The effects of book disinfection to the airborne microbiological community in a library environment

The management of fungal contaminants inside libraries and archives has become a big challenge for librarians, restorers and scientists. Several disinfection treatments have been developed in recent years, using both chemical and physical approaches on book collections and indoor environments. However, there is a lack of knowledge about the temporal efficiency of these cleanings, especially in relation with the preservation environments. The aim of this study was to determine the long-term effect of a chemical disinfection that interested a previous-contaminated book collection inside a University library. The monitoring after 6 months and 1 year from the cleaning confirmed any fungal growth on the disinfected books and the reduction of 90% of the airborne fungal load, highlighting anyway the presence of high fungal diversity. Sixty-eight different airborne fungal entities were isolated, in particular Aspergillus vitricola, Bulleromyces albus, Cladosporium cladosporioides, Cladosporium pseudocladosporioides, Cladosporium sphaerospermum, Penicillium brevicompactum, Rodothorula mugillaginosa and Sporodiobolus pararoseus. Several fungal species were sampled from the disinfected books, in particular Aspergillus penicillioides and Penicillium chrysogenum. The presence of these fungi both as airborne and as settled particles highlights the importance to maintain clean the preservation environments in order to prevent further microbial contaminations.     

New <Emphasis Type="Italic">Penicillium</Emphasis> and <Emphasis Type="Italic">Talaromyces</Emphasis> species from honey,pollen and nests of stingless bees

Penicillium and Talaromyces species have a worldwide distribution and are isolated from various materials and hosts, including insects and their substrates. The aim of this study was to characterize the Penicillium and Talaromyces species obtained during a survey of honey, pollen and the inside of nests of Melipona scutellaris. A total of 100 isolates were obtained during the survey and 82% of those strains belonged to Penicillium and 18% to Talaromyces. Identification of these isolates was performed based on phenotypic characters and β-tubulin and ITS sequencing. Twenty-one species were identified in Penicillium and six in Talaromyces, including seven new species. These new species were studied in detail using a polyphasic approach combining phenotypic, molecular and extrolite data. The four new Penicillium species belong to sections Sclerotiora (Penicillium fernandesiae sp. nov., Penicillium mellis sp. nov., Penicillium meliponae sp. nov.) and Gracilenta (Penicillium apimei sp. nov.) and the three new Talaromyces species to sections Helici (Talaromyces pigmentosus sp. nov.), Talaromyces (Talaromyces mycothecae sp. nov.) and Trachyspermi (Talaromyces brasiliensis sp. nov.). The invalidly described species Penicillium echinulonalgiovense sp. nov. was also isolated during the survey and this species is validated here.     

The influence of meteorological parameters on <Emphasis Type="Italic">Alternaria</Emphasis> and <Emphasis Type="Italic">Cladosporium</Emphasis> fungal spore concentrations in Beja (Southern Portugal): preliminary results

Introduction Fungal spores constitute an important fraction of bioaerosols in the atmosphere. Objectives To analyse the content of Alternaria and Cladosporium spores in the atmosphere of Beja and the effect of meteorological conditions on their concentrations. Methodology The daily and hourly data of Alternaria and Cladosporium fungal spores concentration in the atmosphere of Beja were monitored from April 12, 2012 to July 30, 2014, based on the Portuguese Aerobiology Network methodology. The influence of meteorological conditions on the studied types of fungal spore concentrations was assessed through Spearman’s correlation analysis. Results During the study period, 20,741 Alternaria spores and 320,862 Cladosporium spores were counted. In 2013, there were 5,822 Alternaria spores and 123,864 Cladosporium spores. The absolute maximum concentrations of Alternaria and Cladosporium spores were recorded on November 8, 2013, with 211 and 1301 spores/m³, respectively. Temperature, insolation and wind direction parameters showed a positive correlation with Alternaria and Cladosporium spore levels, while relative humidity and precipitation presented a negative correlation, which is statistically significant. Wind speed only showed a statistically significant positive correlation in terms of Alternaria spore levels. Conclusion Alternaria and Cladosporium spores are present in the atmospheric air of Beja throughout the year, with the highest concentration period occurring during spring and autumn. There was a clear effect of meteorological parameters on airborne concentrations of these fungal spores.     

