Novel species of fungi described in this study include those from various countries as follows: Antarctica, Cladosporium arenosum from marine sediment sand. Argentina, Kosmimatamyces alatophylus (incl. Kosmimatamyces gen. nov.) from soil. Australia, Aspergillus banksianus, Aspergillus kumbius, Aspergillus luteorubrus, Aspergillus malvicolor and Aspergillus nanangensis from soil, Erysiphe medicaginis from leaves of Medicago polymorpha, Hymenotorrendiella communis on leaf litter of Eucalyptus bicostata, Lactifluus albopicri and Lactifluus austropiperatus on soil, Macalpinomyces collinsiae on Eriachne benthamii, Marasmius vagus on soil, Microdochium dawsoniorum from leaves of Sporobolus natalensis, Neopestalotiopsis nebuloides from leaves of Sporobolus elongatus, Pestalotiopsis etonensis from leaves of Sporobolus jacquemontii, Phytophthora personensis from soil associated with dying Grevillea mccutcheonii. Brazil, Aspergillus oxumiae from soil, Calvatia baixaverdensis on soil, Geastrum calycicoriaceum on leaf litter, Greeneria kielmeyerae on leaf spots of Kielmeyera coriacea. Chile, Phytophthora aysenensis on collar rot and stem of Aristotelia chilensis. Croatia, Mollisia gibbospora on fallen branch of Fagus sylvatica. Czech Republic, Neosetophoma hnaniceana from Buxus sempervirens. Ecuador, Exophiala frigidotolerans from soil. Estonia, Elaphomyces bucholtzii in soil. France, Venturia paralias from leaves of Euphorbia paralias. India, Cortinarius balteatoindicus and Cortinarius ulkhagarhiensis on leaf litter. Indonesia, Hymenotorrendiella indonesiana on Eucalyptus urophylla leaf litter. Italy, Penicillium taurinense from indoor chestnut mill. Malaysia, Hemileucoglossum kelabitense on soil, Satchmopsis pini on dead needles of Pinus tecunumanii. Poland, Lecanicillium praecognitum on insects' frass. Portugal, Neodevriesia aestuarina from saline water. Republic of Korea, Gongronella namwonensis from freshwater. Russia, Candida pellucida from Exomias pellucidus, Heterocephalacria septentrionalis as endophyte from Cladonia rangiferina, Vishniacozyma phoenicis from dates fruit, Volvariella paludosa from swamp. Slovenia, Mallocybe crassivelata on soil. South Africa, Beltraniella podocarpi, Hamatocanthoscypha podocarpi, Coleophoma podocarpi and Nothoseiridium podocarpi (incl. Nothoseiridium gen. nov.) from leaves of Podocarpus latifolius, Gyrothrix encephalarti from leaves of Encephalartos sp., Paraphyton cutaneum from skin of human patient, Phacidiella alsophilae from leaves of Alsophila capensis, and Satchmopsis metrosideri on leaf litter of Metrosideros excelsa. Spain, Cladophialophora cabanerensis from soil, Cortinarius paezii on soil, Cylindrium magnoliae from leaves of Magnolia grandiflora, Trichophoma cylindrospora (incl. Trichophoma gen. nov.) from plant debris, Tuber alcaracense in calcareus soil, Tuber buendiae in calcareus soil. Thailand, Annulohypoxylon spougei on corticated wood, Poaceascoma filiforme from leaves of unknown Poaceae. UK, Dendrostoma luteum on branch lesions of Castanea sativa, Ypsilina buttingtonensis from heartwood of Quercus sp. Ukraine, Myrmecridium phragmiticola from leaves of Phragmites australis. USA, Absidia pararepens from air, Juncomyces californiensis (incl. Juncomyces gen. nov.) from leaves of Juncus effusus, Montagnula cylindrospora from a human skin sample, Muriphila oklahomaensis (incl. Muriphila gen. nov.) on outside wall of alcohol distillery, Neofabraea eucalyptorum from leaves of Eucalyptus macrandra, Diabolocovidia claustri (incl. Diabolocovidia gen. nov. ) from leaves of Serenoa repens, Paecilomyces penicilliformis from air, Pseudopezicula betulae from leaves of leaf spots of Populus tremuloides. Vietnam, Diaporthe durionigena on branches of Durio zibethinus and Roridomyces pseudoirritans on rotten wood. Morphological and culture characteristics are supported by DNA barcodes.
