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Cocoa pod-opening delay and bean fermentation promote the organoleptic quality of chocolate. The present research investigated the changes in the volatile fingerprint of cocoa harvested at a traditional plantation. Cocoa beans extracted from 2-days pod-opening delay were simultaneously fermented for 5 days using container and then sun-dried to 7-8% moisture content at five different locations: Akoupé, San Pedro, Soubré, Djekanou and Daloa. The aromatic analysis were done on cocoa using the HS-SPME-GC/MS technique. Professional panelists evaluated the sensory perceptions of the chocolate. The results shows that cocoa fermented in both Daloa and Soubré regions were differentiated by 2,3-butanediol while those processed in other regions presented highest acetoin content. However, fermented cocoa from Soubré region exhibited most amount of 2,3-butanediol, diacetate A whereas 2,3,5,6-tetramethylpyrazine differentiated those from Daloa region. Sensory properties of chocolate were not linked to the aromatic compound precursors profile of beans. The fermentation performed in San Pédro region promote both the generation of more desirable aromatic compounds of cocoa and sensory attributes of the finished chocolate. The fermentation location generates a greater differentiation of the volatile fingerprint of cocoa and the sensory perceptions of the finished chocolate.

Wild yeasts isolated from the surface of apples were screened for antagonistic activity against Penicillium expansum, the main producer of the mycotoxin patulin. Three antagonistic yeasts (Y33, Y29 and Y24) from a total of 90 were found to inhibit P. expansum growth. Identification by ITS region sequence and characterization showed that three selected isolates of yeast should be different strains of Metschnikowia pulcherrima. Several concentrations of the selected yeasts were used to study their in vitro antifungal effectivity against P. expansum on Petri dishes (plates with 63.6 cm2 surface) whereas their potential activity on patulin reduction was studied in liquid medium. Finally, the BCA that had the best in vitro antifungal capacity against P. and the best patulin degradation capacity was selected to be assessed directly on apples. All the selected strains demonstrated antifungal activity in vitro but the most efficient was the strain Y29. Isolated strains were able to reduce patulin content in liquid medium, Y29 being the only strain that completely reduced patulin levels within 120 h. The application of Y29 as biocontrol agent on the surface of apples inoculated with P. expansum, inhibited fungal growth and patulin production during storage. Therefore, the results shown that this yeast strain could be used for the reduction of P. expansum and its mycotoxin in apples or apple-based products by adapting the procedure application., The authors acknowledge the financial support of the Agence Nationale de la Recherche (France) (PATRISK project: ANR-17-CE21-0008) and the financial support of the European Union (Erasmus grant attributed to Laura Settier-Ramírez).

Ochratoxin A (OTA) is a secondary metabolite produced by fungal pathogens such as Penicillium verrucosum, which develops in food commodities during storage such as cereals, grapes, and coffee. It represents public health concerns due to its genotoxicity, carcinogenicity, and teratogenicity. The objective of this study was to evaluate the ability of actinobacteria and their metabolites to degrade OTA and/or to decrease its production. Sixty strains of actinobacteria were tested for their ability to prevent OTA formation by in vitro dual culture assays or with cell free extracts (CFEs). In dual culture, 17 strains strongly inhibited fungal growth, although it was generally associated with an increase in OTA specific production. Seventeen strains inhibited OTA specific production up to 4% of the control. Eleven actinobacteria CFEs reduced OTA specific production up to 62% of the control, while no substantial growth inhibition was observed except for two strains up to 72% of the control. Thirty-three strains were able to degrade OTA almost completely in liquid medium whereas only five were able to decrease it on solid medium, and two of them reduced OTA to an undetectable amount. Our results suggest that OTA decrease could be related to different strategies of degradation/metabolization by actinobacteria, through enzyme activities and secretion of secondary metabolites interfering with the OTA biosynthetic pathway. CFEs appeared to be ineffective at degrading OTA, raising interesting questions about the detoxification mechanisms. Common degradation by-products (e.g., OT&alpha, or L-&beta, phenylalanine) were searched by HPLC-MS/MS, however, none of them were found, which implies a different mechanism of detoxification and/or a subsequent degradation into unknown products.

