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PubMed2004

Antifungal activities of four fatty acids against plant pathogenic fungi.

The effect of the fatty acids linolenic acid, linoleic acid, erucic acid and oleic acid on the growth of the plant pathogenic fungi Rhizoctonia solani, Pythium ultimum, Pyrenophora avenae and Crinipellis perniciosa were examined in in vitro studies. Linolenic and linoleic acids exhibited activity against all of the fungi. However, whereas linolenic acid reduced mycelial growth of R. solani and C. perniciosa at 100 microM, the concentration had to be increased to 1000 microM before any effect on mycelial growth of P. ultimum and P. avenae was observed. Linoleic acid only reduced mycelial growth of R. solani, P. ultimum and P. avenae at 1000 microM, but led to a significant reduction in growth of C. perniciosa at 100 microM. In contrast, oleic acid had no significant effect on growth of R. solani or P. avenae, but gave significant reductions in mycelial growth of P. ultimum at 100 microM and reduced growth of C. perniciosa significantly at 1000 microM. All of the fatty acids reduced biomass production by all of the fungi significantly in liquid culture when added to the media at 100 microM. Erucic acid had no effect on fungal growth at any concentration examined. The antifungal activities exhibited by linolenic, linoleic and oleic acids may be useful in the search for alternative approaches to controlling important plant pathogens, such as those examined in this study.

Ascomycota↗
PubMed2026

Activation of secondary metabolism in Aspergillus and related filamentous fungi through regulatory engineering.

Filamentous fungi are major contributors to diverse secondary metabolites with broad applications to medicine, agriculture, and biotechnology. Advances in genome sequencing and bioinformatic tools have revealed that fungal genomes encode far more biosynthetic gene clusters (BGCs) than are expressed under normal laboratory conditions, leaving much biosynthetic potential transcriptionally silent. Overcoming this gap between predicted and observed secondary metabolism has become a major challenge in fungal natural product discovery. In this review, we summarize current strategies for activating silent or weakly expressed fungal BGCs through regulatory engineering, with an emphasis on approaches validated in Aspergillus, Penicillium, Monascus, and related filamentous fungi. We focus on genetic and chemical manipulations that enable coordinated activation of multiple biosynthetic pathways through chromatin-level modifiers, global transcriptional regulators, and developmental regulators. By framing these regulators as practical tools rather than solely biological components, we demonstrate their strengths, limitations, and applications in Aspergillus and related filamentous fungi. We further discuss emerging combinatorial and integrative approaches that use regulatory engineering alongside omics technologies and predictive tools, outlining alternatives and future directions for improving the interpretability of silent pathway activation.

Journal Article↗
PubMed1998

'Floral' scent production by Puccinia rust fungi that mimic flowers.

Crucifers (Brassicaceae) in 11 genera are often infected by rust fungi in the Puccinia monoica complex. Infection causes a 'pseudoflower' to form that is important for attracting insect visitors that sexually outcross the fungus. 'Pollinator' attraction is accomplished through visual floral mimicry, the presence of a nectar reward and floral fragrances. Here we used gas chromatography and mass spectrometry to identify and quantify fragrance production by these rust fungi on several Arabis hosts, and by co-occurring true flowers that share insect visitors. Fungal pseudoflowers produced distinctive floral fragrances composed primarily of aromatic alcohols, aldehydes and esters. Pseudoflower fragrances were chemically similar to noctuid-moth-pollinated flowers, such as Cestrum nocturnum and Abelia grandiflora, but were very different from host flowers, host vegetation and the flowers of coblooming, nonhost angiosperms. There was variation in the quantity and composition of fragrance profiles from different fungal species as well as within and among hosts. The evolution of scent chemistry is relatively conservative in these fungi and can be most parsimoniously explained in three steps by combining chemical data with a previously determined rDNA ITS sequence-based phylogeny. Pseudoflower scent does not appear to represent a simple modification of host floral or vegetative emissions, nor does it mimic the scent of coblooming flowers. Instead, we suspect that the unique fragrances, beyond their function as pollinator attractants, may be important in reducing gamete loss by reinforcing constancy among foraging insects.

Basidiomycota↗
PubMed2001

Prevalence of yeasts and mycelial fungi in bovine parasitic otitis in the State of Minas Gerais, Brazil.

