Search

Purpose: The identification of anaerobes, Mycobacterium and Nocardia species, and moulds by MALDI-TOF-MS remains a challenge. This study aimed to evaluate the performance of MALDI-TOF in the identification of these organisms., Methods: A total of 382 strains, comprising 128 (33.5 %) anaerobes, 126(33.0 %) mycobacterial, 113(29.6 %), mycelial fungi, and 15(3.9 %) Nocardia species were evaluated by VITEK MS Version 3.0. The results were compared with the identification of the isolates by DNA sequence analysis. The DNA sequences used for analysis were the 16S rRNA for anaerobic bacteria, hsp65 gene for mycobacteria, whereas both 16S rRNA and hsp65 gene for Nocardia species, and internal transcribed spacer (ITS) and 28S rRNA gene's D1/D2 regions of fungi., Results: The VITEK-MS accurately identified 78.3 % (299/382) of the strains at the species, and 9.4 % (36/382) at the genus level. Misidentifications were observed in 3.9 % (15/382) isolates. Of isolates tested, 8.4 % (32/382) were not identified by the system, and 7.06 % (27/382) were not included in the IVD database., Conclusion: An upgraded VITEK MS V3.0 database provides reasonably accurate and rapid identification of clinically relevant anaerobes, mycobacteria, Nocardia species, and moulds to the species level., Competing Interests: Declaration of competing interest The author(s) declare that there are no conflicts of interest., (Copyright © 2024. Published by Elsevier Inc.)

Dermatophytic pseudomycetoma (DPM) is a rarely reported invasive fungal infection of humans and animals, especially cats. This study aimed to identify dermatophytes, breed associations, and the frequency of extracutaneous (EC) involvement in feline DPM. Electronic records and formalin-fixed paraffin-embedded tissue (FFPET) from 32 suspected DPM cases in 30 cats were retrieved from a diagnostic laboratory between 2018 and 2024. To confirm DPM and molecular identity, DNA was extracted from FFPET for ITS2 sequencing, and immunohistochemistry was performed on PCR-negative cases. All cases were confirmed as DPM. Microsporum canis was the only dermatophyte identified. The sensitivity and specificity of ITS2 sequencing for M. canis identification in FFPET were 22/32 (68.8%) and 21/22 (95.5%), respectively. Exotic (36.7%) and Persian (23.3%) but not British breeds (26.3%) were over-represented compared to feline admissions at an affiliated veterinary hospital (8.5%, p < 0.001; 3%, p < 0.001; 21.6%, p = 0.817, respectively). Five cases (16.7%) had EC lesions; two had intra-abdominal masses; two had oral cavity masses, including one which extended into the cranial vault; and one had superficial cervical lymph node invasion. Exotic and Persian breeds are over-represented for DPM and M. canis is the primary cause. EC lesions of DPM may occur more commonly than previously thought.

Background: The use of industrial Cannabis sativa L. for recreational, cosmeceutical, nutraceutical, and medicinal purposes has gained momentum due to its rich content of valuable phytochemicals, such as cannabidiol (CBD) and cannabigerol (CBG). However, there are concerns regarding the risk of microbial contamination in plants grown outside controlled environments. Microbes associated with hemp can be either epiphytes or endophytes and may pose a risk of infectious illness for humans., Methods: Seven Italian hemp genotypes, including Bernabeo, Carmagnola, Carmaleonte, Codimono, CS, Eletta Campana, and Fibranova, were cultivated in two distinct geographic locations, Catania and Rovigo, for three consecutive years from 2019 to 2021. Total aerobic microbes (TAMC), total combined yeasts/moulds (TYMC), the presence of bile-tolerant Gram-negative bacteria, and the absence of Escherichia coli and Salmonella spp. were evaluated and compared. The main phytocannabinoid content was measured and correlated with microbial contamination., Results: Most samples analyzed in this study did not meet the European Pharmacopoeia microbiological limits. The detection of potential pathogens, such as E. coli and Salmonella spp., in the samples indicates that the use of inflorescences may represent a possible source of infection. Microbial contamination varied among harvesting seasons and production sites, with agroclimatic conditions influencing microbial load and composition. The presence of potentially pathogenic bacteria was less associated with seasonal climate variability and more likely affected by sporadic contamination from external sources. CBD concentration exhibited a negative correlation with bile-tolerant Gram-negative bacteria and total yeasts/moulds levels. Samples with lower CBD content were more contaminated than those with higher CBD levels, suggesting a potential protective effect of this phytochemical on the plant., Conclusions: The threshing residues (inflorescences, floral bracts, and leaves) of industrial hemp varieties represent a valuable product and a source of beneficial phytochemicals that warrants further exploration. While post-harvest sterilization methods may reduce microbiological risks, they may also degrade heat- and light-sensitive bioactive phytochemicals. The most promising strategy involves implementing best agronomic practices to maintain healthy and uncontaminated cultures. Rigorous monitoring and quality certification protocols are essential to mitigate the microbiological risk associated with the consumption of hemp-derived products., (© 2024. The Author(s).)

Background: Data on mixed mould infection with COVID-19-associated pulmonary aspergillosis (CAPA) and COVID-19-associated pulmonary mucormycosis (CAPM) are sparse., Objectives: To ascertain the prevalence of co-existent CAPA in CAPM (mixed mould infection) and whether mixed mould infection is associated with early mortality (≤7 days of diagnosis)., Methods: We retrospectively analysed the data collected from 25 centres across India on COVID-19-associated mucormycosis. We included only CAPM and excluded subjects with disseminated or rhino-orbital mucormycosis. We defined co-existent CAPA if a respiratory specimen showed septate hyphae on smear, histopathology or culture grew Aspergillus spp. We also compare the demography, predisposing factors, severity of COVID-19, and management of CAPM patients with and without CAPA. Using a case-control design, we assess whether mixed mould infection (primary exposure) were associated with early mortality in CAPM., Results: We included 105 patients with CAPM. The prevalence of mixed mould infection was 20% (21/105). Patients with mixed mould infection experienced early mortality (9/21 [42.9%] vs. 15/84 [17.9%]; p = 0.02) and poorer survival at 6 weeks (7/21 [33.3] vs. 46/77 [59.7%]; p = 0.03) than CAPM alone. On imaging, consolidation was more commonly encountered with mixed mould infections than CAPM. Co-existent CAPA (odds ratio [95% confidence interval], 19.1 [2.62-139.1]) was independently associated with early mortality in CAPM after adjusting for hypoxemia during COVID-19 and other factors., Conclusion: Coinfection of CAPA and CAPM was not uncommon in our CAPM patients and portends a worse prognosis. Prospective studies from different countries are required to know the impact of mixed mould infection., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Fungi are important contaminants of historic canvas paintings worldwide. They can grow on both sides of the canvas and decompose various components of the paintings. They excrete pigments and acids that change the visual appearance of the paintings and weaken their structure, leading to flaking and cracking. With the aim of recognizing the most dangerous fungal species to the integrity and stability of paintings, we studied 55 recently isolated and identified strains from historic paintings or depositories, including 46 species from 16 genera. The fungi were categorized as xero/halotolerant or xero/halophilic based on their preference for solutes (glycerol or NaCl) that lower the water activity (a w ) of the medium. Accordingly, the a w value of all further test media had to be adjusted to allow the growth of xero/halophilic species. The isolates were tested for growth at 15, 24 °C and 37 °C. The biodeterioration potential of the fungi was evaluated by screening their acidification properties, their ability to excrete pigments and their enzymatic activities, which were selected based on the available nutrients in paintings on canvas. A DNase test was performed to determine whether the selected fungi could utilize DNA of dead microbial cells that may be covering surfaces of the painting. The sequestration of Fe, which is made available through the production of siderophores, was also tested. The ability to degrade aromatic and aliphatic substrates was investigated to consider the potential degradation of synthetic restoration materials. Xerotolerant and moderately xerophilic species showed a broader spectrum of enzymatic activities than obligate xerophilic species: urease, β-glucosidase, and esterase predominated, while obligate xerophiles mostly exhibited β-glucosidase, DNase, and urease activity. Xerotolerant and moderately xerophilic species with the highest degradation potential belong to the genus Penicillium , while Aspergillus penicillioides and A. salinicola represent obligately xerophilic species with the most diverse degradation potential in low a w environments.

