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Mycotoxin and brewing technology (APEH Daniel O.)


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Mycotoxin and brewing technology (APEH Daniel O.)

  2. 2. INTRODUCTION Beer refers to beverages resulting from the germination and fermentation of starches mainly derived from cereal (Gutcho, 1976). The third most popular drink overall after water and tea ( Africa is the third largest beer consuming continent (WHO, 2004). Nigeria is among the first 30 top world consumers (FAO, 2003; WHO, 2004) High demand for beer across various continents equals high risk of consumption of beer associated toxins (Karolína et al., 2012). Mycotoxins are major cereal contaminants (Reviewed in Makun, et al., 2009; Chandrashekar et al., 2000). Processed cereals in form of beer may be contaminated with mycotoxins.
  3. 3. MYCOTOXINS Over 300 fungi produced toxic compounds that contaminate a wide variety of agricultural commodities (Nielson and Smedsgaard, 2003) Produced as secondary metabolites (Pitt, 2000). At Pre-harvest, Post-harvest or During storage (Reviewed in Kumar et al., 2008) Ingestion cause a range of toxic responses, from acute to chronic health disorders (Roger, 1993) Affect trade Major producers Aspergillus species Penicillium species Fusarium species
  4. 4. Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Aflatoxin M1 Zearalenone Fumonisin Patulin Citrulin Ergot AlkaloidFIGURE 1: STRUCTURAL REPRESENTATION OF SOME MYCOTOXINS
  5. 5. HEALTH IMPACT OF MYCOTOXINS Diseases resulting from mycotoxin exposure are called mycotoxicosisThey elicit short or long term impacts(Wayne, 2007)  Inhibition of metabolic pathways  Impairment of growth and development  Immunosupression  Carcinogenicity, mutagenicity and teratogenicity T-2 toxin was responsible for the deaths of several Russians from 1942-1947 (Gao and Yoshizawa, 1997). Aflatoxin intake relates to high incidence of liver cancer is S.A, Kenya etc. (Fabio, 1999); Aflatoxicosis
  6. 6. ECONOMIC IMPACT OF MYCOTOXINS Mycotoxin contamination affects up to 25% of global food and feed (FAO, 2009).  Crop losses and Reduced animal productivity Costs from improving technologies for production, storage and transport of crops  Member states of the African Groundnut Council—The Gambia, Mali, Niger, Nigeria, Senegal, and Sudan—have calculated the annual cost of implementing a program to reduce aflatoxin contamination at US$7.5 million (Atanda, 2011). The cost of analytical testing, especially as regulations become more stringent  EU regulation on aflatoxins cost Africa US$750 million each year in exports of cereals, dried fruit and nuts (World bank study; retrieved from
  7. 7. BREWING TECHNOLOGY Refers to the processes involved in beer production (Ertan Anli and Mert, 2010) The most commonly used cereal grain is barley; others are wheat, maize (corn), sorghum and rice (Gutcho, 1976).
