Mould Strains Research Articles

Potential Production of Mycotoxins by the Fungal Flora Isolated from "Garba" (Côte D'ivoire)

Garba, an Ivorian dish composed of attiéké (a fermented cassava couscous), fried tuna, and condiments, is increasingly consumed by the population, particularly in the streets of Abidjan. However, garba could pose a health risk to consumers. This study, aimed at preserving public health, sought to identify the toxigenic fungal flora present in garba sold on the streets of Abidjan. To this end, 300 garba samples were collected from four districts of Abidjan. Fungal strains contaminating the garba were enumerated and identified using classical mycological methods. Mycotoxins (aflatoxins and ochratoxins) were quantified using High-Performance Liquid Chromatography (HPLC). Results showed that the average mold counts ranged from 0 to 1.8 x 10^3 CFU/g. The isolated mold strains belonged to ten species grouped into four genera: Aspergillus, Mucor, Penicillium and Paecilomyces, with Mucor spp. (36.89%) and Aspergillus niger (30.09%) being the most predominant. The mycotoxins detected in garba were aflatoxins (B1, B2, G1, and G2) and ochratoxin A, with average levels ranging from 0.42 to 8.07 µg/kg. Approximately 37% and 23% of garba samples had total aflatoxin and aflatoxin B1 levels exceeding regulatory limits. The presence of potentially mycotoxigenic fungal strains in garba could pose a health risk to consumers. Compliance with good hygiene practices during the production and sale of garba will therefore be necessary to considerably reduce contamination and protect consumer health.

New Refined Experimental Analysis of Fungal Growth in Degraded Bio-Based Materials

When exposed to different building environmental conditions, bio-composite materials, such as hemp mortars, represent a risk of mold proliferation. This later plays a critical role in the biodeterioration of the materials when their physical properties are locally modified by the natural aging process. The primary objectives of the present work are first to assess the evolution of the surface of contaminated mortar; second, to investigate an accurate DNA extraction method that could be used for both bio-composite mortars and their fiber sources collected in situ; then, to understand the process of the proliferation of mold strains on both hemp shives and hemp mortar; and finally, to compare mold strains present in these phases to show their relationship to mold contamination and their impact on human health. In situ hemp mortar contamination behavior was investigated in the region of Pau (France) two months after hemp mortar application in extreme conditions (high humidity, low temperature, no aeration), which did not match the standard conditions under which hemp mortar must be used. The SEM observations and FTIR and pH analyses highlighted the decrease in pH level and the presence of organic matter on the mortar surface. DNA sequencing results showed that hemp shives were the main source of fungal contamination of hemp mortar. A mold population analysis showed that the most dominant phylum was Ophistokonta, which represented 83.6% in hemp shives and 99.97% in hemp mortar. The Acrostalagmus genus representatives were the most abundant, with 42% in hemp shives and 96% in hemp mortar. The interconnection between the mold strain characteristics (particularly the ability to grow in extreme environments) and the presence of hemp mortar was emphasized.

Solid-state cultivation of multiple industrial strains of koji mold on different Thai unpolished rice cultivars: biotransformation of phenolic compounds and their effects on antioxidant activity.

Colored rice is abundant in polyphenols, and koji molds have potential for biotransformation. This study aimed to produce Thai-colored rice koji to study its polyphenolic biotransformation. Four industrial koji mold strains: Aspergillus oryzae 6001, A. oryzae 6020, A. sojae 7009, and A. luchuensis 8035, were cultivated on unpolished Thai-colored rice (Riceberry and Sangyod), unpolished Thai white rice (RD43), and polished Japanese white rice (Koshihikari). We discovered that koji molds grew on all the rice varieties. Methanol extracts of all rice kojis exhibited an approximately 2-fold or greater increase in total phenolic content and DPPH antioxidant activity compared to those of steamed rice. Moreover, quercetin, quercetin-3-O-glucoside, isorhamnetin-3-O-glucoside, ferulic acid, caffeic acid, protocatechuic acid, vanillic acid, (+)-catechin, and (-)-epicatechin content increased in Riceberry and Sangyod koji samples. Consequently, Aspergillus solid-state cultivation on unpolished Thai-colored rice exhibited higher functionalization than the cultivation of unpolished Thai white rice and polished Japanese white rice.

