Heat-resistance of Hamigera avellanea and Thermoascus crustaceus isolated from pasteurized acid products.

Products containing sugar or fruit derivatives are usually subjected to a pasteurization process that can anyway be ineffective to kill ascospores from heat-resistant molds. Although the most occurring and economically relevant heat-resistant species belong to Byssochlamys, Neosartorya, Talaromyces, and Eupenicillium genera, an increasing number of uncommon heat-resistant isolates have been recently detected as spoiling microorganisms in such products. Since Hamigera spp. and Thermoascus spp. were those more frequently isolated at SSICA, heat resistance of Hamigera avellanea and Thermoascus crustaceus strains from pasteurized acid products was studied in apple juice, in blueberry and grape juice and in a buffered glucose solution. Data obtained from thermal death curves and statistical elaboration of raw data showed that D values of H. avellanea may vary between 11.11 and 66.67 min at 87°C, between 4.67 and 13.51 at 90°C, and between 0.43 and 1.52 min at 95°C. Similarly, D values of T. crustaceus may vary between 18.52 and 90.91 min at 90°C, between 2.79 and 19.23 at 93°C, and between 1.11 and 2.53 min at 95°C. For both strains studied, the z-values calculated from the decimal reduction time curves did not prove to be significantly influenced by the heating medium, that being 4.35°C, 5.39°C or 5.27°C for H. avellanea and 4.42°C, 3.69°C or 3.37°C for T. crustaceus, respectively in apple juice, in blueberry and grape juice or in the buffered glucose solution. Considering the pasteurization treatments industrially applied to fruit-based foods, the variation of thermal parameters does not seem to be a possible way to avoid product spoilage by these two species and only good practices applied to reduce the original load of heat-resistant fungi can help producers to prevent losses in contaminated finished products, as usually happens for other heat resistant molds.

Rasamsonia argillacea species complex: taxonomy, pathogenesis and clinical relevance.

Since 2010, colonizations/infections by Rasamsonia argillacea species complex, previously known as Geosmithia argillacea, have been regularly reported in literature. We reviewed all available cases focusing on pathogenesis and clinical relevance. The number of cases may be underestimated, as these fungi are frequently misidentified as Penicillium or Paecilomyces species. Major underlying conditions that predispose for infections by the R. argillacea species complex include cystic fibrosis (CF) and chronic granulomatous disease (CGD). While the pathogenic role of the colonization of CF lungs is still under debate, these molds are the causative agent of pneumonia and/or invasive infections in CGD patients. Given their thermotolerance and their resistance to various antifungals, especially the azole drugs, a special attention should be paid to the chronic colonization of the airways by these fungi in CF and CGD patients.

Rasamsonia, a new genus comprising thermotolerant and thermophilic Talaromyces and Geosmithia species.

The phylogenetic relationship among Geosmithia argillacea, Talaromyces emersonii, Talaromyces byssochlamydoides and other members of the Trichocomaceae was studied using partial RPB2 (RNA polymerase II gene, encoding the second largest protein subunit), Tsr1 (putative ribosome biogenesis protein) and Cct8 (putative chaperonin complex component TCP-1) gene sequences. The results showed that these species form a distinct clade within the Trichocomaceae and Trichocoma paradoxa is phylogenetically most closely related. Based on phenotypic and physiological characters and molecular data, we propose Rasamsonia gen. nov. to accommodate these species. This new genus is distinct from other genera of the Trichocomaceae in being thermotolerant or thermophilic and having conidiophores with distinctly rough walled stipes, olive-brown conidia and ascomata, if present, with a scanty covering. Species within the genus Rasamsonia were distinguished using a combination of phenotypic characters, extrolite patterns, ITS and partial calmodulin and ß-tubulin sequences. Rasamsonia brevistipitata sp. nov. is described and five new combinations are proposed.

The extreme xerophilic mould Xeromyces bisporus--growth and competition at various water activities.

