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Toxins
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Ochratoxins 2011-2012
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Ochratoxins 2011-2012

A special issue of Toxins (ISSN 2072-6651). This special issue belongs to the section "Mycotoxins".

Deadline for manuscript submissions: closed (31 December 2012) | Viewed by 86214

Special Issue Editors

Prof. Dr. Richard A. Manderville

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Guest Editor
Department of Chemistry and Toxicology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada
Interests: DNA damage by phenolic toxins including ochratoxin A; Modified DNA bases as fluorescent probes
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Annie Pfohl-Leszkowicz

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Guest Editor
Department Bioprocess & Microbial Systems, Laboratory Chemical Engineering, INP/ENSA Toulouse, University of Toulouse, UMR 5503 CNRS/INPT/UPS, Auzeville-Tolosane 31320, France
Interests: mycotoxin; ochratoxin; fumonisin; zearalenone; biomarker; risk evaluation; environmental toxicology; polycyclic aromatic compounds; genotoxicity; DNA adduct; balkan endemic nephropathy; kidney cancer; biotransformation
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

In 2010 a special issue of Toxins entitled “Ochratoxins” published 31 papers concerned with detection of ochratoxins in feed and human foodstuff, occurrence and estimation of dietary intake, and understanding mechanisms of toxicity and carcinogenicity for the development of detoxification processes. In terms of ochratoxin A (OTA) carcinogenicity, highlights from the special issue include the findings by Stoev that OTA induces renal tumors in chicks. Schwartz and coworkers demonstrated that in utero exposure to OTA causes adducts in the testicular DNA of male offspring in support of a possible role for OTA in testicular cancer. 2010 also saw Mantle and coworkers publish convincing evidence that part of the mechanism of OTA carcinogenesis involves direct covalent interaction with DNA.  In the special issue Mantle and Nolan raised new questions about a difference between young adults and mature adults in sensitivity of male rats to the ochratoxin A-induced DNA damage necessary for renal carcinogenesis.  Several papers that include those by Fusi, Varga, Abrunhosa, and Perez examined control strategies for reducing or preventing OTA-induced toxicoses. For OTA detection, Yu and Lai outlined recent advances of molecularly imprinted polymers (MIPs) for the extraction and analysis of ochratoxins. Rapid and reliable visual tests for OTA detection was reviewed by Bazin and coworkers, while a new ELISA test for OTB detection was described by Dietrich coworkers. Numerous other studies demonstrated the wide-spread occurrence of OTA, highlighting that problematic exposure to the toxin is not limited to the Balkan region. Given the level of participation and interest in the special issue “Ochratoxins”, the editorial board of Toxins was eager to run this special issue again. To the many authors that contributed to ‘Ochratoxins” we wish to thank you for sharing your research findings that made the special issue such a success. We hope that this second special issue of Toxins entitled “Ochratoxins 2011” further highlights the ongoing interdisciplinary research on the ochratoxins and provides the readership with a further understanding of the key issues being addressed at the present time.

Prof. Dr. Richard A. Manderville
Prof. Dr. Annie Pfohl-Leszkowicz
Guest Editors

Keywords

  • ochratoxins
  • mycotoxin
  • carcinogen
  • Balkan endemic nephropathy
  • DNA damage
  • genotoxic
  • intake
  • biotransformation
  • risk assessment

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Published Papers (11 papers)

