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Enterococcus faecium SE920, Debaryomyces hansenii FHSCC 253H, Penicillium chrysogenum CECT 20922, producer of the antifungal protein PgAFP, and this protein itself have previously been proposed to control toxigenic molds in dry-cured meat products. However, their effects on the usual microbial population, and the sensory characteristics of these foods, have not yet been evaluated. The aim of this study was to assess the viability of the inoculation of these protective cultures, and their impact on the quality of dry-cured fermented sausages. These microorganisms were co-inoculated with a native desirable population (Penicillium nalgiovense, P. chrysogenum, D. hansenii, and Staphylococcus vitulinus) in a dry-cured fermented sausage (salchichón)-based medium in the presence and absence of PgAFP. Macroscopically, the biocontrol candidates did not produce relevant changes in the growth of the native population, enabling their coexistence. However, PgAFP causes the alteration of the hyphae structure in desirable molds. Thus, PgAFP was discarded for use on the surface of raw dry-cured fermented sausages (salchichón) in the pilot plant. The used biocontrol agents did not negatively affect the physico-chemical parameters of the dry-cured fermented sausages (salchichón) after ripening, which showed the typical volatile profile and odor. Thus, the application of E. faecium SE920, D. hansenii FHSCC 253H, and P. chrysogenum CECT 20922 as protective cultures against toxigenic molds during the ripening of dry-cured fermented sausages does not modify their typical sensorial quality.
Micaela Álvarez; María J. Andrade; Carmen García; Juan J. Rondán; Félix Núñez. Effects of Preservative Agents on Quality Attributes of Dry-Cured Fermented Sausages. Foods 2020, 9, 1505 .
AMA StyleMicaela Álvarez, María J. Andrade, Carmen García, Juan J. Rondán, Félix Núñez. Effects of Preservative Agents on Quality Attributes of Dry-Cured Fermented Sausages. Foods. 2020; 9 (10):1505.
Chicago/Turabian StyleMicaela Álvarez; María J. Andrade; Carmen García; Juan J. Rondán; Félix Núñez. 2020. "Effects of Preservative Agents on Quality Attributes of Dry-Cured Fermented Sausages." Foods 9, no. 10: 1505.
Toxigenic moulds can develop on the surface of dry-cured meat products during ripening due to their ecological conditions, which constitutes a risk for consumers. A promising strategy to control this hazard is the use of antifungal microorganisms usually found in these foods. However, to date, the effectiveness of gram-positive catalase-positive cocci (GCC+) has not been explored. The aim of this work was to select GCC+ isolates with antifungal activity to study its effectiveness in a dry-cured ham model system at the environmental conditions reached during the ripening. Forty-five strains of GCC+ were evaluated and the isolate Staphylococcus xylosus Sx8 was selected to assess its efficacy at two different concentrations (106 and 104 cfu/mL) against Penicillium nordicum, Aspergillus flavus, Aspergillus parasiticus, and Penicillium griseofulvum at 15, 20, and 25 °C. The results showed that the inoculation of 106 cfu/mL of S. xylosus completely inhibited the growth of most fungi. In addition, in the presence of this strain at 104 cfu/mL, a significant reduction in fungal growth and mycotoxins production was observed at the three temperatures studied. In conclusion, S. xylosus Sx8 possesses great potential as a biological agent to control toxigenic moulds in dry-cured meat products.
Eva Cebrián; Félix Núñez; Fernando J. Gálvez; Josué Delgado; Elena Bermúdez; Mar Rodríguez. Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System. Microorganisms 2020, 8, 793 .
AMA StyleEva Cebrián, Félix Núñez, Fernando J. Gálvez, Josué Delgado, Elena Bermúdez, Mar Rodríguez. Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System. Microorganisms. 2020; 8 (6):793.
Chicago/Turabian StyleEva Cebrián; Félix Núñez; Fernando J. Gálvez; Josué Delgado; Elena Bermúdez; Mar Rodríguez. 2020. "Selection and Evaluation of Staphylococcus xylosus as a Biocontrol Agent against Toxigenic Moulds in a Dry-Cured Ham Model System." Microorganisms 8, no. 6: 793.
Contamination of foods and agricultural commodities by various types of toxigenic fungi is a concerning issue for human and animal health
Mar Rodríguez; Félix Núñez. Novel Approaches to Minimizing Mycotoxin Contamination. Toxins 2020, 12, 216 .
AMA StyleMar Rodríguez, Félix Núñez. Novel Approaches to Minimizing Mycotoxin Contamination. Toxins. 2020; 12 (4):216.
Chicago/Turabian StyleMar Rodríguez; Félix Núñez. 2020. "Novel Approaches to Minimizing Mycotoxin Contamination." Toxins 12, no. 4: 216.