Phenotypic and genotypic diversity of airborne fungal spores in Demänovská Ice Cave (Low Tatras,Slovakia)

This paper is the first aero-mycological report from Demänovská Ice Cave. Fungal spores were sampled from the internal and external air of the cave in June, 2014, using the impact method with a microbiological air sampler. Airborne fungi cultured on PDA medium were identified using a combination of classical phenotypic and molecular methods. Altogether, the presence of 18 different fungal spores, belonging to 3 phyla, 9 orders and 14 genera, was detected in the air of the cave. All of them were isolated from the indoor samples, and only 9 were obtained from the outdoor samples. Overall, airborne fungal spores belonging to the genus Cladosporium dominated in this study. However, the spores of Trametes hirsuta were most commonly found in the indoor air samples of the cave and the spores of C. herbarum in the outdoor air samples. On the other hand, the spores of Alternaria abundans, Arthrinium kogelbergense, Cryptococcus curvatus, Discosia sp., Fomes fomentarius, Microdochium seminicola and T. hirsuta were discovered for the first time in the air of natural and artificial underground sites. The external air of the cave contains more culturable airborne fungal spores (755 colony-forming units (CFU) per 1 m³ of air) than the internal air (from 47 to 273 CFU in 1 m³), and these levels of airborne spore concentration do not pose a threat to the health of tourists. Probably, the specific microclimate in the cave, including the constant presence of ice caps and low temperature, as well as the location and surrounding environment, contributes to the unique species composition of aeromycota and their spores in the cave. Thus, aero-mycological monitoring of underground sites seems to be very important for their ecosystems, and it may help reduce the risk of fungal infections in humans and other mammals that may arise in particular due to climate change.     

Diversity of the microscopic fungi in the littoral sands of the White Sea

For the first time, the study of the mycobiota of littoral sands of the cold-water White Sea was conducted. Twenty-four samples were collected in July–August 2015 in the Onega Bay, the Kandalaksha Bay, and on the Terskii coast. Culture methods were used to detect the number of colony forming units (CFU) and the fungi diversity. The number of CFUs is small; it ranges from 0 to 57 per 1 cm³. A total of 54 morphotypes were detected, of which 22 were identified to the species, seven to the genus level, and 25 were not identified, because they did not form sporulation. Only two species, Cladosorium sphaerospermum and Paradendryphiella salina, were characterized by the relative abundance of more than 10% and the frequency of occurrence of more than 50%. Another five species were found in more than one sample: Penicillium chrysogenum, Cadophora fastigiata, Acremonium fuci, anamorph of Pseudogymnoascus pannorum, and teleomorph of Pseudeurotium hygrophilum. All other morphotypes were found in a small number in single samples. In general, darkcolored forms predominated in the mycobiota both in diversity and in number.     

Isolation and molecular characterization of the fungal endophytic microbiome from conventionally and organically grown avocado trees in South Florida

Fungal endophytes are the most ubiquitous and highly diverse microorganisms that inhabit the interior of healthy plants. They are important in plant ecology and offer untapped potential to improve plant health and productivity in agroecosystems. The endophytic assemblage of avocado is poorly understood; therefore, surveys of fungal endophytes of Persea americana Mill. (Avocado) in South Florida organic and conventional orchards were conducted. A total of 17 endophytic fungal species were recovered from healthy avocado terminal branches. Endophytic fungal species were identified by rDNA sequencing of the internal transcribed spacer (ITS) region, using UNITE Species Hypotheses to reliably assign a taxon name, and determined as belonging to the genera Alternaria, Cladosporium, Colletotrichum, Corynespora, Diaporthe, Lasiodiplodia, Neofusicoccum, Neopestalotiopsis, Phyllosticta, and Strelitziana. Endophyte community assemblage differed between organic and conventional agroecosystems. This is the first report of Alternaria eichhorniae, Cladosporium tenuissimum, Corynespora cassiicola, Colletotrichum alatae, Diaporthe fraxini-angustifoliae, Lasiodiplodia gonubiensis, Neofusicoccum algeriense, Neofusicoccum andinum, Neopestalotiopsis foedans, Phyllosticta capitalensis, and Strelitziana africana as endophytes of avocado. Evaluation using pathogenicity tests on avocado leaves and terminal branches showed that endophytic fungal isolates did not cause disease symptoms.     