Clinical Relevance: Invasive fungal infections (IFIs) and oomycoses (hereafter termed invasive fungal-like infections [IFLIs]) are characterised by penetration of tissues by fungal elements. The environment is the most common reservoir of infection. IFIs and IFLIs can be frustrating to treat because long treatment times are usually required and, even after attaining clinical cure, there may be a risk of relapse. Owner compliance with medication administration and recheck examinations can also decline over time. In addition, some antifungal drugs are expensive, have variable interpatient pharmacokinetic properties, can only be administered parenterally and/or have common adverse effects (AEs). Despite these limitations, treatment can be very rewarding, especially when an otherwise progressive and fatal disease is cured., Aim: In the second of a two-part article series, the spectrum of activity, mechanisms of action, pharmacokinetic and pharmacodynamic properties, and AEs of antifungal drugs are reviewed, and the treatment and prognosis of specific IFIs/IFLIs - dermatophytic pseudomycetoma, cryptococcosis, sino-orbital aspergillosis, coccidioidomycosis, histoplasmosis, sporotrichosis, phaeohyphomycosis, mucormycosis and oomycosis - are discussed. Part 1 reviewed the diagnostic approach to IFIs and IFLIs., Evidence Base: Information on antifungal drugs is drawn from pharmacokinetic studies in cats. Where such studies have not been performed, data from 'preclinical' animals (non-human studies) and human studies are reviewed. The review also draws on the wider published evidence and the authors' combined expertise in feline medicine, mycology, dermatology, clinical pathology and anatomical pathology., Abbreviations for Antifungal Drugs: AMB (amphotericin B); FC (flucytosine); FCZ (fluconazole); ISA (isavuconazole); ITZ (itraconazole); KCZ (ketoconazole); PCZ (posaconazole); TRB (terbinafine); VCZ (voriconazole)., Competing Interests: Conflict of interestThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objectives: Invasive Aspergillus infections during the early phase of childhood acute lymphoblastic leukemia (ALL) treatment come with morbidity and mortality. The interaction with vincristine hampers first-line azole prophylaxis. We describe the efficacy of an alternative twice-a-week micafungin regimen for Aspergillus prophylaxis., Methods: Newly diagnosed paediatric patients with ALL treated according to the ALL-11 protocol received micafungin twice-a-week (9 mg/kg/dose [max. 300 mg]) during the induction course (first 35 days of treatment) as part of routine care. A historical control cohort without Aspergillus prophylaxis was used. During the first consolidation course (day 36-79), standard itraconazole prophylaxis was used in both groups. The percentage of proven/probable Aspergillus infections during the induction/first consolidation course was compared between the cohorts. The cumulative incidence of proven/probable Aspergillus infections was estimated using a competing risk model. For safety evaluation, liver laboratory chemistry values were analysed., Results: A total of 169 and 643 paediatric patients with ALL were treated in the micafungin cohort (median age: 4 years [range 1-17]) and historical cohort (median age: 5 years [range 1-17]). The percentage of proven/probable Aspergillus infections was 1·2% (2/169) in the micafungin cohort versus 5·8% (37/643) in the historical cohort (p=0.013; Fisher's exact test). The differences in estimated cumulative incidence were assessed (p=0·014; Gray's test). Although significantly higher ALT/AST values were reported in the micafungin cohort, no clinically relevant side effects were observed., Conclusions: Twice-a-week micafungin prophylaxis during the induction course significantly reduced the occurrence of proven/probable Aspergillus infections in the early phase of childhood ALL treatment., (Copyright © 2023. Published by Elsevier Ltd.)

Colour defects can affect the appearance of cheese, its flavour, the safety of its consumption, and the price it can demand. This work reports the identification of five fungal isolates from a dairy plant where the surface of most cheeses was affected by patent, reddish-to-brown stains. One of these isolates was obtained from cheese, two from brine, and two from a bulk tank containing ewe milk. Molecular identification by partial amplification, sequencing, and database comparison of the concatenated sequence of the genes coding for the largest subunit of RNA polymerase II (RPB2), β-tubulin (β-TUB), and the large subunit of the rRNA molecule (LSU), plus the internal transcribed sequence (ITS) regions, assigned the isolates to Epicoccum layuense, Epicoccum italicum, and Epicoccum mezzettii. Features of the growth of these different species on different agar-based media, and of the morphology of their conidia following sporulation, are also reported. The strain isolated from cheese, E. layuense IPLA 35011, was able to recreate the reddish-brown stains on slices of Gouda-like cheese, which linked the fungus with the colour defect. In addition, two other strains, E. italicum IPLA 35013 from brine and E. italicum IPLA 35014 from milk, also produced stains on cheese slices. Epicoccum species are widely recognized as plant pathogens but have seldom been reported in the dairy setting, and never as human or animal pathogens., Competing Interests: Declaration of competing interest The research was conducted in the absence of any conflict interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The main component of an air conditioning system is air filters. Over time, the filters of an air conditioning system in cars can turn into sources of emission of microbiological hazards. The aim of this study was to quantitatively and qualitatively assess the presence of infectious and toxic fungi in the AC filters in passenger cars. The studied non-woven filters were removed from passenger cars during the "winter"/"summer" seasons. The taxonomic identification of the fungi isolated from the filters was performed using both the culture-based and molecular methods. RT-PCR was applied to assess the presence of gene fragments regulating aflatoxin biosynthesis in the isolates obtained from fungal cultures. The average fungal concentrations in the filter samples collected during the summer/winter season were 5.4 × 10 4 cfu/m 2 and 2.4 × 10 4 cfu/m 2 , respectively. Most of the filter samples, collected in both the studied seasons, revealed the presence of Aspergillus species including A. fumigatus, A. niger, A. terreus and/or A. flavus. The recorded levels of fungal contamination of AC filters in passenger cars indicate the necessity for more frequent filter replacement in this type of vehicle. Occupational exposure to moulds and the resulting health problems that may be experienced by professional drivers should be properly recognised in order to undertake effective preventive measures., (© 2023. The Author(s).)

Launch of in-house sensitive cell-free deoxyribonucleic acid (cfDNA) mould polymerase chain reaction (PCR) assays increased detection of moulds meeting suspected healthcare-associated infection (HAI) criteria. Definition was based on time from admission and mould detection in culture or via molecular methods. We created a modified mould HAI algorithm incorporating clinical context into the case definition, which allowed for better capture of possible mould HAIs, decreased number of investigations, and improved utilization of Infection Prevention and Control (IPC) resources., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2023.)

