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Successful control of systemic Aspergillus niger infections in two patients with acute leukemia.

The diagnosis and successful control of systemic Aspergillus niger infection in 2 adult patients with acute leukemia is reported. During induction therapy, the first patient developed pulmonary infiltrates, skin lesions and abnormal liver function tests. Aspergillus niger was found on skin and liver biopsy. This patient was successfully treated with Amphotericin B and granulocyte transfusions and he remains in remission. The second patient developed a pneumonitis and adynamic ileus with positive sputum and stool cultures for Aspergillus niger. The infection only responded to Amphotericin B and granulocyte transfusions and the leukemia to cytoreductive chemotherapy. The patient later relapsed and died after a febrile illness. Fungi morpholocially consistent with Aspergillus were found in the liver at autopsy. Infection with A. niger is rare even in this patient population; however fungal infections have become an increasing problem. The need for a high index of suspicion, especially when an infection is unresponsive to antibacterial antibiotics, the various diagnostic tools, and the need for aggressive therapy are stressed. Amphotericin B is the chemotherapy of choice but may be insufficient in a severely neutropenic host where the simultaneous use of granulocyte transfusions might be lifesaving.

Acute Disease

The black aspergilli (Aspergillus niger complex) and their role in human, animal, and plant diseases.

SUMMARYThe Aspergillus niger complex, also known as the black aspergilli or section Nigri, comprises a diverse group of filamentous fungi with wide-ranging ecological, industrial, and pathogenic significance. While traditionally associated with food spoilage and industrial fermentation, black aspergilli have emerged as opportunistic pathogens affecting humans, animals, and plants. This review provides a comprehensive synthesis of the taxonomy, ecology, pathogenicity, and antifungal resistance of the A. niger complex. Advances in phylogenetics and whole-genome sequencing have clarified the taxonomy of section Nigri, now comprising six core species in series Nigri. Clinically, A. niger complex is implicated in various conditions, including otomycosis, keratitis, cutaneous infections, onychomycosis, chronic pulmonary aspergillosis, and, less commonly, invasive aspergillosis. In animals, black aspergilli have been isolated from respiratory, cutaneous, and systemic infections, particularly in immunocompromised or stressed hosts. Plant pathogenicity is significant, with A. niger complex contributing to pre- and post-harvest spoilage and producing mycotoxins such as ochratoxin A and oxalic acid. The common finding of elevated minimum inhibitory concentrations (MICs) to triazoles, particularly in both environmental and clinical isolates, raises concern, with underlying mechanisms differing from those characterized in A. fumigatus. Reduced susceptibility is potentially driven by efflux pumps and environmental exposure to azole fungicides. Due to commonly higher MICs, antifungal therapy with itraconazole and isavuconazole may have reduced efficacy, and alternatives such as voriconazole or posaconazole should be considered, guided by susceptibility testing where available. This review emphasizes the need for a One Health approach to managing black aspergilli, integrating surveillance, diagnostics, and targeted interventions across human, veterinary, and agricultural sectors.

Humans

Optimization of protoplast based DNA isolation and genome analysis in a gamma-irradiated Aspergillus niger mutant strain.

Aspergillus niger is an important industrial fungus widely used for citric acid production and a range of biotechnological applications. In this study, a protoplast-based DNA isolation protocol was optimized for a gamma-irradiated A. niger AN-L103_M1 mutant strain, followed by whole-genome sequencing and functional genome analysis. Protoplast yield was strongly influenced by enzyme concentration and the molarity of the osmotic stabilizer. The highest yield was achieved at an enzyme concentration of 50&#xa0;mg/mL (2.487&#x2009;&#xb1;&#x2009;0.04&#x2009;&#xd7;&#x2009;10&#x2078; cells/mL) and 0.8&#xa0;M KCl (2.550&#x2009;&#xb1;&#x2009;0.06&#x2009;&#xd7;&#x2009;10&#x2078; cells/mL), with both factors showing significant effects (p&#x2009;<&#x2009;0.0001) in GraphPad Prism 11.0.0. Whole-genome sequencing performed using an Illumina NovaSeq 6000 platform yielded a 37.06&#xa0;Mb draft genome assembled into 537 contigs, with an N50 of 363,084&#xa0;bp and a GC content of 48.2%. BUSCO 14 analysis showed high completeness (97.95% complete BUSCOs). Functional annotation and KEGG pathway mapping identified genes involved in glycolysis, the tricarboxylic acid cycle, and citrate biosynthesis, while biosynthetic gene cluster analysis revealed diverse potential for secondary metabolite production. These findings provide an optimized workflow for protoplast-based DNA isolation and genome-scale functional analysis in A. niger, proposing a basis for future comparative genomics, transformation studies, and experimentally validated metabolic engineering.