Diversity and enzyme activity of <Emphasis Type="Italic">Penicillium</Emphasis> species associated with macroalgae in Jeju Island

A total of 28 strains of 19 Penicillium species were isolated in a survey of extracellular enzyme-producing fungi from macroalgae along the coast of Jeju Island of Korea. Penicillium species were identified based on morphological and β-tubulin sequence analyses. In addition, the halo-tolerance and enzyme activity of all strains were evaluated. The diversity of Penicillium strains isolated from brown algae was higher than the diversity of strains isolated from green and red algae. The commonly isolated species were Penicillium antarcticum, P. bialowiezense, P. brevicompactum, P. crustosum, P. oxalicum, P. rubens, P. sumatrense, and P. terrigenum. While many strains showed endoglucanase, β-glucosidase, and protease activity, no alginase activity was detected. There was a positive correlation between halo-tolerance and endoglucanase activity within Penicillium species. Among 19 Penicillium species, three species–P. kongii, P. olsonii, and P. viticola–have not been previously recorded in Korea.     

Bioaerosol assessment in the library of Istanbul University and fungal flora associated with paper deterioration

Health problems in people who are in indoor environments with poor ventilation have resulted in an increase in the number of studies regarding air quality. Microorganisms and inferior indoor air-climatic conditions not only affect human health but also cause decay of invaluable materials present in libraries. Therefore, this study aimed to assess the culturable bioaerosol composition and concentration in the library of Istanbul University. The culturable fungal flora, a biodeterioration agent, of the damaged archival materials was also examined. The air was sampled for a year, as were the surfaces of 207 biologically damaged books. The fungal colonies’ range was between 235 and 1850 CFU/m³, and the bacterial colonies range between 270 and 1920 CFU/m³. This is the first volumetric record in Turkey. Although the microbial contamination was not very high, molds such as Aspergillus versicolor, Cladosporium sphaerospermum, Penicillium chrysogenum, Alternaria alternata, Chaetomium globosum, Aspergillus flavus, and Aspergillus ochraceus might cause harm to human beings or to books as biodeterioration agents. The highest amount of fungus was determined in Archive 3, which contained damaged books. The fungal species isolated from the air and books were essentially the same. Therefore, it is important to determine not only the numeric values but also the microbiological composition of fungal colonies because the variety of fungal species is indicative of a deterioration process in effect over a long period. The deterioration of books must be remedied.     

Airborne pollen and fungal spores in Garki,Abuja (North-Central Nigeria)

The ambient atmosphere is dominated with pollen and spores, which trigger allergic reactions and diseases and impact negatively on human health. A survey of pollen and fungal spores constituents of the atmosphere of Garki, Abuja (North-Central Nigeria) was carried out for 1 year (June 1, 2011–May 31, 2012). The aim of the study was to determine the prevalence and abundance of pollen and fungal spores in the atmosphere and their relationship with meteorological parameters. Airborne samples were trapped using modified Tauber-like pollen trap, and the recipient solutions were subjected to acetolysis. Results revealed the abundance of fungal spores, pollen, fern spores, algal cysts and diatoms in decreasing order of dominance. The atmosphere was qualitatively and quantitatively dominated by pollen during the period of late rainy/harmattan season than the rainy season. Numerous fungal spores were trapped throughout the sampling periods among which Alternaria spp., Fusarium spp., Cladosporium spp. and Curvularia spp. dominated. These fungi have been implicated in allergic diseases and are dermatophytic, causing diverse skin diseases. Other pathogenic fungi found in the studied aeroflora were Dreschlera spp., Helminthosporium spp., Torula spp., Pithomyces spp., Tetraploa spp., Nigrospora ssp., Spadicoides spp., Puccinia spp. and Erysiphe graminis. Total pollen and fungal spores counts do not show significant correlation with meteorological parameters.     