In a sustainable development concern, the agricultural practices notably those to control the pathogenic and/or mycotoxigenic fungi that maintain the quality and the abundance of crops with respecting the ecosystems and public health are indispensable. Our study focuses on evaluating the biocontrol potential of the actinomycete microflora of some organic amendments against the phytopathogenic and mycotoxigenic Fusarium verticillioides. The inhibitory effect was assessed in term of growth and mycotoxin production (fumonisin FB1 and FB2) of F. verticillioides in the presence of amended soils and selected antagonist agents from the amendments. Results demonstrated that the amended soils with different organic amendments exhibited a higher inhibition rate of Fusarium verticillioides growth than the reference soil. They were able to inhibit the fungal pigment and spore productions which are the major mean of survival and dispersion of this fungus. The fumonisin production was strongly reduced by the metabolites produced by the amended soils’ microbiota, up to 68.7% and 92.5% for FB1 and FB2 respectively. Actinomycete strains isolated from these amendments were identified as Streptomyces and were also tested for their antifungal potential. The fungal growth was reduced up to more than 60% in the presence of extracellular metabolites of several Streptomyces strains. In confrontation assays, most of the Streptomyces strains inhibited strongly both fungal growth and fumonisin production. The FB1 and FB2 amounts were decreased at best, by up to 87.5% and 98.2% respectively, depending on the actinomycete strains and fungal culture time. These results are promising for the use of these biocontrol agents, and especially some actinomycete strains, as tools to reduce fumonisin contamination in cereal crops and derived products.

Cereals products enriched with leguminous or oleaginous are used in Burkina Faso as food complement or to avoid infant malnutrition. Infant formulas from cereals are consumed as food supplements after the weaning age. They are produced in Burkina Faso with a mixture of several cereals (maize, millet, rice, or sorghum), leguminous (bean), oleaginous (peanut, soya or sesame), sugar or salt and sometimes milk powder. The production is artisanal or semi-industrial. These infant foods from cereals should be free from mycotoxins and pathogenic bacteria. The objective of this work was to determine the occurrence of mycotoxins like aflatoxins, ochratoxin A and fumonisins in infant formulas and grains in Ouagadougou (Burkina Faso). The mycotoxins (aflatoxins, ochratoxin A and fumonisins) were determined by HPLC-FLD in 248 samples of infant formulas produced by the Recovery and Nutrition Education Centers (CRENs) and sold on the market places in Burkina Faso. Results showed that the majority of samples of infant formulas presented high levels of mycotoxins. The frequency of contaminated samples by aflatoxin B1, ochratoxin A and fumonisins in analyzed samples were 83.9% (167/199), 7.5% (15/199) and 1.5% (3/199), respectively. The highest values registered in infant formulas were 900, 6 and 3 times higher for aflatoxin B1 (EU limit: 0.1 μg/kg), ochratoxin A (EU limit: 0.5 μg/kg) and fumonisins (EU limit: 200 μg/kg), respectively, than the EU regulation limits (1881/2006). This study presents the first results concerning the safety assessment of infant formulas regarding mycotoxins in Burkina Faso.

'Kankankan' is a popular spice powder used to season roasted meat in Côte d'Ivoire. However, produced in a traditional way, the conditions of production and storage of kankankan favour the proliferation of mycotoxin-producing fungal strains. The aim of this study was to carry out an inventory of mycotoxin contamination of this spice powder and to assess risk exposure to consumers. In total, 75 samples of kankankan were collected from wholesalers (6), sellers of kankankan in the markets (35) and sellers of roasted meat (34) across three municipalities of Abidjan, the economic capital of Côte d'Ivoire. Ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to detect and quantify nine different mycotoxins. Dietary exposure was calculated by using estimated daily intake (EDI), whereas risk characterisation was assessed using the margin of exposure (MOE) approach. Aflatoxins and fumonisins were found in 99% of samples assessed, while contamination with beauvericin was proportionally lowest (28%). At all the three types of actors within the food production chain (wholesalers, kankankan sellers and roasted meat sellers) the mean concentrations of aflatoxin B1 (AFB1) in samples exceeded the European standard for spice mixtures, with concentrations reaching up to 502 µg/kg. The estimated daily intakes of aflatoxins observed in the different populations were above the recommended level of 0.017 ng kg−1 b.w. day−1. The MOES values for adolescents and adults were 8.10 and 12.78, respectively, well below the safe margin of 10,000. The co-occurrence of mycotoxins in kankankan samples together with high aflatoxin exposure to consumers represent a potential risk to public health, calling for immediate risk management and education of kankankan producers and consumers.