Infestations by rhabditiform nematodes and acarids of the genus Raillietia are considered the primary causes of external otitis in cattle in tropical regions. Recently, yeasts of the genus Malassezia have been associated with a relatively high percentage of otitis cases, but the occurrence of other yeasts and mycelial fungi has not yet been reported in the literature. This work studied the presence of fungi in the ear canal of 45 cattle with external parasitic otitis. The results were positive for yeasts of the genus Malassezia in 31 (68.9%) of the 45 cultures in Mycosel medium supplemented with olive oil. The 45 cultures in Sabouraud dextrose medium revealed the growth of seven (15.5%) yeasts of the genus Candida, five (11.1%) Rhodotorula mucilaginosa, two (4.4%) fungi of the genus Aspergillus and eight 'Micelia sterilia'. Future studies may confirm and elucidate the importance of these agents in the aetiology of bovine otitis.

Animals↗
PubMed2004

Pulmonary defenses against fungi.

Pulmonary immunity to fungal pathogens requires both innate and adaptive immune responses. Alveolar macrophages, dendritic cells, and neutrophils are the phagocytic cells of the lung innate system. These cells produce early inflammatory mediators (i.e., reactive oxygen species, cytokines, and chemokines) in response to fungal infection. The production of early cytokines by innate cells, namely tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-12, plays a central role in the development of protective cell-mediated immunity against fungi. T helper 1 (Th1) cell-mediated immunity is essential for limiting a pulmonary fungal infection. Virulence factors produced by the fungi can also modulate the host immune response. Fungal virulence factors include the production of prostaglandins and a polysaccharide capsule. The type of adaptive immune response (T1 vs T2) generated determines whether the fungi are cleared from the lungs or a chronic fungal infection prevails.

Journal Article↗
PubMed2004

Dematiaceous fungi.

Dematiaceous fungi are the etiologic agents of phaeohyphomycosis and are increasingly recognized as causing disease in humans. A wide variety of infectious syndromes are seen, from local infections due to trauma to widely disseminated infection in immunocompromised patients. Pulmonary disease may be divided into allergic bronchopulmonary and nonallergic syndromes, depending on the species. These fungi have unique pathogenic mechanisms owing to the presence of melanin in their cell walls, which imparts the characteristic dark color to their spores and hyphae. Melanin is a known virulence factor in certain fungi, including Cryptococcus neoformans and Wangiella dermatitidis. Therapy depends upon the clinical syndrome. Local infection may be cured with excision alone, whereas systemic disease is often refractory to therapy. Azoles such as itraconazole and voriconazole have the most consistent in vitro activity, though there is more clinical experience with itraconazole. Further studies are needed to better understand the pathogenesis and treatment of these uncommon infections.

Journal Article↗
PubMed1998

Evidence for balancing selection operating at the het-c heterokaryon incompatibility locus in a group of filamentous fungi.

In filamentous fungi, het loci (for heterokaryon incompatibility) are believed to regulate self/nonself-recognition during vegetative growth. As filamentous fungi grow, hyphal fusion occurs within an individual colony to form a network. Hyphal fusion can occur also between different individuals to form a heterokaryon, in which genetically distinct nuclei occupy a common cytoplasm. However, heterokaryotic cells are viable only if the individuals involved have identical alleles at all het loci. One het locus, het-c, has been characterized at the molecular level in Neurospora crassa and encodes a glycine-rich protein. In an effort to understand the role of this locus in filamentous fungi, we chose to study its evolution by analyzing het-c sequence variability in species within Neurospora and related genera. We determined that the het-c locus was polymorphic in a field population of N. crassa with close to equal frequency of each of the three allelic types. Different species and even genera within the Sordariaceae shared het-c polymorphisms, indicating that these polymorphisms originated in an ancestral species. Finally, an analysis of the het-c specificity region shows a high occurrence of nonsynonymous substitution. The persistence of allelic lineages, the nearly equal allelic distribution within populations, and the high frequency of nonsynonymous substitutions in the het-c specificity region suggest that balancing selection has operated to maintain allelic diversity at het-c. Het-c shares this particular evolutionary characteristic of departing from neutrality with other self/nonself-recognition systems such as major histocompatibility complex loci in mammals and the S (self-incompatibility) locus in angiosperms.

Alleles↗
PubMed2001

Dominant fungi in the rhizosphere of established tea bushes and their interaction with the dominant bacteria under in situ conditions.