Assessment of the Traditional Processing Methods in the Reduction of Aflatoxin Levels in Maize and Maize Products in Kenya

Aflatoxins are fungal secondary toxic metabolites produced predominantly by certain strains of the Aspergillus molds. The objective of this study was to investigate the effectiveness of Kenyan traditional processing methods in reduction of aflatoxin in maize. The traditional methods used included; alkaline cooking using Magadi soda and maize cob ash, decortication by pounding wet maize with mortar and pestle, dehulling in mechanical mill and exposure to solar radiation. Standardization of alkali cooking was done before actual treatment of the contaminated samples. Naturally contaminated samples of maize were sampled and used in the experiment. Aflatoxin analysis before and after each of the treatments was done using ELISA method and total aflatoxins expressed in parts per billion (ppb) (dry weight basis). The results were subjected to statistical analysis at 5% significance levels (p≤0.05). The reduction of aflatoxin was found to be highest when maize was boiled in maize cob ash solution which resulted in loss of aflatoxin from 83.1±0.3 ppb to 7.0±3.9 ppb. Dry decortication reduced aflatoxin from 51.3±15.3 to 9.6±0.8 ppb, boiling in Magadi soda led to a drop from 59.5±3.818 ppb to 13.4±0.424 ppb, solar irradiation caused a drop from 60.8±1.8 ppb to 13.7±0.1 ppb and pounding in Magadi soda resulted in least loss of aflatoxin from 81.5±0.3 to 72.7±0.2ppb. Aflatoxin loss in all the treatments was significant except for dry decortication. Only dry decortication and boiling in maize cob ash solution brought down the aflatoxin levels to below the maximum allowable limit as set by the Kenya Bureau of Standards (10ppb)

Chitosan as an Antimicrobial, Anti-Insect, and Growth-Promoting Agent for Potato (Solanum tuberosum L.) Plants.

A growing trend in plant protection is replacing chemical preparations with environmentally friendly biological compositions. Chitosan, due to its biocompatibility, biodegradability, and bioactivity, is an effective agent against plant diseases. The purpose of the study was to evaluate chitosan as a potential biopesticide for potato plants. Three variants of chitosan were tested: high (310-375 kDa, >75% deacetylated), medium (190-310 kDa, 75-85% deacetylated), and low (50-190 kDa, 75-85% deacetylated) molecular weight. The chitosan variants were dissolved in lactic and succinic acids and tested for antibacterial and antifungal properties against eight strains of mould and two strains of bacteria responsible for potato diseases. The possible cytotoxicity of chitosan was evaluated against different cell lines: insect Sf-9, human keratinocyte HaCaT, and human colon carcinoma Caco-2. The bioprotective activities of the chitosan were also evaluated in situ on potato tubers. Chitosan inhibited the growth of almost all the selected phytopathogens. The most active was medium molecular chitosan in lactic acid. This formula was characterized by low toxicity towards human cells and high toxicity towards Sf-9 cells. It was also found to have positive effects on the growth of stems and roots, gas exchange, and chlorophyll index in potato plants. Selected chitosan formulation was proposed as a functional biopesticide for potato protection against phytopathogens.

Screening, identification, and characterization of molds for brewing rice wine: Scale-up production in a bioreactor.

