scholarly journals Quantitative Monitoring of Mycelial Growth of Aspergillus fumigatus in Liquid Culture by Optical Density

2022 ◽  
Author(s):  
Ken Miyazawa ◽  
Takashi Umeyama ◽  
Yasutaka Hoshino ◽  
Keietsu Abe ◽  
Yoshitsugu Miyazaki
Keyword(s):  
Aspergillus Fumigatus ◽  
Filamentous Fungi ◽  
Optical Density ◽  
Mycelial Growth ◽  
Liquid Culture ◽  
Quantitative Monitoring ◽  
Unicellular Organisms

Filamentous fungi generally form hyphal pellets in liquid culture. This property prevents filamentous fungi to apply the methods used for unicellular organisms such as yeast and bacteria.

scholarly journals Selection of filamentous fungi that are resistant to the herbicides atrazine, glyphosate and pendimethalin

2021 ◽  
Vol 43 ◽  
pp. e51656
Author(s):  
Nara Priscila Barbosa Bravim ◽  
Anatércia Ferreira Alves ◽  
José Fábio França Orlanda ◽  
Patricia Barbosa Rodrigues Silva
Keyword(s):  
Filamentous Fungi ◽  
Mycelial Growth ◽  
Culture Medium ◽  
Agricultural Soils ◽  
Culture Media ◽  
Fusarium Verticillioides ◽  
Fungal Growth ◽  
Relative Growth ◽  
Maximum Inhibition ◽  
Selection Of

The objective of the present study was to isolate fungi from agricultural soils and evaluate fungal growth in culture medium contaminated with atrazine, glyphosate and pendimethalin. Filamentous fungi were isolated from agricultural soils and cultured in a modified culture medium containing 0, 10, 20, 50, and 100 μg mL-1 atrazine, glyphosate and pendimethalin for 14 days at 28°C. The fungi that presented optimal and satisfactory growth were plated in Sabouraud culture medium with 4% dextrose and containing the herbicides at concentrations of 0, 10, 20, 50, and 100 μg mL-1 for seven days at 28°C. The mean mycelial growth values were submitted to analysis of variance and the Tukey test (p < 0.05%) for comparison and relative growth determination, and maximum inhibition rates were calculated. The isolated fungi Aspergillus fumigatus, Fusarium verticillioides and Penicillium citrinum were shown to be resistant to atrazine, glyphosate and pendimethalin. F. verticillioides showed higher mean mycelial growth in the culture media contaminated with atrazine and glyphosate than the other two fungi. In the culture medium contaminated with pendimethalin, F. verticillioides, and A. fumigatus presented the highest mean mycelial growth values.

Enhanced production of Fumigaclavine C in liquid culture of Aspergillus fumigatus under a two-stage process

2014 ◽  
Vol 152 ◽  
pp. 162-168 ◽  
Author(s):  
Yi-Xiang Zhu ◽  
Ling-Yun Yao ◽  
Rui-Hua Jiao ◽  
Yan-Hua Lu ◽  
Ren-Xiang Tan
Keyword(s):  
Aspergillus Fumigatus ◽  
Liquid Culture ◽  
Two Stage ◽  
Enhanced Production ◽  
Stage Process

Recherche de nouvelles activités saccharifiantes thermostables chez les champignons filamenteux

1990 ◽  
Vol 36 (9) ◽  
pp. 625-630 ◽  
Author(s):  
P. Vanacker ◽  
B. Bacle ◽  
G. Vidal ◽  
L. Lacoste
Keyword(s):  
Aspergillus Niger ◽  
Aspergillus Fumigatus ◽  
Key Words ◽  
Filamentous Fungi ◽  
Half Life ◽  
The Other ◽  
Thermostable Enzymes ◽  
Life Measure ◽  
Industrial Use ◽  
Better Than

We have searched for producers of a saccharifying activity with improved thermostability compared with industrial amyloglucosidases. These producers were chosen among thermophilic, thermotolerant, or even mesophilic fungi. Among the 846 isolated strains, five species (two Thermoascus spp., a member of the Aspergillus fumigatus group, and two members of the Aspergillus niger group) showed an amyloglucosidasic complex with the required property. Whereas the first three of these were thermophilic or thermotolerant strains, the latter two strengthen the idea that mesophilic strains can produce thermostable enzymes. The thermostability of the saccharifying complex of the Thermoascus spp., established with a half-life measure, was found to be far better than the other ones. The industrial use of these strains was discussed, and Thermoascus crustaceus seems to be the most advantageous one. Key words: filamentous fungi, amyloglucosidases, thermostability, Thermoascus.

