The Evolution of the Satratoxin and Atranone Gene Clusters of Stachybotrys chartarum

Stachybotrys chartarum is frequently isolated from damp building materials or improperly stored animal forage. Human and animal exposure to the secondary metabolites of this mold is linked to severe health effects. The mutually exclusive production of either satratoxins or atranones defines the chemotypes A and S. Based upon the genes (satratoxin cluster, SC1-3, sat or atranone cluster, AC1, atr) that are supposed to be essential for satratoxin and atranone production, S. chartarum can furthermore be divided into three genotypes: the S-type possessing all sat- but no atr-genes, the A-type lacking the sat- but harboring all atr-genes, and the H-type having only certain sat- and all atr-genes. We analyzed the above-mentioned gene clusters and their flanking regions to shed light on the evolutionary relationship. Furthermore, we performed a deep re-sequencing and LC-MS/MS (Liquid chromatography–mass spectrometry) analysis. We propose a first model for the evolution of the S. chartarum genotypes. We assume that genotype H represents the most ancient form. A loss of the AC1 and the concomitant acquisition of the SC2 led to the emergence of the genotype S. According to our model, the genotype H also developed towards genotype A, a process that was accompanied by a loss of SC1 and SC3

Influence of silver nanoparticles on metabolism and toxicity of moulds*

The unique antimicrobial features of silver nanoparticles (AgNPs) are commonly applied in innumerable products. The lack of published studies on the mechanisms of AgNPs action on fungi resulted in identification of the aim of this study, which was: the determination of the influence of AgNPs on the mould cytotoxicity for swine kidney cells (MTT test) and the production of selected mycotoxins, organic acids, extracellular enzymes by moulds. The conducted study had shown that silver nanoparticles can change the metabolism and toxicity of moulds. AgNPs decrease the mycotoxin production of Aspergillus sp. (81-96%) and reduce mould cytotoxicity (50-75%). AgNPs influence the organic acid production of A. niger and P. chrysogenum by decreasing their concentration (especially of the oxalic and citric acid). Also, a change in the extracellular enzyme profile of A. niger and P. chrysogenum was observed, however, the total enzymatic activity was increased

Occupational Exposure to Mycotoxins in Swine Production: Environmental and Biological Monitoring Approaches

Free PMC article: https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/30717100/The authors are grateful to the swine farms employers and workers that collaborate in this research project. R.A. and C.M. are grateful to INSA and to CESAM (UID/AMB/50017/2013) through national funds (FCT), and the co-funding by the Fundo Europeu de Desenvolvimento Regional (FEDER) (POCI-01-0145-FEDER-00763), within the PT2020 Partnership Agreement and Compete 2020.Swine production workers are exposed simultaneously to multiple contaminants. Occupational exposure to aflatoxin B1 (AFB1) in Portuguese swine production farms has already been reported. However, besides AFB1, data regarding fungal contamination showed that exposure to other mycotoxins could be expected in this setting. The present study aimed to characterize the occupational exposure to multiple mycotoxins of swine production workers. To provide a broad view on the burden of contamination by mycotoxins and the workers’ exposure, biological (urine) samples from workers (n = 25) and 38 environmental samples (air samples, n = 23; litter samples, n = 5; feed samples, n = 10) were collected. The mycotoxins biomarkers detected in the urine samples of the workers group were the deoxynivalenol-glucuronic acid conjugate (60%), aflatoxin M1 (16%), enniatin B (4%), citrinin (8%), dihydrocitrinone (12%) and ochratoxin A (80%). Results of the control group followed the same pattern, but in general with a lower number of quantifiable results (<LOQ). Besides air samples, all the other environmental samples collected presented high and diverse contamination, and deoxynivalenol (DON), like in the biomonitoring results, was the most prominent mycotoxin. The results demonstrate that the occupational environment is adding and contributing to the workers’ total exposure to mycotoxins, particularly in the case of DON. This was confirmed by the biomonitoring data and the high contamination found in feed and litter samples. Furthermore, the followed multi-biomarker approach allowed to conclude that workers and general population are exposed to several mycotoxins simultaneously. Moreover, occupational exposure is probably described as being intermittent and with very high concentrations for short durations. This should be reflected in the risk assessment process.This research was funded by Instituto Politécnico de Lisboa, Lisbon, Portugal: Project “Bacterial Bioburden assessment in the context of occupational exposure and animal health of swine productions (IPL/2016/BBIOR_ESTeSL)” and also by FCT—Fundação para Ciência e Tecnologia: Project “EXPOsE – Establishing protocols to assess occupational exposure to microbiota in clinical settings (02/SAICT/2016 – Project nº 23222)”.info:eu-repo/semantics/publishedVersio