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Research

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203 KiB  
Article
First Evidence of Placental Transfer of Ochratoxin A in Horses
by Fiorenza Minervini, Alessandra Giannoccaro, Michele Nicassio, Giuseppe Panzarini and Giovanni Michele Lacalandra
Toxins 2013, 5(1), 84-92; https://doi.org/10.3390/toxins5010084 - 11 Jan 2013
Cited by 23 | Viewed by 6613
Abstract
Ochratoxin A (OTA) is a renal mycotoxin and transplacental genotoxic carcinogen. The aim of this study was to evaluate the natural occurrence of OTA in equine blood samples and its placental transfer. For the assessment of OTA levels, serum samples were collected from [...] Read more.
Ochratoxin A (OTA) is a renal mycotoxin and transplacental genotoxic carcinogen. The aim of this study was to evaluate the natural occurrence of OTA in equine blood samples and its placental transfer. For the assessment of OTA levels, serum samples were collected from 12 stallions, 7 cycling mares and 17 pregnant mares. OTA was found in 83% of serum samples (median value = 121.4 pg/mL). For the assessment of placental transfer, serum samples were collected from the 17 mares after delivery and from the umbilical cords of their foals, after foaling. Fourteen serum samples from pregnant mares contained OTA (median value = 106.5 pg/mL), but only 50% of their foals were exposed (median values = 96.6 pg/mL). HPLC analysis carried out on four serum samples (collected from two mares and their respective foals) supported the ELISA results on OTA placental transfer. This is the first report on the natural occurrence of OTA in horse serum samples and placental transfer in horses. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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583 KiB  
Article
Ochratoxin A Management in Vineyards by Lobesia botrana Biocontrol
by Giuseppe Cozzi, Stefania Somma, Miriam Haidukowski and Antonio F. Logrieco
Toxins 2013, 5(1), 49-59; https://doi.org/10.3390/toxins5010049 - 02 Jan 2013
Cited by 29 | Viewed by 6905
Abstract
Grape berries attacked by Lobesia botrana larvae are more easily infected by Aspergillus section Nigri (black aspergilli) ochratoxigenic species. Two-year field trials were carried out in Apulia (Italy) to evaluate a bioinsecticide control strategy against L. botrana and the indirect effect on reducing [...] Read more.
Grape berries attacked by Lobesia botrana larvae are more easily infected by Aspergillus section Nigri (black aspergilli) ochratoxigenic species. Two-year field trials were carried out in Apulia (Italy) to evaluate a bioinsecticide control strategy against L. botrana and the indirect effect on reducing ochratoxin A (OTA) contamination in vineyards. A commercial Bacillus thuringiensis formulate and an experimental Beauveria bassiana (ITEM-1559) formulate were tested in two vineyards cultivated with the same grape variety, Negroamaro, but with two different training systems (espalier and little-arbor techniques). In both years and training systems the treatments by B. bassiana ITEM-1559 significantly controlled L. botrana larvae attacks with effectiveness similar to B. thuringensis (more than 20%). A significant reduction of OTA concentrations (up to 80% compared to untreated controls) was observed only in the first year in both training systems, when the metereological parameters prior to harvest were more favorable to the insect attack. Results of field trials showed that B. bassiana ITEM-1559 is a valid bioinsecticide against L. botrana and that grape moth biocontrol is a strategy to reduce OTA contamination in vineyard in seasons with heavy natural infestation. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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1818 KiB  
Article
Apoptosis Induction by OTA and TNF-α in Cultured Primary Rat Hepatocytes and Prevention by Silibinin
by Ebtisam Essid, Yousef Dernawi and Ernst Petzinger
Toxins 2012, 4(11), 1139-1156; https://doi.org/10.3390/toxins4111139 - 02 Nov 2012
Cited by 24 | Viewed by 7005
Abstract
In cultures of primary rat hepatocytes, apoptosis occurred after application of 20 ng/mL tumor necrosis factor alpha (TNF-α). However, this was only in the presence of 200 ng/mL of the transcriptional inhibitor actinomycin D (ActD). This toxic effect was completely prevented in the [...] Read more.