scholarly journals Role of laeA in the Regulation of alb1, gliP, Conidial Morphology, and Virulence in Aspergillus fumigatus

2007 ◽  
Vol 6 (9) ◽  
pp. 1552-1561 ◽  
Author(s):  
Janyce A. Sugui ◽  
Julian Pardo ◽  
Yun C. Chang ◽  
Arno Müllbacher ◽  
Kol A. Zarember ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Survival Data ◽  
Mycelial Growth ◽  
Mrna Level ◽  
Nutritional Requirements ◽  
Wild Type ◽  
Conidial Morphology ◽  
Culture Filtrates ◽  
Data Culture

ABSTRACT The alb1 (pksP) gene has been reported as a virulence factor controlling the pigmentation and morphology of conidia in Aspergillus fumigatus. A recent report suggested that laeA regulates alb1 expression and conidial morphology but not pigmentation in the A. fumigatus strain AF293. laeA has also been reported to regulate the synthesis of secondary metabolites, such as gliotoxin. We compared the role of laeA in the regulation of conidial morphology and the expression of alb1 and gliP in strains B-5233 and AF293, which differ in colony morphology and nutritional requirements. Deletion of laeA did not affect conidial morphology or pigmentation in these strains, suggesting that laeA is not involved in alb1 regulation during conidial morphogenesis. Deletion of laeA, however, caused down-regulation of alb1 during mycelial growth in a liquid medium. Transcription of gliP, involved in the synthesis of gliotoxin, was drastically reduced in B-5233laeAΔ, and the gliotoxin level found in the culture filtrates was 20% of wild-type concentrations. While up-regulation of gliP in AF293 was comparable to that in B-5233, the relative mRNA level in AF293laeAΔ was about fourfold lower than that in B-5233laeAΔ. Strain B-5233laeAΔ caused slower onset of fatal infection in mice relative to that with B-5233. Histopathology of sections from lungs of infected mice corroborated the survival data. Culture filtrates from B-5233laeAΔ caused reduced death in thymoma cells and were less inhibitory to a respiratory burst of neutrophils than culture filtrates from B-5233. Our results suggest that while laeA is not involved in the regulation of alb1 function in conidial morphology, it regulates the synthesis of gliotoxin and the virulence of A. fumigatus.

scholarly journals Comparison of Fractional Inhibitory Concentration Index with Response Surface Modeling for Characterization of In Vitro Interaction of Antifungals against Itraconazole-Susceptible and -Resistant Aspergillus fumigatus Isolates

2002 ◽  
Vol 46 (3) ◽  
pp. 702-707 ◽  
Author(s):  
D. T. A. Te Dorsthorst ◽  
P. E. Verweij ◽  
J. F. G. M. Meis ◽  
N. C. Punt ◽  
J. W. Mouton
Keyword(s):  
Aspergillus Fumigatus ◽  
Response Surface ◽  
Filamentous Fungi ◽  
Inhibitory Concentration ◽  
Surface Modeling ◽  
Drug Combinations ◽  
Surface Model ◽  
Response Surface Modeling ◽  
Fractional Inhibitory Concentration

