Cyclopiazonic acid (CPA)-producing Penicillium griseofulvum is usually found on the dry-cured ham surface during its ripening. The objective of this work was to evaluate the effect of temperature and water activity (aw) of dry-cured ham processing on growth, CPA production, and temporal relative expression of genes involved in CPA biosynthesis on dry-cured meat-based media. P. griseofulvum CECT 2919 grew faster than P. griseofulvum IBT 14319 in all conditions tested, although no growth occurred at 0.85 aw. Besides, the dry-cured ham-based medium favoured CPA synthesis for both strains compared to the meat-based medium. For the strain CECT 2919, the expression of the mfs-1 and pks-nrps genes were stimulated at 0.90 and 0.95 aw, respectively, while the dmaT gene expression was inhibited during the incubation time. By contrast, the strain IBT 14319 showed that the dmaT gene expression was stimulated at 0.90 aw, while the pks-nrps and mfs-1 genes were repressed throughout incubation time. In conclusion, it is necessary to reduce aw on the surface of the hams below 0.85 during ripening before to increase temperature to reduce growth of P. griseofulvum and CPA production. This information may be useful to design preventive and corrective actions to minimise risks associated with the presence of CPA in dry-cured ham.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Alapont C, López-Mendoza MC, Gil JV, Martínez-Culebras PV (2014) Mycobiota and toxigenic Penicillium species on two Spanish dry-cured ham manufacturing plants. Food Addit Contam Part A 3:93–104. https://doi.org/10.1080/19440049.2013.849007
Ansari P, Häubl G (2016) Determination of cyclopiazonic acid in white mould cheese by liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) using a novel internal standard. Food Chem 221:978–982. https://doi.org/10.1016/j.foodchem.2016.05.063
Astoreca A, Vaamonde G, Dalcero A, Marín S, Ramos A (2014) Abiotic factors and their interactions influence on the co-production of aflatoxin B1 and cyclopiazonic by Aspergillus flavus isolated from corn. Food Microbiol 38:276–283. https://doi.org/10.1016/j.fm.2013.07.012
Bailly JD, Tabuc C, Quérin A, Guerre P (2005) Production and stability of patulin, ochratoxin A, citrinin, and cyclopiazonic acid on dry-cured ham. J Food Prot 68:1516–1520. https://doi.org/10.4315/0362-028X-68.7.1516
Banani H, Marcet-Houben M, Ballester AR, Abbruscato P, González-Candelas L, Gabaldón T, Spadaro D (2016) Genome sequencing and secondary metabolism of the postharvest pathogen Penicillium griseofulvum. BMC Genomics 17(19). https://doi.org/10.1186/s12864-015-2347-x
Baquião AC, Lopes EL, Corrêa B (2016) Molecular and mycotoxigenic biodiversity of Aspergillus flavus isolated from Brazil nuts. Food Res Int 89:266–271. https://doi.org/10.1016/j.foodres.2016.08.005
Berni E, Cacchioli C, Diaferia C, Spotti E (2007) Microbial surface colonization in Nebrodi salame. Proceedings of the 6th International Symposium on the Mediterranean Pig. Messina, Capo d’Orlando, pp 253–257
Berni E, Cacchioli C, Diaferia C (2012) Characterization of surface mycoflora in Nebrodi hams. De Pedro E.J. (ed.), Cabezas A.B. (ed.) 7th International Symposium on the Mediterranean Pig. Zaragoza. CIHEAM (pp. 437–440). (Options Méditerranéennes: Série A. Séminaires Méditerranéens; n. 101)
Chang PK, Ehrlich KC, Fujii I (2009a) Cyclopiazonic acid biosynthesis of Aspergillus flavus and Aspergillus oryzae. Toxins 1:74–99. https://doi.org/10.3390/toxins1020074
Chang PK, Horn BW, Dorner JW (2009b) Clustered genes involved in cyclopiazonic acid production are next to the aflatoxin biosynthesis gene cluster in Aspergillus flavus. Fungal Genet Biol 46:176–182. https://doi.org/10.1016/j.fgb.2008.11.002
Córdoba JJ, Antequera T, García C, Ventanas J, López-Bote C, Asensio MA (1994) Evolution of free amino acids and amines during ripening of Iberian cured ham. J Agric Food Chem 42:2296–2301. https://doi.org/10.1021/jf00046a040
