بهینه سازی فعالیت کنترل زیستی جدایه های مخمرهای کم تخمیر در برابر آسپرژیلوس نایجر به منظور سم زدایی از آب انگور
محورهای موضوعی : میکروب شناسی غذاییآرش بابایی 1 , محسن دوزبخشان 2 , حدیث طوافی 3
1 - استادیار، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه ملایر، ملایر، ایران
2 - مربی، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه ملایر، ملایر، ایران.
3 - استادیار، گروه زیست شناسی، دانشکده علوم پایه، دانشگاه ملایر، ملایر، ایران
کلید واژه: آسپرژیلوس نایجر, آب انگور, کنترل زیستی, اوخراتوکسین A,
چکیده مقاله :
سابقه و هدف: انگورهای آلوده به آسپرژیلوس می‌توانند موجب تولید اوخراتوکسین A در فراوری نوشیدنی‌هایی مانند شراب یا آب انگور شوند. هدف از این پژوهش ارزیابی توانایی زیستی ضد قارچی دو جدایه بومی مخمری با میزان تخمیر کم (A01 و G01) و سه سویه استاندارد ساکارومیسس سرویزیه، کاندیدا گیلرموندی، مچنیکوویا آگاوس علیه آسپرژیلوس نایجر و توانایی آن در حذف اوخراتوکسین A در انگور و محصولات آن بدون تولید الکل در حین فرایند بود.مواد و روش‌ها: دو جدایه مخمری(A01 و G01) بومی به ترتیب از سیب‌ها و انگورهای بومی منطقه جداسازی شدند و بر روی محیط PDA کشت داده شدند. جدایه‌های بومی با روش تعیین توالی نواحی D1 و D2 و همچنین ITS1 و ITS2 ازDNA ریبوزومی شناسایی شدند.یافته‌ها: نتایج حاصل از تعیین توالی ژنوم مربوط به دو جدایه بومی نشان داد که هر دو توالی مربوط به ساکارومیسس می‌باشند. تمامی سویه‌ها توانایی قابل توجهی در ممانعت از رشد آسپرژیلوس نایجر بر روی دانه‌های انگور و نیز محیط کشت از خود نشان دادند. از طرف دیگر میزان تولید الکل مخمرها نیز بسیار ناچیز بود. نتیجه گیری: کنترل زیستی آسپرژیلوس نایجر و گندزدایی اوخراتوکسین A با استفاده از مخمرهای کم تخمیر پیشنهاد دهنده روشی مطابق موازین اسلامی و نوشیدنی‌های حلال می‌باشد.
Background & Objectives: Grapes infected with Aspergillus can produce ochratoxin A (OTA) in the processing of beverages such as wine or grape juice. This study aimed to evaluate the antifungal biological potential of two low-fermenting native yeast isolates (A01 G01) and three standards (Saccharomyces cerevisiae, Candida guilliermondii, Metschnikowia agaves) yeast isolates against Aspergillus niger and their ability to remove OTA in grape juice and its products, without any considerable alcohol production during the process. Material & Methods: Two native yeast isolates (A01 and G01) were obtained from Malayer apples and grapes, respectively, and inoculated on the PDA culture medium. Native isolates were identified by sequencing D1 and D2 and ITS 1 and ITS2 regions of the ribosomal DNA gene.Results: The results of DNA sequencing identified both native isolates as Saccharomyces. All strains showed a significant ability in inhibition of A. niger growth both on grape berries and in culture media. Meanwhile, yeast isolates produced a trace amount of alcohol.Conclusion: Biological control of A. niger and OTA-decontamination using yeast is proposed as an approach to meet the Islamic dietary laws regarding the absence of alcohol in halal beverages.
table grapes using Bacillus subtilis. Food Control. 2014; 35(1): 41-48.
2. Abeer H, Abd-Allah E, Al-Obeed R, Mridha M, Al-Huqail AA. Non-chemical strategies to
control postharvest losses and extend the shelf life of table grape fruits. J Biol Agr Horticulture.
2013; 29(2): 82-90.
3. Romanazzi G, Smilanick JL, Feliziani E, Droby S. Integrated management of postharvest gray
mold on fruit crops. Postharvest Biol Technol. 2016; 113: 69-76.
4. Türkel S, Korukluoğlu M, Yavuz M. Biocontrol activity of the local strain of Metschnikowia
pulcherrima on different postharvest pathogens. Biotech Res Int. 2014; 24: 123-134.
5. Khlangwiset P, Shephard GS, Wu F. Aflatoxins and growth impairment: a review.
Crit Rev Toxicol. 2011; 41(9): 740-755.