Rice wine, well known for its unique flavor, rich nutritional value, and health benefits, has potential for extensive market development. Rhizopus and Aspergillus are among several microorganisms used in rice wine brewing and are crucial for determining rice wine quality. The strains were isolated via Rose Bengal and starch as a combined separation medium, followed by oenological property and sensory evaluation screening. The strain exhibiting the best performance can be screened using the traditional rice wine Qu. The strains YM-8, YM-10, and YM-16, which exhibited strong saccharification and fermentation performance along with good flavor and taste, were obtained from traditional rice wine Qu. Based on ITS genetic sequence analysis, the YM-8, YM-10, and YM-16 strains were identified as Rhizopus microsporus, Rhizopus arrhizus, and Aspergillus oryzae. The optimum growth temperature of each of the three strains was 30°C, 32°C, and 30°C, and the optimum initial pH was 6.0, 6.5, and 6.5, respectively. The activities of α-amylase, glucoamylase, and protease of YM-16 were highest at 220.23±1.88, 1,269.04±30.32, and 175.16±1.81 U/g, respectively. The amino acid content of rice wine fermented in a 20-L bioreactor with the three mold strains was higher than that of the control group, except for arginine, which was significantly lower than that of the control group. The total amino acid content and the total content of each type of amino acid were ranked as YM-16 > YM-8 > YM-10 > control group, and the amino acid content varied greatly among the strains. The control group had a higher content, whereas YM-8 and YM-16 had lower contents of volatile aroma components than the control group and had the basic flavor substances needed for rice wine, which is conducive to the formation of rice wine aroma. This selected strain, YM-16, has strong saccharification and fermentation ability, is a rich enzyme system, and improves the flavor of rice wine, thereby demonstrating its suitability as a production strain for brewing.

Identification of Food Spoilage Fungi Using MALDI-TOF MS: Spectral Database Development and Application to Species Complex.

Fungi, including filamentous fungi and yeasts, are major contributors to global food losses and waste due to their ability to colonize a very large diversity of food raw materials and processed foods throughout the food chain. In addition, numerous fungal species are mycotoxin producers and can also be responsible for opportunistic infections. In recent years, MALDI-TOF MS has emerged as a valuable, rapid and reliable asset for fungal identification in order to ensure food safety and quality. In this context, this study aimed at expanding the VITEK® MS database with food-relevant fungal species and evaluate its performance, with a specific emphasis on species differentiation within species complexes. To this end, a total of 380 yeast and mold strains belonging to 51 genera and 133 species were added into the spectral database including species from five species complexes corresponding to Colletotrichum acutatum, Colletotrichum gloeosporioides, Fusarium dimerum, Mucor circinelloides complexes and Aspergillus series nigri. Database performances were evaluated by cross-validation and external validation using 78 fungal isolates with 96.55% and 90.48% correct identification, respectively. This study also showed the capacity of MALDI-TOF MS to differentiate closely related species within species complexes and further demonstrated the potential of this technique for the routine identification of fungi in an industrial context.

Evaluation of Cold Plasma–Activated water “…Enriched Metal…” Cations as an Antifungal Agent for Controlling of Penicillium italicum and Penicillium digitatum Molds

The utilization of Cold Plasma (CP) technology for decontamination and disinfection has garnered considerable attention across diverse industries. This study aims to investigate the interaction between pH and electrical conductivity (EC) (μS/cm) in Cold Plasma-Activated Water (CPAW) enriched with metal cations and its potential as an antifungal agent against two Penicillium (P.) mold strains. The investigation focuses on elucidating the augmented chemical interactions induced by plasma between radicals, charged particles, and microorganisms' cell membranes within an aqueous environment. Our findings demonstrate a positive correlation between the inactivation potential of CPAW (operating at 10 kV voltage, 2.5 kHz high frequency, and 500 mA current intensity) and pH and EC(μS/cm) values. Notably, the relative chemical reactivity and solubility of calcium oxide emerge as significant factors, highlighting the pronounced link between P. Italicum and Plasma-Activated Water containing Copper cations (CPAW + Cu2+) (p < 0.05). Our study distinctly emphasizes (1) the substantial impact of both activated water type and mold species on CFU/mL values (p < 0.05); (2) the mold-specific effect of activated water on CFU/mL; and (3) the noteworthy EC(μS/cm) enhancement and pH decrease with prolonged activation time, attaining statistical significance (p < 0.01).

