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The study aimed to evaluate the slaughter value and meat characteristics of ten ostrich females reared and slaughtered at the age of 18 months in Hungary. The ratio of selected body parts, the main organs and the lean meat parts were examined. The nutritive composition, the colour, the technological and organoleptic characteristics of five valuable meat parts (outside strip—M. flexor cruris lateralis, oyster—M. iliofemoralis externus, tip—M. femorotibialis medius, outside leg—M. gastrocnemius pars externa, medal—M. ambiens) and the amino acid, fatty acid and mineral composition of outside strip (M. flexor cruris lateralis) were also evaluated. The ratio of body parts and the main organs as the percentage of live weight, and the lean meat part as the percentage of carcass weight showed 16.74 ± 0.01%, 6.16 ± 0.01% and 57.29 ± 0.59%, respectively. The dry matter content of the examined valuable meat parts ranged between 24.89 ± 0.08 and 26.23 ± 0.13%, the protein ratio took on values between 18.40 ± 0.09 and 20.62 ± 0.16%, the fat content showed values between 2.36 ± 0.07 and 4.50 ± 1.09% and the hydroxyproline content ranged between 0.01 ± 0.001 and 0.08 ± 0.001%. The amino acid content of the outside strip showed a range between 0.15 and 3.33%. The ratio of SFA, MUFA and PUFA was 35.10 ± 0.53, 37.37 ± 1.52 and 27.54 ± 1.01. The n-6/n-3 ratio showed 3.91 ± 0.43 and the SFA/UFA ratio was 0.54 ± 0.02. Among the examined minerals, the content of Ca, K, Mg, Na and P was the highest in the meat. In the case of the colour, regarding L* value, we could reveal no significant difference between the examined meat parts. For a* and b* values, the outside leg had the lowest data of all. We could not reveal a significant difference between the pH values of the meat parts. Regarding technological parameters, meat differed only in thawing loss. The significantly lowest thawing loss could be detected in the outside leg (2.72 ± 0.01%) and in the medal (2.32 ± 0.01%). The results of the organoleptic evaluation showed that the outside strip and the tip had the best flavour and tenderness. In comparison with the younger birds (10–14 months of age) in the literature, the 18-month-old ostriches in our study showed similar or slightly lower slaughter weight, skin weight and head ratio, greater liver weight, lighter meat, lower protein and higher fat content, higher essential amino acid and lower non-essential amino acid content and higher SFA content in some cases. However, data on nutrition and population size were not always available. In comparison with other ratites (emu and rhea), ostrich meat has lower dry matter and protein, but higher fat, SFA, MUFA and PUFA content and lower n-6/n-3 ratio.

Aflatoxins (AFs) are toxic secondary metabolites produced mostly by Aspergillus species. AF contamination entering the feed and food chain has been a crucial long-term issue for veterinarians, medicals, agroindustry experts, and researchers working in this field. Although different (physical, chemical, and biological) technologies have been developed, tested, and employed to mitigate the detrimental effects of mycotoxins, including AFs, universal methods are still not available to reduce AF levels in feed and food in the last decades. Possible biological control by bacteria, yeasts, and fungi, their excretes, the role of the ruminal degradation, pre-harvest biocontrol by competitive exclusion or biofungicides, and post-harvest technologies and practices based on biological agents currently used to alleviate the toxic effects of AFs are collected in this review. Pre-harvest biocontrol technologies can give us the greatest opportunity to reduce AF production on the spot. Together with post-harvest applications of bacteria or fungal cultures, these technologies can help us strictly reduce AF contamination without synthetic chemicals.

