Assessment of the adaptation capabilities of the bivalves Modiolus modiolus (Linnaeus, 1758) and Crenomytilus grayanus (Dunker, 1853) under increased levels of heavy metals in the environment

The active biomonitoring method was used to examine the changes in heavy-metal (Fe, Zn, Cu, Cd, Mn, Pb, and Ni) contents in the organs (muscle, gonads, mantle, gills, digestive gland, and kidney) of the mussels Crenomytilus grayanus and Modiolus modiolus. The dynamics of trace element concentrations in mussel organs during the experiment were compared with their subcellular distribution. The defense strategy of M. modiolus consisted of the threshold accumulation of toxic metals in all organs followed by their excretion, whereas the strategy of C. grayanus involved short-term isolation from adverse environmental influence. Under chronic pollution, in C. grayanus the main loads occurred in the digestive gland and kidney. Under acute changes in environmental conditions, the processes of regulation (detoxification/excretion) of Fe, Mn, and Pb in this species were impaired substantially.     

Comparison of fungal complexes of Japanese scallop Mizuhopecten yessoensis (Jay, 1856) from different areas in the Peter the Great Bay of the Sea of Japan

Mycological investigation of the Japanese scallop Mizuhopecten yessoensis (Jay) (Bivalvia) from different areas of the Peter the Great Bay (Sea of Japan) was conducted. Isolates from internal organs of M. yessoensis scallop comprise 72 species of filamentous fungi from 30 genera of ascomycetes, anamorphic fungi, and zygomycetes. Species richness of filamentous fungi—fungi of the genera Aspergillus, Penicillium, Cladosporium, and Chaetomium—in the internal organs of bivalve mollusks increases in polluted coastal waters.     

Effects of Environmental Factors on Byssal Thread Formation in Some Members of the Family Mytilidae from the Sea of Japan

The ability to attach repeatedly to a substrate (glass, boulders, sand) in three common mussel species of the upper sublittoral zone of the Sea of Japan, Grayan's mussel Crenomytilus grayanus, the Korean mussel Mytilus coruscus, and northern horse mussel Modiolus modiolus, was studied under experimental conditions. It was found that during 120 h of the experiment C. grayanus and M. modiolus produced more byssal threads than M. coruscus. A decrease in the water temperature from 20 to 0°C slowed the rate of production of byssal threads down to full passivity in some experimental mollusks. This was more typical of M. coruscus and less typical of C. grayanus. Renewed threads differed in their length, thick, size of the adhesive plate, and strength. M. coruscus formed the shortest, thickest, and strongest threads with rather a large adhesive disk. The observed differences are discussed from the position of morphophysiological adaptations of species for colonization of different natural substrata under contrasting conditions of the upper sublittoral zone.     

Incidence of seed-borne fungi and aflatoxins in Sudanese lentil seeds

Thirteen seed samples of lentil (Lens esculenta) were incubated on agar plate and moist filter papers (Moist Chambers) at 28 ± 2 °C for determination of the incidence of seed-borne fungi. Aflatoxins content of the seeds was measured using the bright greenish-yellow fluorescence test (BGYF) and thin-layer chromatography (TLC). Sixty-nine species and seven varieties, which belong to 24 genera of fungi, were isolated from this crop. Of these fungi, 51 species and two varieties are considered new for this crop, whereas seven genera and 13 species are new to the mycoflora of the Sudan. The genus Aspergillus (13 species and 6 varieties) which comprising 44% of the total colony count was the most prevalent genus followed by Rhizopus (2 species, 19%), Penicillium (6 species) and Fusarium (8 species) (12%), Chaetomium (3 species) and Cladosporium (5 species) (6%), where the 18 genera (1–4 species) showed very low level of incidence (19%). Of the possible pathogens of lentil plants, F. oxysporum the main cause of vascular wilt was recovered from seeds of this crop. Thin layer chromatographic analysis of chloroform extracts of 13 seed samples showed that only one samples was naturally contaminated with aflatoxins B₁, B₂, G₁ and G₂ (14.3 μg/kg). This revised version was published online in June 2006 with corrections to the Cover Date.     