ABSTRACT Although the fractional inhibitory concentration (FIC) index is most frequently used to define or to describe drug interactions, it has some important disadvantages when used for drugs against filamentous fungi. This includes observer bias in the determination of the MIC and no agreement on the endpoints (MIC-0, MIC-1, or MIC-2 [≥95, ≥75, and ≥50% growth inhibition, respectively]) when studying drug combinations. Furthermore, statistical analysis and comparisons are troublesome. The use of a spectrophotometric method to determine the effect of drug combinations yields quantitative data and permits the use of model fits to the whole response surface. We applied the response surface model described by Greco et al. (W. R. Greco, G. Bravo, and J. C. Parsons, Pharmacol. Rev. 47:331-385, 1995) to determine the interaction coefficient alpha (ICα) using a program developed for that purpose and compared the results with FIC indices. The susceptibilities of amphotericin B (AM), itraconazole (IT), and terbinafine (TB) were tested either alone or in combination against 10 IT-susceptible (IT-S) and 5 IT-resistant (IT-R) clinical strains of Aspergillus fumigatus using a modified checkerboard microdilution method that employs the dye MTT [3-(4,5-dimethyl-2-thiazyl)2,5-diphenyl-2H-tetrazolium bromide]. Growth in each well was determined by a spectrophotometer. FIC indices were determined and ICα values were estimated for each organism strain combination, and the latter included error estimates. Depending on the MIC endpoint used, the FIC index ranged from 1.016 to 2.077 for AM-IT, from 0.544 to 1.767 for AM-TB, and from 0.656 to 0.740 for IT-TB for the IT-S strains. For the IT-R strains the FIC index ranged from 0.308 to 1.767 for AM-IT, from 0.512 to 1.646 for AM-TB, and from 0.403 to 0.497 for IT-TB. The results indicate that the degree of interaction is not only determined by the agents themselves but also by the choice of the endpoint. Estimates of the ICα values showed more consistent results. Although the absolute FIC indices were difficult to interpret, there was a good correlation with the results obtained using the ICα values. The combination of AM with either IT or TB was antagonistic in vitro, whereas the combination of IT and TB was synergistic in vitro for both IT-S and IT-R strains. The use of response surface modeling to determine the interaction of drugs against filamentous fungi is promising, and more consistent results are obtained by this method than by using FIC indices.

scholarly journals Both galactosaminogalactan and α-1,3-glucan contribute to aggregation of Aspergillus oryzae hyphae in liquid culture

2019 ◽  
Author(s):  
Ken Miyazawa ◽  
Akira Yoshimi ◽  
Motoaki Sano ◽  
Fuka Tabata ◽  
Asumi Sugahara ◽  
...  
Keyword(s):  
Filamentous Fungi ◽  
Aspergillus Oryzae ◽  
Double Mutant ◽  
Liquid Culture ◽  
Bacterial Cells ◽  
Wild Type ◽  
Hydrogen Bond Formation ◽  
Relative Contribution ◽  
In The Wild ◽  
Increase Productivity

AbstractFilamentous fungi generally form aggregated hyphal pellets in liquid culture. We previously reported that α-1,3-glucan-deficient mutants of Aspergillus nidulans did not form hyphal pellets and their hyphae were fully dispersed, and we suggested that α-1,3-glucan functions in hyphal aggregation. Yet, Aspergillus oryzae α-1,3-glucan-deficient (AGΔ) mutants still form small pellets; therefore, we hypothesized that another factor responsible for forming hyphal pellets remains in these mutants. Here, we identified an extracellular matrix polysaccharide galactosaminogalactan (GAG) as such a factor. To produce a double mutant of A. oryzae (AG-GAGΔ), we disrupted the genes required for GAG biosynthesis in an AGΔ mutant. Hyphae of the double mutant were fully dispersed in liquid culture, suggesting that GAG is involved in hyphal aggregation in A. oryzae. Addition of partially purified GAG fraction to the hyphae of the AG-GAGΔ strain resulted in formation of mycelial pellets. Acetylation of the amino group in galactosamine of GAG weakened GAG aggregation, suggesting that hydrogen bond formation by this group is important for aggregation. Genome sequences suggest that α-1,3-glucan, GAG, or both are present in many filamentous fungi and thus may function in hyphal aggregation in these fungi. We also demonstrated that production of a recombinant polyesterase, CutL1, was higher in the AG-GAGΔ strain than in the wild-type and AGΔ strains. Thus, controlling hyphal aggregation factors of filamentous fungi may increase productivity in the fermentation industry.ImportanceProduction using filamentous fungi is an important part of the fermentation industry, but hyphal aggregation in these fungi in liquid culture limits productivity compared with that of yeast or bacterial cells. We found that galactosaminogalactan and α-1,3-glucan both function in hyphal aggregation in Aspergillus oryzae, and that the hyphae of a double mutant deficient in both polysaccharides become fully dispersed in liquid culture. We also revealed the relative contribution of α-1,3-glucan and galactosaminogalactan to hyphal aggregation. Recombinant protein production was higher in the double mutant than in the wild-type strain. Our research provides a potential technical innovation for the fermentation industry that uses filamentous fungi, as regulation of the growth characteristics of A. oryzae in liquid culture may increase productivity.