The environmental conditions during the ripening of dry-cured meats and their nutritional composition promote the colonisation of their surface by Penicillium spp., including Penicillium nordicum producer of ochratoxin A (OTA). The objective of this work was to study the competitiveness of three potential biocontrol candidates (Debaryomyces hansenii FHSCC 253H, Enterococcus faecium SE920 and Penicillium chrysogenum CECT 20922) against the ochratoxigenic P. nordicum FHSCC4 under environmental and nutritional conditions simulating the ripening of dry-cured meat products. For this, the nutritional utilisation pattern, niche overlap index (NOI), interactions by dual-culture assays and OTA production were determined. The number of carbon sources (CSs) metabolised depended on the microorganism and the interacting water activity (aw) x temperature conditions. The number of CSs utilised by both filamentous fungi was quite similar and higher than those utilised by D. hansenii and E. faecium. The yeast isolate metabolised a number of CSs much larger than the bacterium. The NOI values showed that, in general, P. nordicum nutritionally dominated E. faecium and D. hansenii regardless of the environmental conditions evaluated. The relationship between the toxigenic and non-toxigenic fungal isolates depended on the aw x temperature combinations, although in none of the conditions a dominance of P. nordicum was observed. According to the interaction assays, both D. hansenii and P. chrysogenum decreased the growth of P. nordicum. The effect of D. hansenii could be attributed to the production of some extra-cellular compounds, while the action of P. chrysogenum is likely related to nutritional competition. In addition, both P. chrysogenum and D. hansenii reduced the OTA levels produced by P. nordicum. The effect of the yeast was more pronounced decreasing the concentration of OTA at quantities lower than the limit established by the Italian legislation. Therefore, P. chrysogenum and D. hansenii can be suggested as biocontrol candidates in the manufacture of dry-cured meat products.
María Micaela Álvarez; Félix Núñez; Josué Delgado; María J. Andrade; Mar Rodríguez; Alicia Rodríguez. Competitiveness of three biocontrol candidates against ochratoxigenic Penicillium nordicum under dry-cured meat environmental and nutritional conditions. Fungal Biology 2020, 125, 134 -142.
AMA StyleMaría Micaela Álvarez, Félix Núñez, Josué Delgado, María J. Andrade, Mar Rodríguez, Alicia Rodríguez. Competitiveness of three biocontrol candidates against ochratoxigenic Penicillium nordicum under dry-cured meat environmental and nutritional conditions. Fungal Biology. 2020; 125 (2):134-142.
Chicago/Turabian StyleMaría Micaela Álvarez; Félix Núñez; Josué Delgado; María J. Andrade; Mar Rodríguez; Alicia Rodríguez. 2020. "Competitiveness of three biocontrol candidates against ochratoxigenic Penicillium nordicum under dry-cured meat environmental and nutritional conditions." Fungal Biology 125, no. 2: 134-142.
Penicillium nordicum, an ochratoxin A (OTA) producer, widely contaminates the surface of dry-cured fermented sausages. Although the meat industry uses antifungal synthetic compounds during ripening, consumers' preferences currently stand up for natural preservatives. The aim of this study was to evaluate the antifungal effect of spices commonly added to dry-cured fermented sausages during their processing on P. nordicum for establishing their value as an alternative strategy to synthetic compounds. The mould strain was grown on a dry-cured fermented sausage-based medium containing oregano, rosemary or thyme. Moreover, an antifungal commercial preparation containing potassium sorbate and natamycin was tested alone and in combination with the spices. The growth rate, OTA accumulation and relative expression of genes involved in OTA biosynthesis (otapks and otanps) and some stress pathways (Hog1 and Rho1) were evaluated. In the growth assessment, the oregano alone or in combination with the antifungal preparation significantly reduced the growth rate. Different patterns were observed at both sampling times for the OTA production and gene expression analyses, and even differences between treatments at the same incubation period were observed. Regarding OTA accumulation, significant reductions were found when adding oregano or rosemary in the presence and absence of the antifungal compounds. While a stimulation of the expression of most of the tested genes was obtained at the initial stages, a repression was generally found at the end of the incubation. This makes sense since the mycotoxin accumulation was usually higher at the initial stages than at the end of the incubation. Accordingly, such spices would allow satisfying the current consumers’ demand for natural preservatives minimising the hazard associated with the OTA presence in cured meats at the same time.
María Micaela Álvarez; Alicia Rodríguez; Félix Núñez; Antonio Silva; María J. Andrade. In vitro antifungal effects of spices on ochratoxin A production and related gene expression in Penicillium nordicum on a dry-cured fermented sausage medium. Food Control 2020, 114, 107222 .
AMA StyleMaría Micaela Álvarez, Alicia Rodríguez, Félix Núñez, Antonio Silva, María J. Andrade. In vitro antifungal effects of spices on ochratoxin A production and related gene expression in Penicillium nordicum on a dry-cured fermented sausage medium. Food Control. 2020; 114 ():107222.
Chicago/Turabian StyleMaría Micaela Álvarez; Alicia Rodríguez; Félix Núñez; Antonio Silva; María J. Andrade. 2020. "In vitro antifungal effects of spices on ochratoxin A production and related gene expression in Penicillium nordicum on a dry-cured fermented sausage medium." Food Control 114, no. : 107222.
