin that contaminates grape-based products and is extremely harmful to the health of the host. It is effectively removed by yeast during the fermentation of wine, whereas the removal mechanism of OTA remains unclear. Therefore, the present study aimed to investigate the removal mechanism of ochratoxin A by yeast and to evaluate the safety of its degradation products. Cryptococcus albidus (20-G) with better effect on ochratoxin A (OTA) was screened out in the main fermentation stage of wine. The results showed that 20-G removed OTA through biosorption and biodegradation. Intracellular enzymes played the main role (18.44%) and yeast cell walls adsorbed a small amount of OTA (8.44%). Furthermore, the identification of proteins in 20-G revealed that the decrease in OTA content was mainly a result of the action of peroxidase, and validation tests were carried out. By analyzing the degradation products of OTA, OTα and phenylalanine with lower toxicity were obtained. Animal experiments showed that the intervention of yeast 20-G reduced the damage and adverse effects caused by OTA toxicity to the mice. The present study demonstrates the mechanism of OTA removal by 20-G and the toxicity of OTA was reduced by peroxidase in 20-G. © 2023 Society of Chemical Industry.

Extensive literature searches on bromine and ochratoxin A in feed

Two Extensive Literature Searches (ELSs) by using PubMed, Web of Science (WoS) and SciFinder for relevant studies on Ochratoxin A (OTA) and bromine, bromate and bromide (BRs) in feed were performed. Search queries allowed to obtain a total number of 8,953 and 34,183 references for OTA and BRs, respectively. The relevance analysis of the documents was performed by screening titles and abstracts against inclusion and exclusion criteria. Relevant references accounted in each Area, for OTA and BRs, were respectively: Area 1 (Information on analytical techniques for quantification) 117 and 117 documents, respectively; Area 2 (Information on occurrence/concentrations and formation) 176 and 176 documents, respectively; Area 3 (Information on exposure of farm and companion animals) 31 and 3 documents, respectively; Area 4 (Information on toxicokinetics (absorption, distribution, metabolism, excretion) in farm and companion animals) 37 and 8 documents, respectively; Area 5 (Information on toxicity of OTA/BRs in farm and companion animals) 249 and 23 documents, respectively; Area 6 (Information on the transfer of OTA/BRs from feed to animal derived food) 18 and 4 documents. A total number of 618 and 100 relevant non-redundant documents was found for OTA and BRs, respectively. Those results were obtained after the full text analysis of all the document previously labelled as relevant.

Protective effect of selenomethionine on kidney injury induced by ochratoxin A in rabbits.

The purpose of this study was to investigate the protective effect and mechanism of selenomethionine (SeMet) on ochratoxin A (OTA)-induced nephrotoxicity in rabbits. In total, sixty Ira rabbits were randomly divided into 5 groups (the control group, OTA group, 0. 2mg/kg SeMet + OTA group, 0. 4mg/kg SeMet + OTA group, and 0. 6mg/kg SeMet + OTA group). The rabbits were fed diets supplemented with different doses of SeMet for 21days and given 0. 2mg/kg OTA starting on day 15 for a week. The results showed that the SeMet supplementation could improve the changes in blood physiological indices and renal function decline caused by OTA poisoning, and alleviate pathological kidney injury in the rabbits. SeMet also increased the activities of total antioxidant capacity, superoxide dismutase, and glutathione peroxidase, and decreased the contents of malondialdehyde and reactive oxygen species and the expression of interleukin-1β, interleukin-6, and tumor necrosis factor-α in the damaged kidneys of the rabbits. In addition, the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream gene heme oxygenase 1 (HO-1) was also inhibited after OTA poisoning, while SeMet activated the Nrf2 signaling pathway and enhanced the expression of Nrf2 and the downstream gene HO-1. In conclusion, SeMet protected against kidney injury caused by OTA in rabbits, and the mechanism may be the activation of the Nrf2 signaling pathway.