Glutathione (GSH) is an abundant tripeptide that plays a crucial role in shielding cellular macromolecules from various reactive oxygen and nitrogen species in fungi. Understanding GSH metabolism is of vital importance for deciphering redox regulation in these microorganisms. In the present study, to better understand the GSH metabolism in filamentous fungi, we investigated functions of the dugB and dugC genes in the model fungus Aspergillus nidulans. These genes are orthologues of dug2 and dug3, which are involved in cytosolic GSH degradation in Saccharomyces cerevisiae. The deletion of dugB, dugC, or both resulted in a moderate increase in the GSH content in mycelia grown on glucose, reduced conidium production, and disturbed sexual development. In agreement with these observations, transcriptome data showed that genes encoding mitogen-activated protein (MAP) kinase pathway elements (e.g., steC, sskB, hogA, and mkkA) or regulatory proteins of conidiogenesis and sexual differentiation (e.g., flbA, flbC, flbE, nosA, rosA, nsdC, and nsdD) were downregulated in the ΔdugB ΔdugC mutant. Deletion of dugB and/or dugC slowed the depletion of GSH pools during carbon starvation. It also reduced accumulation of reactive oxygen species and decreased autolytic cell wall degradation and enzyme secretion but increased sterigmatocystin formation. Transcriptome data demonstrated that enzyme secretions—in contrast to mycotoxin production—were controlled at the posttranscriptional level. We suggest that GSH connects starvation and redox regulation to each other: cells utilize GSH as a stored carbon source during starvation. The reduction of GSH content alters the redox state, activating regulatory pathways responsible for carbon starvation stress responses. IMPORTANCE Glutathione (GSH) is a widely distributed tripeptide in both eukaryotes and prokaryotes. Owing to its very low redox potential, antioxidative character, and high intracellular concentration, GSH profoundly shapes the redox status of cells. Our observations suggest that GSH metabolism and/or the redox status of cells plays a determinative role in several important aspects of fungal life, including oxidative stress defense, protein secretion, and secondary metabolite production (including mycotoxin formation), as well as sexual and asexual differentiations. We demonstrated that even a slightly elevated GSH level can substantially disturb the homeostasis of fungi. This information could be important for development of new GSH-producing strains or for any biotechnologically relevant processes where the GSH content, antioxidant capacity, or oxidative stress tolerance of a fungal strain is manipulated.

The fungus

FvatfA from the maize pathogen Fusarium verticillioides putatively encodes the Aspergillus nidulans AtfA and Schizasaccharomyces pombe Atf1 orthologous bZIP-type transcription factor, FvAtfA. In this study, a ΔFvatfA deletion mutant was constructed and then genetically complemented with the fully functional FvatfA gene. Comparing phenotypic features of the wild-type parental, the deletion mutant and the restored strains shed light on the versatile regulatory functions played by FvAtfA in (i) the maintenance of vegetative growth on Czapek-Dox and Potato Dextrose agars and invasive growth on unwounded tomato fruits, (ii) the preservation of conidiospore yield and size, (iii) the orchestration of oxidative (H2O2, menadione sodium bisulphite) and cell wall integrity (Congo Red) stress defences and (iv) the regulation of mycotoxin (fumonisins) and pigment (bikaverin, carotenoid) productions. Expression of selected biosynthetic genes both in the fumonisin (fum1, fum8) and the carotenoid (carRA, carB) pathways were down-regulated in the ΔFvatfA strain resulting in defected fumonisin production and considerably decreased carotenoid yields. The expression of bik1, encoding the polyketide synthase needed in bikaverin biosynthesis, was not up-regulated by the deletion of FvatfA meanwhile the ΔFvatfA strain produced approximately ten times more bikaverin than the wild-type or the genetically complemented strains. The abolishment of fumonisin production of the ΔFvatfA strain may lead to the development of new-type, biology-based mycotoxin control strategies. The novel information gained on the regulation of pigment production by this fungus can be interesting for experts working on new, Fusarium-based biomass and pigment production technologies.Key points• FvatfA regulates vegetative and invasive growths of F. verticillioides.• FvatfA also orchestrates oxidative and cell wall integrity stress defenses.• The ΔFvatfA mutant was deficient in fumonisin production.• FvatfA deletion resulted in decreased carotenoid and increased bikaverin yields. Electronic supplementary material The online version of this article (10.1007/s00253-020-10717-6) contains supplementary material, which is available to authorized users.

The fungus Aspergillus oryzae could be shown to be a viable alternative for biosorption of valuable metals from solution. Fungal biomass can be obtained easily in high quantities as a waste of biofermentation processes, and used in a complex, multi-phase solution mimicking naturally occurring, mining-affected water samples. With test solution formulated after natural conditions, formation of secondary Al and Fe phases co-precipitating Ce was recorded in addition to specific biosorption of rare earth elements. Remarkably, the latter were removed from the solution despite the presence of high concentrations of interfering Fe and Al. The biomass was viable even after prolonged incubation in the metal solution, and minimal inhibitory concentrations for single metals were higher than those in the test solution. While precipitation/biosorption of Ce (maximal biosorption efficiency was 58.0 ± 22.3% after 6 h of incubation) coincided with the gross removal of Fe from the metal solution, Y (81.5 ± 11.3% efficiency, 24 h incubation) and Nd (87.4 ± 9.1% efficiency, 24 h incubation) were sequestered later, similarly to Ni and Zn. The biphasic binding pattern specific to single metals could be connected to dynamically changing pH and NH4+ concentrations, which were attributed to the physiological changes taking place in starving A. oryzae biomass. The metals were found extracellularly in minerals associated with the cell wall, and intracellularly precipitated in the vacuoles. The latter process was explained with intracellular metal detoxification resulting in metal resistance.