Occupational exposure to microbially contaminated metal working fluids (MWF) can cause hypersensitivity pneumonitis (HP). An important step in the diagnosis of HP is to identify the triggering antigen by detection of corresponding specific IgG antibodies (sIgG). As commercial sIgG tests are currently not available, protein antigens were prepared from MWF-workplace samples and from MWF-typical bacterial isolates. In 57 % of suspected HP-cases (n = 30) elevated sIgG concentrations were measured to at least one MWF-relevant antigen, of which Mycobacterium immunogenum was most prominent (88 %), followed by Pseudomonas oleovorans and Pseudomonas spec (82 % each), MWF-antigen mix and Pseudomonas alcaliphila (65 % each). Elevated sIgG concentrations to other microorganisms were measured to Micropolyspora faeni (82 %) and Aureobasidium pullulans (77 %). Correlation of sIgG values of all tested microbial antigens showed a significant relationship of MWF-antigen mixture to Pseudomonas antigens, but a low correlation to moulds. These newly prepared MWF-antigens are useful tools for the diagnosis of patients with suspected MWF-HP and are available for further investigations., Competing Interests: Declaration of Competing Interest All authors declare that there is no conflict of interest regarding this work., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Chemical composition and in vitro antimicrobial activity of parsley [ Petroselinum crispum (Mill.) Fuss] and ginger ( Zingiber officinale Roscoe) essential oils from store-bought parsley plants and ginger rhizomes were evaluated. A sensory analysis was performed on aged cheese slices treated with a mix of each essential oil and seed oil. Parsley essential oil was selected as the best compromise between sensory influence and antimicrobial activity and tested to evaluate its antifungal effects on cheese slices experimentally contaminated with Aspergillus flavus . Monoterpenes and α-zingiberene were the most abundant compounds in parsley and ginger essential oil, respectively. Both essential oils showed in vitro antimicrobial activity against different fungal species while only a bacterial strain ( Enterococcus faecalis ) was sensible to ginger essential oil. Only the cheese slices treated with parsley essential oil were appreciated by the panellists and its use has been effective in preventing the A. flavus growth in cheese.

During the last decade, scientists have given increasingly frequent warnings about global warming, linking it to mycotoxin-producing moulds in various geographical regions across the world. In the future, more pronounced climate change could alter host resilience and host-pathogen interaction and have a significant impact on the development of toxicogenic moulds and the production of their secondary metabolites, known as mycotoxins. The current climate attracts attention and calls for novel diagnostic tools and notions about the biological features of agricultural cultivars and toxicogenic moulds. Since European climate environments offer steadily rising opportunities for Aspergillus flavus growth, an increased risk of cereal contamination with highly toxic aflatoxins shall be witnessed in the future. On top of that, the profile (representation) of certain mycotoxigenic Fusarium species is changing ever more substantially, while the rise in frequency of Fusarium graminearum contamination, as a species which is able to produce several toxic mycotoxins, seen in northern and central Europe, is becoming a major concern. In the following paper, a high-quality approach to a preventative strategy is tailored to put a stop to the toxicogenic mould- and mycotoxin-induced contamination of foods and feeds in the foreseeable future.

This work describes a novel approach for Al-7.5Si/Al-18Si liquid-liquid bimetal casting in sand and metallic moulds. The aim of the work is to facilitate and develop a simple procedure to produce an Al-7.5Si/Al-18Si bimetallic material with a smooth gradient interface structure. The procedure involves the theoretical calculation of total solidification time (TST) of the first liquid metal (M1), pouring the liquid metal (M1), and allowing it to solidify; then, before complete solidification, the second liquid metal (M2) is introduced into the mould. This novel approach has been proven to produce Al-7.5Si/Al-18Si bimetal materials using liquid-liquid casting. The optimum time interval of Al-7.5Si/Al-18Si bimetal casting with modulus of cast M c ≤ 1 was estimated based on subtracting 5-15 s or 1-5 s from TST of M1 for sand and metallic moulds, respectively. Future work will involve determining the appropriate time interval range for castings having modulus ≥ 1 using the current approach.

While plant-specialized metabolites can affect mammal health, their fate during the aerobic deterioration of crop silage remains poorly understood. In this study, we investigated the metabolization of benzoxazinoids (BXs) in silages of two maize genotypes (W22 wild type and bx1 mutant line) during aerobic deterioration. In W22 plants, concentrations of the aglucone BXs DIMBOA and HMBOA in silage decreased over time upon air exposure, while concentrations of MBOA and BOA increased. Mutant plants had low levels of BXs, which did not significantly vary over time. Aerobic stability was BX-dependent, as pH and counts of yeasts and molds were higher in W22 compared to that in bx1 silage. The nutrient composition was not affected by BXs. These preliminary results may be used to estimate the amounts of BXs provided to farm animals via silage feeding. However, further research is warranted under different harvest and storage conditions.

The aim of the present study was to develop meat-based media with compositions similar to those of dry-fermented meat products and to evaluate their use in studying the growth of Aspergillus parasiticus and the kinetics of aflatoxin B1 (AFB1) production. In our previous experiments, we found that the strain A. parasiticus ŽMJ7 produced a high amount of AFB1. Cooked meat agar (CMA2) was used as a novel complex meat-based medium with four variations: CMA2G (CMA2 supplemented with 1% glucose), CMA2YE (CMA2 supplemented with 0.2% yeast extract), and CMA2GYE (CMA2 supplemented with 1% glucose and 0.2% yeast extract). Media were inoculated with an A. parasiticus spore suspension (10 5 spores/mL) and incubated at 25 °C for up to 15 days. The A. parasiticus lag phase lasted less than 1 day, irrespective of the growth medium, with the exception of control medium CMA1 (cooked meat agar) as an already known meat-based medium. The highest mean colony growth rate was observed on CMA2 and CMA2G. Reversed-phase UPLC-MS/MS analysis was performed to determine the AFB1 concentration in combination with solid phase extraction (SPE). The highest AFB1 concentration in meat-based media was detected in CMA2GYE after 15 days of incubation (13,502 ± 2367 ng/mL media). The results showed that for studying AFB1 production in dry-fermented meat products, novel suitable media such as CMA2-based media are required. This finding could represent a potential concern with regard to the production of dry-fermented meat products.

