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IMPORTANCE: Saprophytic fungi can cause nosocomial infections in high-risk patients. These infections are related to high mortality and cost. In the current study, different species of filamentous fungi and yeast were isolated from the environment of the studied hospitals. Some species were resistant to antifungal drugs. We suggest that the future work concentrates on the relationship between the level/quantification of saprophytic contamination in the environment of hospitals and fungal infections in patients.

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Accurate mold identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is dependent on robust organism representation in available databases. The Mass Spectrometry Identification (MSI) platform has proven successful for mold identification in clinical and veterinary settings but has yet to be studied with a large set of environmental isolates. Here, we performed a retrospective study using spectra collected by the Bruker MALDI Biotyper (MBT) v4.1 microflex LT instrument to evaluate the MSI-2 database alongside the combined use of the Bruker MBT (including the MBT Filamentous Fungi Library) and the National Institutes of Health (NIH) mold database (MBT/NIH databases). Analysis was performed for 462 environmental fungal isolates (representing 73 different fungi) cultured from the hospital pharmacy and cellular therapy suites as part of the current good manufacturing practices (cGMP) environmental monitoring program at the NIH. When used alone, MSI-2 identified 237 spectra (51.3%) at its higher score threshold (index A), while the MBT/NIH databases identified only 183 spectra (39.6%; P < 0.001) at the equivalent threshold of ≥2.0. The combination of all three databases at the respective high thresholds improved identification sensitivity to 327 spectra (70.8%). The combination of MSI-2 with the MBT/NIH databases at a lowered threshold (index B or ≥1.7, respectively) identified 400/462 environmental spectra (86.6%). Our results show that the MSI-2 database, in combination with existing databases, may be useful for environmental surveillance, particularly by clinical or industry laboratories involved in cGMP or current good tissue practices (cGTP) applications, such as cellular therapy manufacturing facilities and sterile compounding pharmacies.

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The distribution and availability of microbes in the environment has an important effect on the composition of the gut microbiome of wild vertebrates. However, our current knowledge of gut-environmental interactions is based principally on data from the host bacterial microbiome, rather than on links that establish how and where hosts acquire their gut mycobiome. This complex interaction needs to be clarified. Here, we explored the relationship between the gut fungal communities of Tibetan macaques (Macaca thibetana) and the presence of environmental (plant and soil) fungi at two study sites using the fungal internal transcribed spacer (ITS) and next generation sequencing. Our findings demonstrate that the gut, plant and soil fungal communities in their natural habitat were distinct. We found that at both study sites, the core abundant taxa and ASVs (Amplicon Sequence Variants) of Tibetan macaques' gut mycobiome were present in environmental samples (plant, soil or both). However, the majority of these fungi were characterized by a relatively low abundance in the environment. This pattern implies that the ecology of the gut may select for diverse but rare environmental fungi. Moreover, our data indicates that the gut mycobiome of Tibetan macaques was more similar to the mycobiome of their plant diet than that present in the soil. For example, we found three abundant ASVs (Didymella rosea, Cercospora, and Cladosporium) that were present in the gut and on plants, but not in the soil. Our results highlight a relationship between the gut mycobiome of wild primates and environmental fungi, with plants diets possibly contributing more to seeding the macaque's gut mycobiome than soil fungi.

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Asthma is a group of inflammatory conditions that compromises the airways of a continuously increasing number of people around the globe. Its complex etiology comprises both genetic and environmental aspects, with the intestinal and lung microbiomes emerging as newly implicated factors that can drive and aggravate asthma. Longitudinal infant cohort studies combined with mechanistic studies in animal models have identified microbial signatures causally associated with subsequent asthma risk. The recent inclusion of fungi in human microbiome surveys has revealed that microbiome signatures associated with asthma risk are not limited to bacteria, and that fungi are also implicated in asthma development in susceptible individuals. In this review, we examine the unique properties of human-associated and environmental fungi, which confer them the ability to influence immune development and allergic responses. The important contribution of fungi to asthma development and exacerbations prompts for their inclusion in current and future asthma studies in humans and animal models.