Archaea and Bacteria constitute a majority of life systems on Earth but have long been considered inferior to Eukarya in terms of solute tolerance. Whereas the most halophilic prokaryotes are known for an ability to multiply at saturated NaCl (water activity (aw) 0.755) some xerophilic fungi can germinate, usually at high-sugar concentrations, at values as low as 0.650–0.605 aw. Here, we present evidence that halophilic prokayotes can grow down to water activities of <0.755 for Halanaerobium lacusrosei (0.748), Halobacterium strain 004.1 (0.728), Halobacterium sp. NRC-1 and Halococcus morrhuae (0.717), Haloquadratum walsbyi (0.709), Halococcus salifodinae (0.693), Halobacterium noricense (0.687), Natrinema pallidum (0.681) and haloarchaeal strains GN-2 and GN-5 (0.635 aw). Furthermore, extrapolation of growth curves (prone to giving conservative estimates) indicated theoretical minima down to 0.611 aw for extreme, obligately halophilic Archaea and Bacteria. These were compared with minima for the most solute-tolerant Bacteria in high-sugar (or other non-saline) media (Mycobacterium spp., Tetragenococcus halophilus, Saccharibacter floricola, Staphylococcus aureus and so on) and eukaryotic microbes in saline (Wallemia spp., Basipetospora halophila, Dunaliella spp. and so on) and high-sugar substrates (for example, Xeromyces bisporus, Zygosaccharomyces rouxii, Aspergillus and Eurotium spp.). We also manipulated the balance of chaotropic and kosmotropic stressors for the extreme, xerophilic fungi Aspergillus penicilloides and X. bisporus and, via this approach, their established water-activity limits for mycelial growth (∼0.65) were reduced to 0.640. Furthermore, extrapolations indicated theoretical limits of 0.632 and 0.636 aw for A. penicilloides and X. bisporus, respectively. Collectively, these findings suggest that there is a common water-activity limit that is determined by physicochemical constraints for the three domains of life.
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).
The effect of atmosphere containing 80% CO(2) and 20% O(2) on growth of Mucor plumbeus, Fusarium oxysporum, Byssochlamys fulva, Byssochlamys nivea, Penicillium commune, Penicillium roqueforti, Aspergillus flavus, Eurotium chevalieri and Xeromyces bisporus was investigated. Production of aflatoxin by A. flavus, patulin by B. nivea, roquefortine C by P. roqueforti, and cyclopiazonic acid by P. commune was also studied. Fungal growth was evaluated by three methods: colony diameter, hyphal length or mycelium dry weight and ergosterol content. Among the nine fungal species examined, two E. chevalieri and X. bisporus, did not grow under these conditions. In this study, fungi differed in their response to modified atmospheres in biomass, ergosterol content, mycotoxin production and morphology. Reductions of 57.8-96.9%, 73.7-99.6% and 91.5-99.9% were obtained in colony diameter, hyphal length and ergosterol content, respectively, under this atmosphere compared to air. Ergosterol content was more affected in most species than other measurements. Patulin, cyclopiazonic acid and roquefortine C were produced in this atmosphere, although levels were very low and aflatoxin was not produced at all. Growth was quite extensive as measured by colony diameters, but hyphal lengths were low and ergosterol production was also affected in all species of this study.
I would like to preface my remarks by saying how honoured I feel to be the recipient of the 2009 AIFST Keith Farrer Award of Merit and to thank those colleagues who nominated me for the Award. I aim to take you on a journey through more than 50 years of Australian food mycology research, most of which has been done at the laboratories of CSIRO Food Research at North Ryde - a Division which has seen many name changes in that period.
Reduction in water activity (aw) is used as a microbiological hurdle to prevent food spoilage. To minimize the levels of salt and sugar, which are commonly used to reduce aw, the potential of food structure as a microbiological hurdle needs to be assessed. The concept of matric potential (Ψm) is used to measure the effect of food structure on water movement. This study reports the effect of reduced aw and reduced Ψm on the germination of xerophilic fungi (represented by Eurotium herbariorum) and non-xerophilic fungi (represented by Aspergillus niger) on model glycerol agar media. Germination curves were plotted with the percentage of germinated spores against time. The germination time (tG), which is defined as the time at which 50% of the total viable spores have germinated, was estimated using the Gompertz model. Total viable spores was defined as those spores that were able to germinate under the optimum aw and Ψm conditions for each species, i.e. 0.95 aw and 2.5% agar for E. herbariorum and 0.98 aw and 2.5% agar for A. niger. As aw decreased from 0.90 to 0.85 aw, tG increased significantly for both the xerophilic fungi and non-xerophilic species at equivalent matric potential values. When matric potential was reduced from −12kPa (2.5% agar) to −38kPa (12.5% agar), tG of A. niger was significantly extended at 0.90 aw; however, tG remained the same for A. niger at 0.85 aw, and for E. herbariorum at 0.80, 0.85 and 0.90 aw. This study demonstrated that the germination time for non-xerophilic and xerophilic fungi was extended by reduced aw, however the effect of reduced Ψm was limited.