The ecological conditions during the ripening of dry-cured ham favour the development of moulds on its surface, being frequently the presence of Penicillium nordicum, a producer of ochratoxin A (OTA). Biocontrol using moulds and yeasts usually found in dry-cured ham is a promising strategy to minimize this hazard. The aim of this work is to evaluate the effect of previously selected Debaryomyces hansenii and Penicillium chrysogenum strains on growth, OTA production, and relative expression of genes involved in the OTA biosynthesis by P. nordicum. P. nordicum was inoculated against the protective cultures individually and combined on dry-cured ham for 21 days at 20 °C. None of the treatments reduced the growth of P. nordicum, but all of them decreased OTA concentration. The lower production of OTA could be related to significant repression of the relative expression of otapksPN and otanpsPN genes of P. nordicum. The efficacy of the combined protective cultures was tested in 24 dry-cured hams in industrial ripening (an 8 month-long production). OTA was detected in nine of the 12 dry-cured hams in the batch inoculated only with P. nordicum. However, in the batch inoculated with both P. nordicum and the combined protective culture, a considerable reduction of OTA contamination was observed. In conclusion, although the efficacy of individual use P. chrysogenum is great, the combination with D. hansenii enhances its antifungal activity and could be proposed as a mixed protective culture to control the hazard of the presence of OTA in dry-cured ham.
Eva Cebrián; Mar Rodríguez; Belén Peromingo; Elena Bermúdez; Félix Núñez. Efficacy of the Combined Protective Cultures of Penicillium chrysogenum and Debaryomyces hansenii for the Control of Ochratoxin A Hazard in Dry-Cured Ham. Toxins 2019, 11, 710 .
AMA StyleEva Cebrián, Mar Rodríguez, Belén Peromingo, Elena Bermúdez, Félix Núñez. Efficacy of the Combined Protective Cultures of Penicillium chrysogenum and Debaryomyces hansenii for the Control of Ochratoxin A Hazard in Dry-Cured Ham. Toxins. 2019; 11 (12):710.
Chicago/Turabian StyleEva Cebrián; Mar Rodríguez; Belén Peromingo; Elena Bermúdez; Félix Núñez. 2019. "Efficacy of the Combined Protective Cultures of Penicillium chrysogenum and Debaryomyces hansenii for the Control of Ochratoxin A Hazard in Dry-Cured Ham." Toxins 11, no. 12: 710.
Moulds positively contribute to the development of typical characteristic flavour and aroma of dry-fermented sausages. However, some mould species, such as Penicillium nordicum and Penicillium verrucosum, may contaminate this product with ochratoxin A (OTA). For this reason, the control of toxigenic moulds is needed. Strategies based on the use of antifungal microorganisms present in the native microbial population in the dry-fermented sausage processing could be a promising strategy. The aim of this work was to study the effect of Enterococcus faecium strains on P. nordicum and P. verrucosum growth and OTA production in a dry-fermented sausage-based medium at conditions of temperature and water activity similar to those occurring during the ripening of these meat products. Six strains were screened to evaluate their growth capacity and antifungal activity against P. nordicum and P. verrucosum at three fixed temperatures related to the sausage ripening. The two E. faecium strains that decreased growth of both species were chosen to further evaluate their effect on growth of P. verrucosum and P. nordicum and their mycotoxin production under conditions simulating the dry-fermented sausage ripening. The presence of E. faecium SE920 significantly reduced OTA production of P. nordicum although it did not affect P. verrucosum. E. faecium SE920, isolated from dry-fermented sausages, could be a good candidate to reduce OTA production by P. nordicum in dry-fermented sausages.
Micaela Álvarez; Alicia Rodríguez; Belén Peromingo; Félix Núñez; Mar Rodríguez. Enterococcus faecium: a promising protective culture to control growth of ochratoxigenic moulds and mycotoxin production in dry-fermented sausages. Mycotoxin Research 2019, 36, 137 -145.
AMA StyleMicaela Álvarez, Alicia Rodríguez, Belén Peromingo, Félix Núñez, Mar Rodríguez. Enterococcus faecium: a promising protective culture to control growth of ochratoxigenic moulds and mycotoxin production in dry-fermented sausages. Mycotoxin Research. 2019; 36 (2):137-145.
Chicago/Turabian StyleMicaela Álvarez; Alicia Rodríguez; Belén Peromingo; Félix Núñez; Mar Rodríguez. 2019. "Enterococcus faecium: a promising protective culture to control growth of ochratoxigenic moulds and mycotoxin production in dry-fermented sausages." Mycotoxin Research 36, no. 2: 137-145.
Environmental conditions during ripening of dry-cured meat products favour growth of fungal population on their surface. Some of these moulds can produce mycotoxins. Paprika is one of the ingredients usually used in the formulation of raw-cured sausages, and its addition could influence the growth and production of mycotoxins of the moulds present in these products. In this work the effect of Spanish smoked paprika "Pimentón de la Vera" on growth of Aspergillus parasiticus and Penicillium nordicum and production of aflatoxins B1 (AFB1), G1 (AFG1) and ochratoxin A (OTA) respectively, was evaluated. Moulds were grown in a culture medium made from lyophilized fresh pork meat added with 4% salt and different concentrations of Spanish smoked paprika (1, 2 and 3%) at several water activity values (0.98, 0.94 and 0.87) and temperature (20-25 °C), to simulate conditions usually found during ripening of dry-cured meat products. Mould growth was evaluated by measuring the diameter of the colony every 24 h, and the production of mycotoxins by UHPLC-MS/MS every 2 days, during 10 days of incubation. Addition of paprika favours growth of the two mould species tested. However, the synthesis of mycotoxins was reduced at 0.94 and 0.98 aw when at least a 2% of paprika was added. Therefore, the addition of Spanish smoked paprika at 2-3% in the formulations may help to minimize AFs and OTA production in dry-cured meat products such as loins or "chorizo" sausages.