Species of Penicillium and Trichoderma were found to dominate the rhizosphere of established tea bushes in a detailed study conducted from various tea growing locations in India. Penicillium erythromellis, P. janthinellum, P. raistrickii, Trichoderma pseudokoningii and T. koningii were found to be closely associated with tea roots. While seasonal fluctuation was observed in the case of Penicillium spp., the population of Trichoderma spp. showed less variation during the year. Both species were sensitive to low temperatures. In general, fungi associated with the tea rhizosphere were found to prefer a mesophillic temperature range (15 degrees C to 35 degrees C). The dominant species of Penicillium and Trichoderma also exhibited tolerance to lower temperatures, i.e., 5 to 10 degrees C on agar plates. Most fungi were able to grow in a wide range of pH (4 to 12). Lowering of soil pH in the rhizosphere of tea bushes was positively correlated with the age of the bush and may have affected the development of a specific microbial community in the rhizosphere. The populations of Penicillium and Trichoderma species were inversely correlated with the populations of two most dominant rhizosphere bacteria, Bacillus subtilis and B. mycoides. Both Bacillus species have been shown to have antagonistic activity against these two fungi under in vitro conditions. The present study demonstrates the existence of a similar antagonism under in situ conditions in the rhizosphere of established tea bushes.

Antibiosis↗
PubMed1981

The isolation of pathogenic fungi and actinomycetes from soil in Egypt.

140 soil samples from the Assiut governorate were examined for pathogenic fungi and actinomycetes. The paraffin-bait technique and a modified sodium citrate-utilizing procedure were used for recovery of Nocardia strains. Five isolations of N. brasiliensis were made by the sodium citrate method while 3 isolates were obtained by the other method. N. gypsoides was isolated for the first time, only the the modified method. It appears that the latter method suppresses to a great extent the growth of saprophytic fungi, hence increasing the possibility of the isolation of Nocardia spp. The effect of keratin enrichment on the isolation of dermatophytes from soil by the hair-bait technique was also investigated. Pretreatment of samples with dog hair or feathers increased the yield of isolates of Microsporum gypseum, Trichophyton terrestre and to a lesser extent, T. mentagrophytes, M. canis was recovered from soil only after its enrichment with dog hair. It is suggested that enrichment with other forms of keratinous material, either separately or in combination, might increase the recovery of keratinophilic fungi from soil.

Egypt↗
PubMed2002

Assessment of health and growth of ring-necked pheasants following consumption of infected insects or conidia of entomopathogenic fungi, Metarhizium anisopliae var. acridum and Beauveria bassiana, from Madagascar and North America.

Isolates of two fungi (Beauveria bassiana and Metarhizium anisopliae var. acridum) from Madagascar are being developed for control of grasshoppers and locusts, as part of a search for alternatives to environmentally detrimental chemical insecticides. The probable effects of these entomopathogens on nontarget species must be determined before operational use. Birds may become exposed to these fungi either directly, by consuming spores deposited on their food items, or secondarily, by consuming grasshoppers or locusts that have died from infection by these biocontrol agents. This article presents the results of per os challenge from fungus-infected food items. Male and female ring-necked pheasants (Phasianus colchicus) were exposed at 4 d of age and again at 9 d of age to challenge treatments, or 2 control treatments (18 male and 18 female birds per treatment group). Pheasants were weighed at 9, 17, and 25 d of age, tarsal length was measured at 25 d of age, and they were observed daily for signs of adverse effects of the experimental treatments. At the time of euthanasia (25 d of age), 2 or 3 randomly selected birds from each of the groups exposed to infected grasshoppers, plus the 2 control groups, underwent complete necropsies and histopathological examination of 16 tissues from each bird. Results show that in both sexes, weight gain at both 17 and 25 d was not significantly different between challenge groups and control groups. Tarsal length at 25 d of age, an indication of structural growth, was also not markedly different among challenged and control groups. Histopathological changes were generally undetectable, mild, or moderate, and not consistently associated with any treatment. Based on these findings, there is little indication that birds are susceptible to detrimental health effects from direct or secondary exposure to the two entomopathogenic fungi studied.

Animal Feed↗
PubMedDate not supplied

Ultrastructural alterations produced by sertaconazole on several opportunistic pathogenic fungi.

Sertaconazole is a new synthetic antifungal which has shown promising activity against numerous fungi. The morphological changes induced by the drug at two concentrations, the MIC and 10 times the MIC, during 24 and 48 h on the filamentous opportunistic fungi Aspergillus fumigatus, Chaetomium atrobrunneum and Scedosporium prolificans indicated that the intensity of the ultrastructural breakdown depended on the dose and not on the time of exposure. Common severe alterations of many fungal cells, which could be considered beyond repair, were seen at the cell wall, plasmalemma and cytoplasm levels. The results obtained underline the drug efficacy for those fungi and suggest its applicability as therapeutic agent in the human infections produced by them.