Mitigation potential of individual and combined dietary supplementation of local Bentonite Clay and Distillery Sludge against Ochratoxin-A induced toxicity in broilers

BackgroundThis study aimed to evaluate the ameliorative effects of dietary supplementation of local bentonite clay (BN) and distillery sludge (DS) alone and in combination on ochratoxin-A (OTA) induced toxicity in broilers. For this purpose, day-old-broiler chicks (n = 270) were procured from the local market and reared under standard management conditions. After 7 days of acclimatization, birds were divided into 2 main groups A and B with respect to OTA inclusion level in feed, each with four sub-groups viz. A1-A4, each challenged with OTA at a dietary inclusion level of 250 µg/kg feed and B1-B4, each challenged with OTA at the level of 500 µg/kg feed and a common control group that was fed with basal feed throughout the experiment. In groups A and B, BN and DS were administered with feed at the rate of 10 g/kg of feed and 5 g/kg of feed alone and in combination, respectively.ResultsResults showed that OTA administration alone resulted in poor feed conversion ratio (FCR) and immunological responses along with increased serum levels of alanine transaminase (ALT), Aspartate transaminase (AST), urea and creatinine (P < 0.05). A significant decrease (P < 0.05) in serum protein levels (albumin, globulin and total protein) was also observed in OTA-fed groups in a dose-dependent manner. The addition of BN at 10 g/kg of OTA-contaminated feed resulted in better FCR and immunological responses as compared to those fed OTA only. The BN supplementation also conferred protection against elevation of serum biochemical parameters when compared with OTA-fed groups. However, the addition of DS could not provide significant protection (P > 0.05) on alteration of serum biochemical parameters in response to the OTA induced toxicity. The combined supplementation of BN and DS resulted in amelioration of OTA-induced toxicity and showed improved FCR, immunological, hematological and serum biochemical parameters (P < 0.05) when compared with other groups. Similarly, BN and DS resulted in a significant decline (P < 0.05) in the OTA tissue residues compared with other groups and control.ConclusionIn conclusion, combined dietary supplementation of BN (10 mg/kg) and DS (05 mg/kg) in feed reduced the toxic effects of OTA contamination at levels of 250 and 500 µg/kg of feed in broilers. So, the combination products of BN and DS may be successfully developed for use in poultry for protection against OTA-induced toxicity in broilers.

Activity of essential oils from Syzygium aromaticum and Rosmarinus officinalis against growth and ochratoxin A production by Aspergillus tubingensis and Aspergillus luchuensis from Moroccan grapes

Essential oils have been used since ancient times in traditional medicine and agri-food science to preserve food, and to combat human diseases. Essential oils (EOs) from clove and rosemary, obtained by hydro-distillation, were analyzed by GC/MS and evaluated for their antifungal activity against strains of Aspergillus tubingensis and A. luchuensis and their effect on ochratoxin A (OTA) production by these fungi. The major constituent of clove EO was eugenol (86.4%), and of rosemary EO was eucalyptol (35.9%). Mycelium growth inhibition assays showed that the EOs had dose-depend inhibition effects, which reached 36.6% for rosemary EO, and 100% for clove EO, at EO concentration of 200 μL L-1. These reductions were different for different Aspergillus strains, the essential oil type, and the strain/oil type interaction. Analyses of OTA in the culture medium extracts of Aspergillus strains was carried out using HPLC-FLD, and was confirmed by LC-MS/MS for positive controls of two OTA-producing strains of A. tubingensis. In general, OTA was reduced (from 45 to 100% reduction), except for two strains of A. tubingensis, where OTA production was stimulated.

Prevalence of toxigenic fungi and mycotoxins in Arabic coffee (Coffea arabica): Protective role of traditional coffee roasting, brewing and bacterial volatiles.