Da Motta S, Valente Soares L (2000) Analytical, nutritional and clinical methods section: simultaneous determination of tenuazonic and cyclopiazonic acids in tomato products. Food Chem 71:111–116. https://doi.org/10.1016/S0308-8146(00)00040-6
Díaz GJ, Thompson W, Martos PA (2010) Stability of cyclopiazonic acid in solution. World Mycotoxin J 3:25–33. https://doi.org/10.3920/WMJ2009.1170
Fernández-Pinto V, Patriarca A, Locani O, Vaamonde G (2001) Natural co-occurrence of aflatoxin and cyclopiazonic acid in peanuts grown in Argentina. Food Addit Contam 18:1017–1020. https://doi.org/10.1080/02652030110057125
Ferrara M, Magistà D, Lippolis V, Cervellieri S, Susca A, Perrone G (2016) Effect of Penicillium nordicum contamination rates on ochratoxin A accumulation in dry-cured salami. Food Control 67:235–239. https://doi.org/10.1016/j.foodcont.2016.03.010
Frisvad JC, Smedsgaard J, Larsen TO, Samson RA (2004) Mycotoxins, drugs and other extrolites produced by species in Penicillium subgenus Penicillium. Stud Mycol 49:201–241
Galvalisi U, Lupo S, Piccini J, Bettucci L (2012) Penicillium species present in Uruguayan salami. Rev Argent Microbiol 44:36–42. https://doi.org/10.1590/S0325-75412012000100008
García D, Ramos AJ, Sanchís V, Marín S (2009) Predicting mycotoxins in foods: a review. Food Microbiol 26:757–769. https://doi.org/10.1016/j.fm.2009.05.014
Gqaleni N, Smith JE, Lacey J (1996) Coproduction of aflatoxins and cyclopiazonic acid in isolates of Aspergillus flavus. Food Addit Contam 13:677–685. https://doi.org/10.1080/02652039609374453
Kato N, Tokuoka M, Shinohara Y, Kawatani M, Uramoto M, Seshime Y, Fujii I, Kitamoto K, Takahashi T, Takahashi S, Koyama Y, Osada H (2011) Genetic safeguard against mycotoxin cyclopiazonic acid production in Aspergillus oryzae. Chem Bio Chem 12:1376–1382. https://doi.org/10.1002/cbic.201000672
Lazzaro I, Susca A, Mula G, Ritieni A, Ferracane R, Marruecos A, Battilani P (2012) Effects of temperature and water activity on FUM2 and FUM21 gene expression and fumonisin B production in Fusarium verticillioides. Eur J Plant Pathol 134:685–695. https://doi.org/10.1007/s10658-012-0045-y
Le Bars J (1979) Cyclopiazonic acid production by Penicillium camemberti Thom and natural occurrence of this mycotoxin in cheese. Appl Environ Microbiol 38:1052–1055
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2 (−ΔΔCT) method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Ludemann V, Pose G, Pollio ML, Segura J (2004) Determination of growth characteristics and lipolytic and proteolytic activities of Penicillium strains isolated from Argentinean salami. Int J Food Microbiol 96:13–18. https://doi.org/10.1016/j.ijfoodmicro.2004.03.003
Medina A, Gilbert MK, Mack BM, O’Brian GR, Rodríguez A, Bhatnagar D, Payne G, Magan N (2017) Interactions between water activity and temperature on the Aspergillus flavus transcriptome and aflatoxin B1 production. Int J Food Microbiol 256: 36–44. https://doi.org/10.1016/j.ijfoodmicro.2017.05.020
Núñez F, Rodríguez MM, Bermúdez E, Córdoba JJ, Asensio MA (1996) Composition and toxigenic potential of the mould population on dry-cured Iberian ham. Int J Food Microbiol 32:185–197. https://doi.org/10.1016/0168-1605(96)01126-9
Núñez F, Westphal CD, Bermúdez E, Asensio MA (2007) Production of secondary metabolites by some terverticillate Penicillia on carbohydrate-rich and meat substrates. J Food Prot 70:2829–2836. https://doi.org/10.4315/0362-028X-70.12.2829
Oliveira CA, Rosmaninho J, Rosim R (2006) Aflatoxin Ml and cyclopiazonic acid in fluid milk traded in São Paulo, Brazil. Food Addit Contam 23:196–201. https://doi.org/10.1080/02652030500398379
Ostry V, Polster M (1989) Detection of cyclopiazonic acid and its producers in food. Vet Med 34:421–430
Peromingo B, Rodríguez A, Bernáldez V, Delgado J, Rodríguez M (2016) Effect of temperature and water activity on growth and aflatoxin production by Aspergillus flavus and Aspergillus parasiticus on cured meat model systems. Meat Sci 122:76–83. https://doi.org/10.1016/j.meatsci.2016.07.024
Peromingo B, Rodríguez M, Delgado J, Andrade MJ, Rodríguez A (2017) Gene expression as a good indicator of aflatoxin contamination in dry-cured ham. Food Microbiol 67:31–40. https://doi.org/10.1016/j.fm.2017.05.008