In this work, we determined the degree of filamentous fungi contamination in cocoa fermented seeds and the fungal potential to produce enzymes that could contribute to the cocoa quality. Internal transcribed spacer amplicon sequencing (ITS) showed 11 different species with Byssochlamys spectabilis, Aspergillus niger, Aspergillus flavus, Aspergillus carbonarius, Circinella muscae and Penicillium citrinum as the most abundant species. Our results evidenced intra and inter-species differences in the enzymes production. Cellulases, β-glucosidase, ornithine decarboxylase and phenylalanine decarboxylase were the most diffused enzymes expressed in the 53 strains here studied. Moreover, A. niger (6/12), A. carbonarius (2/3), and P. citrinum (3/3) showed high pectinolytic activity. Remarkable was the amino decarboxylase activity of P. citrinum, and A. flavus strains. For the first time we reported the presence of B. spectabilis in cocoa fermented beans, which could play an important role in the biogenic amines formation. In addition, we explored the capability of the Aspergillus section Nigri strains, to produce ochratoxin A (OTA) in a cacao model system (CPMS) and in malt extract medium (MEL). We observed that CPMS, but not MEL, stimulated the OTA production in 6 out 15 strains of Aspergillus section Nigri, reaching values ranging between 1.70 and 4995 μg OTA kg -1 dry mycelium., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The processing of dried figs in the industry involves a number of stages that present a significant risk of filamentous fungal infection of the fruit and subsequent mycotoxin contamination, due to the changes in temperature and water activity (a w ) to which dried figs are exposed. In this study, the environmental conditions and the physicochemical parameters of dried figs at different processing stages were evaluated in 3 different industries, and were associated with fungal counts and the presence of toxigenic moulds and their mycotoxins. For this, dried figs at 5 relevant stages of industrial processing (curing, sizing, blanching, storage, and final product) in 3 industries located in Extremadura (Spain) were sampled. Changes in moisture content and a w of dried figs during processing were observed and they influenced the mycological quality of figs. Among the fungal genera, Aspergillus spp. predominated in most stages except blanching, where Penicillium spp. prevailed. About 10% of the dried fig samples were contaminated with aflatoxins (AFs) and 6% with ochratoxin A (OTA). Based on findings, longer drying times are necessary after blanching to reduce a w and to avoid the development of toxigenic moulds. In addition, all stages covering industry processing, final storage, and retailing of dried figs are advisable to be conducted at refrigeration conditions and controlled relative humidity to avoid mycotoxin production. The enumeration of AFs- and OTA- producing moulds by real-time PCR seems to be a good indicator for integration into prevention strategies to control filamentous fungal hazards and subsequent mycotoxin synthesis during the processing of dried figs., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Traditionally, KOH microscopy and fungal culture are the two preferred tests as gold standard for diagnosis of onychomycosis. Recently, other diagnostic methods have been developed to improve the microbiological diagnosis. The EUROArray dermatomycosis kit is a PCR-based microarray test system for the detection and direct identification of species that are most frequently involved in skin and nail infections., Objectives: Our primary aim was to evaluate the real-life applicability of the EUROArray dermatomycosis kit in the diagnosis of onychomycoses. In addition, we compared the aetiology of onychomycoses found in our patients with those described in the literature., Patients/methods: We prospectively studied consecutive 100 patients with suspected onychomycoses. Samples of suspect toenails were taken as part of routine medical management. Nail specimens were evaluated by means of three diagnostic methods: KOH preparation, culture and EUROArray dermatomycosis kit., Results: Onychomycosis was diagnosed in 47/100 patients who proved positive on at least one reference diagnostic test and in 49/100 patients who proved positive on PCR. The combination of microscopy and PCR had better sensitivity than microscopy (p = .0397), fungal culture (p = .0061) and PCR alone (p = .0117). Moulds were more frequently positive in culture than in PCR (p = .033). Dermatophytes proved positive more frequent than moulds and yeasts in both culture and PCR; in particular, Trichophyton interdigitale was the most frequent pathogen., Conclusions: In conclusion, introducing EUROArray dermatomycosis kit into the diagnostic algorithm of onychomycosis increases the sensitivity of direct microscopy and yields more rapid results than culture., (© 2021 Wiley-VCH GmbH.)

Microwave (MW) irradiation is a non-destructive method that can be applied as an alternative method to inhibit the growth of microorganisms. The present study evaluated the effect of MW irradiation on the occurrence of moulds in nuts and almonds. Samples of unshelled natural almonds, pistachios, and in-shell peanuts were treated with different doses of MW irradiation (2400-4000 W). The effect of MW irradiation on mould counts was evaluated by cultivating immediately after irradiation and after 3 and 6 months of storage. The most represented genera in all analysed samples were Aspergillus (68%), Penicillium (21%), and a small amount of Cladosporium (3%). Mould numbers significantly decreased after MW treatment. The treatments with MW irradiations at 3000 and 4000 W significantly reduced the mould colony counts, and their effect persisted during storage; irradiation at 2400 W was partially effective. The strongest effect of MW irradiation was observed in in-shell peanuts. MW irradiation seems to be a promising method for maintaining the microbiological quality of nuts.

Background: Moulds are often wrongly considered contaminants, not very sensitive to conventional antifungal treatments, but they may cause ungual hyphomycosis, particularly Aspergillus. Due to the lack of precise diagnostic criteria, their real impact is underestimated., Objectives: Retrospective descriptive analysis of all ungual hyphomycosis cases diagnosed at Montpellier Hospital from 1991 to 2019 to: (i) determine the incidence of onychomycosis by pseudo-dermatophytes and moulds; (ii) perform an epidemiological analysis of nail aspergillosis; and (iii) provide simple criteria for mould involvement in onychopathy., Patients/methods: Data concerning 4053 patients were collected: age, sex, onychomycosis location, direct examination results, species(s) identified and fungal co-infections. Moreover, clinical data of patients with nail aspergillosis were analysed to identify potential contributing factors, and the classical criteria for mould involvement in onychopathy were critically reviewed., Results: A pseudo-dermatophyte or a mould was involved in nail colonisation in 17.25% of patients (men/women ratio: 0.70; mean age: 53.1 years). The identified hyphomycetes belonged mainly to the genera Fusarium (45.68%), Scopulariopsis (30.23%) and Aspergillus (16.94%). Analysis of the clinical reports of 102 patients with ungual aspergillosis (men/women ratio: 0.67; mean age: 56.3 years) identified cardiovascular (43.9%), endocrine (25.8%), cancer (19.7%) and skin (18.2%) diseases as contributing factors., Conclusions: The adoption of simple and reliable criteria (ie, characteristic filaments on direct microscopic examination after periodic acid-Schiff staining, growth at seeding points in culture) allows determining the formal involvement of a mould in chronic onychopathies and avoiding possible side effects and costs of empirical or inappropriate and repetitive treatments., (© 2021 Wiley-VCH GmbH.)

The aim of this study was to identify and compare surface mycobiota of traditional and industrial Croatian dry-fermented sausage Kulen , especially toxicogenic species, and to detect contamination with mycotoxins recognized as the most important for meat products. Identification of mould species was performed by sequence analysis of beta- tubulin and calmodulin gene, while the determination of mycotoxins aflatoxin B 1 (AFB 1 ), ochratoxin A (OTA), and cyclopiazonic acid (CPA) was carried out using the LC-MS/MS (liquid chromatography-tandem mass spectrometry) method. The results showed a significantly higher number of mould isolates and greater species (including of those mycotoxigenic) diversity in traditional Kulen samples in comparison with the industrial ones. P. commune , as a potential CPA-producer, was the most represented in traditional Kulen (19.0%), followed by P. solitum (16.6%), which was the most represented in industrial Kulen samples (23.8%). The results also showed that 69% of the traditional sausage samples were contaminated with either CPA or OTA in concentrations of up to 13.35 µg/kg and 6.95 µg/kg, respectively, while in the industrial samples only OTA was detected (in a single sample in the concentration of 0.42 µg/kg). Mycotoxin AFB 1 and its producers were not detected in any of the analysed samples (

Background: There are important advances in the management of bacterial infection in patients with cystic fibrosis (CF), but there are many gaps in the field of fungal infections., Aims: The aim of this study was to analyse whether chronic respiratory filamentous fungal colonization had clinical impact and whether antifungal treatment can change the disease., Methods: The prospective, bicentric and descriptive study was carried out within a 3-year follow-up period, with four-month periodicity medical controls. Adult patients from two CF units of tertiary hospitals were included. Clinical, microbiological, analytical and spirometric variables were collected. Quality of life was evaluated in a subgroup, using the Spanish version of the Revised Cystic Fibrosis Quality of Life Questionnaire (CFQ-R). To statistically analyze the evolution of forced expiratory along time (volume of air blown out in 1 second -FEV 1 -) and the forced vital capacity (FVC), mixed linear models were carried out., Results: From the ninety-eight patients under study, 40 suffered chronic filamentous fungal colonization. The presence of filamentous fungi in airway was associated to an annual fall of FEV 1 and FVC of 0.029 and 0.017 litres, respectively (p<0.001). In addition, worse quality of life based on CFQ-R, significant when concerning physical condition and emotional state, was also linked with the fungal colonization. Protocolized antifungal therapy, nebulized or oral, improved FEV 1 in 0.023 and 0.024 litres per year, respectively (p<0.001)., Conclusions: Chronic filamentous fungal colonization in patients with CF is associated with a significant annual decline of lung function that persists over time. Chronic antifungal therapy slows down this progression, mainly in the patient with more advanced disease., (Copyright © 2021 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Hypersensitivity pneumonitis (HP) is an interstitial lung disease (ILD) caused by the inhalation of antigens. Antigen-specific IgG antibodies (sIgG) are used as biomarkers of exposure when diagnosing HP, but little is known about the longitudinal relation between antibody levels and risk of HP or other ILD. In a follow-up design, we explored the relationship between sIgG antibodies against Aspergillus fumigatus and the diagnosis of HP in 647 subjects suspected of HP. We showed that IgG levels above the reference value resulted in a hazard ratio of 9.5 for subsequent HP. Our findings support a relationship between high levels of sIgG against A. fumigatus and risk of HP.