Aspergillus niger

Relation of lipids to the action of streptomycin on Bacillus subtilis and Escherichia coli and griseofulvin and fungizone on Aspergillus niger.

An attempt was made to reveal the relation between the lipid content and components of B. subtilis and E. coli and the action of streptomycin, and those of A. niger and the action of griseofulvin and fungizone (Amphotericin B). Total lipids were extracted in CO2 atmosphere, dried, and weighed. Lipid components were analyzed by thin-layer chromatography. Complements to the growth medium of A. niger with extracted total lipids and phospholipids were made to verify the obtained results. It has been found that the action of streptomycin and griseofulvin was not correlated with lipids. On the other hand, fungizone action was manifested by the decrease in total lipids, free sterols, sterol esters, and triglycerides. Supplementation studies gave evidence of the role of total lipids and phospholipids in protecting A. niger cells against fungizone. Supplementation of 1 g of phospholipid per litre medium raised the M.I.D. of fungizone from 2.5 up to 25 microgram/ml. It was suggested to keep the fat level controlled at a minimum if it is required to increase fungizone activity.

Amphotericin B

[Localised aspergillosis and oxalosis of the lung caused by aspergillus niger: soil of ornamental plants as a reservoir of aspergilli (case report) (author's transl)].

Haemoptysis occurred in a 64-year-old man with a history of pulmonary tuberculosis, negative for M. tuberculosis. Precipitating antibodies to Aspergillus niger were found in serum and the fungus found in sputum. Autopsy revealed aspergilloma, colonization of the pulmonary cavity by A. niger and local oxalosis if the cavity wall. Ornamental plant soil containing A. niger conidia, as well as the air in the patient's living-room, was the probable source of infection.

Aspergillosis, Allergic Bronchopulmonary

Late onset asthma due to inhalation of Aspergillus niger.

Clinical observations and immunological evidence are presented to support a diagnosis of late onset asthma due to exposure to Aspergillus niger in a 70-year-old man. Asthma occurred on only three occasions, each time approximately 6 hr after exposure to an area containing Aspergillus niger. His sputum culture contained the same organism. Immunological correlates using the Aspergillus niger extract included positive immediate and late skin tests, histamine release from leukocytes, and serum precipitins.

Aged

Degradation of the plant flavonoid phellamurin by Aspergillus niger.

We have previously described the structure of phellamurin, a plant flavonoid, as 3,4',5,7-tetrahydroxy-8-isoprenylflavanone-7-O-glucoside (17). Degradation of phellamurin by Aspergillus niger using modified Czapek-Dox medium as well as phellamurin or one of its degradation products as a sole carbon source, is reported here. Eleven compounds are identified from phellamurin degradation products. A. niger apparently decomposes phellamurin by first removing glucose with beta-glucosidase; neophellamuretin is the first degradation product. Fission of the heterocyclic ring of (5''-hydroxyisopropyl-4'',5''-dihydrofurano)[2'',3''-h]3,4',5-trihydroxyflavanone, which is obtained from neophellamuretin through a few alterations of the side chain, is followed by cleavage of a C--C bond between C=O and carbon at alpha-position and conversion of (5''-hydroxyisopropyl-4'',5''-dihydrofurano)[2'',3''-d]-2',4,6',alpha-tetrahydroxychalcone to rho-hydroxymandelic acid (B-ring) and 2,4,6-trihydroxy-5-carboxyphenylacetic acid (A-ring). It is suggested that rho-hydroxymandelic acid is oxidized to rho-hydroxybenzoic acid. 2,4,6-Trihydroxy-5-carboxyphenylacetic acid is metabolized to phloroglucinol carboxylic acid, which subsequently is decarboxylated to phloroglucinol. These results provided new information on the isoprene unit metabolism of the side chain of phellamurin and firmly established the degradation pathway of phellamurin by A. niger.