Fungal infection and synthesis of mycotoxins in coffee leads to significant economic losses. This study aimed to investigate the prevalence of toxigenic fungi, their metabolites, and the effect of traditional roasting and brewing on ochratoxin A (OTA) and aflatoxins (AFs) contents of naturally contaminated coffee samples. In addition, in vivo biocontrol assays were performed to explore the antagonistic activities of Bacillus simplex 350–3 (BS350-3) on the growth and mycotoxins synthesis of Aspergillus ochraceus and A. flavus. The relative density of A. niger, A. flavus, Penicillium verrucosum and A. carbonarius on green coffee bean was 60.82%, 7.21%, 3.09% and 1.03%, respectively. OTA contents were lowest in green coffee beans (2.15 μg/kg), followed by roasted (2.76 μg/kg) and soluble coffee (8.95 μg/kg). Likewise, AFs levels were highest in soluble coffee (90.58 μg/kg) followed by roasted (33.61 μg/kg) and green coffee (9.07 μg/kg). Roasting naturally contaminated coffee beans at three traditional methods; low, medium and high, followed by brewing resulted in reduction of 58.74% (3.50 μg/kg), 60.88% (3.72 μg/kg) and 64.70% (4.11 μg/kg) in OTA and 40.18% (34.65 μg/kg), 47.86% (41.17 μg/kg) and 62.38% (53.73 μg/kg) AFs contents, respectively. Significant inhibitions of AFs and OTA synthesis by A. flavus and A. carbonarius, respectively, on infected coffee beans were observed in presence of Bacillus simplex BS350-3 volatiles. Gas chromatography mass spectrochemistry (GC-MS/MS) analysis of head-space BS350-3 volatiles showed quinoline, benzenemethanamine and 1-Octadecene as bioactive antifungal molecules. These findings suggest that marketed coffee samples are generally contaminated with OTA and AFs, with a significant level of roasted and soluble coffee contaminated above EU permissible limits for OTA. Further, along with coffee roasting and brewing; microbial volatiles can be optimized to minimize the dietary exposure to mycotoxins.

Risk assessment of ochratoxin A in food.

The European Commission asked EFSA to update their 2006 opinion on ochratoxin A (OTA) in food. OTA is produced by fungi of the genus Aspergillus and Penicillium and found as a contaminant in various foods. OTA causes kidney toxicity in different animal species and kidney tumours in rodents. OTA is genotoxic both invitro and invivo; however, the mechanisms of genotoxicity are unclear. Direct and indirect genotoxic and non-genotoxic modes of action might each contribute to tumour formation. Since recent studies have raised uncertainty regarding the mode of action for kidney carcinogenicity, it is inappropriate to establish a health-based guidance value (HBGV) and a margin of exposure (MOE) approach was applied. For the characterisation of non-neoplastic effects, a BMDL 10 of 4.73μg/kg body weight (bw) per day was calculated from kidney lesions observed in pigs. For characterisation of neoplastic effects, a BMDL 10 of 14.5μg/kg bw per day was calculated from kidney tumours seen in rats. The estimation of chronic dietary exposure resulted in mean and 95th percentile levels ranging from 0.6 to 17.8 and from 2.4 to 51.7 ng/kg bw per day, respectively. Median OTA exposures in breastfed infants ranged from 1.7 to 2.6 ng/kg bw per day, 95th percentile exposures from 5.6 to 8.5 ng/kg bw per day in average/high breast milk consuming infants, respectively. Comparison of exposures with the BMDL 10 based on the non-neoplastic endpoint resulted in MOEs of more than 200 in most consumer groups, indicating a low health concern with the exception of MOEs for high consumers in the younger age groups, indicating a possible health concern. When compared with the BMDL 10 based on the neoplastic endpoint, MOEs were lower than 10,000 for almost all exposure scenarios, including breastfed infants. This would indicate a possible health concern if genotoxicity is direct. Uncertainty in this assessment is high and risk may be overestimated.

Protective Effect of Astaxanthin on Ochratoxin A-Induced Kidney Injury to Mice by Regulating Oxidative Stress-Related NRF2/KEAP1 Pathway.

