Occurrence, Toxicity, and Analysis of Major Mycotoxins in Food

Mycotoxins are toxic secondary metabolites produced by certain filamentous fungi (molds). These low molecular weight compounds (usually less than 1000 Daltons) are naturally occurring and practically unavoidable. They can enter our food chain either directly from plant-based food components contaminated with mycotoxins or by indirect contamination from the growth of toxigenic fungi on food. Mycotoxins can accumulate in maturing corn, cereals, soybeans, sorghum, peanuts, and other food and feed crops in the field and in grain during transportation. Consumption of mycotoxin-contaminated food or feed can cause acute or chronic toxicity in human and animals. In addition to concerns over adverse effects from direct consumption of mycotoxin-contaminated foods and feeds, there is also public health concern over the potential ingestion of animal-derived food products, such as meat, milk, or eggs, containing residues or metabolites of mycotoxins. Members of three fungal genera, Aspergillus, Fusarium, and Penicillium, are the major mycotoxin producers. While over 300 mycotoxins have been identified, six (aflatoxins, trichothecenes, zearalenone, fumonisins, ochratoxins, and patulin) are regularly found in food, posing unpredictable and ongoing food safety problems worldwide. This review summarizes the toxicity of the six mycotoxins, foods commonly contaminated by one or more of them, and the current methods for detection and analysis of these mycotoxins.

[1]  S. Sumner,et al.  Stability of fumonisins in thermally processed corn products. , 1998, Journal of food protection.

[2]  Ewald Usleber,et al.  Rapid Detection of Fumonisin B1 in Corn-Based Food by Competitive Direct Dipstick Enzyme Immunoassay/Enzyme-Linked Immunofiltration Assay with Integrated Negative Control Reaction , 1995 .

[3]  Haibo Yao,et al.  Developments in detection and determination of aflatoxins , 2015 .

[4]  J. Robens,et al.  Aflatoxins in animal and human health. , 1992, Reviews of environmental contamination and toxicology.

[5]  N. Magan,et al.  Detection and differentiation between mycotoxigenic and non‐mycotoxigenic strains of two Fusarium spp. using volatile production profiles and hydrolytic enzymes , 2000, Journal of applied microbiology.

[6]  J. Richard,et al.  Some major mycotoxins and their mycotoxicoses--an overview. , 2007, International journal of food microbiology.

[7]  Dafeng Jiang,et al.  Fumonisins B1, B2 and B3 in corn products, wheat flour and corn oil marketed in Shandong province of China , 2015, Food additives & contaminants. Part B, Surveillance.

[8]  C. V. Van Peteghem,et al.  Development of a solid-phase cleanup and portable rapid flow-through enzyme immunoassay for the detection of ochratoxin a in roasted coffee. , 2002, Journal of agricultural and food chemistry.

[9]  A. Visconti,et al.  Optimization of a fluorescence polarization immunoassay for rapid quantification of deoxynivalenol in durum wheat-based products. , 2006, Journal of food protection.

[10]  Anton Simeonov,et al.  Fluorescence polarization assays in small molecule screening , 2011, Expert opinion on drug discovery.

[11]  C. Maragos,et al.  Challenges in the analysis of multiple mycotoxins , 2016 .

[12]  Itziar Ruisánchez,et al.  Qualitative method for determination of aflatoxin B1 in nuts. , 2004, Journal of AOAC International.

[13]  E. Stigter,et al.  Rapid surface plasmon resonance-based inhibition assay of deoxynivalenol. , 2003, Journal of agricultural and food chemistry.

[14]  S. Chulze,et al.  Strategies to reduce mycotoxin levels in maize during storage: a review , 2010, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[15]  M. Jiménez,et al.  Comparison of different sample treatments for the analysis of ochratoxin A in must, wine and beer by liquid chromatography. , 2004, Journal of chromatography. A.

[16]  Giorgio Sberveglieri,et al.  Detection of toxigenic strains of Fusarium verticillioides in corn by electronic olfactory system , 2005 .

[17]  Jian Zhong Shen,et al.  The development of a fluorescence polarization immunoassay for aflatoxin detection. , 2014, Biomedical and environmental sciences : BES.