Aspergillus niger

Distribution and conformation of crystalline nigeran in hyphal walls of Aspergillus niger and Aspergillus awamori.

Hyphal walls of Aspergillus awamori containing increased amount of the alpha-glucan, nigeran, became correspondingly more opaque when viewed in the electron microscope as shadowed preparations. However, increased polymer deposition was not accompanied by any significant change in wall thickness. The nigeran of both A. awamori and Aspergillus niger occurred in situ in a crystalline conformation identical to that of single crystals prepared with pure polysaccharide. Furthermore, this polymer was the dominant crystalline material in the hyphae whether or not they were enriched in nigeran. Enzymic digestion of nigeran in A. niger and A. awamori revealed that the bulk of the polymer was exposed to the cell's exterior. However, a certain fraction was accessible to enzymic attack only after the wall was treated with boiling water. A third portion, detectable only by x-ray diffraction, was associated with other components and could not be extracted, even with prolonged boiling. It was removed by hot, dilute alkali and was associated in the wall with another glucan fraction. Dry heating of A. niger walls altered their susceptibility to enzymic digestion of nigeran in situ. It is proposed that this treatment introduces interstices in the crystal surface that facilitate attack.

Aspergillus

[Effect of pH on the enzymic activity of the fungi Trichothecium roseum and Aspergillus niger hydrolyzing nonstarch polysaccharides].

The purpose of the study was to determine optimal pH values for the enzymic activity of the fungi Trichothecium roseum and Aspergillus niger hydrolyzing nonstarch polysaccharides of barley and disrupting cell walls (cytolysis) of grain, the so called cytolytic enzymes. The effect of the acidity of the medium on the stability of these enzymes was also investigated. In this connection total cytolytic activity (i. e. total activity of the enzymes hydrolyzing nonstarch polysaccharides of cell walls of barley) and hemicellulase activity of the fungi at different pH values were measured. The activity of these enzymes in aqueous extracts from the fungal culture at optimal pH after preincubation was determined at different acidity levels. The optimum of the hemicellulase activity of both fungi was at 4.6, the optimum of the total cytolytic activity of Tr. roseum at pH 5.6 and of Asp. niger at pH 3.0. The enzymes of the fungus Asp. niger showed a far higher acid stability than those of Tr. roseum.

Aspergillus

Cadmium and zinc sensitivity and tolerance in Bacillus subtilis subsp. niger and in a Pseudomonas sp.

The action of Cd2+ and Zn2+ on Bacillus subtilis subsp. niger ATCC 9372, and on a Pseudomonas sp. (possibly Pseudomonas fluorescens), isolated from cadmium-polluted soil has been determined and compared with results obtained previously with Klebsiella aerogenes. In liquid medium the lag and the mean generation time of Bacillus subtilis subsp, niger increased with increasing Cd2+ or Zn2+ concentrations whereas only the total biomass of the Pseudomonas sp. was affected. Nevertheless, the responses of both species indicated a specific action at low concentrations and a more general toxic action at high concentrations. The survival on Cd2+ - or Zn2+ - agar depended on the state of the metal ions with regard to chelating ligands and on the nutritional stage of the organisms. In admixture, the metal ions acted synergistically, particularly on the Pseudomonas sp. Resistance to both metal ions developed. It was graded to the training concentration and reciprocal cross-resistance occurred with Bacillus subtilis. subsp. niger but not with the Pseudomonas sp.

Bacillus subtilis

The effect of osmotic pressure and salinity of the medium on the growth and sporulation of Aspergillus niger and Paecilomyces lilacinum species.

The effect of osmotic pressure and salinity on the production of conidia and growth of Aspergillus niger and Paecilomyces lilacinum, isolated from the soil, have been studied in this investigation. The maximum production of conidia in both species was observed in the nutrient medium containing 1% NaCl wheras maximum growth was recorded in the nutrient medium containing 3% NaCl. It has been thus observed that the salinity and the osmotic pressure related to it effect the reproductive as well as the vegetative development of both species, and these effects have been observed to be different. The sensitivity of both fungal species to the salt was also investigated. Conidia were not observed in the P. lilacinum species grown in nutrient medium with 5% NaCl whereas conidial development did take place in the case of Aspergillus niger grown in the nutrient medium containing 5% NaCl.