The current study assesses the natural occurrence of mycotoxins and fungal metabolites in Tunisian Pearl millet. Investigations using a multi-mycotoxin liquid chromatography/electrospray ionization-tandem mass spectrometric (LC/ESI–MS/MS) method were carried out on a total of 220 pearl millet samples collected from farmers (n = 144) located in four different agroecological zones in Tunisia and from retail shops (n = 76). Results revealed fifty-seven regulated and emerging metabolites contaminating 91.4% of grain samples. Among major mycotoxins, both aflatoxin B1 (AFB1) and ochratoxin A (OTA) were the most prevalent at rate of 8.6% each and occurring at an average level of 106 and 69.4 μg/kg, respectively. All positives samples were significantly exceeding the European thresholds (5 and 3 μg/kg, respectively). Lower incidence (5.9%) of Zearalenone (ZEA) contaminated samples at levels below the authorized limits (100 μg/kg) was recorded. Pearl millet grains from the south coastal zone (CT.S) had significantly the highest mean AFB1 contamination (p < 0.05). Meanwhile, OTA was encountered only in CT.S zone. Furthermore, commercial samples showed higher OTA and ZEA mean concentrations than farmers samples (p < 0.05). As regards emerging mycotoxins, Fusarium mycotoxins including beauvericin (BEA), equisetin (EQUS), monoacetoxyscirpenol (MAS) and diacetoxyscirpenol (DAS) were detected in 43.2%, 43.2%, 11.4% and 10.5% of positive samples, respectively. Furthermore, 12.7%, 50.9%, 53.2% and 14.1% of grains were contaminated with Alternaria mycotoxins namely alternariol (AOH), alternariolmonomethyl ether (AME), tentoxin (TEN) and tenuazonic acid (TA), respectively. The majority of emerging Fusarium and Alternaria mycotoxins mean levels were significantly higher in grains from north coastal area (p < 0.01). The present study highlighted a wide range of mycotoxins in Tunisian pearl millet with a different mycotoxin patterns within the agroecological zones.

The effects of light at different wavelengths and photoperiod on growth and ochratoxin A production of Aspergillus carbonarius and Aspergillus westerdijkiae were studied: far-red (740 nm), red (625 nm), blue (445 nm), and UV-A (366 nm). Fungal growth was not significantly affected by photoperiod or light wavelength; the only exception was A. westerdijkiae which showed reduced growth under UV-A light (366 nm). Short-wavelength blue light (445 nm) and UV-A light caused a reduction in ochratoxin A production of both fungal species. However, long-wavelength red light (625 nm) and far-red light (740 nm) reduced ochratoxin A production only in A. westerdijkiae but not in A. carbonarius. It is believed that this difference in reactivity to light is due to differences in the melanin content of the two fungal species: A. carbonarius is a black fungus with higher melanin content than A. westerdijkiae, a yellow fungus. Other possible explanations for the reduction of ochratoxin A production by light were also discussed.