The present study aimed to investigate the effects of astaxanthin (ASX) on ochratoxin A (OTA)-induced renal oxidative stress and its mechanism of action. Serum kidney markers, histomorphology, ultrastructural observation, and oxidative stress indicators were assessed. Meanwhile, quantitative real-time reverse transcription PCR and western blotting detection of NRF2 (encoding nuclear factor, erythroid 2 like) and members of the NRF2/KEAP1 signaling pathway (KEAP1 (encoding Kelch-like ECH-associated protein), NQO1 (encoding NAD(P)H quinone dehydrogenase), HO-1 (encoding heme oxygenase 1), γ-GCS (gamma-glutamylcysteine synthetase), and GSH-Px (glutathione peroxidase 1)) were performed. Compared with the control group, the OTA-treated group showed significantly increased levels of serum UA (uric acid) and BUN (blood urea nitrogen), tubular epithelial cells were swollen and degenerated, and the levels of antioxidant enzymes decreased significantly, and the expression of NRF2 (cytoplasm), NQO1, HO-1, γ-GCS, and GSH-Px decreased significantly. More importantly, after ASX pretreatment, compared with the OTA group, serum markers were decreased, epithelial cells appeared normal; the expression of antioxidant enzymes increased significantly, NQO1, HO-1, γ-GCS and GSH-Px levels increased significantly, and ASX promoted the transfer of NRF2 from the cytoplasm to the nucleus. These results highlight the protective ability of ASX in renal injury caused by OTA exposure, and provide theoretical support for ASX’s role in other mycotoxin-induced damage.

Red orange and lemon extract prevents the renal toxicity induced by ochratoxin A in rats.

In this work, we investigated the effects of red orange and lemon extract (RLE) on ochratoxin A (OTA)-induced nephrotoxicity. In particular, we analyzed the change in renal function and oxidative stress in Sprague-Dawley rats treated with OTA (0.5 mg/kg body weight, b.w.) and with RLE (90 mg/kg b.w.) by oral administration. After OTA treatment, we found alterations of biochemical and oxidative stress parameters in the kidney, related to a severe decrease of glomerular filtration rate. The RLE treatment normalized the activity of antioxidant enzymes and prevented the glomerular hyperfiltration. Histopathological examinations revealed glomerular damages and kidney cortex fibrosis in OTA-rats, while we observed less severe fibrosis in OTA plus RLE group. Then, we demonstrated that oxidative stress could be the cause of OTA renal injury and that RLE reduces this effect.

Effects of Gamma Irradiation on Ochratoxin A Stability and Cytotoxicity in Methanolic Solutions and Potential Application in Tunisian Millet Samples

Gamma irradiation is a useful technology for degrading mycotoxins. The purpose of this study was to investigate the effect of irradiation on ochratoxin A (OTA) stability under different conditions. OTA was irradiated in methanolic solution and on millet flour at doses of 2 and 4 kGy. Residual OTA concentrations and possible degradation products in irradiated samples were analyzed by high-performance liquid chromatography with fluorescence detection and liquid chromatography coupled to mass spectrometry. The extent of in vitro cytotoxicity of OTA to HepG2 cells, with and without irradiation treatment, was assessed with an MTT assay. OTA was more sensitive to gamma radiation on Tunisian millet flour than in methanolic solutions. After irradiation of naturally contaminated millet flour, the OTA concentration was significantly reduced by 48 and 62% at a dose of 2 and 4 kGy, respectively. However, in the methanolic solution, OTA at concentrations of 1 and 5 μg mL-1 was relatively stable even at a dose of 4 kGy, with no degradation products detected in the chemical analysis. Analytical results were confirmed by cell culture assays. The remaining cytotoxicity (MTT assay) of OTA following irradiation was not significantly affected compared with the controls. These findings indicate that gamma irradiation could offer a solution for OTA decontamination in the postharvest processing chain of millet flour. However, the associated toxicological hazard of decontaminated food matrices needs more investigation.

Effects of selected natural preservatives on the mycelial growth and ochratoxin A production of the food-related moulds Aspergillus westerdijkiae and Penicillium verrucosum

ABSTRACTThe present study examines the influence of the natural preservatives carvacrol, eugenol, trans-cinnamaldehyde and the essential oil (EO) Origanum vulgare on ochratoxin A (OTA) production and the mycelial growth of two food-related moulds, Penicillium verrucosum and Aspergillus westerdijkiae, by broth macro-dilution assay for 21 days. With the addition of ½ minimum inhibitory concentration (MIC) carvacrol, eugenol and O. vulgare EO, the mycelial dry weight of both moulds decreased significantly over the whole incubation period of 7, 14 and 21 days. Trans-cinnamaldehyde slightly stimulated the growth of A. westerdijkiae and P. verrucosum at 14 and 21 days of incubation. Growth inhibition did not accompany inhibition of OTA production. Although the growth of both moulds was inhibited after the addition of ½ MIC carvacrol, eugenol and O. vulgare EO, the OTA production of the strong mycotoxin producer A. westerdijkiae was stimulated. Only trans-cinnamaldehyde inhibited the production of OTA with the addition of ½ MIC. P. verrucosum produced significantly less OTA than A. westerdijkiae, and its mycotoxin production was almost completely inhibited by the addition of ½ MIC of the natural preservatives.