scholarly journals The Siderophore Transporters Sit1 and Sit2 Are Essential for Utilization of Ferrichrome-, Ferrioxamine- and Coprogen-Type Siderophores in Aspergillus fumigatus

2021 ◽  
Vol 7 (9) ◽  
pp. 768
Author(s):  
Mario Aguiar ◽  
Thomas Orasch ◽  
Matthias Misslinger ◽  
Anna-Maria Dietl ◽  
Fabio Gsaller ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Filamentous Fungi ◽  
Fungal Pathogens ◽  
Fungal Infections ◽  
The Novel ◽  
Human Pathogen ◽  
Substrate Specificities ◽  
Triacetylfusarinine C ◽  
Opportunistic Human Pathogen ◽  
Siderophore Transporter

Siderophore-mediated acquisition of iron has been shown to be indispensable for the virulence of several fungal pathogens, the siderophore transporter Sit1 was found to mediate uptake of the novel antifungal drug VL-2397, and siderophores were shown to be useful as biomarkers as well as for imaging of fungal infections. However, siderophore uptake in filamentous fungi is poorly characterized. The opportunistic human pathogen Aspergillus fumigatus possesses five putative siderophore transporters. Here, we demonstrate that the siderophore transporters Sit1 and Sit2 have overlapping, as well as unique, substrate specificities. With respect to ferrichrome-type siderophores, the utilization of ferrirhodin and ferrirubin depended exclusively on Sit2, use of ferrichrome A depended mainly on Sit1, and utilization of ferrichrome, ferricrocin, and ferrichrysin was mediated by both transporters. Moreover, both Sit1 and Sit2 mediated use of the coprogen-type siderophores coprogen and coprogen B, while only Sit1 transported the bacterial ferrioxamine-type xenosiderophores ferrioxamines B, G, and E. Neither Sit1 nor Sit2 were important for the utilization of the endogenous siderophores fusarinine C and triacetylfusarinine C. Furthermore, A. fumigatus was found to lack utilization of the xenosiderophores schizokinen, basidiochrome, rhizoferrin, ornibactin, rhodotorulic acid, and enterobactin. Taken together, this study characterized siderophore use by A. fumigatus and substrate characteristics of Sit1 and Sit2.

scholarly journals Effects of Two Salts Compounds on Mycelial Growth, Sporulation, and Spore Germination of Six Isolates ofBotrytis cinereain the Western North of Algeria

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Boualem Boumaaza ◽  
Mohamed Benkhelifa ◽  
Moulay Belkhoudja
Keyword(s):  
Sodium Chloride ◽  
Calcium Chloride ◽  
Mycelial Growth ◽  
Liquid Culture ◽  
Germination Percentage ◽  
Germination Capacity ◽  
Calcium Salts ◽  
Sodium Salts ◽  
High Tunnels ◽  
Three Stages

Six isolates ofBotrytis cinereawere isolated from leaves and stems of different tomato varieties taken from four areas in the northwest of Algeria where tomato is mostly grown in greenhouses and high tunnels. The purpose of this research was to determine the effect of two salts, NaCl and CaCl2, on three stages ofBotrytis cinerea’slife cycle. All isolates tested were stimulated in 50 to 150 ppm; NaCl was the most effective treatment to increase mycelial growth at two tested concentrations. However, at 300 ppm concentration, CaCl2completely inhibited the growth of mycelium; they reach 34.78% for the isolate TR46 and 26.72% for isolate F27. The sodium and calcium salts stimulated conidia production in liquid culture. We noticed that the effect of calcium chloride on sporulation was average while sodium chloride. In the medium containing 50 ppm, calcium chloride and sodium chloride increased the germination capacity of most isolates compared with the control. Other calcium salts, at 100 or 300 ppm, decreased the germination percentage of the conidia. With the exception of sodium salts, the inhibitions of germination reduce at 150 or 300 compared with the control. Conidial germination was slightly inhibited by sodium chloride only when the concentration was over 300 ppm.

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