[18]  S. Piletsky,et al.  Analytical methods for determination of mycotoxins: a review. , 2009, Analytica chimica acta.

[19]  G. Shephard Current Status of Mycotoxin Analysis: A Critical Review. , 2016, Journal of AOAC International.

[20]  T. Whitaker Detecting mycotoxins in agricultural commodities , 2003, Molecular biotechnology.

[21]  A. G. Frenich,et al.  Multi-mycotoxin analysis in eggs using a QuEChERS-based extraction procedure and ultra-high-pressure liquid chromatography coupled to triple quadrupole mass spectrometry. , 2011, Journal of chromatography. A.

[22]  S. Pepeljnjak,et al.  Studies of ochratoxin A-induced inhibition of phenylalanine hydroxylase and its reversal by phenylalanine. , 2000, Toxicology and applied pharmacology.

[23]  Rudolf Krska,et al.  Mycotoxin analysis: An update , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[24]  W. Ragab,et al.  Mycotoxins in fruits, fruit juices, and dried fruits. , 2003, Journal of food protection.

[25]  Selamat Jinap,et al.  Qualitative and Quantitative Analysis of Mycotoxins. , 2009, Comprehensive reviews in food science and food safety.

[26]  S. Stoev Food Safety and Increasing Hazard of Mycotoxin Occurrence in Foods and Feeds , 2013, Critical reviews in food science and nutrition.

[27]  N. Thiex,et al.  Multiresidue mycotoxin analysis in corn grain by column high-performance liquid chromatography with postcolumn photochemical and chemical derivatization: single-laboratory validation. , 2009, Journal of AOAC International.

[28]  Steven J Lehotay,et al.  Fast and easy multiresidue method employing acetonitrile extraction/partitioning and "dispersive solid-phase extraction" for the determination of pesticide residues in produce. , 2003, Journal of AOAC International.

[29]  S. Cunha,et al.  Mycotoxins in cereals and related foodstuffs: A review on occurrence and recent methods of analysis , 2014 .

[30]  C. V. Van Peteghem,et al.  Dipstick enzyme immunoassay to detect Fusarium T-2 toxin in wheat , 1996, Applied and environmental microbiology.

[31]  R. Krska,et al.  Improving methods of analysis for mycotoxins: molecularly imprinted polymers for deoxynivalenol and zearalenone , 2003, Food additives and contaminants.

[32]  J. Gilbert,et al.  Bioactive compounds in foods , 2008 .

[33]  C. Baggiani,et al.  Mycotoxin detection. , 2016, Current opinion in biotechnology.

[34]  I. Jajić,et al.  Comparison of ELISA, HPLC-FLD and HPLC-MS/MS Methods for Determination of Aflatoxin M1 in Natural Contaminated Milk Samples. , 2016, Acta chimica Slovenica.

[35]  V. Lattanzio,et al.  Determination of trichothecenes in cereals and cereal-based products by liquid chromatography–tandem mass spectrometry , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[36]  Karolina Kowalska,et al.  Zearalenone as an endocrine disruptor in humans. , 2016, Environmental toxicology and pharmacology.

[37]  W. Carlton,et al.  Patulin mycotoxicosis in the rat: toxicology, pathology and clinical pathology. , 1982, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[38]  J. Miller,et al.  A Concise History of Mycotoxin Research. , 2017, Journal of agricultural and food chemistry.

[39]  C. Abnet Carcinogenic Food Contaminants , 2007, Cancer investigation.

[40]  Wolfgang Lindner,et al.  Towards ochratoxin A selective molecularly imprinted polymers for solid-phase extraction. , 2002, Journal of chromatography. A.

[41]  M. Zheng,et al.  A Review of Rapid Methods for the Analysis of Mycotoxins , 2006, Mycopathologia.

[42]  Chulze Sn Strategies to reduce mycotoxin levels in maize during storage: a review. , 2010 .

[43]  S. Yazar,et al.  Fumonisins, Trichothecenes and Zearalenone in Cereals , 2008, International journal of molecular sciences.