Effect of the essential oils from Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on the synthesis of ochratoxin A by Aspergillus niger and Aspergillus carbonarius isolated from tropical wine grapes.

The influence of essential oils (EOs) extracted from the leaves of Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on ochratoxin A (OTA) synthesis by fungi was studied. The extraction of EOs was performed by hydrodistillation (Clevenger apparatus) over a 2-h period and subsequently analyzed by GC-MS and GC-FID. The toxigenic activity of the essential oils (31.25; 15.62 and 7.81µgmL-1) was evaluated by inhibiting the production of OTA by Aspergillus niger and Aspergillus carbonarius in Czapek agar medium culture. The quantification of the toxin was performed by HPLC. The production of OTA was dependent on the fungal species, incubation temperature (15 and 25°C) and the presence of the essential oils. In tests carried out at 15°C, the oils caused a reduction in OTA synthesis that ranged from 57.60 to 76.93% and from 54.78 to 98.68% for the fungal species A. carbonarius and A. niger, respectively. At 25°C, reductions ranged from the 38.66 to 75.93% and from 17.94 to 71.79% for the respective fungi. The study concluded that natural products could be potential biocontrol agents against OTA contamination in food.

Alkali-based pre-treatment may prevent ochratoxin A in grapes

In traditional raisin production process in Turkey, one of the major raisin producers in the world, grapes undergo an alkali treatment prior to drying. This procedure involves dipping grapes in a potassium carbonate solution for 10-15 seconds to remove the wax layer on the surface to accelerate the drying process. In this study, we aimed to investigate the effect of alkali treatment on ochratoxin A (OTA), an important risk factor for grapes and grape-derived products. First, the stability of the OTA was examined under variable pH (4-12), ion type (Na+, K+), ionic strength (0.1-10%) and temperature (4-60 °C). Then,Aspergillus ochraceus contaminated grapes were treated with an alkaline solution and a subsequent OTA analysis was performed through a high performance liquid chromatography-fluorescence detection. OTA started to decompose at pH values exceeding 10.0. The degradation followed a first order kinetics. Although both sodium carbonate and potassium carbonate solutions were at the same alkali pH and they both triggered the OTA degradation at increasing concentrations, potassium carbonate was more effective even at low concentrations. Elevated temperatures were also found to effect degradation. The degradation rate constant at 40 °C was approximately 5 times greater than the one at 25 °C. Furthermore, OTA levels dropped up to 50% in contaminated grapes after the alkali treatment. Our results confirmed that the amide bond of OTA rapidly hydrolysed to a non-toxic ochratoxin-alpha and phenylalanine under strong alkaline conditions. The results suggest that the alkaline pre-treatment unintentionally offers a great advantage to reduce OTA levels in raisins and it could be a viable approach for other dried fruits through the application of good manufacturing practice.

Reduction of ochratoxin A in chicken feed using probiotic.

Mycotoxins present in fodders may evoke health problems of animals and people. The data published by FAO in 2001 show that 25% of raw materials are contaminated with mycotoxins, while their type and concentration are to a great extent dependable on the climatic zone. Biological detoxification of mycotoxins by the use of microorganisms is one of the well-known strategies for the management of mycotoxins in foods and feeds. The aim of this study was to determine the influence of spontaneous fermentation and that with the use of probiotic bacteria and yeast on ochratoxin A (OTA) concentration and the microbiota pattern during fermentation. The probiotic preparation is a natural product containing bacteria resistant to gastric juice and bile: Lactobacillus paracasei LOCK 0920, Lactobacillus brevis LOCK 0944, Lactobacillus plantarum LOCK 0945, as well as live yeasts Saccharomyces cerevisiae LOCK 0140 of high fermenting capacity. After 6-hour fermentation with the probiotic, in feed with a low concentration of ochratoxin A (1 mg/kg) the amount of ochratoxin A decreased by 73%. In the case of high a concentration (5 mg/kg) the decrease in ochratoxin A was lower at about 55%. This tendency was sustained during the following hours of incubation (12th and 24th hours). The application of probiotic bacteria and yeasts resulted in the reduction of aerobic spore forming bacteria. It can be concluded that the probiotic preparation containing bacteria of Lactobacillus strains and yeasts Saccharomyces cerevisiae used in the study was conducive to detoxification of ochratoxin A added to a feed.