Aspergillus niger

Purification and characterization of an extracellular exo-D-galacturonanase of Aspergillus niger.

A D-galacturonanase (EC 3.2.1.67) catalyzing the degradation of D-galacturonans by terminal action pattern was purified from a culture filtrate of Aspergillus niger by a procedure including the salting-out with ammonium sulfate, precipitation by ethanol, chromatography on DEAE-cellulose, and gel chromatography on Sephadex G-100. The obtained preparation was slightly contaminated by an enzymically inactive protein fraction. Maximum activity and stability of the enzyme was observed at pH 5.2. The enzyme degrades digalacturonic acid, p-nitrophenyl-alpha-D-galactopyranuronide, as well as oligogalacturonides containing at the nonreducing end 4-deoxy-L-threo-hexa-4-enopyranosyluronate. It differs from all A. niger enzymes so far described which degrade D-galaturonans by the terminal action pattern, in not clearly preferring low-molecular substrates. It is therefore classified as an exo-D-galacturonanase.

Aspergillus

Infection of a burn wound by Aspergillus niger. Gross appearance simulating ecthyma gangrenosa.

Growth of Aspergillus niger on a burn wound clinically simulated the early (hemorrhagic) phase of an invasive infection by Pseudomonas aeruginosa or ecthyma gangrenosa. Wound biopsy for histologic examination and culture readily yielded a definitive diagnosis of noninvasive mycotic infection of the burn wound. Pigmentation surrounding mycotic hyphae (otherwise typical for Aspergillus spp.) strongly suggested Aspergillus niger. Cultural data confirmed the diagnosis.

Adult

Partial purification and some properties of oxalacetase from Aspergillus niger.

1. Oxalacetase from Asperigillus niger was found to be an inducible enzyme, the induction being dependent not only on neutralisation of the acidic growth medium but also on the presence of carbonate. An explanation is proposed. 2. Three methods were established for the quantitative determination of oxalacetase activity. These are based on the determination of the product acetate, on the absorbance of oxaloacetate and on coupling the hydrolysis of oxaloacetate to the oxidation of malate by NAD in the presence of malate dehydrogenase. 3. Oxalacetase was purified about 50-fold from cell-free extracts of A. niger and used to determine some of its properties such as kinetic constants. 4. 2S-[U-14C, 3-2H2] Malate in the presence of oxalacetase, NAD and malate dehydrogenase was partially converted to acetate and oxalate. The 3H/14C ratio of the isolated acetate was nearly twice as high as that of the malate used initially. The result demonstrates that the keto form of oxaloacetate, not the enol, is the substrate of the enzyme. 5. Equimolecular mixtures of 2S, 3S-[3-2H1] malate + 2S-[2-2H1] malate (mixture 1) and 2S, 3R-[3-2H1, 3H1] malate + 2S, 3R-[2-2H1, 3-3H1] malate (mixture 2) were prepared from 2S-[3-3H2] malate by incubation with fumarase in normal and tritiated water, respectively. The isolated mixture 1, in the presence of oxalacetase, NAD and malate dehydrogenase was incubated in tritiated water for formation of acetate and oxalate; the isolated mixture 2 was treated likewise in normal water. 6. The mixtures of symmetrically labelled [3H1] acetate and chiral acetates thus produced were isolated and the configuration of the [3H1, 3H1] acetate specimens was determined in the sequence acetate leads to malate leads to fumarate, as usual. The [2H1, 3H1] acetate derived from 2S, 3S-[3-2H1] malate (present in mixture 1( yielded a malate which on incubation with fumarase retained 65.0% of its total tritium content. This chiral acetate, therefore, had the R configuration. The [2H1, 3H1] acetate derived from 2S, 3R-[2-2H1, 3-3H1] malate produced a malate which retained 35% of its total tritium content, and therefore had the S configuration. 7. It was concluded that the detachment of the oxaloyl residue from oxaloacetate and its replacement by a proton proceed with inversion of configuration at the methylene group which becomes methyl during the hydrolysis.

Acetates

beta-Galactosidase from Aspergillus niger. Separation and characterization of three multiple forms.