Glass bead system to study mycotoxin production of Aspergillus spp. on corn and rice starches

Mycotoxin production by aflatoxin B1 (AFB1) -producing Aspergillus flavus Zt41 and sterigmatocystin (ST) -hyperproducer Aspergillus creber 2663 mold strains on corn and rice starch, both of high purity and nearly identical amylose-amylopectin composition, as the only source of carbon, was studied. Scanning electron microscopy revealed average starch particle sizes of 4.54 ± 0.635 µm and 10.9 ± 2.78 µm, corresponding to surface area to volume ratios of 127 1/µm for rice starch and 0.49 1/µm for corn starch. Thus, a 2.5-fold difference in particle size correlated to a larger, 259-fold difference in surface area. To allow starch, a water-absorbing powder, to be used as a sole food source for Aspergillus strains, a special glass bead system was applied. AFB1 production of A. flavus Zt41 was determined to be 437.6 ± 128.4 ng/g and 90.0 ± 44.8 ng/g on rice and corn starch, respectively, while corresponding ST production levels by A. creber 2663 were 72.8 ± 10.0 µg/g and 26.8 ± 11.6 µg/g, indicating 3–fivefold higher mycotoxin levels on rice starch than on corn starch as sole carbon and energy sources.Key points• A glass bead system ensuring the flow of air when studying powders was developed.• AFB1 and ST production of A. flavus and A. creber on rice and corn starches were studied.• 3–fivefold higher mycotoxin levels on rice starch than on corn starch were detected.

Extraction Kinetics and Antifungal Activities of Essential Oil from the Aerial Parts of Gros Baume (Hyptis suaveolens) Against Three Fungal Strains in Côte d'Ivoire

Introduction: Microscopic fungi colonize many food substrates resulting in considerable economic losses. Their development leads to an alteration in the organoleptic quality of the food, and produces mycotoxins. The aim of the present work was to evaluate the extraction kinetics and in vitro antifungal activity of the essential oil of Hyptis suaveolens on the growth of three toxigenic fungal strains. Methodology: The essential oil was extracted by steam distillation using a Clevenger-type device. It was tested for in vitro antifungal activity against three mold strains (Aspergillus flavus, Aspergillus niger and Fusarium sp) using the agar dilution method. Results: The results of the extraction kinetics evaluation show that in fifty minutes of distillation, almost all the essential oil contained in the aerial parts of H. suaveolens is recovered. In vitro antifungal tests showed a slight slowdown in mycelial growth for the three strains studied. However, the antifungal parameters determined show that these strains are sensitive to 50 μL/mL with a CMF of 100 μL/mL. This is just a simple fungistatic effect of this extract on the mycelial growth of the strains tested. Aspergillus strains seem to be slightly more affected than Fusarium strains. Conclusion: The antifungal potential of H. suaveolens essential oil does not offer a novel approach to the control of these three fungal strains.

Antifungal Activity, Total Phenolic Content and Antioxidant Activity Properties of Some Spices Extracts as Alternative Natural Antimicrobial Agents

In this study, extracts were obtained from rosemary, anise, cinnamon, ginger, peppermint, turmeric, fennel, clove, laurel leaves and thyme. The total phenolic content amount, antioxidant activity value and antifungal properties of these extracts were aimed to determine the extracts. Among the extracts, clove, cinnamon, turmeric and ginger were superior in terms of total phenolic content values, clove, cinnamon, turmeric, ginger, laurel leaves and rosemary extracts were superior in terms of antioxidant activity. The highest inhibition zone diameters among mold strains were determined by the use of extracts of cinnamon, turmeric, ginger, clove and laurel leaves against Aspergillus oryzae, Penicillium digitatum and Aspergillus niger strains. The results suggested the potential use of cinnamon and clove extracts as natural agents.

Specific Antimicrobial Activities Revealed by Comparative Evaluation of Selected Gemmotherapy Extracts.

