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Aims: Contamination with filamentous fungi reduces the quality of cocoa beans and poses a health risk for consumers due to the potential accumulation of mycotoxins. The aim of this study was to develop antifungal Bacillus cultures for cocoa beans fermentation. Methods and Results: Antifungal activity of 7 Bacillus isolates was investigated using overlay technique. Solid culture media inoculated with bacterial and fungal isolates were cultivated under aerobic conditions at 30 ºC for 5 days. The Bacillus strain with strong antifungal activity was studied and inoculated to 18 kg box fermentations cocoa beans in Cote d'Ivoire in three time-independent replications each including a spontaneous control fermentation. The comparison of inoculated and spontaneous fermentation processes revealed that the Bacillus thuringiensis strain ATCC 10792 don't affected the fermentation process and cocoa bean quality. The anti-fungal in vivo assays revealed that the Bacillus thuringiensis strain ATCC 10792 completely inhibited growth of all fungi on the surface of cocoa beans. The result showed that sensory evaluation of chocolates is not significant differences for all treatment. The sensory evaluation of control and chocolates made from the inoculated cocoa beans with Bacillus showed excellent sensory quality, demonstrating that Bacillus thuringiensis strain ATCC 10792 did not affect aroma, texture and appearance of chocolate. Conclusion: From this study, it is revealed that the addition of Bacillus thuringiensis strain ATCC 10792 during fermentation cocoa beans suppress fungal growth and improve the quality of cocoa beans. Therefore, the anti-fungal culture Bacillus thuringiensis strain ATCC 10792 is recommended for future applications and its capacity to limit fungal growth during industrial-scale cocoa bean fermentation should be investigated in further studies.

Aims: Mold contamination of foods especially by mycotoxin producing fungi is not only a global food quality concern for food manufacturers, but it also constitutes a high risk for human and animal health resulting in massive economic losses globally. This study investigated the effect of Lactic Acid Bacteria (LAB) on the growth of Aspergillus carbonarius strains and their production of ochratoxin A (OTA). Methodology: Seven fresh coffee cherry and 9 dry coffee cherry samples were collected from Man, Daloa and Akoupé 3 main coffee producing regions in Côte d’Ivoire. LAB were isolated from fresh coffee cherries while mold strains were from both fresh and dry coffee cherry. The inhibitory effect against mold growth and the ability for OTA removal of selected LAB strains were tested successively in vitro and then during coffee cherry primary postharvest processing before evaluating their influence on sensory quality of beverage. OTA production ability of molds strains are studied using both solid (CYA) and liquid (CYB) Cazapeck Yeast medium. Results: About 34 fungal isolates belonging to Aspergillus and Penicillium genus were studied for their OTA production using the agar plug technique and HPLC-FD. Five A. carbonarius strains were capable of OTA production between 15.9 and 83mg.kg-1.Out of seven isolates of Lactobacillus plantarum, two were successful in inhibition of mycelial growthproduced fungicidal activity; five were successful in retarding it produced fungistatical activity All of L. plantarum isolates exhibited OTA reduction ability at about 99 %. The inoculation of two highest anti-ochratoxigenic LAB to fermenting coffee cherries resulted in great inhibition of mold growth and OTA contents reduction varying from 63.2 to 82.2%. The addition of LAB to coffee cherries did not influence the sensory attributes of the beverages produced thereof. Conclusion: This study highlighted that LAB are very promising biological candidates for reduction of mold contamination and removal OTA from coffee cherry during primary postharvest processing.

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.

The aim of this study was to evaluate the interactions between wheat plant (spikelets and straws), a strain of mycotoxigenic pathogen Fusarium graminearum and commercial biocontrol agents (BCAs). The ability of BCAs to colonize plant tissue and inhibit the pathogen or its toxin production was observed throughout two phases of the life cycle of pathogens in natural conditions (colonization and survival). All evaluated BCAs showed effective reduction capacities of pathogenic traits. During establishment and the expansion stage, BCAs provoked an external growth reduction of F. graminearum (77–93% over the whole kinetic studied) and mycotoxin production (98–100% over the whole kinetic studied). Internal growth of pathogen was assessed with digital droplet polymerase chain reaction (ddPCR) and showed a very strong reduction in the colonization of the internal tissues of the spikelet due to the presence of BCAs (98% on average). During the survival stage, BCAs prevented the formation of conservation perithecia of the pathogen on wheat straw (between 88 and 98% of perithecia number reduction) and showed contrasting actions on the ascospores they contain, or perithecia production (−95% on average) during survival form. The mechanisms involved in these different interactions between F. graminearum and BCAs on plant matrices at different stages of the pathogen’s life cycle were based on a reduction of toxins, nutritional and/or spatial competition, or production of anti-microbial compounds.

The aim of this study was to develop a set of experiments to screen and decipher the mechanisms of biocontrol agents (BCAs), isolated from commercial formulation, against two major mycotoxigenic fungi in cereals, Fusarium graminearum and Fusarium verticillioides. These two phytopathogens produce mycotoxins harmful to human and animal health and are responsible for the massive use of pesticides, for the protection of cereals. It is therefore essential to better understand the mechanisms of action of alternative control strategies such as the use of BCAs in order to optimize their applications. The early and late stages of interaction between BCAs and pathogens were investigated from germination of spores to the effects on perithecia (survival form of pathogen). The analysis of antagonist activities of BCAs revealed different strategies of biocontrol where chronological, process combination and specialization aspects of interactions are discussed. Streptomyces griseoviridis main strategy is based on antibiosis with the secretion of several compounds with anti-fungal and anti-germination activity, but also a mixture of hydrolytic enzymes to attack pathogens, which compensates for an important deficit in terms of spatial colonization capacity. It has good abilities in terms of nutritional competition. Trichoderma asperellum is capable of activating a very wide range of defenses and attacks combining the synthesis of various antifungal compounds (metabolite, enzymes, VOCs), with different targets (spores, mycelium, mycotoxins), and direct action by mycoparasitism and mycophagy. Concerning Pythium oligandrum, its efficiency is mainly due to its strong capacity to colonize the environment, with a direct action via microbial predation, stimulation of its reproduction at the contact of pathogens and the reduction of perithecia formation.