  8. 8. Grain (Barley, Sorghum, Wheat) Moisture content Microorganisms Malting (3 stages) Gibberellic Acid Hydrolytic Enzymes (Gutcho, 1976).Adjunct Malt(Rice, Wheat, Ground malt + AdjunctsMaize, Barley Amylases and proteases hydrolysis & extractione.t.c) Mashing Increase in particle size (Milling) Separation Wort is boiled sometimes with hopsPre-treatment Boiling; Concentrate and sterilize the wort Inactivate enzymes Boiling Coagulate and precipitate protein Hop Caramelize the sugar slightly in order Cooling to develop the color of the beerSyrups Hops contribute antiseptic compoundsSugars Wort Yeast is inoculated in Wort Fermentation Yeast converts sugar into organic compounds Yeast C6H12O6+2Pi+2ADP 2C2H5OH+2CO2+ 2ATP Glucose Ethanol + 2H2O Maturation BEER Lactic acid bacteria is inoculated in Wort Results in souring (Lerio, 1993)FIGURE 2: GENERAL CHART OF THE BREWING PROCESS
  9. 9. MYCOTOXIN AND BREWING Some mycotoxins seems to survive major beer production processes namely malting, mashing, boiling and fermentation into beer (Scott, 1996) Mycotoxins in beer could either be developed or detoxified at various brewing processes (Mably et al., 2005)
  10. 10. Grain (Sorghum, Wheat) Steeping resulted in about 45% Malting AFL lost (Yahl, 1971; Romer, 1984) MaltAdjunct Adjunct is a source of AFL(Rice, Wheat, Maize, B Mashingarley e.t.c) contamination (Karolína et al., 2012) Separation Pre-treatment At 100°C and 250°C for 30min Boiling showed 10.4% and 99% reduction Hop in AFB1 (Oluwafemi and Ikeowa, 2005) CoolingSyrupsSugars Wort For 72hrs showed 50% reduction of AFB1 (Yuan et al., 2008) Yeast Fermentation Lactic acid fermentations lead to Maturation opening of the lactone ring (Nout 1994). BEERUpto 89% AFB1 was removed during Lager beer processing (Oluwafemi, 2004) FIGURE 3: AFLATOXIN AND BREWING
  11. 11. Grain (Sorghum, Wheat) Almost completely lost Malting ( Krogh et al., 1974). Malt Adjunct (Rice, Wheat, Maize, B Mashing 40–89% was lost (Sylvie et arley e.t.c) al., 2011) Separation Pre-treatment Boiling Most probably due to Hop proteolytic degradation Cooling (Ertan Mert, 2010).Syrups WortSugars Fermentation Destroyed in the fermentation Yeast process (Kostecki et al., 1991; Maturation SCOOP, 2010) BEERHigher in the ‘‘non-alcoholic’’ beer (Tangni and Lanrondelle, 2003)Upto 70% of the OTA was degraded in the production steps beer(Nip et al., 1975 FIGURE 4: OCHRATOXIN AND BREWING
  12. 12. Grain (Sorghum, Wheat) Increase in Fusarium Malting spp, and fumonisin Malt Brewing adjuncts e.gAdjunct (Rice, Wheat,Maize, Barley e.t.c) Mashing corn contributes fumonisin (Scott and Lawrence, 1995). Separation Pre-treatment Boiling Hop Heat-stable (Alberts et al., 1990) CoolingSyrups Wort Level change notSugars Fermentation significant (Scott et al., 1993). Yeast Maturation BEERLevels above maximum limit has been reported in severel countries(Drager, 1996; Mbugua and Gathumbi) e.g 150±24 ng/ml in Cameroonian beer(Roger, 2011).Gushing increases with fumonisin in beer (Casey,1996;). FIGURE 5: FUMONISIN AND BREWING
  13. 13. Grain (Sorghum, Wheat) Increased by 18–114% of Malting that present on the Malt original barley in 5 dayAdjunct malts, (Scott, 1996; Schwarz et(Rice, Wheat, Maize, Ba Mashing al., 2005, Lancova et al., 2008)rley e.t.c) Separation Pre-treatment Boiling Stable after 7-9 days of Hop alcoholic fermentation with S. CoolingSyrups cerevisiae (Scott, 1992) Wort Detected at high levels in bothSugars Fermentation the solid residue and the Yeast fermented liquid (Bennet and Maturation Richard 1996). BEERTrichothecenes are stable to brewing process; occurring in commercialbeer in many countries (Shim et al., 1997; Molto et al., 2000; Baxter et al., 2006 ).After brewing, 80–93% of DON present on the malt grist was detected inthe beer (Schwarz et al., 2005). FIGURE 6: DEOXYNIVALENOL (DON) AND BREWING