[44]  Leilei Chen,et al.  Quantitative Analysis of 10 Mycotoxins in Wheat Flour by Ultrahigh Performance Liquid Chromatography-Tandem Mass Spectrometry with a Modified QuEChERS Strategy. , 2016, Journal of food science.

[45]  K. Kallela,et al.  The oestrogenic Fusarium toxin (zearalenone) in hay as a cause of early abortions in the cow. , 1984, Nordisk veterinaermedicin.

[46]  A. Mueller,et al.  Mycotoxin Analysis Using Imprinted Materials Technology: Recent Developments. , 2016, Journal of AOAC International.

[47]  Zhaowei Zhang,et al.  Advanced hyphenated chromatographic-mass spectrometry in mycotoxin determination: current status and prospects. , 2013, Mass spectrometry reviews.

[48]  Karsten Haupt,et al.  Imprinted polymers-tailor-made mimics of antibodies and receptors. , 2003, Chemical communications.

[49]  Toshitsugu Tanaka,et al.  Worldwide contamination of cereals by the Fusarium mycotoxins nivalenol, deoxynivalenol, and zearalenone. 1. Survey of 19 countries , 1988 .

[50]  C. Lino,et al.  A review on ochratoxin A occurrence and effects of processing of cereal and cereal derived food products. , 2010, Food microbiology.

[51]  C. Maragos Fluorescence polarization for mycotoxin determination , 2006, Mycotoxin Research.

[52]  R. Krska,et al.  Development of a method for the determination of Fusarium fungi on corn using mid-infrared spectroscopy with attenuated total reflection and chemometrics. , 2003, Analytical chemistry.

[53]  K. Scudamore Prevention of ochratoxin A in commodities and likely effects of processing fractionation and animal feeds , 2005, Food additives and contaminants.

[54]  A Prandini,et al.  On the occurrence of aflatoxin M1 in milk and dairy products. , 2009, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[55]  N. Leepipatpiboon,et al.  Evaluation of a modified QuEChERS method for analysis of mycotoxins in rice. , 2014, Food chemistry.

[56]  R. Krska,et al.  Portable Infrared Laser Spectroscopy for On-site Mycotoxin Analysis , 2017, Scientific Reports.

[57]  M. Pascale,et al.  Detection methods for mycotoxins in cereal grains and cereal products , 2009 .

[58]  H. Chun,et al.  A fluorescence polarization immunoassay for the detection of zearalenone in corn. , 2009, Analytica chimica acta.

[59]  A. Pier Major biological consequences of aflatoxicosis in animal production. , 1992, Journal of animal science.

[60]  C. Maragos,et al.  Capillary electrophoresis of the mycotoxin zearalenone using cyclodextrin-enhanced fluorescence. , 2007, Journal of chromatography. A.

[61]  Y. Grosse,et al.  Mycotoxins as human carcinogens—the IARC Monographs classification , 2017, Mycotoxin Research.

[62]  V. Dell’Orto,et al.  Application of an electronic nose to detection of aflatoxins in corn , 2009, Veterinary Research Communications.

[63]  V. Lattanzio,et al.  Analysis of T-2 and HT-2 toxins in cereal grains by immunoaffinity clean-up and liquid chromatography with fluorescence detection. , 2005, Journal of chromatography. A.

[64]  Jiujiang Yu,et al.  Understanding the genetics of regulation of aflatoxin production and Aspergillus flavus development , 2006, Mycopathologia.

[65]  T B Whitaker,et al.  Sampling Foods for Mycotoxins , 2006, Food additives and contaminants.

[66]  J. Schnürer,et al.  Detection and quantification of ochratoxin A and deoxynivalenol in barley grains by GC-MS and electronic nose. , 2002, International journal of food microbiology.

[67]  A. Pittet,et al.  Modern methods and trends in mycotoxin analysis , 2005 .

[68]  G. Osweiler Mycotoxins. , 2000, The Veterinary clinics of North America. Equine practice.

[69]  C. Magnoli,et al.  Occurrence of ochratoxin A and ochratoxigenic mycoflora in corn and corn based foods and feeds in some South American countries , 2007, Mycopathologia.