There are extensive data on effects of antifungal agents on the plant pathogens, especially on Fusariums spp. species. However, investigations on the interaction of chemicals and the treated cultivars are rare. The aim of the study was to test two types of fungicide mixtures, azoxystrobin-propiconazole, and prothioconazole-tebuconazole, which are applied in wheat cultivars intensively, on six fodder maize hybrids that were infected with Fusarium proliferatum in the R1 growth stage in a field trial. The effect of the fungicide treatment was tested on the starch content and antifungal, antioxidant polyphenols of the kernels in the R3–R4 and R6 stage of the cultivars. The level of the fungal presence and the fumonisin concentration of the kernels were increased significantly under the artificial infection. The fumonisin concentration was variable at the R6 stage of the hybrid maize kernels. The treatment with prothioconazole and tebuconazole was found to be suitable when it was done before flowering, while the azoxystrobin-propiconazole treatments were equally successful before and after maize flowering considering the decreasing fumonisin concentration of the kernels. Both fungicide mixtures, when they were applied after maize flowering, affected the starch biosynthesis to the R3–R4 stage significantly. Meanwhile, azoxystrobin-propiconazole also significantly affected the antioxidant flavone/flavanol contents from the R3–R4 stage to the R6 stage.

The effect of different supplementary diets containing humic substances and probiotics for common carp, based on growth rate, specific growth rate %, protein and fat content was investigated. Humic acid was mixed with different mineral elements and the gram-positive bacteria combined as probiotics supplementary diets. These supplements were added to the experimental basal diet (control) at a rate of 0.5%, 1% and 2%. Four groups of fingerling with an average mass of 0.5 g were fed on the experimental diets, three times per day for 11 weeks. 30 fish per tank were weighed randomly every week to measure their growth rate and at the end of the experiment, 5 fish from each treatment were weighed and their fat and protein content was analyzed individually. The results showed that 1% and 2% probiotics increased the growth rate while all treatments have less specific growth rate % than control. In relation to fat and protein contents, the results showed that fish fed with probiotics 1% had high fat and protein contents compared to other groups in general. All fish fed with different levels of supplemented diets showed a decrease in fat in compare with control group. However, protein contents in fish fed with HS 0.5% and probiotics 1% were significantly higher than control and other group of fish fed with different level of supplemented diets. These results suggest that humic acid substances cannot improve fish growth rate, but fat and protein contents could be changed slightly, while probiotic supplementary diets can improve growing rates, having a significant effect on protein content of fish meat.

Cornmeal agar (CMA) is a good to model natural conditions (low C and N, high antioxidants, crude fat) for phytopathogenic fungi. Different CMA media was prepared to model the maize kernel as growth...

The application of yeasts has great potential in reducing the economic damage caused by toxigenic fungi in the agriculture. Some yeasts may act as biocontrol agents inhibiting the growth of filamentous fungi. These species may also gain importance in the preservation of agricultural products and in the reduction of their mycotoxin contamination, yet the extent of mycotoxin production in the presence of biocontrol agents is relatively less understood. The application of yeasts in various technological processes may have a direct inhibitory effect on the toxin production of certain molds, which is independent of their growth suppressing effect. Furthermore, several yeast species are capable of accumulating mycotoxins from agricultural products, thereby effectively decontaminating them. Probiotic yeasts or products containing yeast cell wall are also applied to counteract mycotoxicosis in livestock. Several yeast strains are also able to degrade toxins to less-toxic or even non-toxic substances. This intensively researched field would greatly benefit from a deeper knowledge on the genetic and molecular basis of toxin degradation. Moreover, yeasts and their biotechnologically important enzymes may exhibit sensitivity to certain mycotoxins, thereby mounting a considerable problem for the biotechnological industry. It is noted that yeasts are generally regarded as safe; however, there are reports of toxin degrading species that may cause human fungal infections. The aspects of yeast-mycotoxin relations with a brief consideration of strain improvement strategies and genetic modification for improved detoxifying properties and/or mycotoxin resistance are reviewed here.