Identification and Pathogenicity of Fungal Pathogens Causing Black Point in Wheat on the North China Plain

Fungi associated with black point were isolated from three highly susceptible wheat genotypes in the North China Plain. The 21 isolates represented 11 fungal genera. The most prevalent genera were Alternaria (isolation frequency of 56.7%), Bipolaris (16.1%), and Fusarium (6.0%). The other eight genera were Curvularia, Aspergillus, Cladosporium, Exserohilum, Epicoccum, Nigrospora, Penicillium, and Ulocladium; their isolation frequencies ranged from 0.8 to 4.8%. The pathogenicity of the isolates was individually assessed in the greenhouse by inoculating wheat plants with spore suspensions. Ten of the 21 isolates caused significantly higher incidences of black point than that the controls. These isolates belonged to eight fungal species (A. alternata, B. sorokiniana, B. crotonis, B. cynodontis, C. spicifera, F. equiseti, E. rostratum, and E. sorghinum) based on morphological traits and phylogenetic analysis. The average incidences of black point in the eight fungal species were 32.4, 54.3, 43.0, 41.9, 37.2, 38.8, 50.1, and 34.1%, respectively. B. sorokiniana and A. alternata were determined to be the most important pathogens in the North China Plain based on fungal prevalence and symptom severity. This study is the first to identify E. rostratum as a major pathogen causing black point in wheat.     

Potential impact of climate change on fungal distributions: analysis of 2 years of contrasting weather in the UK

The impact of climate change on fungal growth and spore production is less well documented than for allergenic pollen grains, although similar implications for respiratory tract diseases in humans occur. Fungal spores are commonly described as either “dry” or “wet” according to the type of weather associated with their occurrence in the air. This study examined the distribution of selected fungal spores (Alternaria spp., Cladosporium spp., Didymella spp., Epicoccum spp., Leptosphaeria spp. and rusts) occurring in the West Midlands of UK during 2 years of contrasting weather. Spore specimens were collected using a 7-day volumetric air sampler and then analysed with the aid of light microscopy. Distributions of spores were then studied using normality tests and Mann–Whitney U test, while relationships with meteorological parameters were investigated using Spearman’s rank test and angular-linear correlation for wind direction analysis. Our results showed that so-called wet spores were more sensitive to the weather changes showing statistically significant differences between the 2 years of study, in contrast to “dry” spores. We predict that in following years we will observe accelerated levels in allergenic fungal spore production as well as changes in species diversity. This study could be a starting point to revise the grouping system of fungal spores as either “dry” or “wet” types and their response to climate change.     

Persistence of ecto- and ectendomycorrhizal fungi associated with <Emphasis Type="Italic">Pinus montezumae</Emphasis> in experimental microcosms

Ectomycorrhizal (ECM) and ectendomycorrhizal fungal species associated with Pinus montezumae were recorded in 8 year-old trees established in microcosms and compared with those associated with 2 year-old trees, in order to determine their persistence over the long-term. Mycorrhizal root tips were morphologically and anatomically characterized and sequenced. The extension of extramatrical mycelium of ECM fungi with long exploration strategies was evaluated. In total, 11 mycorrhizal species were registered. Seven mycorrhizal species were detected on both 2 and 8 year-old pines: Atheliaceae sp., Rhizopogon aff. fallax, R. aff. occidentalis, Suillus pseudobrevipes, Tuber separans, Wilcoxina mikolae and Wilcoxina rehmii. One species, Thelephora terrestris, was exclusively associated with two year–old seedlings, while Cenococcum geophilum, Pezizaceae sp. and Pyrenomataceae sp. were exclusively found on 8 year-old trees. Atheliaceae sp. was the ECM fungal species that presented the most abundant mycelium. Finally, we report one new fungal species of Pezizaceae occurring as a symbiont of P. montezumae.     