6. Nally MC, Pesce VM, Maturano YP, Toro ME, Combina M, de Figueroa LC, Vazqueza F.
Biocontrol of fungi isolated from sour rot infected table grapes by Saccharomyces and other
yeast species. Postharvest Biol Technol. 2013; 86: 456-462.
7. Battilani P, Pietri A, Logrieco A. Risk assessment and management in practice: ochratoxin in
grapes and wine. Int Food Technol. 2004: 244.
8. Battilani P, Logrieco A, Giorni P, Cozzi G, Bertuzzi T, Pietri A. Ochratoxin A production by
Aspergillus carbonarius on some grape varieties grown in Italy. J Sci Food Agr. 2004; 84(13):
1736-1740.
9. Rodriguez H, Reveron I, Doria F, Costantini A, De Las Rivas B, Munoz R, Garcia-Moruno E.
Degradation of ochratoxin A by Brevibacterium species. J Agric Food Chem. 2011; 59(19):
10755-10760.
10. Gallo A, Knox BP, Bruno KS, Solfrizzo M, Baker SE, Perrone G. Identification and
characterization of the polyketide synthase involved in ochratoxin A biosynthesis in
Aspergillus carbonarius. Int J Food Microbiol. 2014; 179: 10-17.
11. Fiori S, Urgeghe PP, Hammami W, Razzu S, Jaoua S, Migheli Q. Biocontrol activity of four
non-and low-fermenting yeast strains against Aspergillus carbonarius and their ability to
remove ochratoxin A from grape juice. Int J Food Microbiol. 2014; 189: 45-50.
12. Espinel-Ingroff A. Mechanisms of resistance to antifungal agents: yeasts and filamentous
fungi. Rev Iberoam Micol. 2008; 25(2): 101.
13. Szmedra P. Banning 2, 4-D and the phenoxy herbicides: potential economic impact. Weed
Sci. 1997: 592-598.
14. Fisher MC, Hawkins NJ, Sanglard D, Gurr SJ. Worldwide emergence of resistance to
antifungal drugs challenges human health and food security. Sci. 2018; 360(6390): 739-742.
15. Droby S. Mode of action of biocontrol agents of postharvest diseases. Bio Ctrl Post Dis
Theo Prac. 1994.
16. Droby S, Wisniewski M, Macarisin D, Wilson C. Twenty years of postharvest biocontrol
research: is it time for a new paradigm? Post Bio Techno. 2009; 52(2): 137-145.
17. Meng XH, Qin GZ, Tian SP. Influences of preharvest spraying Cryptococcus laurentii
combined with postharvest chitosan coating on postharvest diseases and quality of table grapes
in storage. LWT Food Sci Technol. 2010; 43(4): 596-601.
18. Petruzzi L, Sinigaglia M, Corbo MR, Beneduce L, Bevilacqua A. Ochratoxin A removal by
Saccharomyces cerevisiae strains: effect of wine‐related physicochemical factors. J Sci Food
Agric. 2013; 93(9): 2110-2115.
19. Cubaiu L, Abbas H, Dobson AD, Budroni M, Migheli Q. A Saccharomyces cerevisiae wine
strain inhibits growth and decreases ochratoxin A biosynthesis by Aspergillus carbonarius and
Aspergillus ochraceus. Toxins. 2012; 4(12): 1468-1481.
20. Armando MR, Dogi CA, Poloni V, Rosa C, Dalcero AM, Cavaglieri LR. In vitro study on
the effect of Saccharomyces cerevisiae strains on growth and mycotoxin production by
Aspergillus carbonarius and Fusarium graminearum. Int J Food Microbiol. 2013; 161(3):
182-188.
21. Pfliegler WP, Pusztahelyi T, Pócsi I. Mycotoxins–prevention and decontamination by yeasts.
J Basic Microbiol. 2015; 55(7): 805-818.
22. Regenstein J, Chaudry M, Regenstein C. The kosher and halal food laws. Comp Rev Food
Sci Food Saf. 2003; 2(3): 111-127.
23. Zailani S. Halal traceability and halal tracking systems in strengthening halal food supply
chain for food industry in Malaysia. J Food Technol. 2010; 8(3): 74-81.
24. Riaz MN, Chaudry MM. Halal food production: CRC press; 2003.
25. Najiha AA, Tajul A, Norziah M, Wan Nadiah W. A preliminary study on halal limits for
ethanol content in food products. Middle East J Sci Res. 2010; 6(1): 45-50.