Quantitative and qualitative seasonal changes in the microbial community from the phyllosphere of sugar beet (Beta vulgaris)

Bacteria, yeasts and filamentous fungi colonizing immature, mature and senescing primary leaves of field grown Beta vulgaris (sugar beet) were analysed over a complete growing season. Greatest microbial numbers were detected on senescing primary leaves and these numbers increased over most of the season. The number of colonizers detected on mature leaves was found to be stable over most of the study.Filamentous fungi and yeasts were identified to the genus level and the communities found to have greatest diversity during the summer months. There was no consistent pattern of diversity according to leaf type. Two genera of filamentous fungi, Cladosporium and Alternaria and two yeast genera, Cryptococcus and Sporobolomyces were the most numerous fungal populations isolated. Only 8 filamentous fungi and 3 yeast genera were commonly isolated on PDA (potato dextrose agar).Bacterial strains (1236) were isolated on Tryptic Soy Broth (TSB) agar and identified to species, or in some cases sub-species level, by analysis of their fatty acid methyl ester (FAME) profiles. Isolated bacteria were grouped into 78 named and 37 unnamed species clusters. Greatest number of bacterial species were isolated from young plants and leaves, sampled during the autumn months. Bacterial community diversity was lowest in mid-summer and winter months. Pseudomonas was the most commonly isolated genus and Erwinia herbicola the most common species. P. aureofaciens was the only species isolated from soil that was also isolated from the phyllosphere of B. vulgaris throughout the season.     

Diversity and decomposition potential of endophytes in leaves of a Cinnamomum camphora plantation in China

This study was carried out to improve our understanding of the diversity and decomposition potential of endophytes in the leaves of Cinnamomum camphora trees grown in a subtropical region of China. We isolated and identified endophytic fungi from senescent leaves of C. camphora and tested their role in decomposition through pure-culture and pre-colonization. A total of 2,861 endophytic fungi isolated from 69 leaves of C. camphora were grouped into 39 taxa comprising 36 Ascomycetes and 3 Basidiomycetes based on sporulation and ITS sequence analysis. Of these, Colletotrichum gloeosporioides was the most common species (69% relative abundance and 96% colonization frequency), followed by Cladosporium sp.1, Colletotrichum sp. and Chaetomium sp. All 39 endophytes had the ability to decompose C. camphora leaf litter in pure culture, and a few exhibited >20% litter mass loss in 2 months. In most cases, single endopyhytic species showed lower mass loss than mixed microbial groups from active soil after 60 or 120 days. In pre-inoculation, endophytic fungi like Chaetomium sp., Cladosporium sp.1, C. gloeosporioides, Colletotrichum sp. and Guignardia sp. exhibited higher abundance and caused greater mass loss, indicating the potential of these groups to enter and significantly accelerate the process of decomposition. This study concludes that, after entering the decomposition process, selected endophytic fungi with high abundance could influence significantly the decomposition process and thus probably affect carbon and nutrient cycling in C. camphora plantations.     

Use of the photo method in the study of the composition and abundance of soft-bottom macrobenthos

The composition and abundance of the macrobenthic fauna associated with soft-bottom beds of the large bivalve mollusks Crenomytilus grayanus and Modiolus modiolus were studied in the inner part of Vostok Bay (Sea of Japan) in spring 2010. More than 30 taxa of animals were found in the mussel beds. Comparison of the data obtained with three different methods of studying muddy-bottom mussel communities showed that 80% of their biomass and only up to 25% of the species composition and 19% of the abundance can be estimated by the photo method. Photo surveys disregard small and/or juvenile animals, as well as infaunal organisms, and thus do not give a complete picture of the species, size, and age composition and abundance of biota.     