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Invasive fungal infections are a growing threat to immunocompromised patients, highlighting the importance of monitoring fungal pathogens. Global warming (including climatic oscillations) may select for environmental species that have acquired thermotolerance, a key step toward pathogenesis to humans. Also, important virulence factors have developed in environmental fungi, because they are essential for yeast survival in the environment. Thus, fungi traditionally regarded as nonpathogenic to humans have virulence factors similar to those of their pathogenic relatives. Here, we highlight the emergence of saprophytic environmental fungi - including species of Cryptococcus, Aspergillus, Penicillium, Candida and Scedosporium - as new human pathogens. Emerging pathogens are, in some cases, resistant to the available antifungals, potentiating the threat of novel fungal diseases.

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　Loop-mediated isothermal amplification (LAMP) is a useful DNA detection method with high specificity and sensitivity. The LAMP reaction is carried out within a short time at a constant temperature without the need for thermal cycling. We developed a LAMP primer set for detecting a wide range of fungi by aligning the sequences of the large subunit ribosomal RNA gene of Candida albicans (Ascomycota), Cryptococcus neoformans (Basidiomycota), and Mucor racemosus (Mucorales). The threshold of C. albicans rDNA as template with our LAMP primer set was in the range of 10-100 copies per a reaction. In this study, we evaluated the correlation between colony forming units (CFU) and LAMP detection rate using the LAMP method for environmental fungi. The LAMP method should be a useful means of detecting fungi in indoor environments, disaster areas, or even in confined manned spacecraft to prevent allergies or infections caused by fungi.

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BACKGROUND: Exposure to molds has been related to asthma risk both positively and negatively, depending on the environmental setting. The pertinent results are based on generic markers or culturing methods although the majority of present fungi cannot be cultured under laboratory conditions. The aim of the present analysis was to assess environmental dust samples for asthma-protective fungal candidates with a comprehensive molecular technique covering also non-cultivable and non-viable fungi. METHODS: Mattress dust samples of 844 children from the GABRIELA study were analyzed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) of the fungus-specific internal transcribed spacer (ITS) region. Known asthma candidate species were tested for their associations with asthma, and further gel positions were sought to explain the above. As a second, data-driven, analysis, we tested the association of each individual gel position with asthma. RESULTS: In the hypothesis-driven approach, Penicillium chrysogenum emerged with an odds ratio of 0.80 (95% confidence interval 0.66-0.96; p = 0.020). The effect size was changed by 39% toward the null when adjusting for the two bands 683 (DNA of Metschnikowia sp., Aureobasidium spp.) and 978 (DNA of Epicoccum spp., Galactomyces spp., uncultured Penicillium). The data-driven approach yielded an additional band (containing DNA of Pseudotaeniolina globosa) with reduced risk of asthma (OR = 0.80 [0.66-0.96], p = 0.012). CONCLUSIONS: A large population-based study revealed several fungal taxa with inverse associations with childhood asthma. Molds produce a variety of bioactive compounds with detrimental but also beneficial immunoregulatory capacities, which renders them promising targets for further asthma research.

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Fungal contamination of indoor air is an issue of increasing public health concern. Essential oils have been demonstrated to have antifungal capabilities, but there are limited studies investigating the efficacy of essential oils against fungi relevant to air quality. This study provides a preliminary screening of the antifungal properties of clove, lavender and eucalyptus essential oils against a range of fungal species isolated from environmental air samples. The ability of the essential oils to inhibit fungal growth was examined using the disk diffusion assay on malt extract agar and was compared with vinegar, bleach and limonene, with phenol as a positive control. Results identified essential oils which demonstrated antifungal potential against species of environmental origin. Clove oil was found to be most efficacious, with eucalyptus and lavender oils showing some antifungal potential albeit less broad spectrum and with less persistence over time in this assay. All essentials oils performed better than traditional cleaning compounds such as vinegar. Clove oil would be a suitable candidate for further research to validate its use in improving indoor air quality. Further research should next take into consideration the practical application method, concentration and long-term persistence of antifungal properties.