26. Alzeer J, Hadeed KA. Ethanol and its Halal status in food industries. Trends Food Sci.
Technol. 2016; 58: 14-20.
27. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: CSHL press;
1989.
28. Heras‐Vazquez L, Javier F, Mingorance‐Cazorla L, Clemente‐Jimenez JM, Rodriguez‐Vico
F. Identification of yeast species from orange fruit and juice by RFLP and sequence analysis of
the 5.8 S rRNA gene and the two internal transcribed spacers. FEMS Yeast Res. 2003; 3(1): 3-9.
29. Jolly N, Augustyn O, Pretorius I. The occurrence of non-Saccharomyces cerevisiae yeast
species over three vintages in four vineyards and grape musts from four production regions of
the Western Cape, South Africa. South Afr J Enol Viticulture. 2017; 24(2): 35-42.
30. Zhao YC, Bassiouni A, Tanjararak K, Vreugde S, Wormald PJ, Psaltis AJ. Role of fungi in
chronic rhinosinusitis through ITS sequencing. Laryngoscope. 2018; 128(1): 16-22.
31. Dakal TC, Giudici P, Solieri L. Contrasting patterns of rDNA homogenization within the
Zygosaccharomyces rouxii species complex. PloS one. 2016; 11(8): e0160744.
32. Takemoto A, Cho O, Morohoshi Y, Sugita T, Muto M. Molecular characterization of the
skin fungal microbiome in patients with psoriasis. Int J Womens Dermatol. 2015; 42(2):
166-170.
33. Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, Chen W. Nuclear
ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for
Fungi. Proc Natl Acad Sci USA. 2012; 109(16): 6241-6246.
34. Hodgeson J, Baumgardner R, Martin B, Rehme K. Stoichiometry in the neutral iodometric
procedure for ozone by gas-phase titration with nitric oxide. Anal Chem. 1971; 43(8):
1123-1126.
35. Aguera E, Bes M, Roy A, Camarasa C, Sablayrolles J-M. Partial removal of ethanol during
fermentation to obtain reduced-alcohol wines. Am J Enol Viticulture. 2010; 61(1): 53-60.
36. Farbo MG, Urgeghe PP, Fiori S, Marceddu S, Jaoua S, Migheli Q. Adsorption of ochratoxin
A from grape juice by yeast cells immobilised in calcium alginate beads. Int J Food Microbiol.
2016; 217: 29-34.
37. Schalchli H, Hormazabal E, Becerra J, Birkett M, Alvear M, Vidal J, Quiroz A. Antifungal
activity of volatile metabolites emitted by mycelial cultures of saprophytic fungi. J Chem Ecol.
2011; 27(6): 503-513.
38. Crous PW, Coertze S, Swart L. The effect of hot-water treatment on fungi occurring in
apparently healthy grapevine cuttings. Phytopathologia Mediterranea. 2001; 40(3): 1000-1003.
39. Raspor P, Miklič-Milek D, Avbelj M, Čadež N. Biocontrol of grey mould disease on grape
caused by botrytis cinerea with autochthonous wine yeasts. Food Technol Biotechnol. 2010; 48
(3): 336-343.
40. Gholampour-Azizi E. Evaluation of the presence of ochratoxin in grape juices and raisins. J
Food Technol Nutr. 2012; 1(33): 95-100.
41. Al‐Mazeedi HM, Regenstein JM, Riaz MN. The issue of undeclared ingredients in halal and
kosher food production: A focus on processing aids. Comp Rev Food Sci Food Saf. 2013; 12(2):
228-233.
42. Martínez-Rodríguez AJ, Carrascosa AV. HACCP to control microbial safety hazards during
winemaking: Ochratoxin A. Food Control. 2009; 20(5): 469-475.
43. Bizaj E, Mavri J, Čuš F, Raspor A. Removal of ochratoxin A in Saccharomyces cerevisiae
liquid cultures. South Afr J Enol Viticulture. 2016; 30(2): 151-155.
44. Droby S, Chalutz E, Wilson C, Wisniewski M. Characterization of the biocontrol activity of
Debaryomyces hansenii in the control of Penicillium digitatum on grapefruit. Can J Microbiol.
1989; 35(8): 794-800
_||_
table grapes using Bacillus subtilis. Food Control. 2014; 35(1): 41-48.
2. Abeer H, Abd-Allah E, Al-Obeed R, Mridha M, Al-Huqail AA. Non-chemical strategies to
control postharvest losses and extend the shelf life of table grape fruits. J Biol Agr Horticulture.
2013; 29(2): 82-90.