Climate changes in Europe, which is characterized by the decrease of rainy days and the higher average temperature at summer, significantly increase the spreading of Aspergillus species and aflatoxin B1 contamination of the staple food and feed materials. The aim of our study was to estimate the possibility of the aflatoxin production of the Aspergilli on crops. From the isolates that were gained from crop samples, higher than 40% of the Aspergillus isolates contained norA, aflR and omtA genes from the aflatoxin genes cluster. Most of these isolates (63%) showed high homology with A. flavus, while three isolates showed high homology to A. tritici/A. candidus, one to A. cristatus/A. amstelodami and one strain showed the highest homology to A. tritici. Six from the A. flavus isolates (85.7%) with norA, aflR and omtA genes could produce aflatoxin B1 on malate extract agar medium. Parallel PCR and toxin measurements are recommended to evaluate the potentiality of aflatoxin production.

Hydrolytic enzyme production is typical of the autolysis in filamentous fungi; however, less attention has been given to the physiological role of the enzymes. Here, the aim was to investigate the possible relation of the chitinolytic enzymes to the changes in the biomass in some filamentous fungi of high importance for pharmaceutical or food industry. In Penicillium and Aspergillus filamentous fungi, which showed different characteristics in submerged cultures, the growth and biomass decline rates were calculated and correlated to the chitinase and N-acetyl-β-D-glucosaminidase enzyme productions. Correlation was found between the biomass decrease rate and the chitinase level at the stationary growth phase; while chitinase production covariates negatively with N-acetyl-β-D-glucosaminidase activities. The chitinase production and the intensive autolysis hindered the production of N-acetyl-β-D-glucosaminidase and, therefore, could hinder the cell death in the cultures.

Aflatoxins due to their toxicity pose significant economic and human health threat; therefore, it is important to avoid this type of contamination in agricultural products. Until now significant aflatoxin contamination occurred mainly in foods of tropical and subtropical origin because the optimal growth of the producer Aspergillus species is between 32–38 ºC. Nowadays the aflatoxin contamination is becoming higher threat in Hungary, due to the imported products, the rising average temperature and the climatic changes. There is a significant knowledge on the genetic and environmental effectors of the aflatoxin production; however, it is remained a great problem to control mold contamination and toxin production in farming and stock-raising. Here we attempted to summarize the knowledge on aflatoxin production and attempts of the elimination.

Knowledge on the functions, cooperation, and interplays of the signaling and regulatory pathways of filamentous fungi is crucial when their industrial performance is improved or when new-type antifungals are developed. Many research groups aim at a deeper understanding of vegetative growth signaling because this cascade also influences other important physiological processes including asexual and sexual developments, autolysis and apoptotic cell death as well as the production of a wide array of important secondary metabolites. This review also focuses on how this signaling pathway is interconnected with other signaling cascades setting up a robust but delicately regulated signaling network in theAspergilli.

Listeria monocytogenesis a foodborne pathogen, and the detection and differentiation of this bacterium from the nonpathogenicListeriaspecies are of great importance to the food industry. Differentiation ofListeriaspecies is very difficult, even with the sophisticated MALDI-TOF MS technique because of the close genetic relationship of the species and the usual gene transfer. The present paper emphasizes the difficulties of the differentiation through the standardized detection and confirmation according to ISO 11290-1:1996 and basic availableL. monocytogenesdetection methods and tests (such as APIListeriatest, MALDI-TOF MS analysis, andhlygene PCR). With the increase of reports on the pathogenesis of atypicalListeriastrains in humans, the significance of species level determination has become questionable, especially in food quality control, and the detection of pathogenic characteristics seems to be more relevant.

Under carbon starvation, Aspergillus nidulans produced a fungal/bacterial type chitinase, ChiB. The chiB gene was cloned and subcloned into pJC40 expression vector containing a 10XHis fusion tag, and the ChiB protein was expressed heterologously in Escherichia coli . Recombinant and native ChiB enzymes shared the same optimal pH ranges and showed similar substrate specificities with endo-acting cleavage patterns.