Lourdes Sánchez-Montero; Juan J. Córdoba; Alberto Alía; Belén Peromingo; Félix Núñez. Effect of Spanish smoked paprika “Pimentón de La Vera” on control of ochratoxin A and aflatoxins production on a dry-cured meat model system. International Journal of Food Microbiology 2019, 308, 108303 .
AMA StyleLourdes Sánchez-Montero, Juan J. Córdoba, Alberto Alía, Belén Peromingo, Félix Núñez. Effect of Spanish smoked paprika “Pimentón de La Vera” on control of ochratoxin A and aflatoxins production on a dry-cured meat model system. International Journal of Food Microbiology. 2019; 308 ():108303.
Chicago/Turabian StyleLourdes Sánchez-Montero; Juan J. Córdoba; Alberto Alía; Belén Peromingo; Félix Núñez. 2019. "Effect of Spanish smoked paprika “Pimentón de La Vera” on control of ochratoxin A and aflatoxins production on a dry-cured meat model system." International Journal of Food Microbiology 308, no. : 108303.
Postharvest fungal diseases are among the main causes of fresh fruit losses. Chemical control is against claims for “natural” or “chemical-free” products. Biocontrol agents, such as antifungal proteins or their producing moulds, may serve to combat unwanted pathogens. Since the effectiveness of these bioprotective agents depends on the food substrate, their effect must be tested on fruits. The objective of this work was to study the effect of the antifungal protein PgAFP and its producer, Penicillium chrysogenum, against Penicillium expansum and Penicillium digitatum growth on apple and oranges respectively, and the PgAFP effect on eleven P. expansum, Penicillium italicum, and P. digitatum strains in vitro, and on patulin production on apple substrate. The sensitivity upon PgAFP was P. digitatum > P. expansum > P. italicum. In oranges, broadly, no inhibitory effect was obtained. PgAFP and P. chrysogenum did not inhibit the P. expansum CMP-1 growth on Golden Delicious apples, however, a successful effect was achieved on Royal Gala apples. On apple substrate, patulin production by P. expansum CMP-1 rose in parallel to PgAFP concentrations, linked with high reactive oxygen species levels. PgAFP cannot be proposed as a bioprotective agent on apple. However, P. chrysogenum is a promising agent to be used on Royal Gala apples.
Josué Delgado; Ana-Rosa Ballester; Félix Núñez; Luis González-Candelas. Evaluation of the activity of the antifungal PgAFP protein and its producer mould against Penicillium spp postharvest pathogens of citrus and pome fruits. Food Microbiology 2019, 84, 103266 .
AMA StyleJosué Delgado, Ana-Rosa Ballester, Félix Núñez, Luis González-Candelas. Evaluation of the activity of the antifungal PgAFP protein and its producer mould against Penicillium spp postharvest pathogens of citrus and pome fruits. Food Microbiology. 2019; 84 ():103266.
Chicago/Turabian StyleJosué Delgado; Ana-Rosa Ballester; Félix Núñez; Luis González-Candelas. 2019. "Evaluation of the activity of the antifungal PgAFP protein and its producer mould against Penicillium spp postharvest pathogens of citrus and pome fruits." Food Microbiology 84, no. : 103266.
Dry-cured meat products are usually contaminated with moulds during ripening. Although fungal development contributes to the desired sensory characteristics, some moulds, such as Penicillium nordicum are able to produce ochratoxin A (OTA) on meat products. Therefore, strategies to prevent OTA contamination in ripened meat products are required. Microorganisms isolated from these meat products can be adequate as biocontrol agents, given that no negative sensory impact is expected. The PgAFP antifungal protein-producer Penicillium chrysogenum (Pc) and Debaryomyces hansenii (Dh) have been shown to successfully inhibit toxigenic moulds. However, scarce information about the mechanism of action of these biocontrol agents on toxigenic mould inhibition is available. Comparative proteomic analysis is a powerful tool to investigate the physiological response of microorganisms to stimuli. Proteomic analysis was carried out on P. nordicum co-cultured with Pc, Dh, PgAFP, and their combinations on a dry-cured ham-based medium. Additionally, OTA production by P. nordicum in the different cultures was measured. The individual inoculation of Pc or Dh repressed OTA production by P. nordicum by 5 and 3.15 fold, respectively. A total of 2844 unique P. nordicum proteins were identified by proteomic analysis. The impact of the biocontrol agents on the proteome of P. nordicum was higher for Pc-containing cultures, followed by Dh-containing treatments. PgAFP alone had minimal impact on the proteome of P. nordicum. Proteomic analyses indicated Pc repressed P. nordicum OTA production through nutrient competition, potentially reducing glucose availability. Data also suggest that Dh and Pc inhibited P. nordicum through cell wall integrity impairment. Both Pc and Dh seem to hamper P. nordicum secondary metabolism (SM) as indicated by lower levels of MAP kinases and SM-associated proteins found in the co-inoculated P. nordicum. This work paves the way to use antifungal agents in the most efficient way to prevent OTA formation in meat products.