Assessment of Abundance and Species Composition of Filamentous Fungi in the Underground Rzeczka Complex in Sowie Mountains (Lower Silesia,Poland)

Our study is the first mycological evaluation of the air and the rocks in the underground Rzeczka complex in Sowie Mountains (Lower Silesia, Poland). The density of filamentous fungi isolated from the air inside and outside the adit ranged between 245.5 to 1332.6 Colony Forming Units in 1 m³ of air. Sixteen species of filamentous fungi were isolated from the internal air sampled and 11 from the outside of the adit. Cladosporium spp. were the fungi most frequently isolated from the internal atmosphere of the adit and from the external air. From the rock walls and from the rock debris on the floor of the adit only seven species of fungi were isolated. The fungi most frequently isolated from the rock walls were the Aspergillus niger group and from the rock debris we also found A. niger group along with species of Mucor. The concentration of airborne fungi in the adit did not exceed official limits and norms present for a health risk to the tourists, but fungi species isolated from the rocks can cause their degradation.     

Diversity and pharmaceutical screening of fungi from benthic mats of Antarctic lakes

During the MICROMAT project, the fungal diversity of microbial mats growing in the benthic environment of Antarctic lakes was accessed for the discovery of novel antibiotics and anticancers. In all, 160 filamentous fungi belonging to fifteen different genera and 171 yeasts were isolated from 11 lakes, classified and cultivated in different media and at different temperatures. Filamentous fungi were then screened to discover novel antimicrobial and cytotoxic compounds. A total of 1422 extracts were prepared by solid phase extraction of the culture broths or by biomass solvent extraction. 47 (29%) filamentous fungi showed antimicrobial activity; most of them inhibited the growth of gram-positive Staphyloccus aureus (14%), gram-negative E. coli (10%), and of yeasts Candida albicans (11%) and Cryptococcus neoformans (8%). Less activity was detected against representatives of enterobacteria and filamentous fungi. The most productive in terms of bioactivities were cold-tolerant cosmopolitan hyphomycetes such as Penicillium, Aspergillus, Beauveria and Cladosporium. Two bioactive bis-anthraquinones (rugulosin and skyrin) were identified by LC–MS as the main products in a strain of Penicillium chrysogenum isolated from a saline lake in the Vestfold Hills. LC–MS fractionation of extracts from two diverse species of Aspergillus, that exhibited relatively potent antimicrobial activities, evidenced a chemical novelty that was further investigated. To our knowledge, this is the first report of new antibiotics produced by fungi from benthic microbial mats from Antarctic lakes. It can be concluded that these microbial assemblages represent an extremely rich source for the isolation of new strains producing novel bioactive metabolites with the potential to be developed as drugs.     

Fungi Isolated from Damaged Flue-cured Tobacco

Species of Aspergillus were the most prevalent fungi isolated from 51 samples of damaged flue-cured tobacco of the 1966 U.S. crop, comprising 57% of the total isolates. Other prevalent fungi were Penicillium (16%), Alternaria (8%), Cladosporium (4%), and Chaetomium (4%). Members of the Aspergillus glaucus group were isolated most frequently from samples with moisture contents ranging from 18 to 28%, whereas Alternaria, Cladosporium, and Penicillium were isolated consistently from samples containing 24 to 32% moisture. Aspergillus niger was prevalent in tobacco ranging in moisture content from 18 to 30%.     

Fungal biodiversity in the air of Turin

The qualitative fungal composition of Turin 's atmospheric environment was surveyed, carrying out a twelve-month study and collecting with a single stage volumetric sieve sampler on Dermasel agar supplemented with 0.4 g l^–1 cycloheximide and 0.05 g l^–1 chloramphenicol. We isolated 165 species and 2 varieties of mesophilic fungi from 58 genera and 26 thermotolerant species from 12 genera. Penicillium, Aspergillus, Acremonium, Chrysosporium, Scopulariopsis, Malbranchea, Paecilomyces, Phialophora and Cladosporium were in sequence the genera most rich in mesophilic species; Aspergillus, Penicillium, Chrysosporium and Scopulariopsis the most rich in thermotolerant species. Many of the species isolated are rarely or never recorded in the atmospheric environment. Cycloheximide can thus be said to select among airborne fungi, giving a characteristic picture.Abbreviation CH Cycloheximide - cfu colony forming unit     

Fungal diversity in ancient documents. A case study on the Archive of the University of Coimbra