Fungi and plants are rich sources of thousands of secondary metabolites. The genetically coded possibilities for secondary metabolite production, the stimuli of the production, and the special phytotoxins basically determine the microscopic fungi-host plant interactions and the pathogenic lifestyle of fungi. The review introduces plant secondary metabolites usually with antifungal effect as well as the importance of signaling molecules in induced systemic resistance and systemic acquired resistance processes. The review also concerns the mimicking of plant effector molecules like auxins, gibberellins and abscisic acid by fungal secondary metabolites that modulate plant growth or even can subvert the plant defense responses such as programmed cell death to gain nutrients for fungal growth and colonization. It also looks through the special secondary metabolite production and host selective toxins of some significant fungal pathogens and the plant response in form of phytoalexin production. New results coming from genome and transcriptional analyses in context of selected fungal pathogens and their hosts are also discussed.

Physiological changes taking place in carbon-starved, autolysing cultures of Aspergillus (Emericella) nidulans strains with mutations in the GanB/RgsA heterotrimeric G protein signalling pathway were studied and compared. Deletion of the ganB, rgsA or both genes did not alter markedly either the autolytic loss of biomass or the extracellular chitinase production. However, they caused a significant decrease in the proteinase formation, which was detected by measuring both extracellular enzyme activity and the transcription of the prtA gene. The deletion mutants also showed significantly higher specific γ -glutamyltranspeptidase activities than the control strain. Deletion of the rgsA gene affected the glutathione peroxidase and catalase formation, as well as the peroxide content of the cells. The concomitant initiations of cell death and developmental genomic programmes may be interconnected via heterotrimeric G-protein signalling and subsequent changes in intracellular ROS levels in ageing A. nidulans. (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

N-Acetyl-D-glucosamine, chito-oligomers and carbon starvation regulated chiA, chiB, and nagA gene expressions in Aspergillus nidulans cultures. The gene expression patterns of the main extracellular endochitinase ChiB and the N-acetyl-beta-D-glucosaminidase NagA were similar, and the ChiB-NagA enzyme system may play a morphological and/or nutritional role during autolysis. Alterations in the levels of reactive oxygen species or in the glutathione-glutathione disulfide redox balance, characteristic physiological changes developing in ageing and autolyzing fungal cultures, did not affect the regulation of either the growth-related chiA or the autolysis-coupled chiB genes although both of them were down-regulated under diamide stress. The transcription of the chiC gene with unknown physiological function was repressed by increased intracellular superoxide concentration.

NADPH is involved in many basically important anabolic processes. For a long time, pentose phosphate pathway (PPS) was regarded as the most important source of NADPH in fungi. Here we present evidence of a metabolic switch to an alternative NADPH-producing pathway in ageing Penicillium chrysogenum cultures, which involves NADP+ -specific isocitrate dehydrogenase (NADP+ -ID) rather than PPS enzymes. Considering the main biochemical functions of NADPH, we propose that NADP+ -ID could have deep impact on many physiological processes switched on glucose deprivation including proteinase production or penicillin biosynthesis. We also demonstrate that although the alternative pathway was inferior to PPS when the fungus was grown on well-utilisable carbon sources yet it could have an important role in fatty acid biosynthesis as well as in the maintenance of high intracellular NADPH/NADP+ ratios.

Fourteen Penicillium strains have been screened on wheat bran–crude chitin mixture medium for extracellular chitinase production in solid-state fermentation. Under the experimental conditions tested, Penicillium aculeatum NRRL 2129 (=ATCC 10409) was selected as the best enzyme producer. The optimum incubation period for chitinase production by the potent organism was found to be 72 h. Chromatofocusing was performed as the first step in the purification scheme, but high amount of contaminating proteins interfered with the method. Hence, ion-exchange chromatography experiments were carried out followed by gel filtration to separate and isolate chitinase isoenzymes. Four major chitinase peaks of molecular weight 82.7, 44.6, 28.2 and 26.9 kDa were observed after gel filtration chromatography while, on SDS-PAGE, three protein bands of molecular weights 82.6, 33.9 and 29.1 kDa were identified. The purified enzyme showed optimal temperature and pH at 50 and 5.5 °C, respectively.