Josué Delgado; Félix Núñez; Miguel A. Asensio; Rebecca A. Owens. Quantitative proteomic profiling of ochratoxin A repression in Penicillium nordicum by protective cultures. International Journal of Food Microbiology 2019, 305, 108243 .
AMA StyleJosué Delgado, Félix Núñez, Miguel A. Asensio, Rebecca A. Owens. Quantitative proteomic profiling of ochratoxin A repression in Penicillium nordicum by protective cultures. International Journal of Food Microbiology. 2019; 305 ():108243.
Chicago/Turabian StyleJosué Delgado; Félix Núñez; Miguel A. Asensio; Rebecca A. Owens. 2019. "Quantitative proteomic profiling of ochratoxin A repression in Penicillium nordicum by protective cultures." International Journal of Food Microbiology 305, no. : 108243.
Dry-cured meat products, such as dry-cured ham or dry-fermented sausages, are characterized by their particular ripening process, where a mould population grows on their surface. Some of these moulds are hazardous to the consumers because of their ability to produce mycotoxins including aflatoxins (AFs). The use of native yeasts could be considered a potential strategy for controlling the presence of AFs in dry-cured meat products. The aim of this work was to evaluate the antagonistic activity of two native Debaryomyces hansenii strains on the relative growth rate and the AFs production in Aspergillus parasiticus. Both D. hansenii strains significantly reduced the growth rates of A. parasiticus when grown in a meat-model system at different water activity (aw) conditions. The presence of D. hansenii strains caused a stimulation of AFs production by A. parasiticus at 0.99 aw. However, at 0.92 aw the yeasts significantly reduced the AFs concentration in the meat-model system. The relative expression levels of the aflR and aflS genes involved in the AFs biosynthetic pathway were also repressed at 0.92 aw in the presence of both D. hansenii strains. These satisfactory results were confirmed in dry-cured ham and dry-fermented sausage slices inoculated with A. parasiticus, since both D. hansenii strains significantly reduced AFs amounts in these matrices. Therefore, both tested D. hansenii strains could be proposed as biocontrol agents within a HACCP framework to minimize the hazard associated with the presence of AFs in dry-cured meat products.
Belén Peromingo; María J. Andrade; Josué Delgado; Lourdes Sánchez-Montero; Félix Núñez. Biocontrol of aflatoxigenic Aspergillus parasiticus by native Debaryomyces hansenii in dry-cured meat products. Food Microbiology 2019, 82, 269 -276.
AMA StyleBelén Peromingo, María J. Andrade, Josué Delgado, Lourdes Sánchez-Montero, Félix Núñez. Biocontrol of aflatoxigenic Aspergillus parasiticus by native Debaryomyces hansenii in dry-cured meat products. Food Microbiology. 2019; 82 ():269-276.
Chicago/Turabian StyleBelén Peromingo; María J. Andrade; Josué Delgado; Lourdes Sánchez-Montero; Félix Núñez. 2019. "Biocontrol of aflatoxigenic Aspergillus parasiticus by native Debaryomyces hansenii in dry-cured meat products." Food Microbiology 82, no. : 269-276.
Ecological conditions during ripening of dry-cured meat products favour the growth of an uncontrolled mould population that could suppose a risk of ochratoxin A (OTA) production. In this work the influence of water activity (aw), temperature and substrate composition on fungal growth and OTA production by Penicillium nordicum and Penicillium verrucosum isolated from dry-cured meat products have been studied. In addition, the relative risk of OTA presence on dry-cured meat products has been evaluated using the Risk Ranger software. Fungal growth was observed in the range of 0.99–0.90 aw and 15–25 °C being mainly temperature-dependent. P. nordicum and P. verrucosum were able to produce OTA in every substrate in these ranges of aw and temperature. The production of OTA by P. verrucosum was mainly influenced by temperature and media composition. However, P. nordicum it is affected mainly by substrate or temperature depending on the strain studied. Both species produce a large amount of OTA on dry-cured ham and on dry-cured fermented sausage “salchichón” in environmental conditions usually found throughout the ripening of these products. The Risk Ranger software reveals that the relative risk of OTA on dry-cured meat products is 75%. Thus, control measures during dry-cured meat products processing to prevent OTA risk should be established.
Lourdes Sánchez-Montero; Juan J. Córdoba; Belén Peromingo; Micaela Álvarez; Félix Núñez. Effects of environmental conditions and substrate on growth and ochratoxin A production by Penicillium verrucosum and Penicillium nordicum: Relative risk assessment of OTA in dry-cured meat products. Food Research International 2018, 121, 604 -611.
AMA StyleLourdes Sánchez-Montero, Juan J. Córdoba, Belén Peromingo, Micaela Álvarez, Félix Núñez. Effects of environmental conditions and substrate on growth and ochratoxin A production by Penicillium verrucosum and Penicillium nordicum: Relative risk assessment of OTA in dry-cured meat products. Food Research International. 2018; 121 ():604-611.