The influence of high carbon dioxide and low oxygen concentrations on growth by the foodborne fungal species, Mucor plumbeus, Fusarium oxysporum, Byssochlamys fulva, Byssochlamys nivea, Penicillium commune, Penicillium roqueforti, Aspergillus flavus, Eurotium chevalieri and Xeromyces bisporus was investigated. Production of aflatoxin by A. flavus, patulin by B. nivea and roquefortine C by P. roqueforti was also studied. Fungal growth was evaluated under atmospheres consisting of 20, 40 and 60% CO2 plus <0.5% O2, on two media, Czapek Yeast Extract agar and Potato Dextrose agar. Several methods for measuring fungal growth were used: colony diameter, ergosterol content, hyphal length and/or mycelium dry weight. Among the nine species, three groups were distinguished with respect to their growth responses under modified atmospheres: (i) species which did not grow in 20% CO2 <0.5% O2 (P. commune, E. chevalieri and X. bisporus); (ii) species which grew in 20% CO2 <0.5% O2, but not 40% CO2 <0.5% O2 (P. roqueforti and A. flavus); (iii) species which grew in 20%, 40% and 60% CO2 <0.5% O2 (M. plumbeus, F. oxysporum, B. fulva and B. nivea). Facultatively anaerobic behaviour was observed in these last four species, which grew under the same conditions as the obligate anaerobe, Clostridium sporogenes. The production of aflatoxin, patulin, and roquefortine C was greatly reduced under all of the atmospheres tested.
The potential efficacy of four weak acids as preservatives in low-acid intermediate moisture foods was assessed using a glycerol based agar medium. The minimum inhibitory concentrations (MIC, % wt./wt.) of each acid was determined at two pH values (pH 5.0, pH 6.0) and two a(w) values (0.85, 0.90) for five food spoilage fungi, Eurotium herbariorum, Eurotium rubrum, Aspergillus niger, Aspergillus flavus and Penicillium roqueforti. Sorbic acid, a preservative commonly used to control fungal growth in low-acid intermediate moisture foods, was included as a reference. The MIC values of the four acids were lower at pH 5.0 than pH 6.0 at equivalent a(w) values, and lower at 0.85 a(w) than 0.90 a(w) at equivalent pH values. By comparison with the MIC values of sorbic acid, those of caprylic acid and dehydroacetic acid were generally lower, whereas those for caproic acid were generally higher. No general observation could be made in the case of capric acid. The antifungal activities of all five weak acids appeared related not only to the undissociated form, but also the dissociated form, of each acid.
It is trite to say that dried foods must be kept dry, heat processed foods must be heated enough to inactivate all relevant spores and preservative concentrations must be high enough to inhibit all fungi. In reality, the science of preserving foods, like so many other disciplines, requires compromise. Really dry foods, i.e. of a safe aw, may be impossible to obtain for climatic or economic reasons or be unacceptable to the consumer; a sufficient heat process may destroy desirable flavours; and permitted preservative levels are set by law. Some fungi, by virtue of specific attributes, simply cannot be processed out of certain types of foods. Of particular importance are Xeromyces bisporus and Zygosaccharomyces rouxii – extreme xerophiles which grow in concentrated foods; Byssochlamys spp., Talaromyces spp. and Neosartorya fischeri with ascospores of very high heat resistance which can survive heat processing and grow in heat processed acid foods; and Zygosaccharomyces bailii, a preservative resistant yeast. Making foods safe from these fungi requires that they be absent from raw materials or destroyed by pasteurisation and then excluded from the processing and packing lines.
Principles underlying fungal classification have been outlined in Chapter 3, including a brief overview of the relevant divisions of the Kingdom Fungi and their principal methods of reproduction. Some further detailed information is necessary in this chapter to assist in the use of the keys which follow.
This paper reports on a survey of the microbiological status of breadcrumbs manufactured in four Australian states. A moisture sorption isotherm of breadcrumbs showed that at a moisture content of 10% the expected a(w) would be 0.45, a safe level for preventing mould growth. The potential for pre-formed mycotoxins in bread returns constitutes a greater risk than fungal growth in the dried product. Breadcrumb manufacturers processing bread returns into crumb need to ensure that mouldy bread is excluded, and that only product within its use-by date is used. They should ensure that breadcrumbs are dried to a moisture content of 10% or less to provide an adequate safety margin to prevent mould growth.