Thirty Trichoderma strains representing 15 species within the genus were screened for extracellular production of chitinolytic enzymes in solid substrate fermentation. Trichoderma longibrachiatum IMI 92027 (ATCC 36838) gave the highest yield (5.0 IU/g of dry matter of substrate) after 3 d of fermentation on wheat bran-crude chitin (9:1 mixture) medium. The optimal moisture content (66.7%), chitin content (20%), initial pH of the medium (2.0-5.0), and time course (5 d) of solid substrate fermentation were determined for strain IMI 92027. Cellulase, xylanase, alpha-amylase, and beta-xylosidase activities were also detected. The pH and temperature optima of the chitinase complex of T. longibrachiatum IMI 92027 were 4.5 and 55 degrees C, respectively. The enzyme totally lost its activity at 70 degrees C in 5 min in the absence of the substrate but retained about 15% of its initial activity even at 70 degrees C after a 60-min incubation in the presence of solid substrate fermentation solids. Purification of protein extract from the solid substrate fermentation material revealed high chitinolytic activities between pI 5.9 and 4.8, where N-acetyl-beta-D-hexosaminidase and chitinase peaks have been found in the same pI range. Two chitinases of 43.5 and 30 kDa were purified at acidic pI.

The morphologic and physiologic effects of vitamin E, a powerful antioxidant, on the autolysis and sporulation of Aspergillus nidulans FGSC26 were studied. In carbon-depleted submerged cultures, reactive oxygen species (ROS) accumulated in the cells and, concomitantly, progressing autolysis was observed, which was characterized by decreasing dry cell masses and pellet diameters as well as by increasing extracellular chitinase activities. Vitamin E supplemented at a concentration of 1 g/L hindered effectively the intracellular accumulation of ROS, the autolytic loss of biomass, the disintegration of pellets, and the release of chitinase activities. In surface cultures, vitamin E inhibited autolysis of both A. nidulans FGSC26 and a loss-of-function FlbA autolytic phenotype mutant. In addition, supplementation of the culture medium with this antioxidant also had a negative effect on the sporulation of strain FGSC26 and the FadAG203R hypersporulating phenotype mutant. These results suggest that accumulation of ROS was involved in the initiation of both sporulation and autolysis in this filamentous fungus, but that FadA/FlbA signaling was not involved in this vitamin E-dependent regulation. Vitamin E can be recommended as a supplement in fermentations in which the disintegration of pellets and gross autolysis should be avoided.

The application of yeasts has great potential in reducing the economic damage caused by toxigenic fungi in the agriculture. Some yeasts may act as biocontrol agents inhibiting the growth of filamentous fungi. These species may also gain importance in the preservation of agricultural products and in the reduction of their mycotoxin contamination, yet the extent of mycotoxin production in the presence of biocontrol agents is relatively less understood. The application of yeasts in various technological processes may have a direct inhibitory effect on the toxin production of certain molds, which is independent of their growth suppressing effect. Furthermore, several yeast species are capable of accumulating mycotoxins from agricultural products, thereby effectively decontaminating them. Probiotic yeasts or products containing yeast cell wall are also applied to counteract mycotoxicosis in livestock. Several yeast strains are also able to degrade toxins to less-toxic or even non-toxic substances. This intensively researched field would greatly benefit from a deeper knowledge on the genetic and molecular basis of toxin degradation. Moreover, yeasts and their biotechnologically important enzymes may exhibit sensitivity to certain mycotoxins, thereby mounting a considerable problem for the biotechnological industry. It is noted that yeasts are generally regarded as safe; however, there are reports of toxin degrading species that may cause human fungal infections. The aspects of yeast-mycotoxin relations with a brief consideration of strain improvement strategies and genetic modification for improved detoxifying properties and/or mycotoxin resistance are reviewed here.

New approaches for treatment of invasive fungal infections are necessary to cope with emerging resistant fungal pathogens of humans. In this paper, three different strategies are presented and evaluated to find new-type antifungal drugs and their targets. While experimental data obtained with potent chitinase inhibitors, e.g. allosamidin, and small-size antifungal proteins of fungal origin are encouraging more efforts are needed to verify and exploit the possible involvement of intracellular thiols, e.g. glutathione, and their metabolic enzymes in the pathogenesis of mycoses caused by dimorphic fungi. Chitinase inhibitors seem to hinder the cell separation of yeasts and the fragmentation of filamentous fungi quite effectively and, hence, they may be implicated in future therapies of systemic mycoses. In addition, small-size antifungal proteins possessing a broad inhibition spectrum may also provide us with promising new agents for the treatment of different kinds of (e.g. cutaneous) fungal infections.