Chicago/Turabian StyleLourdes Sánchez-Montero; Juan J. Córdoba; Belén Peromingo; Micaela Álvarez; Félix Núñez. 2018. "Effects of environmental conditions and substrate on growth and ochratoxin A production by Penicillium verrucosum and Penicillium nordicum: Relative risk assessment of OTA in dry-cured meat products." Food Research International 121, no. : 604-611.
An extraction method and an UHPLC-MS/MS method for the quantification of CPA in dry-cured ham were developed and validated. To optimise detection and quantification of CPA, the composition of mobile phase, flow rates, gradient–related factors and solvents used for resuspension of dry extracts were evaluated. Besides, four extraction methods were tested. The best peak shape and resolution were obtained by eluting the mobile phase consisting in acetic acid-ammonium acetate buffer pH 5.75/methanol in gradient mode at a flow rate of 0.2 mL/min. The method 4 relied on the QuEChERS methodology was the most effective one. Almost half of the 61 dry-cured ham samples examined were contaminated with CPA, with values ranging from 36.1 to 540.1 ng/g. The combination of a QuEChERS-based extraction method and analysis by UHPLC-MS/MS allows highly sensitive, fast, reliable and cheap detection and quantification of CPA for routine analysis in ham.
Belén Peromingo; Mar Rodríguez; Félix Núñez; Antonio Silva; Alicia Rodríguez. Sensitive determination of cyclopiazonic acid in dry-cured ham using a QuEChERS method and UHPLC–MS/MS. Food Chemistry 2018, 263, 275 -282.
AMA StyleBelén Peromingo, Mar Rodríguez, Félix Núñez, Antonio Silva, Alicia Rodríguez. Sensitive determination of cyclopiazonic acid in dry-cured ham using a QuEChERS method and UHPLC–MS/MS. Food Chemistry. 2018; 263 ():275-282.
Chicago/Turabian StyleBelén Peromingo; Mar Rodríguez; Félix Núñez; Antonio Silva; Alicia Rodríguez. 2018. "Sensitive determination of cyclopiazonic acid in dry-cured ham using a QuEChERS method and UHPLC–MS/MS." Food Chemistry 263, no. : 275-282.
Aflatoxigenic molds can grow and produce aflatoxins on dry-fermented meat and cheese. The small, basic, cysteine-rich antifungal protein PgAFP displays a time-limited inhibitory ability against unwanted molds by increasing reactive oxygen species (ROS), which can lead to increased aflatoxin production. However, calcium abolishes the inhibitory effect of PgAFP on certain Aspergillus spp. To maximize the antifungal effect, this protein may be combined with protective cultures. Yeasts and lactic acid bacteria may counteract the impact of calcium on PgAFP fungal inhibition. The objective of this work was to study the effect of PgAFP and different combined treatments with Debaryomyces hansenii and/or Pediococcus acidilactici against growth of and aflatoxin production by an aflatoxigenic strain of Aspergillus parasiticus in both culture media and dry-fermented foods with low or high calcium levels. Aflatoxins production was increased by PgAFP but dramatically reduced by P. acidilactici in low calcium culture medium, whereas in the Ca-enriched culture medium, all treatments tested led to low aflatoxins levels. To study whether PgAFP and the protective microorganisms interfere with ROS and aflatoxin production, the relative expression of genes foxA, which is involved in peroxisomal β-oxidation, and aflP, which is required for aflatoxin biosynthesis, were evaluated. The aflatoxin overproduction induced by PgAFP seems not to be linked to peroxisomal β-oxidation. The combination of PgAFP and D. hansenii provided a successful inhibitory effect on A. parasiticus growth as well as on aflatoxin production on sliced dry-fermented sausage and cheese ripened up to 15 days, whereas P. acidilactici did not further enhance the protective effect of the two former agents. Therefore, the combined treatment of PgAFP and D. hansenii seems to provide a promising protective mean against aflatoxin-producing A. parasiticus on dry-fermented foods.
Josué Delgado; Alicia Rodríguez; Alfredo Garcia-Sanchez; Félix Núñez; Miguel A. Asensio. Inhibitory Effect of PgAFP and Protective Cultures on Aspergillus parasiticus Growth and Aflatoxins Production on Dry-Fermented Sausage and Cheese. Microorganisms 2018, 6, 69 .
AMA StyleJosué Delgado, Alicia Rodríguez, Alfredo Garcia-Sanchez, Félix Núñez, Miguel A. Asensio. Inhibitory Effect of PgAFP and Protective Cultures on Aspergillus parasiticus Growth and Aflatoxins Production on Dry-Fermented Sausage and Cheese. Microorganisms. 2018; 6 (3):69.
Chicago/Turabian StyleJosué Delgado; Alicia Rodríguez; Alfredo Garcia-Sanchez; Félix Núñez; Miguel A. Asensio. 2018. "Inhibitory Effect of PgAFP and Protective Cultures on Aspergillus parasiticus Growth and Aflatoxins Production on Dry-Fermented Sausage and Cheese." Microorganisms 6, no. 3: 69.