Nowadays, unprecedented health challenges are urging novel solutions to address antimicrobial resistance as multidrug-resistant strains of bacteria, yeasts and moulds are emerging. Such microorganisms can cause food and feed spoilage, food poisoning and even more severe diseases, resulting in human death. In order to overcome this phenomenon, it is essential to identify novel antimicrobials that are naturally occurring, biologically effective and increasingly safe for human use. The development of gemmotherapy extracts (GTEs) using plant parts such as buds and young shoots has emerged as a novel approach to treat/prevent human conditions due to their associated antidiabetic, anti-inflammatory and/or antimicrobial properties that all require careful evaluations. Seven GTEs obtained from plant species like the olive (Olea europaea L.), almond (Prunus amygdalus L.), black mulberry (Morus nigra L.), walnut (Juglans regia L.), blackberry (Rubus fruticosus L.), blackcurrant (Ribes nigrum L.) and bilberry (Vaccinium myrtillus L.) were tested for their antimicrobial efficiency via agar diffusion and microbroth dilution methods. The antimicrobial activity was assessed for eight bacterial (Bacillus cereus, Staphylococcus aureus, Salmonella enterica subsp. enterica, Proteus vulgaris, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes), five moulds (Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Penicillium citrinum, Penicillium expansum) and one yeast strain (Saccharomyces cerevisiae). The agar diffusion method revealed the blackberry GTE as the most effective since it inhibited the growth of three bacterial, four moulds and one yeast species, having considered the total number of affected microorganism species. Next to the blackberry, the olive GTE appeared to be the second most efficient, suppressing five bacterial strains but no moulds or yeasts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for each GTE and the microorganisms tested. Noticeably, the olive GTE appeared to feature the strongest bacteriostatic and bactericidal outcome, displaying specificity for S. aureus, E. faecalis and L. monocytogenes. The other GTEs, such as blueberry, walnut, black mulberry and almond (the list indicates relative strength), were more effective at suppressing microbial growth than inducing microbial death. However, some species specificities were also evident, while the blackcurrant GTE had no significant antimicrobial activity. Having seen the antimicrobial properties of the analysed GTEs, especially the olive and black mulberry GTEs, these could be envisioned as potential antimicrobials that might enhance antibiotic therapies efficiency, while the blackberry GTE would act as an antifungal agent. Some of the GTE mixtures analysed have shown interesting antimicrobial synergies, and all the antimicrobial effects observed argue for extending these studies to include pathological microorganisms.

A remarkable step in the aerobic biological treatment of Olive Mill Wastewater (OMW): A combination of selected microbial strains that enhance their decolorization and depollution

The olive industry extracts oil from olives but also generates solid co-products called pomace and liquid wastes known as Olive Mill Wastewater (OMW). With global annual production exceeding 30 million tons and approximately 685,000 tons in Morocco alone, these wastes pose environmental challenges due to their high acidity, organic load, and phenolic compounds. Our research aims to depollute and recycle OMW using aerobic biological treatment methods. Samples were collected from various ecological sites across four Moroccan regions. We isolated and purified several strains of molds, yeasts, and bacteria capable of decolorizing OMW. Decolorization experiments revealed promising results, with a combination of seven selected molds showing significant reductions in chemical oxygen demand (COD) by 71.44%, biochemical oxygen demand (BOD5) by 69.91%, and polyphenols content by 84.22%. Encouraged by these findings, we propose further treatment using sourdoughs composed of combinations of different pure strains, including yeasts and selected bacteria such as Bacillus subtilis and Pseudomonas aeruginosa. This approach demonstrates a practical and cost-effective method for depolluting and recycling OMW, contributing to environmental protection and human health preservation. By mitigating the risks associated with untreated OMW discharge, this study offers a viable solution to the environmental challenges posed by olive processing industries globally, particularly in regions like Morocco where olive cultivation is significant.