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.

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.

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 prevalence of diarrheal diseases in children aged from 6 to 24 months in Burkina Faso is 38%. These diarrheas may be due to the consumption of contaminated weaning food. Therefore, the microbiological quality of follow up infant flours used as supplement foods is a key-point to reduce children diseases. In this study, the microbiological safety of locally-produced infant flours was investigated. One hundred and ninety-nine (199) samples were collected mainly in retails outlets and in Recovery and Nutrition Education Centers. According to the Burkina Faso regulations, microbiological analyses were carried out for Total Aerobic Mesophilic Flora (TAMF), thermotolerant coliforms, Salmonella spp. and yeasts/molds. The bacterial and fungal isolates were identified using phenotypic and genotypic methods and the study of the production of mycotoxins was carried out from the fungal isolates. In collected samples, the TAMF count ranged from 0 to 1.8 × 106 CFU/g with a total average of 6.3 × 104 CFU/g. About 2% of the samples had a microbial load exceeding the standards (105 CFU/g). No Salmonella spp. was isolated in the final infant flours. However, the presence of Enterobacteriaceae (Klebsiella spp. Enterobacter spp. and Cronobacter spp.) was detected and molecular characterization revealed also the presence of fungal species of the genus Aspergillus spp., Penicillium spp. and Fusarium spp. Some of these species were found to produce aflatoxins, ochratoxin A and fumonisins, which are potential carcinogenic toxins. These results demonstrated the need for a preventive approach based on the application of Hazard Analysis Critical Control Point in the food industry to ensure food safety of infant flours in Burkina Faso.

Considering the demand from many industrial sectors, the production of microbial lipids constitutes an attractive topic for research. Oleaginous microorganisms can produce over 60% of their dry matter in single cell oils (SCOs), especially in the form of triglycerides (TAGs). Approximately forty microbial genera, especially yeasts and moulds, are known to be high lipid producers. Many abiotic factors, such as the culture conditions or nutriment availability, influence the production of microbial lipids that are of interest. Among the factors that can influence SCO production, the carbon source, limiting nutriments (especially nitrogen), pH and temperature are cited most often in the literature. Improvement in SCO production through the optimisation of abiotic factors encounters several limitations. Thus, research is currently increasingly oriented toward genetic modifications to optimise the synthesis of lipids by oleaginous microorganisms.

L’ochratoxine A (OTA) est une toxine cancerogene produite par certaines moisissures. Elle contamine de nombreux aliments et notamment le cafe dans lequel elle est retrouvee a toutes les etapes du parcours technologique du champ a la tasse. Cette etude a pour objet de faire le point sur les etapes critiques des procedes de transformation du cafe, que ce soit durant les traitements post-recolte ou pendant la transformation en produit consommable, en ce qui concerne sa contamination par l’OTA. La qualite et le type de traitements post-recolte ont une influence non negligeable sur le niveau de contamination en OTA des grains verts. Les defauts agronomiques (insectes et attaques fongiques) contribuent egalement a un niveau eleve d’OTA dans le cafe vert. Diverses etudes effectuees sur l’impact de la torrefaction mettent en evidence une degradation thermique de l’OTA. Toutefois, la contamination initiale des feves, le procede utilise et le degre de torrefaction influencent fortement cette degradation thermique et donc la concentration residuelle en OTA dans les grains torrefies. Finalement, il apparait que c’est la maitrise de la contamination en amont par de bonnes pratiques agricoles et par les traitements post-recolte qui est le plus sur gage de salubrite pour la boisson.

Dracaena cochinchinensis Lour. is an ethnomedicinally important plant used in traditional Chinese medicine known as dragon’s blood. Excessive utilization of the plant for extraction of dragon’s blood had resulted in the destruction of the important niche. During a study to provide a sustainable way of utilizing the resources, the endophytic Actinobacteria associated with the plant were explored for potential utilization of their medicinal properties. Three hundred and four endophytic Actinobacteria belonging to the genera Streptomyces, Nocardiopsis, Brevibacterium, Microbacterium, Tsukamurella, Arthrobacter, Brachybacterium, Nocardia, Rhodococcus, Kocuria, Nocardioides, and Pseudonocardia were isolated from different tissues of D. cochinchinensis Lour. Of these, 17 strains having antimicrobial and anthracyclines-producing activities were further selected for screening of antifungal and cytotoxic activities against two human cancer cell lines, MCF-7 and Hep G2. Ten of these selected endophytic Actinobacteria showed antifungal activities against at least one of the fungal pathogens, of which three strains exhibited cytotoxic activities with IC50-values ranging between 3 and 33 μg·mL−1. Frequencies for the presence of biosynthetic genes, polyketide synthase- (PKS-) I, PKS-II, and nonribosomal peptide synthetase (NRPS) among these 17 selected bioactive Actinobacteria were 29.4%, 70.6%, and 23.5%, respectively. The results indicated that the medicinal plant D. cochinchinensis Lour. is a good niche of biologically important metabolites-producing Actinobacteria.

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.