The environmental conditions reached during the ripening of dry-cured meat products favour the proliferation of moulds on their surface. Some of these moulds are hazardous to consumers because of their ability to produce ochratoxin A (OTA). Biocontrol using Debaryomyces hansenii could be a suitable strategy to prevent the growth of ochratoxigenic moulds and OTA accumulation in dry-cured meat products. The aim of this work was to evaluate the ability of two strains of D. hansenii to control the growth and OTA production of Penicillium verrucosum in a meat model under water activities (aw) values commonly reached during the dry-cured meat product ripening. The presence of D. hansenii strains triggered a lengthening of the lag phase and a decrease of the growth rate of P. verrucosum in meat-based media at 0.97 and 0.92 aw. Both D. hansenii strains significantly reduced OTA production (between 85.16 and 92.63%) by P. verrucosum in the meat-based medium at 0.92 aw. Neither absorption nor detoxification of OTA by D. hansenii strains seems to be involved. However, a repression of the expression of the non-ribosomal peptide synthetase (otanpsPN) gene linked to the OTA biosynthetic pathway was observed in the presence of D. hansenii. To confirm the protective role of D. hansenii strains, they were inoculated together with P. verrucosum Pv45 in dry-fermented sausage and dry-cured ham slices. Although P. verrucosum Pv45 counts were not affected by the presence of D. hansenii in both meat matrices, a reduction of OTA amount was observed. Therefore, the effect of D. hansenii strains on OTA accumulation should be attributed to a reduction at transcriptional level. Consequently, native D. hansenii can be useful as biocontrol agent in dry-cured meat products for preventing the hazard associated with the presence of OTA.
Belén Peromingo; Félix Núñez; Alicia Rodriguez; Alberto Alía; Maria J. Andrade. Potential of yeasts isolated from dry-cured ham to control ochratoxin A production in meat models. International Journal of Food Microbiology 2018, 268, 73 -80.
AMA StyleBelén Peromingo, Félix Núñez, Alicia Rodriguez, Alberto Alía, Maria J. Andrade. Potential of yeasts isolated from dry-cured ham to control ochratoxin A production in meat models. International Journal of Food Microbiology. 2018; 268 ():73-80.
Chicago/Turabian StyleBelén Peromingo; Félix Núñez; Alicia Rodriguez; Alberto Alía; Maria J. Andrade. 2018. "Potential of yeasts isolated from dry-cured ham to control ochratoxin A production in meat models." International Journal of Food Microbiology 268, no. : 73-80.
The ability of Aspergillus flavus to produce aflatoxins in dairy products presents a potential hazard. The antifungal protein PgAFP from Penicillium chrysogenum inhibits various foodborne toxigenic fungi, including Aspergillus flavus. However, PgAFP did not inhibit A. flavus growth in cheese, which was related to the associated cation content. CaCl increased A. flavus permeability and prevented PgAFP-mediated inhibition in potato dextrose broth (PDB). PgAFP did not elicit any additional increase in permeability of CaCl-incubated A. flavus. Furthermore, PgAFP did not alter metabolic capability, chitin deposition, or hyphal viability of A. flavus grown with CaCl. Comparative proteomic analysis after PgAFP treatment of A. flavus in calcium-enriched PDB revealed increased abundance of 125 proteins, including oxidative stress-related proteins, as determined by label-free mass spectrometry (MS)-based proteomics. Seventy proteins were found at lower abundance, with most involved in metabolic pathways and biosynthesis of secondary metabolites. These changes do not support the blockage of potential PgAFP receptors in A. flavus by calcium as the main cause of the protective role. A. flavus resistance appears to be mediated by calcineurin, G-protein, and γ-glutamyltranspeptidase that combat oxidative stress and impede apoptosis. These findings could serve to design strategies to improve PgAFP activity against aflatoxigenic moulds in dairy products.
Josué Delgado; Rebecca Owens; Sean Doyle; Félix Núñez; Miguel A. Asensio. Quantitative proteomics reveals new insights into calcium-mediated resistance mechanisms in Aspergillus flavus against the antifungal protein PgAFP in cheese. Food Microbiology 2017, 66, 1 -10.
AMA StyleJosué Delgado, Rebecca Owens, Sean Doyle, Félix Núñez, Miguel A. Asensio. Quantitative proteomics reveals new insights into calcium-mediated resistance mechanisms in Aspergillus flavus against the antifungal protein PgAFP in cheese. Food Microbiology. 2017; 66 ():1-10.
Chicago/Turabian StyleJosué Delgado; Rebecca Owens; Sean Doyle; Félix Núñez; Miguel A. Asensio. 2017. "Quantitative proteomics reveals new insights into calcium-mediated resistance mechanisms in Aspergillus flavus against the antifungal protein PgAFP in cheese." Food Microbiology 66, no. : 1-10.
Elena Bermúdez; Félix Núñez; Josué Delgado; Miguel A. Asensio. Penicillium and Talaromyces. Laboratory Models for Foodborne Infections 2017, 555 -575.
AMA StyleElena Bermúdez, Félix Núñez, Josué Delgado, Miguel A. Asensio. Penicillium and Talaromyces. Laboratory Models for Foodborne Infections. 2017; ():555-575.
Chicago/Turabian StyleElena Bermúdez; Félix Núñez; Josué Delgado; Miguel A. Asensio. 2017. "Penicillium and Talaromyces." Laboratory Models for Foodborne Infections , no. : 555-575.