Portuguese coffee: is there a concern regarding mycotoxins contamination?

According to the International Coffee Organization, global coffee production is rising driven by an increase in total demand. However, similar to other crops, coffee cherries, and beans are exposed to contamination and consequent colonization by fungi during different phases of plant development, harvesting, transport, and storage. Consequently, mycotoxins also contaminate coffee beans although the roasting process has an important role in reducing mycotoxins presence, depending upon the combination of time and temperature. Since each country applies different roasting conditions, significant differences in mycotoxins concentrations occur across Europe with respect to contamination levels even when the origin of the coffee is the same. A previous paper showed high contamination by toxigenic mycobiota of coffee beans to be used in Portuguese coffee industries.info:eu-repo/semantics/publishedVersio

Portuguese groceries: a potential indoor environment to be used as public health sentinel for fungal and mycotoxins contamination

This study aimed to characterize microbial contamination in Portuguese grocery stores. The research was carried out in 15 grocery stores in Cascais, Portugal. Electrostatic dust cloths (EDC) and surface swabs were used on 3 sampling sites: checkout, fruits/vegetables, and warehouse/dispenser. Fungal contamination was characterized by culture-based methods, through the inoculation onto two different culture media: malt extract agar (MEA) supplemented with chloramphenicol (0.05 %) and dichloran-glycerol agar (DG18). Screening of azole resistance was performed by inoculation of EDC extracts' in azole-supplemented Sabouraud dextrose agar (SDA) media, according to EUCAST guidelines. A total of 39 samples were screened for mycotoxin contamination. The highest fungal contamination was obtained from fruits/vegetables in swabs (76% MEA; 71% DG18). The most prevalent genera in swabs was Cladosporium sp. (54% MEA). Regarding EDC, besides Penicillium sp. (63%) the most prevalent were Aspergillus sections Circumdati (25%) and Aspergilli (35%). Penicillium sp. was highly prevalent in the checkout (61% SDA; 87% voriconazole), and warehouse/dispenser sectors (73% voriconazole; 52% posaconazole). Circumdati and Fumigati were the most prevalent Aspergillus sections in fruits/vegetables (73% and 64% in voriconazole, respectively). Four mycotoxins (fumonisin B1, B2, B3) were detected simultaneously in most of the 36 samples. Identifying the most critical workplaces in groceries concerning fungal and mycotoxin contamination is of utmost importance to human health (workers and consumers) when using a One Health approach.info:eu-repo/semantics/publishedVersio

Unveiling this Portuguese occupational exposure scenario

Copyright © 2022 Elsevier Ltd. All rights reserved.Previous studies anticipated that microorganisms and their metabolites in waste will increase as a consequence of a decreased collection frequency and due to differences in what kind of waste is bagged before collection leading to an increased exposure of workers handling the waste. This study aim was to investigate the microbial contamination present in the waste collection trucks (WCT) and in the support facilities (waste collection station - WCS). It was applied a multi-approach protocol using active (air sampling by impingement and impaction) and passive (surface swabs, electrostatic dust cloths and settled dust) sampling methods. The screening of azole-resistance, the investigation of mycotoxins and the assessment of the elicited biological responses in vitro were also carried out aiming recognizing the possible health effects of waste collection drivers. SARS-CoV-2 detection was also performed. In WCS only air samples had contamination in all the four sampling sites (canteen, operational removal core, operational removal center, and administrative service). Among all the analyzed matrices from the WCT a higher percentage of total bacterial counts and Gram-was detected in swabs (66.93%; 99.36%). In WCS the most common species were Penicillium sp. (43.98%) and Cladosporium sp. (24.68%), while on WCT Aspergillus sp. (4.18%) was also one of the most found. In the azole resistance screening Aspergillus genera was not observed in the azole-supplemented media. SARS-CoV-2 was not detected in any of the environmental samples collected, but Aspergillus section Fumigati was detected in 5 samples. Mycotoxins were not detected in EDC from WCS, while in WCT they were detected in filters (N = 1) and in settled dust samples (N = 16). In conclusion, our study reveals that a comprehensive sampling approach using active and passive sampling (e.g. settled dust sampling for a representative mycotoxin evaluation) and combined analytic methods (i.e., culture-based and molecular) is an important asset in microbial exposure assessments. Concerning the waste collection exposure scenario, the results of this study unveiled a complex exposure, particularly to fungi and their metabolites. Aspergillus section Fumigati highlight the significance of targeting this section in the waste management industry as an indicator of occupational health risk.publishersversionpublishe