This multidisciplinary research combines knowledge in molecular biology with fungal morphology, aiming at the identification of infecting fungi from historical documents on the Archive of the University of Coimbra. The identification of infecting fungi on several bibliographic documents and support materials was based both on ribosomal DNA loci amplification and sequencing, and morphological identification, using macro- and microscopical traits. A high fungal diversity was found in all types of support: parchment, laid-paper and wood-pulp paper. Fourteen fungal genera were isolated, identified, and kept in culture. The most frequent were Cladosporium, Penicillium and Aspergillus, and other less frequent genera, such as Alternaria, Botrytis, Chaetomium, Chromelosporium, Epicoccum, Phlebiopsys and Toxicocladosporium were also present. Within these genera, 20 different species were identified, from which 15 were found only in a single support type. Cladosporium cladosporioides and Penicillium chrysogenum were the only species present in all support types.     

Comparative Analysis of Subcellular Distribution of Heavy Metals in Organs of the Bivalve Mollusks <Emphasis Type="Italic">Crenomytilus grayanus</Emphasis> and <Emphasis Type="Italic">Modiolus modiolus</Emphasis> in a Continuously Polluted Environment

The distribution of Zn, Fe, Ni, Cu, Mn, Cd, and Pb in subcellular fractions, and of Cd, Zn, and Cu in cytoplasm proteins of the kidney and digestive gland of the mussels Crenomytilus grayanus and Modiolus modiolus, sampled from contaminated and conditionally clean areas, was studied. It was found that, in a contaminated environment, the organs of mussels were more highly enriched with metals. It was shown that essential trace elements were accumulated mostly in the cytosol of organs of both molluscan species from contaminated areas, whereas in the background areas the trace elements were associated mostly with membrane structures in Gray's mussel, C. grayanus, and with the cytosol in M. modiolus, the northern horse mussel. The lead was bound mostly to membrane structures in organs of both mussel species at all stations. The method of gel chromatography enabled us to isolate metallothionein-like proteins from the kidney of the northern horse mussel sampled in contaminated areas, whereas their concentration in the kidney of Gray's mussels was lower than the limiting error of the method. It is supposed that in the kidney of Gray's mussel the synthesis of metallothionein-like proteins was quenched by the integrated effect of the accumulated metals.     

Exploration of cultivable fungal communities in deep coal‐bearing sediments from ∼1.3 to 2.5 km below the ocean floor

Although subseafloor sediments are known to harbour a vast number of microbial cells, the distribution, diversity, and origins of fungal populations remain largely unexplored. In this study, we cultivated fungi from 34 of 47 deep coal‐associated sediment samples collected at depths ranging from 1289 to 2457 m below the seafloor (mbsf) off the Shimokita Peninsula, Japan (1118 m water depth). We obtained a total of 69 fungal isolates under strict contamination controls, representing 61 Ascomycota (14 genera, 23 species) and 8 Basidiomycota (4 genera, 4 species). Penicillium and Aspergillus relatives were the most dominant genera within the Ascomycetes, followed by the members of genera Cladosporium, Hamigera, Chaetomium, Eutypella, Acremonium, Aureobasidium, Candida, Eurotium, Exophiala, Nigrospora, Bionectria and Pseudocercosporella. Four Basidiomycota species were identified as genera Schizophyllum, Irpex, Bjerkandera and Termitomyces. Among these isolates, Cladosporium sphaerospermum and Aspergillus sydowii relatives were isolated from a thin lignite coal‐sandstone formation at 2457 mbsf. Our results indicate that these cultivable fungal populations are indigenous, originating from past terrigenous environments, which have persisted, possibly as spores, through ～20 million years of depositional history.     

Microbial population dynamics on Golden Delicious apples from bud to harvest and effect of fungicide applications