[70]  P. Mantle,et al.  Experimental one year ochratoxin A toxicosis in pigs. , 2002, Experimental and toxicologic pathology : official journal of the Gesellschaft fur Toxikologische Pathologie.

[71]  Rudolf Krska,et al.  Rapid test strips for analysis of mycotoxins in food and feed , 2009, Analytical and bioanalytical chemistry.

[72]  O. Mwanda,et al.  Acute aflatoxicosis: case report. , 2005, East African medical journal.

[73]  S. Saeger,et al.  Natural occurrence of mycotoxins and their masked forms in food and feed products , 2012 .

[74]  S. Feast Potential applications of electronic noses in cereals. , 2001 .

[75]  G. Cano-Sancho,et al.  Mycotoxins: occurrence, toxicology, and exposure assessment. , 2013, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[76]  C. Maragos,et al.  Fluorescence polarization as a means for determination of fumonisins in maize. , 2001, Journal of agricultural and food chemistry.

[77]  S. Piletsky,et al.  Analytical methods for determination of mycotoxins: An update (2009-2014). , 2015, Analytica chimica acta.

[78]  Felicia Wu,et al.  Mycotoxin risk assessment for the purpose of setting international regulatory standards. , 2004, Environmental science & technology.

[79]  A. Marocco,et al.  Rapid detection of kernel rots and mycotoxins in maize by near-infrared reflectance spectroscopy. , 2005, Journal of agricultural and food chemistry.

[80]  Yan Liu,et al.  Global Burden of Aflatoxin-Induced Hepatocellular Carcinoma: A Risk Assessment , 2010, Environmental health perspectives.

[81]  John P. Rheeder,et al.  Production of Fumonisin Analogs by Fusarium Species , 2002, Applied and Environmental Microbiology.

[82]  A. Moretti,et al.  Mycotoxins: An Underhand Food Problem. , 2017, Methods in molecular biology.

[83]  Lena Åberg,et al.  Near infrared spectroscopy for determination of mycotoxins in cereals , 2003 .

[84]  S. Jinap,et al.  Validation of the procedure for the simultaneous determination of aflatoxins ochratoxin A and zearalenone in cereals using HPLC-FLD , 2010, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[85]  F. Fry,et al.  Determination of aflatoxins B1, B2, G1, and G2 and ochratoxin A in ginseng and ginger by multitoxin immunoaffinity column cleanup and liquid chromatographic quantitation: collaborative study. , 2008, Journal of AOAC International.

[86]  O. Padilla-Zakour,et al.  Comprehensive Review of Patulin Control Methods in Foods. , 2005, Comprehensive reviews in food science and food safety.

[87]  B. Salleh,et al.  An overview of plant-derived products on control of mycotoxigenic fungi and mycotoxins. , 2010 .

[88]  G. Kaushik Effect of Processing on Mycotoxin Content in Grains , 2015, Critical reviews in food science and nutrition.

[89]  Roger M. Smith,et al.  3.40 – Sample Preparation for Food Contaminant Analysis , 2012 .

[90]  N. Botsoglou,et al.  Distribution and stability of aflatoxin M1 during processing, ripening and storage of Telemes cheese , 2001, Food additives and contaminants.

[91]  A Logrieco,et al.  DNA arrays, electronic noses and tongues, biosensors and receptors for rapid detection of toxigenic fungi and mycotoxins: A review , 2005, Food additives and contaminants.

[92]  P. Brereton,et al.  Simultaneous determination of aflatoxins and ochratoxin A in food using a fully automated immunoaffinity column clean-up and liquid chromatography-fluorescence detection. , 2004, Journal of chromatography. A.

[93]  Wolfgang Lindner,et al.  Molecularly imprinted polymer-assisted sample clean-up of ochratoxin A from red wine: merits and limitations. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[94]  K. Krishnamachari,et al.  Investigations into an outbreak of hepatitis in parts of western India. , 1975, The Indian journal of medical research.

[95]  C. Hurburgh,et al.  Potential economic losses to the US corn industry from aflatoxin contamination , 2016, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[96]  H. Nelis,et al.  Development of a flow-through enzyme immunoassay and application in screening green coffee samples for ochratoxin A with confirmation by high-performance liquid chromatography. , 2001, Journal of food protection.