Aptamers: A Promising Tool for Ochratoxin A Detection in Food Analysis

The contamination of food and feed by mycotoxins has become an increasingly serious problem. Mycotoxins represent a major risk to human and animal health, as well as economics. Herein, we focus on Ochratoxin A (OTA), which is one of the most common mycotoxins contaminating feed and foodstuffs. OTA is a secondary metabolite produced by various Aspergillus and Penicillium strains. Upon ingestion, OTA has a number of acute and chronic toxic effects. It is nephrotoxic, teratogenic, immunosuppressive, and carcinogenic (group 2B). As a consequence, some regulatory limits have been introduced on the levels of OTA in several commodities. The toxic nature of OTA demands highly sensitive and selective monitoring techniques to protect human and animal health. As alternative to traditional analytical techniques, biochemical methods for OTA analysis have attained great interest in the last few decades. They are mainly based on the integration of antibodies or aptamers as biorecognition elements in sensing platforms. However, aptamers have gained more attention in affinity-based assays because of their high affinity, specificity, stability, and their easy chemical synthesis. In this brief review, we present an overview of aptamer-based assays and their applications in OTA purification and detection, appeared in the literature in the last five years.

In vitroremoval of ochratoxin A by two strains ofSaccharomyces cerevisiaeand their performances under fermentative and stressing conditions

The aim of this research was to study the effect of time, temperature, sugar content and addition of diammonium phosphate (DAP) on ochratoxin A (OTA) removal by two strains of Saccharomyces cerevisiae using a completely randomized design. The strains were grown in a medium containing OTA (2 μg l(-1)), two sugar levels (200 and 250 g l(-1)), with or without DAP (300 mg l(-1)), and incubated at 25-30°C. The yeasts were able to decrease the toxin amount by c. 70%, with the highest removing effect observed after 3 days at 30°C in the presence of 250 g l(-1) of sugars and with DAP; after 10 days, the toxin was partially released into the medium. The strains produced high ethanol and glycerol contents, showed high tolerance to single/combined stress conditions and possessed β-d-glucosidase, pectinase and xylanase activities. Ochratoxin A removal was affected by time, temperature, sugar and addition of DAP. Moreover, the phenomenon was reversible. Ochratoxin A removal could be an interesting trait for the selection of promising strains; however, the strains removing efficiently the toxin could release it back; thus, the selection of the starter should take into account both the removal and the binding ability of OTA.

Direct and indirect contamination with ochratoxin A of ripened pork products

The aim of this study was to evaluate the contribution of direct and indirect contamination on ochratoxin A (OTA) levels in different ripened pork products. A total of 24 Large White pigs were fed naturally OTA contaminated diets (4 different contamination levels, from 0.40 to 171 μg kg−1) for 2 weeks. Typical Italian pork products (dry sausage, dry-cured pork neck, dry-cured streaky bacon and dry-cured ham) were prepared using the contaminated tissues and ripened in 3 dry-cured ham manufacturing plants. Plasma, organs, tissues and ripened pork products were analysed for OTA content. As regards the animals fed at a level slightly less than 50 μg kg−1, the guidance value recommended by the Commission of European Communities, the OTA levels in muscle and in the ripened products were close to 1 μg kg−1 (range of the mean values 0.65–1.62 μg kg−1), the guideline value in meat products recommended by the Italian Ministry of Health. Dry sausage and dry-cured pork neck showed significantly higher OTA concentrations than dry-cured ham and dry-cured streaky bacon. OTA was partially degraded during the long ripening time of dry-cured ham. In dry sausage and dry-cured pork neck direct contamination was low and it was not detected in dry-cured streaky bacon; on the contrary, very high OTA levels were detected in several outer dry-cured ham samples (maximum OTA value: 314 μg kg−1). These results confirmed that direct contamination should be mainly monitored in dry-cured ham, whereas indirect contamination may eventually be relevant in other ripened pork products.