Antifungal Agents↗
PubMed1999

Exposure to biohazards in wood dust: bacteria, fungi, endotoxins, and (1-->3)-beta-D-glucans.

Personal exposure to fungi, bacteria, endotoxin, and (1-->3)-beta-D-glucan was determined at different woodworking sites--logging sites, sawmills, woodchipping sites, and joineries. Exposure levels to fungi at logging sites and sawmills were in the range of 10(3)-10(4) cfu/m3, at the woodchipping mill, 10(3)-10(5) cfu/m3, and at joineries, 10(2)-10(4) cfu/m3. Although mean endotoxin levels were lower than the suggested threshold value of 20 ng/m3, some personal exposures at sawmills and a joinery exceeded the standard. The geometric mean personal (1-->3)-beta-D-glucan exposure level at the woodchipping mill was 2.32 ng/m3, at sawmills, 1.37 ng/m3, at logging sites, 2.02 ng/m3, and at joineries, 0.43 ng/m3. Highly significant associations were found between mean personal inhalable endotoxin exposures and Gram-negative bacteria levels (p < 0.0001), and mean personal inhalable (1-->3)-beta-D-glucan exposures and fungi levels (p = 0.0003). The prevalence of cough, phlegm, chronic bronchitis, nasal symptoms, frequent headaches, and eye and throat irritations was significantly higher among woodworkers than controls. Dose-response relationships were found between personal exposures and work-related symptoms among joinery workers and sawmill and chip mill workers.

Adult↗
PubMed2000

Differential host susceptibility to pulmonary infections with bacteria and fungi in mice deficient in myeloperoxidase.

Myeloperoxidase (MPO), which is located within neutrophils capable of producing hypochlorous acid, is active in vitro against bacteria and fungi. However, MPO-deficient persons are usually healthy. To define the in vivo contribution of MPO to early host defense against pulmonary infections, MPO-deficient and control mice were intranasally infected with various fungi and bacteria, and the number of residual microorganisms in lungs was compared 48 h later. MPO-deficient mice showed severely reduced cytotoxicity to Candida albicans, Candida tropicalis, Trichosporon asahii, and Pseudomonas aeruginosa. However, the mutant mice showed a slight but significantly delayed clearance of Aspergillus fumigatus and Klebsiella pneumoniae and had comparable levels of resistance to the wild type against Candida glabrata, Cryptococcus neoformans, Staphylococcus aureus, and Streptococcus pneumoniae. These results suggest that the MPO-dependent oxidative system is important for host defense against fungi and bacteria, although the effect varies by pathogen species.

Animals↗
PubMed1986

Calcofluor white detection of fungi in cytopathology.

Calcofluor/Cellufluor (CFW) binds to fungal cell walls and causes them to fluoresce blue-green when illuminated with UV light. Retrospective and prospective studies were made to determine if CFW could be added to the Papanicolaou (PAP) stain procedure without altering diagnostic cytopathologic features while still allowing fungi to be identified. The retrospective study included 136 cytology specimens that were designated positive for fungus by PAP stain; these were stained with a 0.1% aqueous solution of CFW and examined by fluorescent microscopy. The overall agreement between the two methods in the detection of fungi was 90.4%. The incorporation of CFW into the PAP stain was tested at various points in the PAP stain sequence; optimum results were obtained when CFW was introduced after acid eosin. A total of 197 random, sequentially accessioned cytology specimens were stained with the PAP/CFW combination in the prospective study. The results indicate that detection of fungi by a combination of regular light and fluorescent microscopy was far more effective than was examination for the organisms by light microscopy alone.

Benzenesulfonates↗
PubMed1997

Infections due to dematiaceous fungi in organ transplant recipients: case report and review.