First volumetric record of fungal spores in the atmosphere of Montevideo City,Uruguay: a 2-year survey

In Uruguay, aeromycological studies are restricted to a gravimetric analysis performed from December 1942 to March 1944 in Montevideo where spores of Pucciniaceae, Alternaria and Helminthosporium were the only specimens identified. Daily monitoring of airborne fungal spores was carried out for the first time in Montevideo, from April 2012 to March 2014, using a Rotorod sampler in order to evaluate the seasonal variation and influence of meteorological parameters. A total of 548,309.68 spores/m³ were recorded which belong to anamorphs of Higher Fungi (69.18 %), Phyla Ascomycota (12.62 %), Basidiomycota (8.01 %), Oomycota (0.37 %) and Myxomycota (0.06 %). Airborne spores occurred in Montevideo throughout the whole year. However, a seasonal pattern was revealed, with the highest concentrations recorded in autumn and summer. The most abundant spore types were Cladosporium (53.22 %), Alternaria (6.62 %), Didymella Group (5.86 %), Leptosphaeria Group (4.37 %) and Coprinus (4.3 %). Temperature appeared to be the most influential meteorological factor correlating significantly and positively with total spore, Cladosporium, Alternaria and Didymella Group abundance. Relative humidity influenced positively total spore, Cladosporium and Didymella Group concentrations while a weak negative association was obtained for Alternaria. Wind speed correlated negatively with total spore, Cladosporium, Alternaria and Didymella Group. Precipitation showed a negative influence on Alternaria, while positive correlations were observed for Didymella Group. For the first time, fungal spores considered allergenic were recorded in Montevideo atmosphere and the risk of exposure would have been high from December to June. However, long-term sampling is needed to define seasonal prevalence patterns and the influence of meteorological conditions on spore abundance.     

Atmospheric concentrations of selected allergenic fungal spores in relation to some meteorological factors,in Timişoara (Romania)

Present investigation was undertaken to study the dynamics of relationships between atmospheric fungal spores and meteorological factors in western Romania. The airborne spore sampling was carried out by employing volumetric sampling. A total of nine meteorological parameters were selected for this investigation. During 2008–2010, it was found the same pattern of behaviour in the atmosphere for selected spore types (Alternaria, Cladosporium, Pithomyces, Epicoccum and Torula). The spores occurred in the air throughout the whole year, but maximum concentrations were reached in summer. Cladosporium and Alternaria peak levels were observed in June. Epicoccum peak value was found in September. The relationships between airborne spore concentrations and environmental factors were assessed using the analysis of Spearman’s rank correlations and multiple linear regressions. Spearman’s rank correlation analysis revealed that maximum, minimum and mean temperature, and number of sunshine hours were strongly (p < 0.01) and directly proportional to the concentration of all analysed fungal spores. Negative and significant correlations were with daily mean relative humidity. The variance explained percentage by regression analyses varied between 30.6 and 39.6 % for Alternaria and Cladosporium airborne spores. Statistical methods used in this study are complementary and confirmed stable dependence of Alternaria and Cladosporium spore concentrations on meteorological factors. The climate change parameters either increased temperatures, changed precipitation regimes or a combination of both affected allergenic fungal spore concentrations in western Romania. This study demonstrates the need for investigations throughout the year, from month to month, regarding the correct interpretation of airborne spore relationships with meteorological parameters.     

Novel fungi from an ancient niche: lachnoid and chalara-like fungi on ferns

A survey was conducted in Brazil to collect fungi on ferns. Based on morphology and inferred phylogeny from DNA sequences of two loci, namely the internal transcribed spacer (ITS) regions and the large subunit nuclear ribosomal RNA gene (LSU), several species belonging to chalara-like genera and lachnoid fungi were recognized. Eighteen fungal isolates, collected from five host species, representing 10 different localities were studied. Three novel genera (Lachnopsis, Scolecolachnum and Zymochalara), and six novel species (Bloxamia cyatheicola, Lachnopsis catarinensis, Lachnopsis dicksoniae, Scolecolachnum pteridii, Zymochalara lygodii and Zymochalara cyatheae) are introduced. Furthermore, two new combinations (Erioscyphella euterpes and Erioscyphella lushanensis) are proposed. Two novel taxa (Lachnopsis catarinensis and Lachnopsis dicksoniae) may be included in the list of potentially endangered fungal species in Brazil, if proven to be restricted to their tree-fern host, Dicksonia sellowiana, which is included in the official list of endangered plant species in Brazil.     