Green process of perillaldehyde modification on regulating hydrophobicity to improve bamboo durability

Bamboo exhibits potential as a viable substitute for wood; nevertheless, its vulnerability to mold poses a significant obstacle to its practical implementation. To address this issue, prevalent techniques, including heat treatment, chemical modification, and compression treatment, alter the characteristics of bamboo and safeguard it against mold. However, these modifications are limited by substantial energy consumption, elevated expenses, and low sustainability. In light of these challenges, perillaldehyde (PAE) emerges as a natural antimicrobial agent capable of impeding the proliferation of diverse mold strains. In this study, the immobilization of PAE in bamboo was achieved for the first time using diethylenetriamine, forming a Schiff base structure with bamboo components. The modified bamboo exhibited three-fold enhanced surface hydrophobicity, which effectively prevented mold infections. Additionally, the modification process improved the dimensional stability of bamboo, albeit at the expense of a slight reduction in elasticity and hardness. The structural analysis of bamboo by FTIR, XPS, and XRD techniques revealed that lignin and hemicellulose served as potential reactive components of PAE. This study presents a novel approach to bamboo preservation utilizing natural components, thereby enhancing the resistance against mold and improving the physical characteristics of bamboo.

Establishment of a Single Temperature Incubation Approach for Environmental Monitoring Samples with Focus on Mold Recoveries.

A robust environmental monitoring program is essential to properly estimate and identify microorganisms in cleanrooms, ensuring that microbial contamination remains acceptably low, and a good state of control is maintained in the manufacturing areas. The incubation conditions are important to support optimal microbial recoveries, considering that there is no single culture medium, temperature and incubation time that can recover all microorganisms. Particularly molds are quite sensitive microorganisms, and some species may have very specific nutritional and environmental needs. In this study, a two-phase approach was used to identify a single incubation-temperature approach that could recover most of the cleanroom microbial flora with focus on molds. Phase 1 included a growth promotion study performed in the laboratory using pharmacopeial and in-house strains, comparing different media (SDA and TSA) at single or dual incubation-temperature approaches for 5 or 6 days. Phase 2 was based on an in situ study where sampling was performed in different areas of a pharmaceutical facility, comparing the recoveries at different incubation conditions. In addition, extension studies of phase 1 and phase 2 were performed to get a better understanding of growth requirements for in-house molds. The results show that an incubation on Tryptone Soy Agar (TSA) at 25-30° for 3-4 days was able to recover most tested microorganisms in phase 1 and a large variety of microorganisms in phase 2, indicating that the single incubation-temperature is an optimal approach for the recovery of microorganisms in cleanrooms. Exceptions were noted for one strain of the species Cutibacterium acnes, a microaerophilic bacterium for which anaerobiosis and higher temperatures were needed, and two mold strains (Sistotrema brinkmannii and Stereum hirsutum), indicating that those molds required a specific media (Sabouraud Dextrose Agar, SDA) for their proliferation. The results showed that TSA incubated at single or dual incubation-temperature approach cannot compensate for the absence of SDA for some environmental molds that may be atypical in cleanrooms. Therefore, in addition to TSA, certain monitoring with SDA at e.g., cleanroom entrance points may be beneficial to recover molds with very specific nutritional requirements.

Phytochemical analysis, antibacterial and antifungal effect of Lavandula dentata L. essential oil and methanol extract

The aim of this study was to analyse the essential oil of Lavandula dentata from Algeria and to test the antioxidant and antimicrobial properties of this plant. The essential oil (EO) (57 constituents) included mainly α-pinene, β-pinene, nopinone, linalool, cryptone, and limonene. The plant polyphenolic contents and the antioxidant activity were determined. The antimicrobial effect of the EO and the methanolic extract (ME) was assessed against referenced and clinical bacterial strains, and also foodborne fungal isolates. The EO minimal inhibitory concentration (MIC) values varied from 0.25 to 4 mg/mL and minimal bactericidal concentrations (MBCs) were less than 8 mg/mL except for S. aureus, clinical Klebsiella, S. epidermidis, and B. subtilis. The mould strains were significantly inhibited by the EO (87.50% to 88.33%). The MIC values were 3.60–15.62 mg/mL and 0.5–4 mg/mL for ME and EO, respectively. The minimal fungicidal concentration (MFC) values ranged from 31 to 125 mg/mL and from 2 to 8 mg/mL for ME and EO, respectively.