[97]  Peiwu Li,et al.  Development of a multiple immunoaffinity column for simultaneous determination of multiple mycotoxins in feeds using UPLC–MS/MS , 2016, Analytical and Bioanalytical Chemistry.

[98]  Masinde Muliro Derivatization Reactions and Reagents for Gas Chromatography Analysis , 2012 .

[99]  P. Mantle Risk assessment and the importance of ochratoxins , 2002 .

[100]  F Bonomi,et al.  Binding of aflatoxin M1 to different protein fractions in ovine and caprine milk. , 2007, Journal of dairy science.

[101]  R. Krska Performance of modern sample preparation techniques in the analysis of Fusarium mycotoxins in cereals. , 1998, Journal of chromatography. A.

[102]  H. S. Hussein,et al.  Toxicity, metabolism, and impact of mycotoxins on humans and animals. , 2001, Toxicology.

[103]  M. Morgan,et al.  Development of surface plasmon resonance-based immunoassay for aflatoxin B(1). , 2000, Journal of agricultural and food chemistry.

[104]  Y. Ueno Trichothecenes : chemical, biological, and toxicological aspects , 1983 .

[105]  M. Spanjer,et al.  LC–MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs , 2008, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[106]  J. Bennett,et al.  Mycotoxins, mycotoxicoses, mycotoxicology andMycopathologia , 1987, Mycopathologia.

[107]  N. Magan,et al.  Volatiles as an indicator of fungal activity and differentiation between species, and the potential use of electronic nose technology for early detection of grain spoilage. , 2000, Journal of stored products research.

[108]  V. Lattanzio,et al.  Multiplex dipstick immunoassay for semi-quantitative determination of Fusarium mycotoxins in cereals. , 2012, Analytica chimica acta.

[109]  J. Pitt,et al.  Toxigenic fungi: which are important? , 2000, Medical mycology.

[110]  M. Busman,et al.  Rapid and advanced tools for mycotoxin analysis: a review , 2010, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[111]  S. Eremin,et al.  Fluorescence polarization immunoassay based on a monoclonal antibody for the detection of ochratoxin A , 2004 .

[112]  A. Dobson,et al.  Mycotoxin production by Aspergillus, Fusarium and Penicillium species. , 1998, International journal of food microbiology.

[113]  T. Whitaker,et al.  Sampling uncertainties for the detection of chemical agents in complex food matrices. , 2005, Journal of food protection.

[114]  Jeanette M Van Emon Bioanalytical methods for food contaminant analysis. , 2010, Journal of AOAC International.

[115]  F. Chen,et al.  Natural Occurrence, Analysis, and Prevention of Mycotoxins in Fruits and their Processed Products , 2014, Critical reviews in food science and nutrition.

[116]  J. Pestka,et al.  Analysis of zearalenone in cereal and Swine feed samples using an automated flow-through immunosensor. , 2005, Journal of agricultural and food chemistry.

[117]  Giuseppe Vasapollo,et al.  Molecularly Imprinted Polymers: Present and Future Prospective , 2011, International journal of molecular sciences.

[118]  W. Kong,et al.  Analysis of zearalenone and α-zearalenol in 100 foods and medicinal plants determined by HPLC-FLD and positive confirmation by LC-MS-MS. , 2013, Journal of the science of food and agriculture.

[119]  Heidi R. C. Dietrich,et al.  Biosensors and multiple mycotoxin analysis , 2003 .

[120]  Chris M. Maragos,et al.  Capillary electrophoresis with laser-induced fluorescence : Method for the mycotoxin ochratoxin A , 1998 .

[121]  Dojin Ryu,et al.  Worldwide Occurrence of Mycotoxins in Cereals and Cereal-Derived Food Products: Public Health Perspectives of Their Co-occurrence. , 2017, Journal of agricultural and food chemistry.

[122]  Ž. Jurjević,et al.  Biology and ecology of mycotoxigenic Aspergillus species as related to economic and health concerns. , 2002, Advances in experimental medicine and biology.