The microbial population dynamics on apples cv. Golden Delicious were analysed every 15 days between bud and harvest in a fully replicated experiment in northern Spain in 1994 and 1995. The total microbial populations varied with developmental stage, and with prevailing climatic conditions. The predominant mycroflora were the filamentous fungi Cladosporium and Alternaria spp. and white and pink yeasts. Other genera isolated included mainly species of Epicoccum, Fusarium and Acremonium. However, the most important post-harvest pathogens Penicillium expansum and Botrytis cinerea were seldom isolated from ripening apples. Maximum total filamentous fungal populations occurred after fruit set and during early ripening [2 × 10⁴cfu (colony-forming units) g^-1 approximately] while those of bacteria were maximum at bud stage (3.5 × 10⁵and 3.0 × 10⁴ cfu g^-1 in 1994 and 1995 respectively). White yeasts were more numerous than pink yeasts. Endophytic infection of apple buds by Alternaria spp., responsible for core rot, was found in almost all bud tissue. By contrast, Cladosporium spp. were initially isolated later from 12.5–50% of tissue samples during blooming and fruit set. The impact of a four-spray fungicide regime during apple development significantly decreased the total filamentous fungal populations in both years, and that of Cladosporium spp. in 1994. However, bacterial populations were often higher on apples from fungicide-treated plots. Fungicide sprays decreased populations of Cladosporium, Alternaria and white yeasts for a maximum of up to 15–30 days after application. Fungicide application had little effect on endophytic infection of apples by Alternaria spp. between bud and harvest.     

Isolation of autochthonous non-white rot fungi with potential for enzymatic upgrading of Venezuelan extra-heavy crude oil

The increasing world demand for fuels makes it necessary to exploit the largest reserve of extra-heavy crude oil (EHCO) of the Orinoco Oil Belt from Venezuela. We propose the use of extracellular oxidative enzymes, in particular, lignin-degrading enzyme systems (LDS) of fungi, for enzymatic improvement of EHCO. Autochthonous non-white rot fungal strains able to use EHCO, and several polycyclic aromatic hydrocarbons (PAHs) as sole carbon source and energy, were isolated from EHCO-polluted soils and identified as belonging to the genera Fusarium, Penicillium, Trichoderma, Aspergillus, Neosartorya, Pseudallescheria, Cladosporium, Pestalotiopsis, Phoma and Paecillomyces. Phenotypic and biochemical assays revealed the ability of these filamentous fungi to synthesize extracellular oxidative enzymes, and suggested a relationship between the LDS and EHCO bioconversion. This work reports, for the first time, the use of o-phenylenediamine dihydrochloride (OPD) as substrate to measure extracellular ligninolytic peroxidases (ELP) in culture broths of filamentous fungi (Fusarium solani HP-1), and constitutes the first formal study of the fungal community associated with the EHCO of the Orinoco Oil Belt.     

A First Record of Obligate Halophilic Aspergilli from the Dead Sea

The isolation of obligate halophilic aspergilli from the Dead Sea and the range of salt tolerance of halophilic fungi isolated, are reported here for the first time. The mycobiota of the Dead Sea isolated in this study, was dominated by Aspergillus and Penicillium species; Cladosporium were found in lesser numbers. All three genera were obtained from the water sample; however, Aspergillus was the only genus obtained from the sediment. There was significant difference in growth of each isolate at different salt concentrations and intraspecies analysis revealed dissimilarity in response of strains to different salt concentrations in the growth medium The isolates were euryhaline, with halotolerance up to 20–25% solar salt, Aspergillus and Penicillium species showing a higher level of halotolerance, as compared to that of Cladosporium. Halophilic fungi were found in greater numbers in the sediment sample as compared to that in the water sample. Penicillium and Cladosporium species were exclusively facultative halophiles, while some species of Aspergillus were facultative halophiles. All the obligate halophiles isolated, belonged to the genus Aspergillus and were identified as A. penicillioides and A unguis, the latter being a first record of the species from the Dead Sea.     