Peromingo B, Rodríguez M, Núñez F, Silva A, Rodríguez A (2018) Sensitive determination of cyclopiazonic acid in dry-cured ham using a QuEChERS method and uHPLC-MS/MS. Food Chem 263:275–282. https://doi.org/10.1016/j.foodchem.2018.04.126
Peromingo B, Sulyok M, Lemmens M, Rodríguez A, Rodríguez M (2019) Diffusion of mycotoxins and secondary metabolites in dry-cured meat products. Food Control 111:144–150. https://doi.org/10.1016/j.foodcont.2019.02.032
Pleadin J, Perši N, Kovačevic D, Vahčić N, Scortichini G, Milone S (2013) Ochratoxin A in traditional dry-cured meat products produced from sub-chronic-exposed pigs. Food Addit Contam: Part A 30:1827–1836. https://doi.org/10.1080/19440049.2013.825817
Pleadin J, Staver MM, Vahčić N, Kovacevi N, Milone S, Saftić L, Scortichini G (2015) Survey of aflatoxin B1 and ochratoxin A occurrence in traditional meat products coming from Croatian households and markets. Food Control 52:71–77. https://doi.org/10.1016/j.foodcont.2014.12.027
Riley RT, Goeger DE, Yoo H, Showker JL (1992) Comparison of three tetramic acids and their ability to alter membrane function in cultured skeletal muscle cells and sarcoplasmic reticulum vesicles. Toxicol Appl Pharmacol 114:261–267. https://doi.org/10.1016/0041-008X(92)90076-5
Rodríguez A, Rodríguez M, Luque MI, Justesen AF, Córdoba JJ (2012a) A comparative study of DNA extraction methods to be used in real-time PCR based quantification of ochratoxin A-producing molds in food products. Food Control 25:666–672. https://doi.org/10.1016/j.foodcont.2011.12.010
Rodríguez A, Rodríguez M, Martín A, Delgado J, Córdoba JJ (2012b) Presence of ochratoxin A on the surface of dry–cured Iberian ham after initial fungal growth in the drying stage. Meat Sci 92:728–734. https://doi.org/10.1016/j.meatsci.2012.06.029
Rodríguez A, Medina A, Córdoba JJ, Magan N (2014) The influence of salt (NaCl) on ochratoxin A biosynthetic genes, growth and ochratoxin a production by three strains of Penicillium nordicum on a dry-cured ham-based medium. Int J Food Microbiol 178:113–119. https://doi.org/10.1016/j.ijfoodmicro.2014.03.007
Rodríguez A, Capela D, Medina A, Córdoba JJ, Magan N (2015) Relationship between ecophysiological factors, growth and ochratoxin A contamination of dry-cured sausage based matrices. Int J Food Microbiol 194:71–77. https://doi.org/10.1016/j.ijfoodmicro.2014.11.014
Sosa MJ, Córdoba JJ, Díaz C, Rodríguez M, Bermúdez E, Asensio MA, Núñez F (2002) Production of cyclopiazonic acid by Penicillium commune isolated from dry-cured ham on a meat extract-based substrate. J Food Prot 65:988–992. https://doi.org/10.4315/0362-028X-65.6.988
Zambonin CG, Monaci L, Aresta A (2001) Determination of cyclopiazonic acid in cheese samples using solid phase microextraction and high performance liquid chromatography. Food Chem 75:249–254. https://doi.org/10.1016/S0308-8146(01)00218-7
The authors acknowledge the technical support provided by the Facility of Innovation and Analysis in Animal Source Foodstuffs of SAIUEx (financed by UEx, Junta de Extremadura, MICINN, FEDER, and FSE).
This work has been funded by the Spanish Ministry of Economy and Competitiveness, Government of Extremadura, and FEDER (AGL2013-45729-P, AGL2016-80209-P, GR15108). B. Peromingo is recipient of a pre-doctoral fellowship (BES-2014-069484) and Dr. A. Rodríguez was supported by a Juan de la Cierva-Incorporación senior research fellowship (IJCI-2014-20666), both from the Spanish Ministry of Economy and Competitiveness.
Conflict of interest
The authors declare that there are no conflicts of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Peromingo, B., Rodríguez, A., Delgado, J. et al. Relationship between cyclopiazonic acid production and gene expression in Penicillium griseofulvum under dry-cured ham processing environmental conditions. Mycotoxin Res 35, 353–361 (2019). https://doi.org/10.1007/s12550-019-00357-9
- Penicillium griseofulvum
- Cyclopiazonic acid
- CPA biosynthetic genes
- Dry-cured ham