3. Romanazzi G, Smilanick JL, Feliziani E, Droby S. Integrated management of postharvest gray
mold on fruit crops. Postharvest Biol Technol. 2016; 113: 69-76.
4. Türkel S, Korukluoğlu M, Yavuz M. Biocontrol activity of the local strain of Metschnikowia
pulcherrima on different postharvest pathogens. Biotech Res Int. 2014; 24: 123-134.
5. Khlangwiset P, Shephard GS, Wu F. Aflatoxins and growth impairment: a review.
Crit Rev Toxicol. 2011; 41(9): 740-755.
6. Nally MC, Pesce VM, Maturano YP, Toro ME, Combina M, de Figueroa LC, Vazqueza F.
Biocontrol of fungi isolated from sour rot infected table grapes by Saccharomyces and other
yeast species. Postharvest Biol Technol. 2013; 86: 456-462.
7. Battilani P, Pietri A, Logrieco A. Risk assessment and management in practice: ochratoxin in
grapes and wine. Int Food Technol. 2004: 244.
8. Battilani P, Logrieco A, Giorni P, Cozzi G, Bertuzzi T, Pietri A. Ochratoxin A production by
Aspergillus carbonarius on some grape varieties grown in Italy. J Sci Food Agr. 2004; 84(13):
1736-1740.
9. Rodriguez H, Reveron I, Doria F, Costantini A, De Las Rivas B, Munoz R, Garcia-Moruno E.
Degradation of ochratoxin A by Brevibacterium species. J Agric Food Chem. 2011; 59(19):
10755-10760.
10. Gallo A, Knox BP, Bruno KS, Solfrizzo M, Baker SE, Perrone G. Identification and
characterization of the polyketide synthase involved in ochratoxin A biosynthesis in
Aspergillus carbonarius. Int J Food Microbiol. 2014; 179: 10-17.
11. Fiori S, Urgeghe PP, Hammami W, Razzu S, Jaoua S, Migheli Q. Biocontrol activity of four
non-and low-fermenting yeast strains against Aspergillus carbonarius and their ability to
remove ochratoxin A from grape juice. Int J Food Microbiol. 2014; 189: 45-50.
12. Espinel-Ingroff A. Mechanisms of resistance to antifungal agents: yeasts and filamentous
fungi. Rev Iberoam Micol. 2008; 25(2): 101.
13. Szmedra P. Banning 2, 4-D and the phenoxy herbicides: potential economic impact. Weed
Sci. 1997: 592-598.
14. Fisher MC, Hawkins NJ, Sanglard D, Gurr SJ. Worldwide emergence of resistance to
antifungal drugs challenges human health and food security. Sci. 2018; 360(6390): 739-742.
15. Droby S. Mode of action of biocontrol agents of postharvest diseases. Bio Ctrl Post Dis
Theo Prac. 1994.
16. Droby S, Wisniewski M, Macarisin D, Wilson C. Twenty years of postharvest biocontrol
research: is it time for a new paradigm? Post Bio Techno. 2009; 52(2): 137-145.
17. Meng XH, Qin GZ, Tian SP. Influences of preharvest spraying Cryptococcus laurentii
combined with postharvest chitosan coating on postharvest diseases and quality of table grapes
in storage. LWT Food Sci Technol. 2010; 43(4): 596-601.
18. Petruzzi L, Sinigaglia M, Corbo MR, Beneduce L, Bevilacqua A. Ochratoxin A removal by
Saccharomyces cerevisiae strains: effect of wine‐related physicochemical factors. J Sci Food
Agric. 2013; 93(9): 2110-2115.
19. Cubaiu L, Abbas H, Dobson AD, Budroni M, Migheli Q. A Saccharomyces cerevisiae wine
strain inhibits growth and decreases ochratoxin A biosynthesis by Aspergillus carbonarius and
Aspergillus ochraceus. Toxins. 2012; 4(12): 1468-1481.
20. Armando MR, Dogi CA, Poloni V, Rosa C, Dalcero AM, Cavaglieri LR. In vitro study on
the effect of Saccharomyces cerevisiae strains on growth and mycotoxin production by
Aspergillus carbonarius and Fusarium graminearum. Int J Food Microbiol. 2013; 161(3):
182-188.
21. Pfliegler WP, Pusztahelyi T, Pócsi I. Mycotoxins–prevention and decontamination by yeasts.
J Basic Microbiol. 2015; 55(7): 805-818.
22. Regenstein J, Chaudry M, Regenstein C. The kosher and halal food laws. Comp Rev Food
Sci Food Saf. 2003; 2(3): 111-127.