Bioburden assessment by passive methods on a clinical pathology service in one central hospital from Lisbon: what can it tell us regarding patients and staff exposure?

Project EXPOsE. Project nº 23222 (02/SAICT/2016).The assessment and control of microbial contamination in health care facilities is presently a mandatory and vital part of strategies to prevent and control hospital-acquired infections. This study aims to assess the bioburden with two passive sampling methods (30 ventilations grids swabs and 16 electrostatic dust collectors (EDCs)) at Clinical Pathology Services. The fungal burden was characterized through molecular tools, antifungal resistance, and the mycotoxins and cytotoxicity profile. Total bacteria presented the highest prevalence in both matrixes, whereas Gram-bacteria presented the lowest. Swabs presented a higher prevalence (27.6%) for fungal burden. Chrysonilia sitophila presented the highest prevalence in swabs, whereas for EDCs, C. sitophila and Mucor sp. were the most prevalent. Concerning Aspergillus genera on swabs, section Flavi was the one with the highest prevalence (58.02%), whereas, for EDCs, section Versicolores was the only section observed (100%). Aspergillus section Fumigati was detected in 10 swabs and 7 EDC samples and Aspergillus section Versicolores were detected in one EDC sample. Fungal growth on azole-supplemented media was observed in eight EDC samples. No mycotoxins were detected in any of the samples. A low cytotoxic effect was observed in two sites upon incubation of collected samples with A549 and SK cells and in two other sites upon incubation of collected samples with SK cells only. A medium cytotoxic effect was observed with one EDC sample upon incubation with A549 cells. This study reinforces the need for determination of the azole resistance profile for fungal species and allowed a preliminary risk characterization regarding the cytotoxicity. An intervention including the use of ultraviolet with a wavelength between 200 nm and 280 nm (UVC)—emitting device and increased maintenance and cleaning of the central heating, ventilation, and air conditioning (HVAC) systems should be ensured to promote the reduction of microbial contamination.info:eu-repo/semantics/publishedVersio

Occupational exposures to organic dust in Irish bakeries and a pizzeria restaurant

Project EXPOsE, nº 23222 (02/SAICT/2016)For decades, occupational exposure to flour dust has been linked to a range of respiratory diseases, including occupational asthma, thought to result from exposure to fungi present in the flour. Antifungal resistance is of increasing prevalence in clinical settings, and the role of occupational and environmental exposures, particularly for specific fungal species, is of concern. Occupational exposure to flour dust can occur in a range of occupational settings, however, few studies have focused on restaurant workers. The objective of this study was to measure occupational exposure to the flour and microbial contamination, including azole resistance screening, in two small commercial bakeries and in a pizzeria. Personal full shift inhalable dust measurements were collected from workers and were analyzed for inhalable dust and fungi, bacteria, azole resistance, and mycotoxins. Samples of settled dust were collected, and electrostatic dust cloths (EDC) were deployed and analyzed for microbial contamination, including azole resistance screening, and mycotoxins. Geometric mean exposures of 6.5 mg m-³ were calculated for inhalable dust, however, exposures of up to 18.30 mg m-³ were measured-70% of personal exposure measurements exceeded the occupational exposure limit for flour dust of 1.0 mg m-³. The air and EDC fungal counts were similar to those reported in previous studies for similar occupational environments. The fungi were dominated by Penicillium genera, however Aspergillus genera, including Fumigati and Flavi sections, were observed using culture-based methods, and the Fumigati section was also observed by molecular tools. Both Aspergillus sections were identified on the azole resistance screening. Mycotoxins were also detected in the settled dust samples, dominated by deoxynivalenol (DON). The role of environmental exposure in both the development of antimicrobial resistance and the total mycotoxin body burden is a growing concern; therefore, the presence of azole-resistant fungi and mycotoxin contamination, although low in magnitude, is of concern and warrants further investigation.info:eu-repo/semantics/publishedVersio