In cultures of primary rat hepatocytes, apoptosis occurred after application of 20 ng/mL tumor necrosis factor alpha (TNF-α). However, this was only in the presence of 200 ng/mL of the transcriptional inhibitor actinomycin D (ActD). This toxic effect was completely prevented in the presence of 25 µg/mL soluble TNF-α receptor I (sTNFR I) in the supernatant of hepatocyte cell cultures. Apoptosis also occurred after application of 12.5 µmol/L ochratoxin A (OTA). However, that was not prevented by up to 500 µg/mL sTNFR I, indicating that TNF-α/TNFR I is not involved in OTA mediated apoptosis in hepatocytes. The antioxidative flavanolignan silibinin in doses from 130 to 260 µmol/L prevented chromatin condensation, caspase-3 activation, and apoptotic DNA fragmentation that were induced by OTA, by 10 mmol/L hydrogen peroxide (H2O2) and by ultraviolet (UV-C) light (50 mJ/cm2), respectively. To achieve protection by silibinin, the drug was applied to the cell cultures for 2 h in advance. OTA stimulated lipid peroxidation on cultured immortalized rat liver HPCT cells, as was revealed by malondialdehyde (MDA) production. Lipid peroxidation occurred further by H2O2 and ActD/TNF-α incubation. These reactions were also suppressed by silibinin pretreatment. We conclude that the anti-apoptotic activity of silibinin against OTA, H2O2 and ActD/ TNF-α is caused in vitro by the antioxidative effects of the flavanolignan. Furthermore, cytotoxicity of the pro-apoptotic toxins was revealed by MTT-test. When applied separately, ActD and TNF-α showed no cytotoxic effects after 24 h, but were cytotoxic if applied in combination. The used concentrations of OTA, H2O2 and the dose of UV-C caused a substantial decrease in viability within 36 h that was prevented mostly by silibinin. We conclude that silibinin is a potent protective compound against apoptosis and cytotoxicity caused by OTA and the investigated compounds. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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897 KiB  
Article
Comparative Immunohistochemical Analysis of Ochratoxin A Tumourigenesis in Rats and Urinary Tract Carcinoma in Humans; Mechanistic Significance of p-S6 Ribosomal Protein Expression
by Patrycja Gazinska, Diana Herman, Cheryl Gillett, Sarah Pinder and Peter Mantle
Toxins 2012, 4(9), 643-662; https://doi.org/10.3390/toxins4090643 - 11 Sep 2012
Cited by 13 | Viewed by 7160
Abstract
Ochratoxin A (OTA) is considered to be a possible human urinary tract carcinogen, based largely on a rat model, but no molecular genetic changes in the rat carcinomas have yet been defined. The phosphorylated-S6 ribosomal protein is a marker indicating activity of the [...] Read more.
Ochratoxin A (OTA) is considered to be a possible human urinary tract carcinogen, based largely on a rat model, but no molecular genetic changes in the rat carcinomas have yet been defined. The phosphorylated-S6 ribosomal protein is a marker indicating activity of the mammalian target of rapamycin, which is a serine/threonine kinase with a key role in protein biosynthesis, cell proliferation, transcription, cellular metabolism and apoptosis, while being functionally deregulated in cancer. To assess p-S6 expression we performed immunohistochemistry on formalin-fixed and paraffin-embedded tumours and normal tissues. Marked intensity of p-S6 expression was observed in highly proliferative regions of rat renal carcinomas and a rare angiosarcoma, all of which were attributed to prolonged exposure to dietary OTA. Only very small OTA-generated renal adenomas were negative for p-S6. Examples of rat subcutaneous fibrosarcoma and testicular seminoma, as well as of normal renal tissue, showed no or very weak positive staining. In contrast to the animal model, human renal cell carcinoma, upper urinary tract transitional cell carcinoma from cases of Balkan endemic nephropathy, and a human angiosarcoma were negative for p-S6. The combined findings are reminiscent of constitutive changes in the rat tuberous sclerosis gene complex in the Eker strain correlated with renal neoplasms, Therefore rat renal carcinogenesis caused by OTA does not obviously mimic human urinary tract tumourigenesis. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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302 KiB  
Article
Dimethylarginine Dimethylaminohydrolase/Nitric Oxide Synthase Pathway in Liver and Kidney: Protective Effect of Cyanidin 3-O-β-D-Glucoside on Ochratoxin-A Toxicity
by Valeria Sorrenti, Claudia Di Giacomo, Rosaria Acquaviva, Matteo Bognanno, Ester Grilli, Nicolantonio D’Orazio and Fabio Galvano
Toxins 2012, 4(5), 353-363; https://doi.org/10.3390/toxins4050353 - 08 May 2012
Cited by 17 | Viewed by 6964
Abstract