  14. 14. TABLE 2 : YEAST AND/OR YEAST CELL WALL MATERIAL AS POTENTIAL MYCOTOXIN DECONTAMINATING AGENTSAGENT Mycotoxin bound Level of Binding ReferenceS. cerevisiae AflatoxinB1 > 60% Shetty and Jespersen, 2006; Devegowda et al., 1996Yeast from West AflatoxinB1 >60% Shetty and Jespersen,African maize 2006Cansida crusei AflatoxinB1 15- 39% Shetty and Jespersen, 2006Mannan- Ochratoxins & High binding Raju & Devegowda,oligosaccharides Zearalenone 2000; Devegowda et DON&FumonisinB1 Little binding al., 1996Modified mannan- AflatoxinB1 77% Devegowda et al., 1996oligosaccharides fromthe S. cerevisiaeMixture of 40% (w/w) Ochratoxin A High binding Grunkemeier, 1990sterilized yeast + 60%(w/w) residue of beerfermentation
  15. 15. TABLE 3 : LACTIC ACID BACTERIA (LAB) AND/OR LAB MATERIAL AS POTENTIAL MYCOTOXIN DECONTAMINATING AGENTSAGENT Mycotoxin bound Level of Reference BindingLactobacillus Aflatoxin B1 40-59% Shetty andplantarum Jespersen, 2006Paralactobacillus Aflatoxin B1 <15% Shetty andserangolensis Jespersen, 2006Lactobacillus Aflatoxin B1 (more than B2, 80% El–Nezami et al.,rhamnosus G1, G2) 1998aLactobacillus Deoxynivalenol, 3- Effective El–Nezami,rhamnosus strains acetyldeoxynivalenol, binding Chrevatidis,LGG and LC 705 nivalenol, fusarenon, Auriola, Salminen, diacetoxyscirpenol, T-2 toxin, & Mykkanen, 2002 and HT-2Lactic Acid Bacteria Aflatoxins Inhibition (Coallier-Ascah & of bio- Idziak, 1985; synthesis Thyagaraja & Hosono, 1994).
  17. 17. Retrieved from www.bioimages.netFIGURE 8: LACTIC ACID BACTERIA CELL WALL COMPONENTS
  19. 19. Dihydrosphingosine N-acyltransferase is one cellulartarget for fumonisin toxicity andcarcinogenicityInhibition of sphingolipidbiosynthesis (Wang et al., 1991).Increases the ratio of sphingoidprecursorsCell deregulationCell death (Riley et al., 1996) TOXICOLOGY Suspected human carcinogen.Toxic to pigs and poultry.Cause equine eucoencepha-lomalacia (ELEM), a fatal diseaseof horsesFIGURE 11: MECHANISM OF ACTION OF FUMONISIN
  20. 20. DON inhibits Translation by1. Interaction with the peptidyl transferase centre on the 605 ribosomal subunit.2. They also prevent release of the polypeptide, by suppressing hydrolysis of the peptidyl—tRNA at termination TOXICOLOGY Feed refusal Reduced weight gain Diarrhea VomitingFIGURE 12: MECHANISM OF ACTION OF DEOXYNIVALENOL
  21. 21. •Disruption Of PhenylalanineMetabolism•Reduced PEPCK atTranslation level•Reduced Glyconeogenesis•Cell DeathFIGURE 10: MECHANISM OF ACTION OF OCHRATOXIN
  22. 22. PREVENTION AND MANAGEMENT OF MYCOTOXIN CONTAMINATION IN COMMODITIES Implementation of GAPs from planting to harvesting Proper storage conditions Biotechnology; Resistant crop varieties (Hell et al., 2011) Coordination between the different stakeholders The establishment of food safety measures needs a legal basis, otherwise no enforcement is possible Public awareness/ University curriculum HACCP and others
  23. 23. REGULATIONS Established in about 100 countries of which 15 are African. Agencies involved; WHO, FAO, WHO-JECFA, EFSA, FSA, SON, ACADEMIC INSTITUTIONS Maximum allowable limits 4 µg/kg aflatoxinB1 for All foods (Nigeria standard)  0.5 µg/kg DON for grain (USA standard)  2-4 µg/kg FumunisinB1 for maize products (EU standard) 5 µg/kg and 3 µg/kg Ochratoxin in cereals and malt respectively (EU standard)
  24. 24. CONCLUSION Brewing technology is a multi-phased process, each phase contributes differently to the final quantity of different mycotoxins that may be found in beer. Grain and/or adjuncts may be source of contamination by fungi and their mycotoxins. Malting increases the risk of fungi and mycotoxin contamination but reduce ochratoxin levels. Fermentums binds mycotoxins on their cell walls. Generally, DON and other tricothecenes are shown to increase across the brewing process, fumonisin is relatively stable while aflatoxin and ochratoxin are generally observed to be reduced across the brewing process