Dematiaceous fungi are being increasingly recognized as pathogens in organ transplant recipients. We describe a case of invasive esophagitis due to Cladophialophora bantiana in a small bowel transplant recipient and review a total of 34 cases of infections due to dematiaceous fungi in organ transplant recipients. The median time to the onset of fungal infection after transplantation was 22 months. Clinically, two distinct patterns of infections were observed: 79% of the patients had skin and/or soft-tissue infections or joint infections (predominantly due to Exophiala species), and 21% had systemic invasive infections (predominantly brain abscesses due to Ochroconis gallopavum [Dactylaria gallopava, Dactylaria constricta var. gallopava]). The overall mortality rate among the patients with skin and/or soft-tissue infections or joint infections and the patients with systemic invasive disease was 7% and 57%, respectively; two of five patients with brain abscesses were cured with antifungal therapy. Recognition of infections due to dematiaceous fungi is important since these infections, unlike invasive aspergillosis, may be more amenable to therapy.

Adult↗
PubMed2003

The bacterial transposon Tn7 causes premature polyadenylation of mRNA in eukaryotic organisms: TAGKO mutagenesis in filamentous fungi.

TAGKO is a Tn7-based transposition system for genome wide mutagenesis in filamentous fungi. The effects of transposon insertion on the expression of TAGKO alleles were examined in Magnaporthe grisea and Mycosphaerella graminicola. Northern analysis showed that stable, truncated transcripts were expressed in the TAGKO mutants. Mapping of the 3'-ends of TAGKO cDNAs revealed that they all contain Tn7 end sequences, regardless of the transposon orientation. Polyadenylation signals characteristic of eukaryotic genes, preceded by stop codons in all frames, are located in both ends of the bacterial transposon. Thus, TAGKO transcripts are prematurely polyadenylated, and truncated proteins are predicted to be translated in the fungal mutants. Depending on the extent of protein truncation, TAGKO mutations in HPD4 (encoding p-hydroxyphenylpyruvate dioxygenase) resulted in tyrosine sensitivity in the two fungi. Similarly, a particular M.grisea CBS1 (encoding cystathionine beta-synthase) TAGKO cDNA failed to complement cysteine auxotrophy in a yeast CBS mutant. TAGKO, therefore, represents a useful tool for in vivo study of truncated gene products in filamentous fungi.

4-Hydroxyphenylpyruvate Dioxygenase↗
PubMed2006

As you reap, so shall you sow: coupling of harvesting and inoculating stabilizes the mutualism between termites and fungi.

At present there is no consensus theory explaining the evolutionary stability of mutualistic interactions. However, the question is whether there are general 'rules', or whether each particular mutualism needs a unique explanation. Here, I address the ultimate evolutionary stability of the 'agricultural' mutualism between fungus-growing termites and Termitomyces fungi, and provide a proximate mechanism for how stability is achieved. The key to the proposed mechanism is the within-nest propagation mode of fungal symbionts by termites. The termites suppress horizontal fungal transmission by consuming modified unripe mushrooms (nodules) for food. However, these nodules provide asexual gut-resistant spores that form the inoculum of new substrate. This within-nest propagation has two important consequences: (i) the mutualistic fungi undergo severe, recurrent bottlenecks, so that the fungus is likely to be in monoculture and (ii) the termites 'artificially' select for high nodule production, because their fungal food source also provides the inoculum for the next harvest. I also provide a brief comparison of the termite-fungus mutualism with the analogous agricultural mutualism between attine ants and fungi. This comparison shows that--although common factors for the ultimate evolutionary stability of mutualisms can be identified--the proximate mechanisms can be fundamentally different between different mutualisms.

Agaricales↗
PubMed2002

PCR identification of dermatophyte fungi Trichophyton rubrum, T. soudanense and T. gourvilii.

Diagnosis of dermatophytosis employing conventional laboratory procedures has been complicated by the slow growth and varied morphological features shown by dermatophytes. After analysis of the nucleotide base sequences of a 1.2-kb fragment amplified from a dermatophyte fungus Trichophyton rubrum by arbitrarily primed PCR with random primer OPD18, a pair of primers (TRIF and TR1R) was designed and evaluated for specific identification of T. rubrum. The sensitivity of the primers TR1F and TR1R was high, as a specific PCR band of c. 600 bp was detected from as little as 7 pg of T. rubrum DNA. By examining 92 dermatophyte strains and clinical isolates, it was found that this pair of primers reacted in PCR with T. rubrum, T. soudanense and T. gourvilii through formation of the specific fragment of 600 bp, but not with any other of the dermatophyte species or varieties, fungi, yeasts or bacteria tested. As T rubrum is one of the most frequently isolated dermatophyte fungi, and T. soudanense and T. gourvilii are relatively uncommon in many parts of the world, these primers can be used for rapid, sensitive and specific identification and differentiation of T. rubrum from other fungi and micro-organisms.

DNA, Fungal↗