The aim of the present study was to evaluate the effect of long-term cyanidin 3-O-β-D-glucoside (C3G) and/or Ochratoxin A (OTA)-exposure on dimethylarginine dimethylamino hydrolase/nitric oxide synthase (DDAH/NOS) pathway in rats. The experiments were performed in rats supplemented with C3G (1 g/kg [...] Read more.
The aim of the present study was to evaluate the effect of long-term cyanidin 3-O-β-D-glucoside (C3G) and/or Ochratoxin A (OTA)-exposure on dimethylarginine dimethylamino hydrolase/nitric oxide synthase (DDAH/NOS) pathway in rats. The experiments were performed in rats supplemented with C3G (1 g/kg feed), OTA (200 ppb), and OTA + C3G. After 4 weeks of daily treatment, liver and kidneys were processed for eNOS, iNOS and DDAH-1 Western blotting, nitrite levels evaluation and DDAH activity determination. Results show that OTA is able to induce iNOS both in kidney and liver, whereas OTA is able to induce eNOS and DDAH-1 overexpression and DDAH activation only in kidney, resulting in increased nitrite levels. In kidney of OTA + C3G fed rats, iNOS, eNOS and DDAH-1 expression were less pronounced compared with those observed in the OTA-treated group. Coherent with the decreased iNOS, eNOS and DDAH-1 expression a decrease in nitrite levels and DDAH activity was observed in the OTA + C3G group. Results demonstrate that C3G is able to counteract the deleterious effects of chronic consumption of OTA and also suggest a possible involvement of iNOS-eNOS-DDAH impairment in OTA nephrocarcinogenity. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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235 KiB  
Article
Mutagenicity of Ochratoxin A and Its Hydroquinone Metabolite in the SupF Gene of the Mutation Reporter Plasmid Ps189
by Steven A. Akman, Marissa Adams, Doug Case, Gyungse Park and Richard A. Manderville
Toxins 2012, 4(4), 267-280; https://doi.org/10.3390/toxins4040267 - 16 Apr 2012
Cited by 19 | Viewed by 7543
Abstract
Ochratoxin A (OTA) is a mycotoxin that enhances renal tumor formation in the outer medulla of male rat kidney. Direct DNA damage and subsequent mutagenicity may contribute to these processes. In this study we have determined whether OTA in the absence or presence [...] Read more.
Ochratoxin A (OTA) is a mycotoxin that enhances renal tumor formation in the outer medulla of male rat kidney. Direct DNA damage and subsequent mutagenicity may contribute to these processes. In this study we have determined whether OTA in the absence or presence of activated rat liver microsomes (RLM) or redox-active transition metals (Fe(III) or Cu(II)) causes promutagenic DNA damage in the supF gene of the mutation reporter plasmid pS189 replicating in human Ad293 cells. In addition, we have assessed the mutagenicity of the hydroquinone metabolite (OTHQ) of OTA in the absence or presence of cysteine without added cofactors. Our results show that oxidation of OTA, either by RLM or by transition metal ions, activates OTA to a directly genotoxic mutagen(s). The Fe(III)/OTA system was the most potent mutagen in our experimental system, causing a 32-fold increase in mutant fraction (MF) above the spontaneous control MF. The Cu(II)/OTA system caused a 9-fold increase in MF, while a 6–10-fold increase in MF was observed for OTA in the presence of RLM. The OTHQ metabolite is also mutagenic, especially in the presence of cysteine, in which a 6-fold increase in MF was observed. Our data provide further insight into OTA bioactivation that may account for its in vivo mutagenicity in male rat kidney. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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525 KiB  
Communication
Sorption of Ochratoxin A from Aqueous Solutions Using β-Cyclodextrin-Polyurethane Polymer
by Michael Appell and Michael A. Jackson
Toxins 2012, 4(2), 98-109; https://doi.org/10.3390/toxins4020098 - 06 Feb 2012
Cited by 35 | Viewed by 6912
Abstract
The ability of a cyclodextrin-polyurethane polymer to remove ochratoxin A from aqueous solutions was examined by batch rebinding assays. The results from the aqueous binding studies were fit to two parameter models to gain insight into the interaction of ochratoxin A with the [...] Read more.