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BACKGROUND: Aspergillosis has been identified as one of the hospital acquired infections but the contribution of water and inhouse air as possible sources of Aspergillus infection in immunocompromised individuals like HIV-TB patients have not been studied in any hospital setting in Nigeria. OBJECTIVE: To identify and investigate genetic relationship between clinical and environmental Aspergillus sp. associated with HIV-TB co infected patients. METHODS: DNA extraction, purification, amplification and sequencing of Internal Transcribed Spacer (ITS) genes were performed using standard protocols. Similarity search using BLAST on NCBI was used for species identification and MEGA 5.0 was used for phylogenetic analysis. RESULTS: Analyses of sequenced ITS genes of selected fourteen (14) Aspergillus isolates identified in the GenBank database revealed Aspergillus niger (28.57%), A. tubingensis (7.14%), A. flavus (7.14%) and A. fumigatus (57.14%). Aspergillus in sputum of HIV patients were Aspergillus niger, A. fumigatus, A. tubingensis and A. flavus. Also, A. niger and A. fumigatus were identified from water and open-air. Phylogenetic analysis of sequences yielded genetic relatedness between clinical and environmental isolates. CONCLUSION: Water and air in health care settings in Nigeria are important sources of Aspergillus sp. for HIV-TB patients.

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The aim of the present study was to assess performance, feasibility and relevance of a Standard Operational Procedure (SOP) for large-scale use in the microbial analysis of children's indoor environments. We analyzed dust settled on Electrostatic Dust Fall Collectors (EDCs) by using qPCR which targeted 6 molds, 3 bacteria and 1 mite, chosen for their involvement in allergic or inflammatory processes. Six types of commercialized electrostatic wipes were tested for their releasing capacity of fungal DNA from fungal spores captured by the wipes. Specificity, repeatability and detection limits of the qPCR procedure were tested using calibrated microbial suspensions. The feasibility and relevance of this sampling and analysis method were assessed in a 75-home pilot study. Our result showed that one specific make of wipe was more effective than the others in releasing fungal DNA. qPCR procedure showed good repeatability. The quantification limit was about 5 fg DNA/µL for all species except Penicillium chrysogenum (0.5 fg DNA/µL) and Dermatophagoïdes pteronyssinus (10 fg DNA/µL). No cross-reactivity was observed. DNA concentrations in the 53/75 homes participating in the pilot study were between 0 and 24 625, 0 and 69 738 equivalent cells per cm(2) for the fungi and bacteria, and between 0 and 1 equivalent mites per cm(2) for D. pteronyssinus. Using the SOP described, we were able to classify the 53 dwellings from the least to the most contaminated according to the quantity of DNA measured for each species. Our SOP measured fungi, bacteria and mites using a cost-efficient, discreet and well-accepted sampling method with just one qPCR tool. The whole procedure can be used for microbial analysis in large cohort studies such as the ELFE study ("Etude Longitudinale Française depuis l'Enfance") and could help improve our understanding of the interactions between the environment, allergic diseases and child development.

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Las enfermedades alérgicas y el asma, sobre todo en la población infantil, son afecciones muy frecuentes en todo el mundo y en Cuba. Con el objetivo de determinar la frecuencia de sensibilización a hongos ambientales y su relación con enfermedades atópicas, se realizó un estudio analítico transversal en 100 escolares de 6 a 7 años de una escuela primaria de San Antonio de los Baños, provincia La Habana, en el período comprendido entre septiembre de 2006 y marzo de 2007. El 27 por ciento de la muestra seleccionada padecía de asma, el 40 por ciento de rinitis alérgica y el 26 por ciento de dermatitis atópica. La sensibilización micótica más frecuente resultó Penicillium, para un 50 por ciento de la muestra. No hubo una asociación estadísticamente significativa entre reactividad cutánea a hongos ambientales y la presencia de enfermedades atópicas; tampoco fue significativa su asociación con la enfermedad alérgica respiratoria ni dermatitis atópica. Se concluye que la mayor sensibilización a hongos anemófilos se apreció al Penicillium, aunque sin asociación con la presencia de enfermedades atópicas(AU)

The allergic diseases and the asthma, mainly in children, are very frequent affections at world level and in Cuba. The objective of present paper was to determine the sensitization frequency to environmental fungi and its relation to atopic diseases, thus, a cross-sectional and analytical study was conducted in 100 school children aged 6 to 7 from a primary school of San Antonio de los Ba±os, La Habana province, from September 2006 to March, 2007. The 27 percent of the selected sample suffered from asthma, the 40 percent of allergic rhinitis, and the 26 percent had Penicillium for the 50 percent of sample. There was not a significant statistic association between the cutaneous reactivity to environmental fungi and the presence of atopic diseases and its association with the respiratory allergic disease as well as the atopic dermatitis was not significant. We conclude that the greater sensitization to anemophilic fungi was to Penicillium, although without an association with the presence of atopic diseases(AU)