Yeasts and filamentous fungi carried by the gynes of leaf-cutting ants

Insect-associated microbes exhibit a wide range of interactions with their hosts. One example of such interactions is the insect-driven dispersal of microorganisms, which plays an essential role in the ecology of several microbes. To study dispersal of microorganisms by leaf-cutting ants (Formicidae: Attini), we applied culture-dependent methods to identify the filamentous fungi and yeasts found in two different body parts of leaf-cutting ant gynes: the exoskeleton and the infrabuccal pocket. The gynes use the latter structure to store a pellet of the ants’ symbiotic fungus during nest founding. Many filamentous fungi (n = 142) and yeasts (n = 19) were isolated from the gynes’ exoskeleton. In contrast, only seven filamentous fungi and three yeasts isolates were recovered from the infrabuccal pellets, suggesting an efficient mechanism utilized by the gynes to prevent contamination of the symbiotic fungus inoculum. The genus Cladosporium prevailed (78%) among filamentous fungi whereas Aureobasidium, Candida and Cryptococcus prevailed among yeasts associated with gynes. Interestingly, Escovopsis, a specialized fungal pathogen of the leaf-cutting ant-fungus symbiosis, was not isolated from the body parts or from infrabuccal pellets of any gynes sampled. Our results suggest that gynes of the leaf-cutter ants Atta laevigata and A. capiguara do not vertically transmit any particular species of yeasts or filamentous fungi during the foundation of a new nest. Instead, fungi found in association with gynes have a cosmopolitan distribution, suggesting they are probably acquired from the environment and passively dispersed during nest foundation. The possible role of these fungi for the attine ant–microbial symbiosis is discussed.     

Seasonal study of the fungal biota of the fur of dogs

During a one year period, 944 dogs from the Municipal kennel of Barcelona were examined to detect animals with suspected dermatophytosis. Only a few animals (1.8%) presented skin lesions but none of them had dermatophytosis. A representative number of dogs without visible skin lesions (n=172), selected at random, were used to carry out a seasonal study of the mycobiota of their fur. Fifteen isolates belonging to the genera Microsporum and Trichophyton were isolated from 14 of the 172 (8.1%) dogs without lesions. The identity of these fungi was Microsporum gypseum (6/15), Trichophyton terrestre (4/15), M. canis (2/15), M. cookei (2/15) and Trichophyton ajelloi (1/15) (one strain each of M. gypseum and T. ajelloi were isolated from one dog). Species of Penicillium (% prevalence=89.5%), Alternaria (86.6%), Cladosporium (84.9%), Aspergillus (77.3%), Scopulariopsis (65.7%) and Chrysosporium (64.5%) were the most prevalent. No significant differences in the fungal biota were observed with respect to age, gender, hair length or between mixed and pure breed dogs. A large number of isolates, including species belonging to the genera Beauveria, Chrysosporium, Malbranchea and Scopulariopsis, that macroscopically and/or microscopically resemble dermatophytes and may be mistaken for them, produced a red color change in Dermatophyte Test Medium. No significant seasonal difference was detected among the isolates belonging to the the most frequently encountered genera, with the exception of Scopulariopsis (higher in summer and autumn) and Chrysosporium (higher in summer). Species from other genera, with lower occurrence also presented significant differences in their seasonal distribution. Arthrinium, Aureobasidium, Chaetomium and Phoma spp. presented maximum prevalence peaks in spring, Fusarium, Paecilomyces, Phoma and Rhizopus spp. in summer and Geotrichum and Mucor spp. in autumn. The Microsporum and Trichophyton species were more frequently isolated in summer.     

Sperm Ultrastructure of Two Mollusk Species Geukensia demissa and Modiolus modiolus (Mytilidae, Bivalvia)

Sperm ultrastructure was studied in two Atlantic species of bivalve mollusks Geukensia demissa and Modiolus modiolus (Mytilidae). These spermatozoa have a classic structure common for species with external fertilization: the head of the spermatozoon of G. demissa and M. modiolus consists of an acrosome, including an acrosomal vesicle and periacrosomal material, a nucleus, and a middle part with two mutually perpendicular centrioles, surrounded by a ring of spherical mitochondria. The spermatozoa of G. demissa and M. modiolus differ in the general shape and size of the head (M. modiolus, 5.8 × 4 m; G. demissa, 4.2 × 2.5 m), as well as the structure of the acrosome and the number of mitochondria (M. modiolus, 8–12; G. demissa, 5–6). The spermatozoa of the Atlantic species M. modiolus and M. modiolus (M. kurilensis) from the Sea of Japan are close in ultrastructure, but differ in the number of mitochondria. This could possibly be a reason to consider M. modiolus from the Sea of Japan as a separate subspecies, M. modiolus kurilensis. In ultrastructure, the spermatozoa of G. demissa differ significantly from the spermatozoa of M. modiolus, but are similar to the spermatozoa of species of the genus Brachidontes.