Little is known about the mould, Xeromyces bisporus, unique in its strong xerophilicity and ability to grow at water activity (a(w)) 0.62, lower than for any other known organism. The linear growth rates of one fast and one slow-growing strain of X. bisporus were assessed at 20, 25, 30 and 37 °C on solid agar media containing a mixture of glucose and fructose to reduce a(w) to 0.94, 0.88, 0.84, 0.80, 0.76 and 0.66. Growth rates of xerophilic species closely related to X. bisporus, viz. Chrysosporium inops, C. xerophilum and Monascus eremophilus, were also assessed. Optimal conditions for growth of both X. bisporus strains were approx. 0.84 a(w) and 30°C, despite FRR 2347 growing two- to five-fold faster than CBS 185.75. X. bisporus FRR 2347 even grew well at 0.66 a(w) (0.48 mm/day). C. inops and C. xerophilum were more tolerant of high a(w) than X. bisporus, and could be differentiated from each other based on: the faster growth of C. xerophilum; its preference for temperatures ≥ 30 °C and a(w) ≥ 0.94 (c.f.≤ 25 °C and ~0.88 a(w) for C. inops); and its ability to grow at 0.66 a(w), which is the lowest a(w) reported to date for this species. M. eremophilus grew slowly (max. 0.4mm/day) even in its optimal conditions of ~0.88 a(w) and 25 °C. To investigate the competitive characteristics of X. bisporus at low a(w), both X. bisporus strains were grown in dual-culture with xerotolerant species Aspergillus flavus and Penicillium roqueforti, and xerophilic species A. penicillioides, C. inops, C. xerophilum and Eurotium chevalieri, on glucose-fructose agar plates at 0.94, 0.84, 0.80 and 0.76 a(w) and at 25 °C. Growth rates and types of interactions were assessed. Excretion of inhibitory substances acting over a long-range was not observed by any species; inhibitors acting over a short-range that temporarily slowed competitors' growth or produced a protective zone around the colony were occasionally observed for A. penicillioides, C. inops and C. xerophilum. Instead, rapid growth relative to the competitor was the most common means of dominance. The xerotolerant species, A. flavus and P. roqueforti were dominant over X. bisporus at 0.94 a(w). E. chevalieri was often dominant due to its rapid growth over the entire a(w) range. At a(w)<0.80, X. bisporus was competitive because it grew faster than the other species examined. This supports the concept that its ideal environmental niche is sugary foods with low a(w).

Sexual reproduction in Aspergillus flavus.

Aspergillus flavus is the major producer of carcinogenic aflatoxins in crops worldwide and is also an important opportunistic human pathogen in aspergillosis. The sexual state of this heterothallic fungus is described from crosses between strains of the opposite mating type. Sexual reproduction occurred between sexually compatible strains belonging to different vegetative compatibility groups. Multiple, indehiscent ascocarps containing asci and ascospores formed within the pseudoparenchymatous matrix of stromata, which places the fungus in genus Petromyces. The teleomorph of P. flavus could not be distinguished from that of P. parasiticus (anamorph = A. parasiticus), another aflatoxin-producing species, based on morphology of the sexual structures. The two species can be separated by anamorph morphology, mycotoxin profile and molecular characters.

Host specificity of Eupenicillium ochrosalmoneum, E. cinnamopurpureum and two Penicillium species associated with the conidial heads of Aspergillus.

The genus Penicillium comprises species that mostly colonize plant matter. However early reports suggest that several species are capable of parasitizing Aspergillus and sporulating on the conidial heads of the host. More recently Eupenicillium ochrosalmoneum and E. cinnamopurpureum, both with Penicillium anamorphs, have been observed sporulating on the heads of Aspergillus species belonging to section Flavi during the colonization of peanut seeds. Little is known about the host specificity underlying these Aspergillus-Penicillium associations. In this study Aspergillus species representing nine taxonomic sections were paired in culture with E. ochrosalmoneum, E. cinnamopurpureum and two unnamed Penicillium species. Eupenicillium ochrosalmoneum, E. cinnamopurpureum and Penicillium sp. 1 sporulated predominantly on the heads of section Flavi species. In contrast Penicillium sp. 2 was restricted to the heads of section Nigri species. All species spread across Aspergillus colonies by means of aerial hyphae that grew from head to head. Additional studies are required to clarify whether Eupenicillium and Penicillium species are parasitic or simply epibiotic on their hosts.

Genomic islands in the pathogenic filamentous fungus Aspergillus fumigatus.

We present the genome sequences of a new clinical isolate of the important human pathogen, Aspergillus fumigatus, A1163, and two closely related but rarely pathogenic species, Neosartorya fischeri NRRL181 and Aspergillus clavatus NRRL1. Comparative genomic analysis of A1163 with the recently sequenced A. fumigatus isolate Af293 has identified core, variable and up to 2% unique genes in each genome. While the core genes are 99.8% identical at the nucleotide level, identity for variable genes can be as low 40%. The most divergent loci appear to contain heterokaryon incompatibility (het) genes associated with fungal programmed cell death such as developmental regulator rosA. Cross-species comparison has revealed that 8.5%, 13.5% and 12.6%, respectively, of A. fumigatus, N. fischeri and A. clavatus genes are species-specific. These genes are significantly smaller in size than core genes, contain fewer exons and exhibit a subtelomeric bias. Most of them cluster together in 13 chromosomal islands, which are enriched for pseudogenes, transposons and other repetitive elements. At least 20% of A. fumigatus-specific genes appear to be functional and involved in carbohydrate and chitin catabolism, transport, detoxification, secondary metabolism and other functions that may facilitate the adaptation to heterogeneous environments such as soil or a mammalian host. Contrary to what was suggested previously, their origin cannot be attributed to horizontal gene transfer (HGT), but instead is likely to involve duplication, diversification and differential gene loss (DDL). The role of duplication in the origin of lineage-specific genes is further underlined by the discovery of genomic islands that seem to function as designated "gene dumps" and, perhaps, simultaneously, as "gene factories".