[123]  I. K. Cigić,et al.  An Overview of Conventional and Emerging Analytical Methods for the Determination of Mycotoxins , 2009, International journal of molecular sciences.

[124]  Rudolf Krska,et al.  A rapid lateral flow test for the determination of total type B fumonisins in maize , 2009, Analytical and bioanalytical chemistry.

[125]  V. Adam,et al.  Deoxynivalenol and its toxicity , 2010, Interdisciplinary toxicology.

[126]  S. de Saeger,et al.  Development and validation of a QuEChERS based liquid chromatography tandem mass spectrometry method for the determination of multiple mycotoxins in spices. , 2013, Journal of chromatography. A.

[127]  Jiujiang Yu,et al.  Toxins of filamentous fungi. , 2002, Chemical immunology.

[128]  Juan Sun,et al.  QuEChERS Purification Combined with Ultrahigh-Performance Liquid Chromatography Tandem Mass Spectrometry for Simultaneous Quantification of 25 Mycotoxins in Cereals , 2016, Toxins.

[129]  L. Stanker,et al.  Recent studies on immunoassays for mycotoxins. , 1996 .

[130]  M. Harrison PRESENCE AND STABILITY OF PATULIN IN APPLE PRODUCTS: A REVIEW , 1988 .

[131]  A. De Girolamo,et al.  Rapid and non-invasive analysis of deoxynivalenol in durum and common wheat by Fourier-Transform Near Infrared (FT-NIR) spectroscopy , 2009, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[132]  P. Mottier,et al.  Combining the quick, easy, cheap, effective, rugged and safe approach and clean-up by immunoaffinity column for the analysis of 15 mycotoxins by isotope dilution liquid chromatography tandem mass spectrometry. , 2014, Journal of chromatography. A.

[133]  P. Battilani,et al.  Field control of Fusarium ear rot, Ostrinia nubilalis (Hübner), and fumonisins in maize kernels. , 2011, Pest management science.

[134]  S. Saeger,et al.  Immunochemical methods for rapid mycotoxin detection: Evolution from single to multiple analyte screening: A review , 2007, Food additives and contaminants.

[135]  S. Jinap,et al.  SIMULTANEOUS DETERMINATION OF AFLATOXINS, OCHRATOXIN A, AND ZEARALENONE IN CEREALS USING A VALIDATED RP-HPLC METHOD AND PHRED DERIVATIZATION SYSTEM , 2013 .

[136]  B. Nicolaï,et al.  Influence of storage conditions of apples on growth and patulin production by Penicillium expansum. , 2007, International journal of food microbiology.

[137]  P. Nelson,et al.  Analysis of naturally occurring mycotoxins in feedstuffs and food. , 1993, Journal of animal science.

[138]  G. Shephard Determination of mycotoxins in human foods. , 2008, Chemical Society reviews.

[139]  G. Perrone,et al.  Toxigenic Fusarium species and Mycotoxins Associated with Head Blight in Small-Grain Cereals in Europe , 2002, European Journal of Plant Pathology.

[140]  R. Göbel,et al.  Simultaneous determination of aflatoxins, ochratoxin A, and zearalenone in grains by new immunoaffinity column/liquid chromatography. , 2004, Journal of AOAC International.

[141]  R. Romero-González,et al.  Simultaneous determination of pesticides, biopesticides and mycotoxins in organic products applying a quick, easy, cheap, effective, rugged and safe extraction procedure and ultra-high performance liquid chromatography-tandem mass spectrometry. , 2011, Journal of chromatography. A.

[142]  P. Galtier,et al.  Biosynthesis and Toxicological Effects of Patulin , 2010, Toxins.

[143]  L. Bingle,et al.  Comparative Ochratoxin Toxicity: A Review of the Available Data , 2015, Toxins.

[144]  J. Miller,et al.  Analysis of wheat extracts for ochratoxin A by molecularly imprinted solid-phase extraction and pulsed elution , 2004, Analytical and bioanalytical chemistry.

[145]  J. Pestka Deoxynivalenol: Toxicity, mechanisms and animal health risks , 2007 .