Ochratoxin A (OTA) is a nephrotoxic, teratogenic, immunotoxic, and carcinogenic mycotoxin which is produced in tropical zones mainly by Aspergillus carbonarius, A. niger, A. ochraceus, and A. westerdijkiae. A. ochraceus and A. westerdijkiae species are phenotypically and genomically very close but A. westerdijkiae produce OTA at a very higher level than A. ochraceus. These species have been differentiated recently. The DNA primer pairs which were drawn so far are not specific and a genomic region of the same size is amplified for both species or they are too specific, and in this case, the DNA of a single species is amplified. To help preventing OTA contamination of foodstuffs, the PCR‐DGGE (Denaturing Gradient Gel Electrophoresis) method was used to discriminate between A. ochraceus and A. westerdijkiae DNA fragments of the same size but with different sequences and thus faster access to a diagnosis of the toxigenic potential of the fungal microflora. The proposed methodology was able to differentiate A. westerdijkiae from A. ochraceus with only one primer pairs in a single run. A calibration based on initial DNA content was obtained from image analysis of the DGGE gels and a method of quantification of the two strains was proposed.

Introduction Ochratoxin A (OTA) is a toxic secondary metabolite produced by fungi of the genera Aspergillus and Penicillium. It has been shown to have carcinogenic and immunotoxic properties in rats and to be responsible for human and animal kidney pathologies. OTA content in coffee was shown to be closely link to harvesting conditions, postharvest processing conditions and especially dry processing, storage and transportation conditions. Purpose Fairly little is known about the conditions for contamination by fungi responsible for OTA production, their propagation and conditions for OTA production. Biodiversity and dynamics of fungal populations linked to OTA production could be studied by PCR-DGGE genetic fingerprinting with the aim to understand the effects of postharvest processing on the microbiota. Results DGGE fingerprints analyzed by multivariate analysis showed an evolution of the fungal microflora of coffee during the different stages of the postharvest treatments (wet and dry processes) by extraction and amplification of 26S (yeasts) and 28S rDNA (filamentous fungi). PCR-DGGE stages were optimized: extraction and amplification, repeatability and sensitivity methodology applied to fungi were tested. Conclusion PCR-DGGE is a rapid molecular technique to monitor the dynamics of coffee microbial populations (fungi and yeast). Significance of study PCR-DGGE is a promising tool in order to investigate OTA production in coffee beans.

Ochratoxin A (OTA) is a mycotoxin that contaminates several foodstuffs, including cocoa. It has nephrotoxic, teratogenic and carcinogenic properties in humans. The effect of post-harvest processing and storage on the OTA contamination of cocoa was studied over three successive cocoa seasons (2005, 2006 and 2007) in Cameroon. The type of fermentation (box or heap) did not significantly influence bean OTA content, which varied from undetectable (

Cocoa beans are the principal raw material for chocolate manufacture. Moulds have an important place in the change in the quality of cocoa beans due to their role in the production of free fatty acids and mycotoxins, namely ochratoxin A (OTA). This study investigated the impact of the key post-harvest treatments, namely the fermentation and drying methods on OTA contamination of raw cocoa beans. Analytical methods for OTA detection were based on solid-liquid extraction, clean-up using an immunoaffinity column, and identification by reversed-phase HPLC with fluorescence detection. Of a total of 104 randomly selected cocoa samples analysed, 32% had OTA contents above 2 µg kg(-1). Cocoa sourced from pods in a bad state of health had a maximum OTA content of 39.2 µg kg(-1), while that obtained from healthy pods recorded 11.2 µg kg(-1). The production of OTA in cocoa beans increased according to the pod-opening delay and reached 39.2 µg kg(-1) after an opening delay of 7 days after harvest, while 6.1 and 11.2 µg kg(-1) were observed when pods were opened after 0 and 4 days. OTA production also seemed to depend considerably to the cocoa fermentation materials. When using plastic boxes for bean fermentation, the OTA production was enhanced and reached an average OTA content of about 4.9 µg kg(-1), while the raw cocoa treated in banana leaves and wooden boxes recorded 1.6 and 2.2 µg kg(-1) on average respectively. In parallel, the OTA production was not really influenced by either the mixing or the duration of the fermentation or the drying materials.