23. Zailani S. Halal traceability and halal tracking systems in strengthening halal food supply
chain for food industry in Malaysia. J Food Technol. 2010; 8(3): 74-81.
24. Riaz MN, Chaudry MM. Halal food production: CRC press; 2003.
25. Najiha AA, Tajul A, Norziah M, Wan Nadiah W. A preliminary study on halal limits for
ethanol content in food products. Middle East J Sci Res. 2010; 6(1): 45-50.
26. Alzeer J, Hadeed KA. Ethanol and its Halal status in food industries. Trends Food Sci.
Technol. 2016; 58: 14-20.
27. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual: CSHL press;
1989.
28. Heras‐Vazquez L, Javier F, Mingorance‐Cazorla L, Clemente‐Jimenez JM, Rodriguez‐Vico
F. Identification of yeast species from orange fruit and juice by RFLP and sequence analysis of
the 5.8 S rRNA gene and the two internal transcribed spacers. FEMS Yeast Res. 2003; 3(1): 3-9.
29. Jolly N, Augustyn O, Pretorius I. The occurrence of non-Saccharomyces cerevisiae yeast
species over three vintages in four vineyards and grape musts from four production regions of
the Western Cape, South Africa. South Afr J Enol Viticulture. 2017; 24(2): 35-42.
30. Zhao YC, Bassiouni A, Tanjararak K, Vreugde S, Wormald PJ, Psaltis AJ. Role of fungi in
chronic rhinosinusitis through ITS sequencing. Laryngoscope. 2018; 128(1): 16-22.
31. Dakal TC, Giudici P, Solieri L. Contrasting patterns of rDNA homogenization within the
Zygosaccharomyces rouxii species complex. PloS one. 2016; 11(8): e0160744.
32. Takemoto A, Cho O, Morohoshi Y, Sugita T, Muto M. Molecular characterization of the
skin fungal microbiome in patients with psoriasis. Int J Womens Dermatol. 2015; 42(2):
166-170.
33. Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, Chen W. Nuclear
ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for
Fungi. Proc Natl Acad Sci USA. 2012; 109(16): 6241-6246.
34. Hodgeson J, Baumgardner R, Martin B, Rehme K. Stoichiometry in the neutral iodometric
procedure for ozone by gas-phase titration with nitric oxide. Anal Chem. 1971; 43(8):
1123-1126.
35. Aguera E, Bes M, Roy A, Camarasa C, Sablayrolles J-M. Partial removal of ethanol during
fermentation to obtain reduced-alcohol wines. Am J Enol Viticulture. 2010; 61(1): 53-60.
36. Farbo MG, Urgeghe PP, Fiori S, Marceddu S, Jaoua S, Migheli Q. Adsorption of ochratoxin
A from grape juice by yeast cells immobilised in calcium alginate beads. Int J Food Microbiol.
2016; 217: 29-34.
37. Schalchli H, Hormazabal E, Becerra J, Birkett M, Alvear M, Vidal J, Quiroz A. Antifungal
activity of volatile metabolites emitted by mycelial cultures of saprophytic fungi. J Chem Ecol.
2011; 27(6): 503-513.
38. Crous PW, Coertze S, Swart L. The effect of hot-water treatment on fungi occurring in
apparently healthy grapevine cuttings. Phytopathologia Mediterranea. 2001; 40(3): 1000-1003.
39. Raspor P, Miklič-Milek D, Avbelj M, Čadež N. Biocontrol of grey mould disease on grape
caused by botrytis cinerea with autochthonous wine yeasts. Food Technol Biotechnol. 2010; 48
(3): 336-343.
40. Gholampour-Azizi E. Evaluation of the presence of ochratoxin in grape juices and raisins. J
Food Technol Nutr. 2012; 1(33): 95-100.
41. Al‐Mazeedi HM, Regenstein JM, Riaz MN. The issue of undeclared ingredients in halal and
kosher food production: A focus on processing aids. Comp Rev Food Sci Food Saf. 2013; 12(2):
228-233.
42. Martínez-Rodríguez AJ, Carrascosa AV. HACCP to control microbial safety hazards during
winemaking: Ochratoxin A. Food Control. 2009; 20(5): 469-475.
43. Bizaj E, Mavri J, Čuš F, Raspor A. Removal of ochratoxin A in Saccharomyces cerevisiae
liquid cultures. South Afr J Enol Viticulture. 2016; 30(2): 151-155.
44. Droby S, Chalutz E, Wilson C, Wisniewski M. Characterization of the biocontrol activity of
Debaryomyces hansenii in the control of Penicillium digitatum on grapefruit. Can J Microbiol.
1989; 35(8): 794-800