Distribution of arbuscular mycorrhizal fungi in four semi-mangrove plant communities

To better understand the diversity and species composition of arbuscular mycorrhizal fungi (AMF) in mangrove ecosystems, the AMF colonization and distribution in four semi-mangrove plant communities were investigated. Typical AMF hyphal, vesicle and arbuscular structures were commonly observed in all the root samples, indicating that AMF are important components on the landward fringe of mangrove habitats. AMF spores were extracted from the rhizospheric soils, and an SSU rDNA fragment from each spore morph-type was amplified and sequenced for species identification. AMF species composition and diversity in the roots of each semi-mangrove species were also analyzed based on an SSU-ITS-LSU fragment, which was amplified, cloned and sequenced from root samples. In total, 11 unique AMF sequences were obtained from spores and 172 from roots. Phylogenetic analyses indicated that the sequences from the soil and roots were grouped into 5 and 14 phylotypes, respectively. AMF from six genera including Acaulospora, Claroideoglomus, Diversispora, Funneliformis, Paraglomus, and Rhizophagus were identified, with a further six phylotypes from the Glomeraceae family that could not be identified to the genus level. The AMF genus composition in the investigated semi-mangrove communities was very similar to that in the intertidal zone of this mangrove ecosystem and other investigated mangrove ecosystems, implying possible fungal adaptation to mangrove conditions.     

Survey of Philippine coffee beans for the presence of ochratoxigenic fungi

In 2012 to 2014, Philippine green coffee beans from Coffea arabica in Benguet and Ifugao; Coffea canephora var. Robusta in Abra, Cavite, and Ifugao; and Coffea liberica and Coffea excelsea from Cavite were collected and assessed for the distribution of fungi with the potential to produce ochratoxin A (OTA). The presence of fungal species was evaluated both before and after surface sterilization. There were remarkable ecological and varietal differences in the population of OTA-producing species from the five provinces. Aspergillus ochraceus, A. westerdijkiae, and Penicillium verruculosum were detected from Arabica in Benguet and Ifugao while Aspergillus carbonarius, Aspergillus niger, and Aspergillus japonicus were isolated in Excelsa, Liberica, and Robusta varieties from Abra, Cavite, and Davao. Contamination by Aspergillus and Penicillium species was found on 59 and 19 %, respectively, of the 57 samples from five provinces. After disinfection with 1 % sodium hypochlorite, the levels of infection by Aspergillus and Penicillium fell to 40 and 17 %, respectively. A total of 1184 fungal isolates were identified to species level comprising Aspergillus sections Circumdati (four species), Clavati (one), Flavi (one), Fumigati (one), Nigri (three), and Terrie (one). Within section Circumdati, 70 % of A. ochraceus produced OTA as high as 16238 ng g^?1 while 40 % of A. westerdijkiae produced maximum OTA of 36561 ng g^?1 in solid agar. Within section Nigri, 16.76 % of A. niger produced OTA at the highest 18439 ng g^?1, 10 % of A. japonicus at maximum level of 174 ng g^?1, and 21.21 % of A. carbonarius yielded maximum OTA of 1900 ng g^?1. Of the 12 species of Penicillium isolated, P. verruculosum was ochratoxigenic, with a maximum OTA production of 12 ng g^?1.     

A New record of four <Emphasis Type="Italic">Penicillium</Emphasis> species isolated from <Emphasis Type="Italic">Agarum clathratum</Emphasis> in Korea

Agarum clathratum, brown algae, play important ecological roles in marine ecosystem, but can cause secondary environment pollution when they pile up on the beach. In order to resolve the environment problem by A. clathratum, we focus to isolate and identify Penicillium because many species are well known to produce extracellular enzymes. A total of 32 Penicillium strains were isolated from A. clathratum samples that collected from 13 sites along the mid-east coast of Korea in summer. They were identified based on morphological characters and phylogenetic analysis using β-tubulin DNA sequences as well as a combined dataset of β-tubulin and calmodulin. A total of 32 strains were isolated and they were identified to 13 Penicillium species. The commonly isolated species were Penicillium citrinum, P. roseomaculatum, and Penicillium sp. Among 13 Penicillium species, four species – P. bilaiae, P. cremeogriseum, P. madriti, and P. roseomaculatum – have not been previously recorded in Korea. For these four new species records to Korea, we provide morphological characteristics of each strain.