The ability of a cyclodextrin-polyurethane polymer to remove ochratoxin A from aqueous solutions was examined by batch rebinding assays. The results from the aqueous binding studies were fit to two parameter models to gain insight into the interaction of ochratoxin A with the nanosponge material. The ochratoxin A sorption data fit well to the heterogeneous Freundlich isotherm model. The polymer was less effective at binding ochratoxin A in high pH buffer (9.5) under conditions where ochratoxin A exists predominantly in the dianionic state. Batch rebinding assays in red wine indicate the polymer is able to remove significant levels of ochratoxin A from spiked solutions between 1–10 μg·L−1. These results suggest cyclodextrin nanosponge materials are suitable to reduce levels of ochratoxin A from spiked aqueous solutions and red wine samples. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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349 KiB  
Article
Biocontrol of Penicillium nordicum Growth and Ochratoxin A Production by Native Yeasts of Dry Cured Ham
by Roberta Virgili, Nicoletta Simoncini, Tania Toscani, Marco Camardo Leggieri, Silvia Formenti and Paola Battilani
Toxins 2012, 4(2), 68-82; https://doi.org/10.3390/toxins4020068 - 01 Feb 2012
Cited by 59 | Viewed by 12162
Abstract
Twelve yeast strains isolated from the surface of Italian typical dry-cured hams, belonging to D. hansenii, D. maramus, C. famata, C. zeylanoides and H. burtonii species, and previously selected for their ability to grow in dry-cured ham-like substrates, were screened [...] Read more.
Twelve yeast strains isolated from the surface of Italian typical dry-cured hams, belonging to D. hansenii, D. maramus, C. famata, C. zeylanoides and H. burtonii species, and previously selected for their ability to grow in dry-cured ham-like substrates, were screened for antagonistic activity against a toxigenic strain of P. nordicum and inhibition of ochratoxin A (OTA) biosynthesis. On average, yeast inhibitory activity was lowered by increasing fungal inoculum and enhanced by NaCl presence. In the assay conditions, H. burtonii and C. zeylanoides were the most effective, both in inhibiting P. nordicum growth and OTA production. D. hansenii was the species with the lowest inhibitory activity, especially in the absence of salt. OTA production dropped from the range < LOD − 5000 ppb in P. nordicum control plates to the range < LOD − 200 ppb in yeast-added plates. OTA production increased in the presence of NaCl in P. nordicum control plates, while salt enhanced inhibition against OTA production in yeast-added plates. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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346 KiB  
Article
Ochratoxigenic Black Species of Aspergilli in Grape Fruits of Northern Italy Identified by an Improved PCR-RFLP Procedure
by Davide Spadaro, Subban Patharajan, Alessia Lorè, Angelo Garibaldi and Maria Lodovica Gullino
Toxins 2012, 4(2), 42-54; https://doi.org/10.3390/toxins4020042 - 30 Jan 2012
Cited by 24 | Viewed by 6559
Abstract
A collection of 356 isolates of Aspergillus spp. collected during 2006 and 2007 from grapevines in northern Italy were identified through Internal Transcribed Spacer based Restriction Fragment Length Polymorphism (ITS-RFLP) and tested for ochratoxin A (OTA) production. Restriction endonuclease digestion of the ITS [...] Read more.
A collection of 356 isolates of Aspergillus spp. collected during 2006 and 2007 from grapevines in northern Italy were identified through Internal Transcribed Spacer based Restriction Fragment Length Polymorphism (ITS-RFLP) and tested for ochratoxin A (OTA) production. Restriction endonuclease digestion of the ITS products using the endonucleases HhaI, HinfI and RsaI, distinguished five different RFLPs. From each pattern, three samples were sequenced and the nucleotide sequences showed different species corresponding to Aspergillus niger, A. carbonarius, A. tubingensis, A. japonicus and A. aculeatus. By comparing the sequences of the ITS regions, also the uniseriate species A. japonicus and A. aculeatus could be differentiated by HinfI digestion of the ITS products. Among the aspergilli, A. niger was the major species associated with grapes during 2006 (57.4%), while A. carbonarius was the major species during 2007 (46.6%). All the strains of Aspergillus were tested for their ability to produce OTA on Yeast extract sucrose medium (YES), as it was tested as an optimal substrate for the evaluation of OTA production by black aspergilli. Out of 356 isolates, 63 (17.7%) isolates produced OTA ranging from 0.05 to 3.0 µg mL−1. Most of the ochratoxigenic isolates were A. carbonarius (46) in both years, but also some strains of A. tubingensis (11) and A. japonicus (6) produced lower amounts of OTA. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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Review