Phosphate solubilization potential and stress tolerance of Eupenicillium parvum from tea soil.

Eupenicillium parvum was recorded for first time during isolation of phosphate-solubilizing microorganisms from the tea rhizosphere. The fungus developed a phosphate solubilization zone on modified Pikovskaya agar, supplemented with tricalcium phosphate. Quantitative estimation of phosphate solubilization in Pikovskaya broth showed high solubilization of tricalcium phosphate and aluminium phosphate. The fungus also solubilized North Carolina rock phosphate and Mussoorie rock phosphate, and exhibited high levels of tolerance against desiccation, acidity, salinity, aluminium, and iron. Solubilization of inorganic phosphates by the fungus was also observed under high stress levels of aluminium, iron, and desiccation, though the significant decline in phosphate solubilization was marked in the presence of aluminium than iron. The fungal isolate showed 100% identity with E. parvum strain NRRL 2095 ITS 1, 5.8S rRNA gene and ITS 2, complete sequence; and 28S rRNA gene, partial sequence.

Effect of intra and interspecific interaction on OTA production by A. section Nigri in grapes during dehydration.

The aim of this study was to assess the colonizing capability and OTA production of different populations of Aspergillus section Nigri spp. in grapes, as affected by the interactions with other fungi, during a simulated in vitro sun-drying. Mature white grapes were divided into two lots of healthy and artificially injured grapes and inoculated with A. section Nigri spp. (A. carbonarius OTA producer, A. niger aggregate OTA producer, A. niger aggregate OTA non-producer), Eurotium amstelodami and Penicillium janthinellum, in different combinations. The drying process was simulated adjusting water activity firstly at 0.98 a(w) and gradually decreasing it to 0.76 a(w) for a total of 20 days. Colonizing grape percentages were recorded after 5, 10, 15 and 20 days of dehydration and OTA content was measured after 5, 7, 10, 12, 15, 17 and 20 days. Colonization of grapes increased with time in all treatments. A. niger aggregate OTA-positive showed the highest colonization percentage, followed by A. carbonarius, and finally their mixed inoculum. When the two OTA-producing strains were combined, addition of any other microorganism increased the percentage of infection by A. section Nigri. A. carbonarius was the highest OTA producer in pure culture, followed by A. niger aggregate OTA-positive. In general, when competing fungi were added to A. carbonarius inoculum, the OTA content was reduced. E. amstelodami was the only competing fungus which increased OTA accumulation. The sun-drying process may be conducive to OTA accumulation in dried grapes. The complex fungal interactions which may take place during this process, may act as an additional control factor, given that the higher presence of A. niger aggregate OTA-negative inhibits OTA accumulation by OTA producing species.

Trichocomaceae in bark survive high temperatures and fire.

Fifty-six species in the Trichocomaceae were recovered from bark of trees and shrubs from hot arid and temperate regions, and following one fire in a temperate region of Australia. Fungi were recovered from dry bark after incubation for up to 1 h at up to 105 degrees C. Fourteen species also regenerated on agar after their conidia were heated for 1 h at 105 degrees C. Anamorphic species were commonly recovered and widespread. Teleomorphic species were only recovered after heating the bark. In addition, anamorphic fungi were recovered from one plant species following a natural fire. The results support the view that both anamorphic and teleomorphic fungi may tolerate extreme temperatures in their environment while dry.

Novel Neosartorya species isolated from soil in Korea.

Phenotypic and genotypic characters of strains of Neosartorya spinosa and related taxa were analysed. N. spinosa, Neosartorya botucatensis and Neosartorya paulistensis had identical partial beta-tubulin and calmodulin gene sequences and could not be differentiated on macro- and micro-morphological characteristics, including by scanning electron microscopy. Based on partial beta-tubulin and calmodulin gene sequences and ascospore morphology, two separate groups are distinguished and are proposed as novel species. Neosartorya laciniosa sp. nov. has microtuberculate ascospores with two bent crests and two distinct equatorial rings of small projections. Neosartorya coreana sp. nov. has rugose to weak reticulate ascospores with two often bent crests, but without the equatorial rings of small projections. The type strain of N. laciniosa is CBS 117721T (=NRRL 35589T=KACC 41657T) and the type strain of N. coreana is CBS 117059T (=NRRL 35590T=KACC 41659T).

Isolation of Neosartorya pseudofischeri from blood: first hint of pulmonary Aspergillosis.

We report a case of invasive pulmonary aspergillosis caused by Neosartorya pseudofischeri S. W. Peterson [anamorph Aspergillus thermomutatus (Paden) S. W. Peterson]. The diagnosis was initially based on a positive blood culture for a strain isolated from a neutropenic patient by means of a BACTEC 9050 blood culture system. The final diagnosis was established based on X-ray and computer tomography scan results as well as the detection of Aspergillus antigen in the patient's serum.