The effects of temperature and relative humidity on ochratoxin A formation in fresh liquorice root

In this study, the effects of temperature and relative humidity (RH) on ochratoxin A (OTA) formation during liquorice root storage were investigated. For this purpose, a real storage procedure in which fresh root was dried in the open air was simulated. Four RH-temperature combinations corresponding to average climate conditions (RH-temperature) of each season in an important liquorice growing and processing region were simulated as follows: A, autumn (T = 15°C, RH = 49%); B, winter (T = 9°C, RH = 51%); C, spring (T = 22°C, RH = 35%); and D, summer (T = 29°C, RH = 27%). The crushed fresh roots were incubated for 60 days, and the OTA content of the roots was determined at 20-day intervals by using inverse ion mobility spectrometry. The results showed that the maximum levels of OTA occurred after 40 days and were 4.3 ± 1.1, 0.9 ± 0.2, 7.3 ± 0.0 and 24.2 ± 2.5 ngg−1 in roots stored under simulated conditions A, B, C and D, respectively. After 40 days, the amount of OTA started to decline in all samples but at different rates. The results seem to indicate that temperature plays a more significant role than RH in producing OTA by moulds in liquorice root. It appears that 22°C could be considered as the critical temperature for OTA formation in liquorice root stored under experimental conditions. It could be concluded that liquorice-processing plants should obtain, dry and store fresh moist root when the temperature is below the critical point. Moreover, the roots stored in the open shade condition should not be covered by plastic films even when it is raining as this increases the root temperature.

Immunological status of the progeny of breeder hens kept on ochratoxin A (OTA)- and aflatoxin B1 (AFB1)-contaminated feeds

This study aimed to evaluate the immunological status of progeny of hens kept on ochratoxin A (OTA)- and aflatoxin B1 (AFB1)-contaminated feed. For this purpose, White Leghorn (WL) layer breeder hens (40-weeks-of-age) were divided into six groups (A–F). Hens in Group A were fed a commercial layer ration while those in Groups B and C were kept on a diet amended with 3 and 5 mg OTA/Kg, respectively. Group D was fed a ration containing 5 mg AFB1/Kg, while hens in Groups E and F were kept on feed amended with OTA and AFB1 each. All feedings were for 1, 2, or 3 weeks. Fertile eggs were set for hatching on a weekly basis to obtain progeny of each week separately. At 14 days-of-age, subsets of progeny were euthanized and the frequency of immunoglobulin(s)-bearing cells in their spleen and bursa of Fabricius assessed; at 16 days-of-age, other chicks in each set were utilized to determine their lymphoblastogenic responses against phytohemagglutinin (PHA-P). At 30 days-of-age, the final sub-set of chicks/group was euthanized and their peritoneal macrophages harvested for measurements of phagocytic potential and nitrite production. Relative weights of the bursa of Fabricius and of the spleen were significantly lower in the progeny of hens fed mycotoxin-contaminated diets for 14 and 21 days. The frequencies of IgA-, IgG-, and IgM-bearing cells were also significantly lower in the bursa of Fabricius and spleen of progeny chicks obtained from hens fed the OTA + AFB1 mixed diet. Feeding contaminated diets to breeder hens also resulted in significantly lower responses to PHA-P. In addition, the percentages of peritoneal macrophages displaying phagocytosis of sheep red blood cells (SRBC), the number of SRBC/macrophage, and nitrite production were each significantly lower in cells from progeny chicks from OTA- and AFB1-fed hens. The findings of the present study indicated there were severe immunosuppressive effects in progeny chicks as a result of exposure of their parent hens to OTA and AFB1 either alone or in combination. These studies provide emphasis for the need for mycotoxin regulation policy with respect to the ingredients used in poultry feed, since it is clear that feeding multi-mycotoxin-contaminated diets to breeder hens will almost certainly result in the hatching of manifestly unhealthy chicks.