In aging carbon-depleted cultures of Penicillium chrysogenum the fragmentation and autolysis of old mycelia were coupled with the production of high levels of extracellular protease and N-acetyl-β-D-hexosaminidase; this could be stopped by the addition of an extra dose of glucose at any incubation time tested, but was not affected by endogenous NH 3 . After the addition of glucose, intracellular enzyme accumulation was observed only for the N-acetyl-β-D-hexosaminidase, and the concomitant decrease in both the extracellular protease and N-acetyl-β-D-hexosaminidase activities was not caused by the action of extracellular proteases. The physiological function of the N-acetyl-β-D-hexosaminidase remains to be elucidated because the P. chrysogenum culture studied did not utilize N-acetyl-D-glucosamine as a carbon source. However, the proteases (mainly serine and, to a much smaller extent, metalloproteases) might provide surviving hyphal fragments with sufficient amino acids to maintain the cryptic growth observed in aging carbon-depleted cultures.

The cell wall integrity (CWI) signaling pathway is responsible for cell wall remodeling and reinforcement upon cell wall stress, which is proposed to be universal in fungal cultures. In Aspergillus nidulans, both the deletion of rlmA encoding the RlmA transcription factor in CWI signaling and low concentrations of the cell wall polymer intercalating agent Congo Red caused significant physiological changes. The gene deletion mutant ΔrlmA strain showed decreased CWI and oxidative stress resistances, which indicated the connection between the CWI pathway and the oxidative stress response system. The Congo Red stress resulted in alterations in the cell wall polymer composition in submerged cultures due to the induction of the biosynthesis of the alkali soluble fraction as well as the hydrolysis of cell wall biopolymers. Both RlmA and RlmA-independent factors induced by Congo Red stress regulated the expression of glucanase (ANID_00245, engA) and chitinase (chiB, chiA) genes, which promoted the autolysis of the cultures and also modulated the pellet sizes. CWI stress and rlmA deletion affected the expression of brlA encoding the early conidiophore development regulator transcription factor BrlA and, as a consequence, the formation of conidiophores was significantly changed in submerged cultures. Interestingly, the number of conidiospores increased in surface cultures of the ΔrlmA strain. The in silico analysis of genes putatively regulated by RlmA and the CWI transcription factors AnSwi4/AnSwi6 in the SBF complex revealed only a few jointly regulated genes, including ugmA and srrA coding for UgmA UDP-galactopyranose mutase and SrrA stress response regulator, respectively.

Under long-term oxidative stress caused by menadione sodium bisulfite, genome-wide transcriptional and proteome-wide translational changes were compared in Aspergillus nidulans vegetative cells. The comparison of proteomic and DNA microarray expression data demonstrated that global gene expression changes recorded with either flip-flop or dendrimer cDNA labeling techniques supported proteome changes moderately with 40% and 34% coincidence coefficients, respectively. Enzyme levels in the glycolytic pathway were alternating, which was a direct consequence of fluctuating gene expression patterns. Surprisingly, enzymes in the vitamin B2 and B6 biosynthetic pathways were repressed concomitantly with the repression of some protein folding chaperones and nuclear transport elements. Under long-term oxidative stress, the peroxide-detoxifying peroxiredoxins and cytochrome c peroxidase were replaced by thioredoxin reductase, a nitroreductase and a flavohemoprotein, and protein degradation became predominant to eliminate damaged proteins.

The formation of N-acetyl-β-D-hexosaminidase is under carbon catabolite repression in growing Penicillium chrysogenum. The enzyme accumulates inside the cells during the stationary phase of growth and is released into the culture fluid when the cell-wall lysis has started and progressed. The time-course of enzyme formation strongly supports the suggestion that fungal N-acetyl-β-D-hexosaminidase may have a role in the degradation of intermediate cell-wall breakdown products during starvation. Regarding that analogous enzymes from other fungi have been shown to be able to liberate GlcNAc monomers from different glycoproteins and glycopeptides a possible participation in the orderly destruction of cellular glycoprotein components cannot be excluded. The physical and enzyme kinetic properties of both intracellular and extracellular N-acetyl-β-D-hexosaminidase are essentially the same indicating that the enzyme is probably released into the culture fluid without modification.