Effectiveness of Ozonation for Improving the Microbiological Safety of Fresh-Cut Parsley (Petroselinum crispum) Leaves

In the last decade, fresh-cut plants have become a more popular flavoring additive in food. It is important to find an effective method for ensuring the safety and quality of plant materials used as food additives. Ozonated water is being considered by the EFSA for approval as a cidal agent for plant protection. The objective of this study was to evaluate the effectiveness of ozonated water in improving the microbial safety of fresh-cut parsley leaves, with a particular focus on mesophilic and psychrotrophic bacteria and fungi. The yeasts and bacteria were identified with the MALDI-TOF MS system. Color changes on the surface of the parsley samples were measured in the CIE L*a*b trichromatic color model. The chemical composition of the essential oil was evaluated via gas chromatography with mass spectrometry (GCMS). The microbial level of the fresh leaves varied and depended on the season. The highest microbial levels were found in the leaves picked in the summer and autumn, at 104 to 106 CFU/g for fungi and 106 to 108 CFU/g for bacteria. Among the isolates with the highest isolation frequency, bacteria belonging to Pseudomonas fluorescens, Staphylococcus warneri, and Bacillus megaterium dominated. The dominant yeasts and molds were Candida sp., Rhodotorula sp., Cladosporium sp., and Fusarium sp. The conditions for water ozonation (ozone dose and time) were established for both mono- and mixed cultures. Time of 3 min, ozone content of 0.5, O3 mg/L or 1 min, and 1.5 mg of O3 mg/L were sufficient for a 90% reduction in the number of living microorganisms. Yeasts and bacteria were sensitive to ozone treatment, excluding P. fluorescens rods. The tested mold strains were the most resistant. However, it was noted that organic matter might reduce the decontamination effect. The effectiveness of ozonation was negatively influenced by organic compound content above 1%. Spectrophotometric measurements of parsley leaves after ozonation, especially after 3 min treatment at 1.5 O3 mg/L, revealed morphological changes. The CIELAB color space (L*a*b*) changed in the direction of lightness and yellowness; however, ΔE showed no statistically significant differences in comparison with the untreated leaves. In preliminary studies, no differences were noted in GLC-MS chromatograms for essential oils of parsley leaves before and after ozonation. The aroma of parsley treated with ozonated water was more intensely herbal than the control sample, probably due to the higher content of α and β phellandrene. The results of this study show that decontamination of parsley leaves by ozonated water containing 1.5 O3 mg/L in a closed 5 min process can effectively ensure the microbiological quality of fresh-cut parsley leaves. It can be concluded that ozone treatments in aqueous form appear to provide promising qualitative and quantitative results for the decontamination of this fresh-cut plant material. However, more work is necessary to study chemical and volatilome changes. Especially the sensory analyses should be conducted before and after ozone treatment.

Antifungal susceptibility patterns for Aspergillus, Scedosporium and Exophiala isolates recovered from cystic fibrosis patients against amphotericin B, and three triazoles and their impact after long term therapies.

In cystic fibrosis (CF) patients, fungal colonization of the respiratory tract is frequently found. Aspergillus fumigatus, Scedosporium genus and Exophiala dermatitidis are the most commonly isolated molds from the respiratory tract secretions of patients with this genetic disorder. The aim of this 5-year surveillance study was to identify trends in species distribution and susceptibility patterns of 212 mold strains identified as Aspergillus spp., Scedosporium spp. and Exophiala spp., isolated from sputum of 63 CF patients who received long term therapy with itraconazole and/or voriconazole. The Aspergillus spp. isolates were identified as members of the sections Fumigati (n=130), Flavi (n=22), Terrei (n=20), Nigri (n=8), Nidulantes (n=1) and Usti (n=1). Among the 16 species of the genus Scedosporium, 9 were S. apiospermum, 3 S. aurantiacum and 4 S. boydii. Among the 14 Exophiala species, all were molecularly identified as E. dermatitidis. Overall, 94% (15/16) of Scedosporium spp., 50% (7/14) of E. dermatitidis, and 7.7% (14/182) of Aspergillus spp. strains, showed high MIC values (≥ 8 µg/ml) for at least one antifungal. Particularly, 8.9% (19/212) of isolates showed high MIC values to amphotericin B, 11.7% (25/212) itraconazole, 4.2% (9/212) to voriconazole and 3.3% (7/212) to posaconazole. In some cases, such as some A. fumigatus and E. dermatitidis isolates recovered from the same patient, susceptibility to antifungal azoles decreased over time. We show that the use of azoles for a long time in CF patients causes the selection/isolation of mold strains with higher MIC values.