The ripening process of dry-cured meat products is characterised by the development of fungi on the product's surface. This population plays a beneficial role, but, uncontrolled moulds represent a health risk, since some of them may produce mycotoxins, such as ochratoxin A (OTA). The aim of the present work is to assess the potential of near-infrared spectroscopy (NIRS) for the detection of OTA-producing mould species on dry-cured ham-based agar. The collected spectra were used to develop Support Vector Machines-Discriminant Analysis (SVM-DA) models by a hierarchical approach. Firstly, an SVM-DA model was tested to discriminate OTA and non-OTA producers; then, two models were tested to discriminate species among the OTA producers and the non-OTA producers. OTA and non-OTA-producing moulds were discriminated with 85% sensitivity and 86% specificity in the prediction. Furthermore, the SVM-DA model could differentiate non-OTA-producing species with a 95% sensitivity and specificity. Promising results were obtained for the prediction of the four OTA-producing species tested, with a 69% and 90% sensitivity and specificity, respectively. The preliminary approach demonstrated the high potential of NIR spectroscopy, coupled with Chemometrics, to be used as a real-time automated routine monitorization of dry-cured ham surfaces.

Purpose: Given tissue inhomogeneity and lack of backscatter media, superficial brachytherapy necessitates more accurate dosimetry than TG-43 formalism. However, the introduction of modern model-based dose calculation algorithms into clinical practice should be carefully evaluated. The aim of this work was to compare dose distributions calculated with TG-43 and advanced collapsed cone engine (ACE) algorithms for individual multi-catheter moulds, and investigate the impact of target size and the lack of bolus to differences between plans., Material and Methods: Eleven treatment plans for individual mould multi-catheter high-dose-rate brachytherapy (IMM HDR) were selected for retrospective analysis. All treatment plans were initially calculated with TG-43 formula and re-calculated using ACE algorithm. Plan re-calculation with ACE was repeated for each plan in order to assess the impact of bolus. To evaluate differences between TG-43 and ACE dose distributions, dose-volume histogram (DVH) parameters for each ROI were compared. D max (maximal point dose), D 0.1cc , and D 2cc were calculated for each risk's organ (OARs) and for external contour. For clinical target volume (CTV), D 98 , D 90 , D 50 , CTV coverage (CTV-V 100 ), and dose delivered to reference point were compared between the plans., Results: A significantly lower values ( p < 0.05) of CTV parameters were observed for treatment plans calculated with ACE algorithm comparing to TG-43. Further analysis showed that differences between CTV-V 100 for ACE and TG-43 plans depended on CTV volume. Dosimetric parameters for OARs were significantly lower in ACE plans than those of TG-43. Only D 2cc for external and D 0.1cc for both eye lenses in ACE plans were insignificantly different comparing to TG-43 plans., Conclusions: Results show that differences between dosimetric parameters are statistically significant. However, their clinical relevance is still undetermined. Careful re-evaluation of the clinical results based on long-term research on TG-43 is necessary to safely introduce modern algorithms to clinical practice., Competing Interests: The authors report no conflict of interest., (Copyright © 2021 Termedia.)

The use of natural compounds as food preservatives is becoming increasingly popular as it is perceived positively by consumers. Among these substances, essential oils have attracted great interest owing to their antioxidant and antimicrobial properties. However, several challenges impair the use of essential oils in food products, such as their degradation or loss during food processing and storage, the strong aroma, even at low concentrations, which may negatively affect the sensory characteristics of food. In this context, the development of nanoformulations able to stabilize essential oils may represent a smart solution to this issue. The aim of the study was to evaluate the efficiency of alginate-based nanoformulations enriched with lemongrass ( Cymbopogon nardus ) essential oil (LEO) and Tween 80 against several fungi namely Penicillium expansus , Aspergillus niger and Rhizopus spp. Firstly, the flow behavior of systems at different concentrations of alginate (1%, 2% and 3% w / w ) were studied. Then, emulsion-based nanoformulations at different concentrations of lemongrass essential oil in the range of 0-2% w / w were stabilized by a fixed amount of Tween 80, characterized and tested for their antifungal activity. Our results showed that the best nanoformulation able to inhibit Rhizopus spp., Penicillium expansum and Aspergillus niger , for at least 10 days, was constituted by 1% alginate/1.5% LEO/1% Tween 80. Hence, the incorporation of essential oil into nanoformulation systems may represent a valid alternative to overcome the disadvantages that limit the commercial application of essential oils.

In the UK and Northern Europe, ripening oats can become contaminated with T-2 and HT-2 mycotoxins, produced mainly by Fusarium langsethiae. There are indicative levels related to the maximum limits for oat grain for these toxins. The objectives of this study were to examine the effect of interacting conditions of temperature (10-30 °C) and water activity (a w , 0.995-0.90) on (a) lag times prior to growth, (b) growth and (c) T-2 and HT-2 toxins by two strains of F. langsethiae isolated from oats in the UK and compare this with the type strain (Fl201059) which has been genomically sequenced, and (d) develop (and validated with published data) a probabilistic models for impacts of temperature × a w on growth and toxin production. All three strains had an optimum a w range and temperature of 0.995-0.98 and 25 °C for growth. For T-2 + HT-2 production these were 0.995 a w and 20 °C. Overall, the type strain produced higher amounts of T-2 + HT-2 with a HT-2/T-2 ratio of up to 76. Using this study data sets and those from the literature, probabilistic models were developed and validated for growth and T-2 + HT-2 toxin production in relation to temperature × a w conditions. These models, when applied in stored oats, will be beneficial in determining the conditions on the relative level of risk of contamination with these two toxins in the context of the EU indicative maximum levels., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Activation of mucosal-associated invariant T cells (MAIT cells) by certain bacteria, viruses, and yeast is well studied, but the activation potential of filamentous moulds from the order Mucorales is not known. Here, we show a rapid response of human MAIT cells against the Mucorales species Mucor circinelloides, Rhizopus arrhizus, and Rhizopus microsporus. This activation included upregulation of CD69 and degranulation marked by increased CD107a expression, while intracellular perforin and granzyme A expression were reduced. Furthermore, blocking of the antigen-presenting molecule major histocompatibility complex class I-related abrogated MAIT cell activation demonstrating a T cell receptor-dependent stimulation by Mucorales., (The Author(s) 2020. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

In this research communication we report on the diversity of yeast and mould species in 69 samples of milk and different dairy products from three plants located in Umbria, central Italy. Isolates were characterised both macroscopically and microscopically and then identified by PCR and genome sequencing of the ITS region and the D1-D2 domain of the large-subunit rRNA gene for filamentous fungi and yeasts, respectively. Out of the 69 samples analysed, 51 (73.9%) tested positive for the presence of yeasts, whereas moulds were detected in 25 (36.2%) samples. A total of 9 yeast species belonging to 8 different genera and 13 mould species belonging to 6 different genera were isolated. The most common genera isolated were Debaryomyces and Kluyveromyces among the yeasts and Penicillium and Galactomyces among the moulds. Microbiota play a key role in the formation of flavour, aroma, texture and appearance of dairy products. This complex microbial ecosystem includes both cultured and external bacteria, yeasts and moulds. Some of them have an important role in the production of cheeses, whereas others are responsible for dairy product spoilage, resulting in significant food waste and economic losses. Some species can produce mycotoxins, representing a potential hazard for the consumer's safety. This study provides interesting information on the diversity of fungi species in dairy products from central Italy that can be of major importance to identify these products and to develop adequate strategies for fungal spoilage control and consumer safety.