Jerusalem artichoke is a favorable substrate for inulin or fructose syrup production. The sugar content and the fructose ratio of inulin depend on various factors, particularly on the date of harvest. Incomplete fermentation of extracts by selected yeasts allows the production of inulin with increased fructose content. The yeast strains (Saccharomyces cerevisiae, S. diastaticus...) are chosen for their ability to ferment sucrose and inulin small polymers, but not easily inulin large polymers. A good increase in the fructose ratio and a good yield in residual sugars can be obtained with the better strains. After fermentation and acid or enzymatic hydrolysis, extracts from “early” and “late” harvested tubers lead to syrups of good quality containing up to 95% and 90% of fructose respectively. This fermentative enrichment process is competitive with others (for example, chromatographic enrichment), is appropriate to raw extracts, simplifies the purification steps, and also permits the simultaneous benefit of production of by-products in the form of ethanol and yeast (in addition to the pulps). Unhydrolyzed inulin polymers with high fructose content can be recovered by this selective fermentation.

Ochratoxin-A (OTA) is a mycotoxin that has nephrotoxic, tetragenic, immunotoxic and carcinogenic effects in the human organism. It contaminates several foodstuffs, notably cocoa. The purpose of our study was to compare the incidence of cocoa pod integrity on the fungal microflora and ochratoxin- A production in Cameroon. Irrespective of pod condition, fermented cocoa beans were contaminated by OTA. The maximum mould content was obtained in beans from damaged pod. However, the fungal microflora was more diversified for beans from pod damaged during harvesting than for beans from intact pods, throughout the post-harvest process. The toxigenic strains isolated belonged to the genera Aspergillus. Aspergillus carbonarius isolated from damaged pods, displayed the greatest OTA production which was 110.7 ng.g-1 on cocoa medium and 2772,0 ng.g-1 on official medium. There was a good correlation between OTA presence in the beans and isolated toxigenic strains.

Yeast flocculation presents a great interest for the industry of fermentation but its mechanism is still not fully understood. In order to enlighten this mechanism, the flocculation of Saccharomyces diastaticus NCYC 625 was studied. As with other Saccharomyces strains, the effects of different factors affecting flocculation and deflocculation (pH, temperature, medium, EDTA, cations…) suggest a lectin-like binding between adjoining cells. The genetic determinism of flocculation is nuclear and not mitochondrial. Although Saccharomyces diastaticus NCYC 625 could be classified in the FLO1 phenotype according to Stratford and Assinder42, allelism tests show that the gene involved in the flocculation control is not allelic with FLO1 or FLO5 and possibly different from FLO8.

At high flow rates, the continuous alcoholic fermentation of sucrose in a laboratory fermenter with internal cell recycle, using a strongly flocculating yeast can be limited by the substrate hydrolysis. This system is sensitive to glucose catabolic repression and to mineral deficiency. The release of invertase activity in the medium is negligible. From theoretical and experimental considerations, the hydrolysis rate is imposed by diffusionnal limitations in the biomass particles. Nevertheless, ethanol productivities as high as 68 g/L.h can be reached, without biomass retention problems. A better understanding of the basic phenomena involved in floc formation and evolution is required to control reactor performances.

Ochratoxin A (OTA) is the main mycotoxin occurring in cocoa. A study was conducted in Cameroon to assess how filamentous fungi and toxigenesis were affected by the type of cocoa post-harvest treatment (boxes or heaps). The filamentous fungi isolated were almost identical when fermentation was carried out in boxes or heaps, with the presence of abundant black Aspergillus filamentous fungi: A. niger and A. carbonarius. Filamentous fungi were more abundant at the end of the harvesting season. Factors affecting bean integrity (poor handling, deferred processing) resulted in a qualitative and quantitative increase in contamination, when the total number of filamentous fungi could reach a maximum value of 5.5+/-1.4x10(7) CFU g(-1) and black Aspergilli a maximum value of 1.42+/-2.2x10(7) CFU g(-1). A toxigenesis study showed that Aspergillus carbonarius was the main OTA-producing strain isolated. Its maximum production could reach 2.77 microg g(-1) on rice medium. Aspergillus niger strains did not always produce OTA and their toxigenesis was much lower. Fermented dried cocoa from poor quality pods was the most contaminated by OTA: up to 48 ng g(-1).