The enzyme beta-galactosidase (EC 3.2.1.23) from Aspergillus niger was purified and resolved into three multiple forms, using molecular sieving, ion-exchange, an hydrophobic chromatography. The isolated enzyme forms accounted for 83%, 8%, and 9% of the total beta-galactosidase activity, respectively. They were glycoproteins with estimated molecular weights of 124,000, 150,000 and 173,000, isoelectric points of about 4.6, and pH optima between 2.5 and 4.0. Amino acid and carbohydrate analyses showed that multiplicity was mainly due to dissimilar carbohydrate contents (about 12.5%, 20.5% and 29% neutral carbohydrates, respectively). The multiple form pattern might depend on the culture conditions. The beta-galactosidase forms were heat-stable up to about 60 degrees C. The Km values for lactose ranged from 85 mM to 125 mM, whereas those for the synthetic substrate o-nitrophenyl-beta-D-galactopyranoside were equal to about 2.4 mM. The V values obtained at 30 degrees C for lactose and o-nitrophenyl-beta-D-galactopyranoside were 104 units/mg enzyme protein and 121 units/mg enzyme protein, respectively (weighted averages for the three enzyme forms). The slight reactional dissimilarities between the three enzyme forms are unlikely to be physiologically relevant. The biological significance of A. niger beta-galactosidase multiplicity might be related to the observed differences in carbohydrate content, as suggested by recent reports on other microbial glycoprotein enzymes.

Amino Acids

Colonization of rye green manure and peanut fruit debris by Aspergillus falvus and Aspergillus niger group in field soils.

Aspergillus flavus and Aspergillus niger group colonization of deep-plowed, decomposing rye green manure cover crops in peanut field soils was studied in four fields during 1972 and 1973; colonization of decomposing peanut fruits was studied in 1972 in two fields. A. flavus colonization of rye and peanut fruits was greater in soils of heavy texture, and an A. flavus population as high as 165 propagules per g of soil was observed in soil adjacent to rye, whereas A. flavus populations in soils not associated with rye were 18 propagules per g of soil or lower. Highest A. flavus populations in soil adjacent to decomposing peanut fruits were usually comparable to populations associated with rye. Little decomposing rye or peanut fruit colonization was generally observed by the A. flavus competitor, A. niger group. A. flavus may maintain or increase its inoculum potential by colonization of these and other moribund plant tissues.

Antibiosis

Effect of respiratory deficiency and temperature on the mitochondrial lipid metabolism of Aspergillus niger.

A comparative study of the mitochondrial lipid composition of a wild strain (V 35) and a respiratory-deficient mutant (rd3) of Aspergillus niger grown at different temperatures (25, 30, 35, and 40 degrees C) has been performed. The lipid spectrum, though qualitatively similar, differs quantitatively in both cases. At the optimum growth temperature (30 degrees C) depletion in ergosterol (40%) and cardiolipin (52%) was observed. This probably indicates the formation of defective mitochondria in the mutant with a resultant impaired respiratory system. Complete depletion of cardiolipin species containing fatty acid (20:5omega3) in the respiratory-deficient mutant suggests a possible role of this lipid in mitochondriogenesis at least in A. niger. The effect of temperature is predominantly on the degree of unsaturation and sterol ester formation. The linoleic acid (18:2omega6) content decreases with a concomitant increase in oleic acid (18:1omega9) content as the growth temperature increases for both cell types. Some morphological changes and effects on the vegetative life cycle have been observed with variation in growth temperature in the wild type and also in the mutant form.

Aspergillus niger

[Conditions for splitting protodioscine--the main glycoside from Tribulus terrestris L. by the enzymatic preparation from Aspergillus niger BKMt-33].

The conditions for splitting protodioscine--the main steroid saponine isolated from Tribulus terrestris L. by the enzymic preparation of Aspergillus niger str. BKMt-33 were investigated. The optimal conditions were found to be as follows: pH 4-5, temperature 30-37 degrees (the substrate concentration--5 mg%, concentration of the enzymic preparation--1%). Under these conditions the enzymolysis continued 24 hours. Mg+2 and K+ ions accelerated the reaction twice. As a result of the enzymic hydrolysis dioscine and trilline were obtained. This indicates beta-glucosidase and alpha-rhamnosidase activities of the enzymic complex isolated from Aspergillus niger str. BKMt-33.

Aspergillus