Josué Delgado; Belén Peromingo; Félix Núñez; Miguel A Asensio. Use of molds and their antifungal proteins for biocontrol of toxigenic molds on dry-ripened cheese and meats. Current Opinion in Food Science 2016, 11, 40 -45.
AMA StyleJosué Delgado, Belén Peromingo, Félix Núñez, Miguel A Asensio. Use of molds and their antifungal proteins for biocontrol of toxigenic molds on dry-ripened cheese and meats. Current Opinion in Food Science. 2016; 11 ():40-45.
Chicago/Turabian StyleJosué Delgado; Belén Peromingo; Félix Núñez; Miguel A Asensio. 2016. "Use of molds and their antifungal proteins for biocontrol of toxigenic molds on dry-ripened cheese and meats." Current Opinion in Food Science 11, no. : 40-45.
Moulds growing on the surface of dry-ripened foods contribute to their sensory qualities, but some of them are able to produce mycotoxins that pose a hazard to consumers. Small cysteine-rich antifungal proteins (AFPs) from moulds are highly stable to pH and proteolysis and exhibit a broad inhibition spectrum against filamentous fungi, providing new chances to control hazardous moulds in fermented foods. The analytical tools for characterizing the cellular targets and affected pathways are reviewed. Strategies currently employed to study these mechanisms of action include 'omics' approaches that have come to the forefront in recent years, developing in tandem with genome sequencing of relevant organisms. These techniques contribute to a better understanding of the response of moulds against AFPs, allowing the design of complementary strategies to maximize or overcome the limitations of using AFPs on foods. AFPs alter chitin biosynthesis, and some fungi react inducing cell wall integrity (CWI) pathway. However, moulds able to increase chitin content at the cell wall by increasing proteins in either CWI or calmodulin-calcineurin signalling pathways will resist AFPs. Similarly, AFPs increase the intracellular levels of reactive oxygen species (ROS), and moulds increasing G-protein complex β subunit CpcB and/or enzymes to efficiently produce glutathione may evade apoptosis. Unknown aspects that need to be addressed include the interaction with mycotoxin production by less sensitive toxigenic moulds. However, significant steps have been taken to encourage the use of AFPs in intermediate-moisture foods, particularly for mould-ripened cheese and meat products.
Josué Delgado; Rebecca A. Owens; Sean Doyle; Miguel A. Asensio; Félix Núñez. Manuscript title: antifungal proteins from moulds: analytical tools and potential application to dry-ripened foods. Applied Microbiology and Biotechnology 2016, 100, 6991 -7000.
AMA StyleJosué Delgado, Rebecca A. Owens, Sean Doyle, Miguel A. Asensio, Félix Núñez. Manuscript title: antifungal proteins from moulds: analytical tools and potential application to dry-ripened foods. Applied Microbiology and Biotechnology. 2016; 100 (16):6991-7000.
Chicago/Turabian StyleJosué Delgado; Rebecca A. Owens; Sean Doyle; Miguel A. Asensio; Félix Núñez. 2016. "Manuscript title: antifungal proteins from moulds: analytical tools and potential application to dry-ripened foods." Applied Microbiology and Biotechnology 100, no. 16: 6991-7000.
Antifungal proteins from molds have been proposed as a valuable tool against unwanted molds, but the resistance of some fungi limits their use. Resistance to antimicrobial peptides has been suggested to be due to lack of interaction with the mold or to a successful response. The antifungal protein PgAFP produced by Penicillium chrysogenum inhibits the growth of various ascomycetes, but not Penicillium polonicum. To study the basis for resistance to this antifungal protein, localization of PgAFP and metabolic, structural, and morphological changes were investigated in P. polonicum. PgAFP bound the outer layer of P. polonicum but not regenerated chitin, suggesting an interaction with specific molecules. Comparative two-dimensional gel electrophoresis (2D-PAGE) and comparative quantitative proteomics revealed changes in the relative abundance of several proteins from ribosome, spliceosome, metabolic, and biosynthesis of secondary metabolite pathways. The proteome changes and an altered permeability reveal an active reaction of P. polonicum to PgAFP. The successful response of the resistant mold seems to be based on the higher abundance of protein Rho GTPase Rho1 that would lead to the increased chitin deposition via cell wall integrity (CWI) signaling pathway. Thus, combined treatment with chitinases could provide a complementary means to combat resistance to antifungal proteins.
Josué Delgado; Rebecca Owens; Sean Doyle; Miguel A. Asensio; Félix Núñez. Increased chitin biosynthesis contributes to the resistance of Penicillium polonicum against the antifungal protein PgAFP. Applied Microbiology and Biotechnology 2015, 100, 371 -383.
AMA StyleJosué Delgado, Rebecca Owens, Sean Doyle, Miguel A. Asensio, Félix Núñez. Increased chitin biosynthesis contributes to the resistance of Penicillium polonicum against the antifungal protein PgAFP. Applied Microbiology and Biotechnology. 2015; 100 (1):371-383.
Chicago/Turabian StyleJosué Delgado; Rebecca Owens; Sean Doyle; Miguel A. Asensio; Félix Núñez. 2015. "Increased chitin biosynthesis contributes to the resistance of Penicillium polonicum against the antifungal protein PgAFP." Applied Microbiology and Biotechnology 100, no. 1: 371-383.