the neglected indoor environment to be tackled in the scope of the One Health approach

Funding Information: H&TRC authors gratefully acknowledge the FCT/MCTES national support through the UIDB/05608/2020 , the UIDP/05608/2020 and the PhD Grant UI/BD/151431/2021 . This work was also supported by national funds through FCT/MCTES/FSE/UE , UI/BD/153746/2022 and CE3C unit UIDB/00329/2020 within the scope of a PhD Grant. Funding Information: This work was supported by the Polish Minister of Science and Higher Education under the program “Regional Initiative of Excellence” in 2019–2022 (Grant No. 008/RID/2018/19 ). Funding Information: All the authors acknowledge the scientific support from the Natural Environment Research Council (NERC) in the scope of BioSkyNet workshop held by University of Essex (Ref: NE/V008293/1 ). Funding Information: ISGlobal authors acknowledge support from the Spanish Ministry of Science and Innovation and State Research Agency through the “Centro de Excelencia Severo Ochoa 2019–2023” Program ( CEX2018-000806-S ) and support from the Generalitat de Catalunya through the CERCA Program”. Publisher Copyright: © 2023 The AuthorsMicrobial contamination in grocery shops (GS) should be evaluated since food commodities are commonly handled by workers and customers increasing the risk of food contamination and disease transmission. The aim of this study was to evaluate the microbial contamination in Portuguese and Spanish GS with a multi-approach protocol using passive (electrostatic dust cloths and surface swabs) sampling methods. The molecular detection of Aspergillus sections, mycotoxin analysis, screening of azole resistance as well as cytotoxicity measurement were conducted to better estimate the potential health risks of exposure and to identify possible relations between the risk factors studied. Fruits/vegetables sampling location was the one identified has being the most contaminated (bacteria and fungi) area in GS from both countries. Aspergillus section Fumigati and Fusarium species were observed in samples from Portuguese groceries with reduced susceptibilities to azoles commonly used in the clinical treatment of fungal infections. Fumonisin B2 was detected in Portuguese GS possible unveiling this emergent threat concerning occupational exposure and food safety. Overall, the results obtained raise concerns regarding human health and food safety and must be surveilled applying a One Health approach.publishersversionpublishe

A new approach to assess fungal burden and mycotoxins occupational exposure in waste trucks workers

Este trabalho foi financiado pelo Concurso Anual para Projetos de Investigação, Desenvolvimento, Inovação e Criação Artística (IDI&CA) 2016 do Instituto Politécnico de Lisboa. Código de referência: IPL/2016/W2E_ESTeSLBackground - Waste application facilities are considered critical regarding occupational exposure to fungal burden since provide optimal conditions for fungal growth (moisture and decaying matter). Aspergillus genera dominance was already reported in Portuguese waste industry. Waste-sorting the most critical setting, presenting not only the highest load but also the wider diversity of Aspergillus sections. It is important to consider the co-occurrence of mycotoxins in this occupational environment. In waste management industry it is common to use forklifts for waste transportation and workers can spend all their working hours in a forklift cab. In cars, the bioburden filtered from the air stream by the automobile air conditioner filter could proliferate under high humidity conditions and, consequently, presenting a potential exposure source to bioaerosols. When the air conditioning system is turned on, the air stream passing through the vehicle filtration system could re-aerosolize the bioburden and subsequently carry them into the vehicle cabinet increasing the exposure.info:eu-repo/semantics/publishedVersio