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166 KiB  
Review
Control of Ochratoxin A Production in Grapes
by María Lorena Ponsone, María Laura Chiotta, Juan Manuel Palazzini, Mariana Combina and Sofía Chulze
Toxins 2012, 4(5), 364-372; https://doi.org/10.3390/toxins4050364 - 14 May 2012
Cited by 29 | Viewed by 7644
Abstract
Ochratoxin A (OTA) is a mycotoxin commonly present in cereals, grapes, coffee, spices, and cocoa. Even though the main objective of the food and feed chain processors and distributors is to avoid the extended contamination of plant-derived foods and animal feeds with mycotoxins, [...] Read more.
Ochratoxin A (OTA) is a mycotoxin commonly present in cereals, grapes, coffee, spices, and cocoa. Even though the main objective of the food and feed chain processors and distributors is to avoid the extended contamination of plant-derived foods and animal feeds with mycotoxins, until now, complete OTA removal from foods and feedstuffs is not feasible. Prevention through pre-harvest management is the best method for controlling mycotoxin contamination. However, in the case that the contamination occurs after this stage, the hazards associated with OTA must be managed through post-harvest strategies. Due to the increasing number of fungal strains resistant to chemical fungicides and the impact of these pesticides on the environment and human health, maximum levels of chemical residues have been regulated in many products. Alternative methods are necessary to substitute or complement treatments with fungicides to control fungi under field or storage conditions. Yeasts are considered one of the most potent biocontrol agents due to their biology and non-toxic properties. Epiphytic yeasts are the major component of the microbial community on the surface of grape berries and they are evolutionarily adapted to this ecological niche. Nowadays, several yeast species included in different genera are considered as potential biocontrol agents to control both, growth of ochratoxigenic Aspergillus species and OTA accumulation. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
238 KiB  
Review
Immunochemical Methods for Ochratoxin A Detection: A Review
by Eline P. Meulenberg
Toxins 2012, 4(4), 244-266; https://doi.org/10.3390/toxins4040244 - 13 Apr 2012
Cited by 116 | Viewed by 10206
Abstract
The safety of food and feed depends to a great deal on quality control. Numerous compounds and organisms may contaminate food and feed commodities and thus pose a health risk for consumers. The compound of interest in this review is ochratoxin A (OTA), [...] Read more.
The safety of food and feed depends to a great deal on quality control. Numerous compounds and organisms may contaminate food and feed commodities and thus pose a health risk for consumers. The compound of interest in this review is ochratoxin A (OTA), a secondary metabolite of the fungi Aspergillus and Penicillium. Due to its adverse health effects, detection and quantification are of utmost importance. Quality control of food and feed requires extraction and analysis, including TLC, HPLC, MS, and immunochemical methods. Each of these methods has its advantages and disadvantages. However, with regard to costs and rapidity, immunochemical methods have gained much interest in the last decade. In this review an introduction to immunochemistry and assay design will be given to elucidate the principles. Further, the application of the various formats to the detection and quantification of ochratoxin will be described, including the use of commercially available kits. Full article
(This article belongs to the Special Issue Ochratoxins 2011-2012)
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