Invasive fungal infections have increased over the last decades and the therapeutic choices to treat them are limited. The antifungal agents currently available are useful and have optimal in vitro activity; however, their activity can be lowered due to the development of fungal resistance. The increase in primary or secondary resistance to some antifungal drugs has led to the search of alternatives such as the combination of drugs or the development of new antifungals. In this paper, the activity of the main families of antifungal drugs, polyenes, azoles, echinocandins, 5-fluorocytosine and other new antifungal drugs, are reviewed. The main resistance mechanisms developed by fungi are also described., (Copyright © 2021 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.)

Dry-fermented sausages are prone to be colonised by Penicillium nordicum, which is one of the main ochratoxin A (OTA)-producing species. Its ability to produce this mycotoxin on dry-fermented sausages has been reported. However, the influence of the conditions of a traditional processing of a Spanish dry-fermented sausage and the intrinsic physicochemical parameters of this product such as water activity (a w ) and pH on OTA production has not been studied yet. Thus, the aim of this study was to evaluate the influence of traditional processing (interaction of relative humidity (RH) x temperature x ripening days) on the evolution of pH and a w during maturation of dry-fermented sausage "salchichón" and its relationship with OTA synthesis by P. nordicum. The expression of otapks and otanps genes, both involved in the biosynthesis of the mycotoxin, was also assessed. For this, 27 raw sausages were inoculated with P. nordicum and ripened for 26 days in a drying chamber (3 days at 5 °C and 84% RH, 17 days at 12 °C and 84% RH, and 6 days at 12 °C and 80% RH). From results, although it seems that the pH slightly influenced on OTA biosynthesis, the a w had a great impact on this mycotoxin production. In fact, the two highest OTA concentrations found coincided with a dramatic rise of the a w value (0.92 a w ) by day 18 of incubation when the RH of the drying chamber was still 84% and at the end of the incubation time when the a w decreased noticeably (0.87 a w ). The expression of the otapks and otanps genes correlated with the OTA produced by P. nordicum. Results from this work confirm that the traditional processing of Spanish dry-fermented sausages favours itself OTA synthesis by P. nordicum. Our findings may help in informed decision-making in relation to RH/temperature of drying chambers and shortening of the ripening process. This may be then effectively incorporated into the hygienic production system in the framework of HACCP together with other measures including the use of Penicillium nalgiovense as protective culture or the monitoring of otapks gene expression, and a w during the processing of dry-fermented sausages. All these strategies together may put ochratoxigenic Penicillia at a disadvantage and minimise OTA contamination risks in dry-fermented sausages., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Olives and their derivatives, in particular olive oil, represent one of the most significant agricultural products in the Mediterranean basin. Storage under inadequate conditions poses serious problems concerning fungal contamination, with consequent defects and potential mycotoxin production in olives and olive oils. Penicillium expansum represents one of the most significant postharvest pathogens in several fruits, including olives. Not only it causes blue mold but also is one of the most relevant patulin producing species of the genus Penicillium. The aim of this research was to evaluate the ecophysiological conditions governing growth and PAT production by P. expansum strains previously isolated from Tunisian olives. For this purpose, four P. expansum isolates were tested in a synthetic medium (Czapek Yeast Autolysate, CYA) and in olive-based medium (OM) for their ability to grow and produce PAT under different temperatures (4 °C, 15 °C and 25 °C) for 10 and 20 d. The mycotoxin was analysed by HPLC-UV. Results showed that all isolates were able to grow on tested media at different temperatures. Different PAT production profiles were found, showing that at 25 °C P. expansum isolates were able to produce PAT on CYA and OM medium. At 15 °C the production of PAT was only detected on CYA medium, while no PAT production was detected at 4 °C for the two media., (Copyright © 2019 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

The aim of the study was to evaluate the fungicidal effect of a H 2 O 2 mist generating system for disinfection of spores of six food-related moulds ( Alternaria alternata , Aspergillus flavus , Geotrichum candidum , Mucor plumbeus , Paecilomyces variotii , and Penicillium solitum ) dried on stainless steel. Exposure to H 2 O 2 mist for 2 or 4 h lead to >3 log reduction in mould spores in the majority of the tests. The presence of the soils 2% skim milk or 3% BSA did not significantly alter the fungicidal effect, while the presence of raw meat juice had an adverse fungicidal effect against Penicillium and Mucor in two out of three tests. Fungicidal suspension tests with liquid H 2 O 2 confirmed the effectiveness of H 2 O 2 on reducing the mould spores. Both the surface test and the suspension test indicated that P. variotii is more resistant to H 2 O 2 compared to the other moulds tested. The study shows the efficiency of H 2 O 2 mist on reducing food-related mould spores on surfaces.

Moulding technologies are remarkably effective for parts requiring high production volumes. Yet cooling the moulds after each injection can cause a significant loss of time. A possibility for reducing the cooling times is to use cooling inserts and conformal cooling strategies. In the present case, the original inserts of a mould must be substituted because the original material cannot be utilized anymore (toxicity). Will it be technically feasible to achieve a proper cooling only by modifying the inserts? Here, the cooling inserts of high production steel moulds utilized to manufacture ribs for swimming pool sinks' plastic cages are redesigned, simulated and manufactured, taking advantage of Selective Laser Melting possibilities and without modifying the geometry of the obtained parts, nor the rest of the moulds. The results reveal a reduction in the mould cooling times of up to 8%, while maintaining the same conformation properties, thus leading to important savings of time and some global costs in the production outcomes. The study also benchmarks the production economic limits of this approach compared to other possible strategies, such as the development of full new conformal cooling moulds or the industrial production of the parts with plastic additive manufacturing (multi jet fusion) technology.

Advances in micro-organism identification techniques have resulted in increased knowledge of the diversity of prokaryotes and eukaryotes in silage. Such knowledge has enhanced the understanding of how fermentation occurs in forage crops with different characteristics and how the process can be improved to enhance silage quality. Undesirable micro-organisms can grow in silage when fermentation does not occur properly. Such micro-organisms may be pathogenic and/or produce toxic metabolic compounds; however, information on the consequences of these metabolites on the health of animals that consume silage is still lacking. The major challenge of ensilage is to produce high-quality feed that is nutritional, sanitary and stable, with a high dry matter recovery rate, in a process involving no interventions during fermentation and considerable variation in the characteristics of the substrates. It is important to note that each substrate has particularities and that we can only improve fermentation if we fully understand microbial diversity. This review is intended to update information related to the fermentation profile of silage, focusing on microbial diversity., (© 2019 The Society for Applied Microbiology.)