Effect of Spoilage by Lipolytic Fungi Strains on Free Fatty Acids (FFA) Formation in Fermented and Dry Cocoa Beans

Aim: High free fatty acids (FFA) content of cocoa beans is high lead to alter quality of butter fraction and chocolate manufactured. During primary post-harvest processing, cocoa beans harbor lipase producing mold speciess could trigger the accumulation of FFA. This study investigated the improvement of the quality of raw cocoa material sourced from Côte d’Ivoire regarding its high FFA concentration.&#x0D; Methods: Fermented and dry cocoa were sampled on-farm level in some main cocoa producing regions of Côte d'Ivoire. Contaminating mold strains were isolated using Sabouraud chloramphenicol agar incubated at 25°C for 7 days. Morphological identification of mold isolates was preformed from younger fungal culture. Biochemical study about lipolytic properties of mold isolates was carried out using Cazpeck-Dox Agar medium enriched with olive oil as carbon source and rhodamine B as indicator of lipase activity. Extraction, PCR amplification using a specific transcribed spacer primer (ITS4/ITS5) and sequencing of DNA were performed for fungal species molecular identification.&#x0D; Results: Approximately 154 mold isolates belonging to 8 genera were found in tested cocoa beans. Only Rhizopus oryzae, Absidia corymbifera, Mucor heimalis, Aspergillus niger, A. tubingensis, A. aculeatus, A. flavus, A. tamari, A. fumigatus, Nigrospora sphaerica, Curvularia geniculate and Penicillium chrysogenum species produced greater lipolytic properties. A. corymbifera, A. niger, A. fumigatus and R. orysae were the most frequent and predominant species. Cocoa beans inoculated with R. oryzae, A. corymbifera or A. niger recorded higher FFA content above 1.75%.&#x0D; Conclusion: Reducing of the mold species growth during cocoa primary post-harvest processing could sustainably produce raw cocoa material with less FFA.

Restoration of Choujiu Koji and evaluation of its brewing performance

Koji is used as a fermentation starter in Choujiu manufacture in northwestern China. Traditional natural Koji (NK) cultivates in open environments, which probably leads to diverse microflora, causes inconsistent quantities in Koji, then results in instable qualities of Choujiu. A native Choujiu producer has been purchasing NK from local workshop, while their Choujiu qualities fluctuate by the stability of NK.As a request from the producer, this study aimed for stable Koji and inheriting sensory traits of current products. Strains of mold, yeast, and LAB, separated from NK, were selected for Restored Koji (RK) making. Brewing performance of RK and NK were compared by analysis of the physicochemical properties and volatile flavor compounds (VFCs) in fermented mash, combined with sensory evaluation in corresponding Choujiu.Pure Koji from individual strains was separately cultured at 28 °C for 48 h, and then reconstituted for RK II at the proportion of mold Koji (M7 Koji): yeast Koji (Y3 Koji: Y4 Koji = 1:1): LAB Koji (L1 Koji) = 1:1:1, where M7 was Rhizopus oryzae, Y3 was Saccharomyces cerevisiae, Y4 was Saccharomycopsis fibuligera, and L1 was Pediococcus pentosaceus. Results showed RK II could meet the desired purpose of this study.