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Las enfermedades alérgicas y el asma, sobre todo en la población infantil, son afecciones muy frecuentes en todo el mundo y en Cuba. Con el objetivo de determinar la frecuencia de sensibilización a hongos ambientales y su relación con enfermedades atópicas, se realizó un estudio analítico transversal en 100 escolares de 6 a 7 años de una escuela primaria de San Antonio de los Baños, provincia La Habana, en el período comprendido entre septiembre de 2006 y marzo de 2007. El 27 por ciento de la muestra seleccionada padecía de asma, el 40 por ciento de rinitis alérgica y el 26 por ciento de dermatitis atópica. La sensibilización micótica más frecuente resultó Penicillium, para un 50 por ciento de la muestra. No hubo una asociación estadísticamente significativa entre reactividad cutánea a hongos ambientales y la presencia de enfermedades atópicas; tampoco fue significativa su asociación con la enfermedad alérgica respiratoria ni dermatitis atópica. Se concluye que la mayor sensibilización a hongos anemófilos se apreció al Penicillium, aunque sin asociación con la presencia de enfermedades atópicas

The allergic diseases and the asthma, mainly in children, are very frequent affections at world level and in Cuba. The objective of present paper was to determine the sensitization frequency to environmental fungi and its relation to atopic diseases, thus, a cross-sectional and analytical study was conducted in 100 school children aged 6 to 7 from a primary school of San Antonio de los Ba±os, La Habana province, from September 2006 to March, 2007. The 27 percent of the selected sample suffered from asthma, the 40 percent of allergic rhinitis, and the 26 percent had Penicillium for the 50 percent of sample. There was not a significant statistic association between the cutaneous reactivity to environmental fungi and the presence of atopic diseases and its association with the respiratory allergic disease as well as the atopic dermatitis was not significant. We conclude that the greater sensitization to anemophilic fungi was to Penicillium, although without an association with the presence of atopic diseases

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In this work, different reactions in vitro between an environmental bacterial isolate and fungal species were related. The Gram-positive bacteria had terminal and subterminal endospores, presented metabolic characteristics of mesophilic and acidophilic growth, halotolerance, positive to nitrate reduction and enzyme production, as caseinase and catalase. The analysis of partial sequences containing 400 to 700 bases of the 16S ribosomal RNA gene showed identity with the genus Bacillus. However, its identity as B. subtilis was confirmed after analyses of the rpoB, gyrA, and 16S rRNA near-full-length sequences. Strong inhibitory activity of environmental microorganisms, such as Penicillium sp, Aspergillus flavus, A. niger, and phytopathogens, such as Colletotrichum sp, Alternaria alternata, Fusarium solani and F. oxysporum f.sp vasinfectum, was shown on co-cultures with B. subtilis strain, particularly on Sabouraud dextrose agar (SDA) and DNase media. Red and red-ochre color pigments, probably phaeomelanins, were secreted by A. alternata and A. niger respectively after seven days of co-culture.

Na presente investigação, nosso objetivo principal foi relatar diferentes interações in vitro de um isolado bacteriano ambiental com espécies fúngicas. Através da identificação clássica, nós verificamos que o bacilo ambiental apresentava endósporos terminais e subterminais, características metabólicas de mesofilia, acidofilia, halotolerância, redução de nitrato e produção de enzimas, como caseinase e catalase. Análise de seqüências parciais do gene 16S RNAr contendo de 400 a 700 bases revelou identidade com gênero Bacillus. No entanto, a espécie Bacillus subtilis foi confirmada somente depois da análise de seqüências dos genes rpoB, gyrA, and 16S RNAr. Intensa atividade inibitória aos fungos ambientais, como Penicillium sp, Aspergillus flavus, A. niger, e fitopatogênicos, como Colletotrichum sp, Alternaria alternata, Fusarium solani e F. oxysporum f.sp vasinfectum, foi observada em coculturas com a cepa bacteriana (B. subtilis), particularmente em ágar Sabouraud dextrose e ágar DNase. Pigmentos de cor avermelhada e vermelho-amarronzado, provavelmente feomelaninas, foram secretados respectivamente por colônias de A. alternata e A. niger depois de sete dias de co-cultivo.