Introduction: Superficial mycosis is one of the most common diseases worldwide, however its epidemiology is changing over time., Aim: To present epidemiological data of the skin fungal infections diagnosed in the years 2011-2016 in Lower Silesia., Material and Methods: A total of 11 004 patients with a clinically suspected superficial mycosis were investigated. Skin scrapings, nail clippings and plucked hair were examined with a direct microscopy, Wood's lamp and culture. Particular species were identified via polymerase chain reaction (PCR) examination. The lesions suspected for pityriasis versicolor were screened for Malassezia with Wood's lamp and direct microscopy., Results: Dermatomycosis was diagnosed in 1653 (15.00%) patients with 1795 fungi identified. 1858 specimens were indicative of fungal infection including dermatophytes, yeasts and moulds. Out of 924 cases of dermatophytic infections (51.48%), Trichophyton rubrum accounted for the majority (71.75%) and was followed by Trichophyton tonsurans (16.77%). Among the yeasts (716; 39.89%), Candida spp. was the most common agent identified (521; 67.66%). The sites affected most often were toenails (956; 51.45%) and fingernails (319; 17.17%). In paediatric population the most common diagnosis was tinea corporis (60, 41.10%)., Conclusions: Our study revealed that toenail onychomycosis remains the most common superficial mycosis and T. rubrum is the most common pathogen. However, in a longer period of observation, a decrease in the number of tinea capitis cases and an increase in infections caused by T. tonsurans were noticed. Observed changes indicate the need for continuing studies to detect the upcoming epidemiological trends., Competing Interests: The authors declare no conflict of interest., (Copyright: © 2018 Termedia Sp. z o. o.)

Exposure to mould is a potential risk factor for asthma in both adults and children. In adult, the relation between exposure, sensitization and symptoms has been demonstrated in severe asthmatics sensitized to Alternaria. For children, exposure to mould in childhood is a risk factor for asthma in both atopic and non-atopic individuals. Exposure or sensitization to moulds are a risk factor for severe asthma and/or exacerbations in children. There appears to be a causal relationship between exposure and asthma. This link seems less significant in adults. However, in adults mould sensitive asthma seems to determine a phenotype of severe asthma associated with more marked obstructive lung disease. Moulds can stimulate either innate or the acquired immunity. They are responsible for a marked Th2 inflammation leading to more severe asthma. Besides the immunological mechanisms, toxic mechanisms can also intervene. It is therefore not correct to reduce the effect of moulds, particularly in respiratory symptoms, to only allergic mechanisms., (Copyright © 2019 SPLF. Published by Elsevier Masson SAS. All rights reserved.)

Objectives: The purpose of this study was to compare the efficiency of mould identification of two matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) systems - Vitek MS (VMS) and Microflex LT (MLT) - and the MSI application., Methods: Moulds were collected retrospectively and prospectively to display epidemiological diversity of a microbiology laboratory. All of them were identified via sequencing. Strains were then identified using the VMS v3.0, the MLT, and the MSI software applied on MLT spectra. Rates of correct identifications to the species, to the complex, and to the genus level were compared with the molecular reference standard., Results: A total of 102 isolates were collected. The rate of correct identification to the species level with the MLT was 42.2% (43/102) with a threshold of 1.7 (vs. 16.7% (17/102) with a threshold of 2.0, p < 0.05). The VMS performed better than the MLT with a threshold of 1.7 for species (49.0% (50/102), p 0.33) and complex level identifications (71.6% (73/102) vs. 54.9% (56/102), p < 0.05). However the highest performances were observed when the MLT spectra were analysed via the Mass Spectrometry Identification (MSI) software reaching 90.2% (92/102) of correct identification to the species, 92.2% (94/102) to the species complex and 94.1% (96/102) to the genus level., Conclusions: The VMS performed better than the MLT for mould identification. However, it remains of utmost importance to expand commercial databases, as performances of the MLT highly improved when using the MSI software and its extended database, reaching far above the VMS system. Thus the VMS could benefit from the use of this online tool., (Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Dominant yeast species in Salsiccia Sarda, a traditional fermented sausage produced in Sardinia (Italy), were evaluated through the monitoring three typical production processes. Six different species were identified by molecular techniques, but Debaryomyces (D.) hansenii proved to be dominant. A D. hansenii strain was selected according to its technological features and used in three experimental sausage productions at farm scale with the aim to evaluate its antifungal effect. In all cases, two batches were inoculated with a previously selected autochthonous starter cultures (Lactobacillus plantarum and Staphylococcus xylosus), whereas two batches were left to spontaneous fermentation. D. hansenii was inoculated on the sausages surface by brushing after the sausages drying, by immersion in a yeast suspension after the stuffing, or, alternatively, casings were dipped in a yeast suspension before the dough stuffing. Microbial counts in the sausages core did not appear to be affected by D. hansenii application, while outcomes obtained for casings appeared soundly diversified. Brushing on the sausages surface at the onset of fermentation proved to be the best approach to treat sausages. Yeast inoculation exerted a noteworthy anti-mould effect, independently of the mode of application and, on the other hand, did not affect the overall quality and typical features of the product., (Copyright © 2019. Published by Elsevier Ltd.)

Objectives: Cannabis allergy has mainly been described following recreational use but some cases also point to cannabis sensitisation as a result of occupational exposure. As a consequence, little is known on the prevalence and clinical phenotype of occupational cannabis allergy. Therefore, this study aims to explore the allergy-associated health risks of occupational cannabis exposure in Belgian police force personnel., Methods: 81 participants, active in the police force, reporting regular occupational cannabis exposure during the past 12 months, were included. History was combined with a standardised questionnaire on allergies and cannabis exposure.Basophil activation tests (BATs) with a crude cannabis extract and rCan s 3 were performed. In addition, specific (s)IgE rCan s 3 as well as sIgE to house dust mite, six pollen and three mould allergens were quantified., Results: Although 42% of the participants reported respiratory and/or cutaneous symptoms on occupational cannabis exposure, all cannabis diagnostics were entirely negative, except one symptomatic case demonstrating a borderline result. Furthermore, there is no significant difference between the groups with and without symptoms on cannabis exposure in terms of allergenic sensitisations., Conclusions: The origins of the reported respiratory and cutaneous symptoms during cannabis exposure remain elusive but are probably due to non-immune reactions. It should be noted that the study was volunteer-based possibly reflecting an excessive number of symptomatic individuals. Nevertheless, as only one participant reported using fully protective gear, much improvement is needed for reducing the number of symptoms reported on duty, independent of their origin., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2019. No commercial re-use. See rights and permissions. Published by BMJ.)

Aims: The production of peptaibols, toxic secondary metabolites of Trichoderma, in the indoor environment is not well-documented. Here, we investigated the toxicity of peptaibols in the guttation droplets and biomass of Trichoderma strains isolated from problematic buildings., Methods and Results: Seven indoor-isolated strains of T. atroviride, T. trixiae, T. paraviridescens and T. citrinoviride were cultivated on malt extract agar, gypsum boards and paperboards. Their biomass extracts and guttation droplets were highly cytotoxic in resting and motile boar sperm cell assays and in inhibition of somatic cell proliferation assays. The toxins were identified with HPLC/ESI-MS/MS as trichorzianines, trilongins, trichostrigocins and trichostrigocin-like peptaibols. They exhibited toxicity profiles similar to the reference peptaibols alamethicin, trilongins, and trichorzianine TA IIIc purified from T. atroviride H1/226. Particular Trichoderma strains emitted the same peptaibols in both their biomasses and exudate droplets. The trilongin-producing T. citrinoviride SJ40 strain grew at 37°C., Conclusions: To our knowledge, this is the first report of indoor-isolated Trichoderma strains producing toxic peptaibols in their guttation droplets., Significance and Impact of the Study: This report proves that indoor isolates of Trichoderma release peptaibols in their guttation droplets. The presence of toxins in these types of exudates may serve as a mechanism of aerosol formation for nonvolatile toxins in the indoor air., (© 2018 The Society for Applied Microbiology.)

The aim of this study was to evaluate antifungal activity in a diverse group of chlorine-containing xanthone and phenoxyethyl amine derivatives - and to select the most promising compounds for further studies. The antifungal efficacy of 16 compounds was tested with qualitative and quantitative methods against both reference and clinical strains of dermatophytes, moulds and yeasts. The disc-diffusion method has demonstrated that from 16 tested compounds, 7 possess good antifungal activity against dermatophytes and/or moulds while none of them has shown good efficacy against yeasts or bacterial strains. The most active compounds (2, 4, 10, 11, 12, 15, 16) were tested quantitatively by broth dilution method to obtain MIC values. The MIC values against dermatophytes ranged from 8 to 64 μg/ml. Compound 2 was the most active one against dermatophytes (MIC 50 and MIC 90 were 8 μg/ml). The MIC values for moulds ranged from 16 to 256 μg/ml. Compound 4 was the most active one against moulds, with MIC 50 and MIC 90 values amounting to 32 μg/ml. Among the tested compounds, compound 4 (derivative of xanthone) was the most active one and expressed good antifungal efficacy against clinical strains of dermatophytes and moulds. However, another xanthone derivative (compound 2) was the most active and selective against dermatophytes., (© 2018 John Wiley & Sons A/S.)

Introduction: Moulds and mite allergens present in indoor environments are well known for their effects on respiratory health., Methods: From 2011 to 2015, the Paris Service for Environmental Health (SPSE) conducted investigations in 293 dwellings following medical referral. These audits included fungal analysis of air (in 12% of dwellings), in mattress surface and floor dust (24%), and mite allergen quantifications in mattresses and carpets (18%)., Results: Indoor air fungal concentrations are not significantly different from those in outdoor air. When there is no ventilation or when the system is malfunctioning, an increase in indoor/outdoor air ratios is observed, indicating mould enrichment in the dwelling's indoor air. With regard to house dust samples, fungal spore concentrations vary according to the media from which samples were collected. Mattress fungal contamination is higher in dwellings where observed surface moulds exceed 1 per square meter. In the same way Der p1 mite allergens levels are greater in mattress dust in dwellings where mould contamination is visible., Conclusions: This study describes the levels of contamination in the dwellings of Parisian patients., (Copyright © 2018 SPLF. Published by Elsevier Masson SAS. All rights reserved.)

Tea is one of the most popular beverages all over the world. Being an everyday drink for almost everyone, for centuries tea was considered safe and healthy. However, fungal contamination of tea at any stage of commodity production can pose a serious health hazard due to the accumulation of toxic secondary metabolites of moulds. Contemporary research revealed incidences of highly contaminated samples. Mycotoxin transfer from naturally contaminated raw tea into beverage was well studied for ochratoxin A only, and the possible leak of other mycotoxins is discussed. The results of several surveys were combined to evaluate aflatoxin B1 and ochratoxin A contamination levels in black tea and Pu-erh. Exposure estimate to aflatoxin B1 and ochratoxin A due to tea consumption was carried out based on these data. Average contamination level corresponds to the exposure of 3⁻40% (aflatoxin B1) and 5⁻24% (ochratoxin A) of mean overall estimates for different cluster diets. Lack of data does not allow the conclusion for the necessity of public health protection measures. It is necessary to perform representative studies of different kinds of tea for regulated mycotoxins at least. Contemporary techniques for analysis of mycotoxins in tea are summarised in the present review.

Matrix-assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS) is commonly used by clinical microbiology laboratories to identify bacterial pathogens and yeasts, but not for the identification of moulds. Recent progress in extraction protocols and the composition of comparative libraries support potential application of MALDI-TOF MS for mould identification in clinical microbiology laboratories. We evaluated the performance of the Bruker Microflex™ MALDI-TOF MS instrument (Billerica, MA, USA) to identify clinical isolates and reference strains of moulds using 3 libraries, the Bruker mould library, the National Institutes of Health (NIH) library and the Mass Spectrometry Identification (MSI) online library, and compared those results to conventional (morphological) and molecular (18S/ITS; gold standard) identification methods. All 3 libraries demonstrated greater accuracy in genus identification (≥94.9%) than conventional methods (86.4%). MALDI-TOF MS identified 73.3% of isolates to species level compared to only 31.7% by conventional methods. The MSI library demonstrated the highest rate of species-level identification (72.0%) compared to NIH (19.5%) and Bruker (13.6%) libraries. Greater than 20% of moulds remained unidentified to species level by all 3 MALDI-TOF MS libraries primarily because of library limitations or imperfect spectra. The overall identification rate of each MALDI-TOF MS library depended on the number of species and the number of spectra representing each species in the library., (© 2018 Blackwell Verlag GmbH.)

The purpose of this study was to investigate the antimicrobial activity and mechanisms of N α -lauroyl arginate ethyl ester (LAE) against Penicillium digitatum and Pectobacterium carotovorum subsp. carotovorum . The minim inhibitory concentrations of LAE against P. digitatum and P. carotovorum were found to be 400 and 25 μg/ml, respectively. Loss of intracellular protein and nucleic acid increased significantly, and membrane permeability reached 76.28, 54.29 and 85.20%, respectively, when 400 μg/ml of LAE was applied to the hyphae and spores of P. digitatum and to P. carotovorum . Flow cytometry showed that LAE reduced the membrane potential, and the depolarization ratios of P. digitatum and P. carotovorum were 98.19 and 97.25% ( P  < 0.05), respectively. Transmission electron microscopy photos revealed that LAE caused a rough surface, irregular cellular organelles, protoplast shrinkage, intracytoplasmic coagulation and empty cavities in all three cell types. These results showed that LAE had notable ability to damage the structure of fungal and bacterial cells, making it a possible alternative chemical for use in the preservation of fruits and vegetables.

In 2013, mould growth on Norwegian ready-to-sell pinnekjøtt (dried and cured lamb ribs) lead to the withdrawal of 200 tons of the product. The aim of this study was to identify the mycobiota at two Norwegian production sites and determine which species present the highest risk for reduced product quality and safety. A total of 485 samples from 2014, 2015 and 2016 were analysed for Penicillium and Aspergillus species. Both production sites showed a persistent mycobiota that remained stable over three seasons. Samples from site A were dominated by P. solitum while samples from site B were equally dominated by P. solitum, P. brevicompactum/bialowiezense and P. nordicum. The presence of P. nordicum was concentrated in one area of the production site where long-time stored hams were produced, but P. nordicum was also found sporadically in other parts of the site. Product samples taken from products with visible mould growth were at both sites dominated by P. solitum, highlighting its importance for product quality. P. nordicum was found frequently in the long-time stored hams, indicating a food safety risk of these products. However, P. nordicum was rarely isolated from pinnekjøtt. Aspergillus spp. were isolated from both sites at all samplings; however, there were no Aspergillus isolated from products, and no sites were repeatedly tested positive for identical species, indicating that Aspergillus is not a part of the persistent mycobiota, but enters the site sporadically. In conclusion, the study showed that a stable mycobiota consisting of few Penicillium species dominated the products and production environments of pinnekjøtt., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Surface-assisted laser desorption/ionization mass spectrometry on gold nanoparticle enhanced target (AuNPET) technique was used for metabolomic analysis and secondary metabolites detection of two mould strains - Aspergillus versicolor and Penicillium chrysogenum in model conditions on microbiological malt extract agar medium. Results obtained with the use of AuNPET-based mass spectrometry technique were compared with traditional matrix-assisted laser desorption/ionization (MALDI) method based on α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB) matrices. Gold nanoparticle enhanced target method enabled effective ionization of microbial cellular extract ingredients without interference from the matrix and also improved calibration of spectra resulting in the detection of much higher amount of characteristic metabolites for studied organisms than MALDI., (Copyright © 2018 Elsevier Inc. All rights reserved.)