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Calcium oxalate crystal deposition in necrotizing otomycosis caused by Aspergillus niger.

Numerous calcium oxalate crystals were present within fruiting heads of Aspergillus niger and among necrotic debris in a case of bilateral invasive otomycosis occurring in a diabetic female with end stage renal disease. This is the first report of in vivo calcium oxalate crystal deposition associated with Aspergillus niger at this anatomic site. The presence of localized oxalate crystals within necrotic tissue from the external auditory canal is presumptive of otomycosis caused by Aspergillus niger, and may serve as a diagnostic clue to the etiologic agent before histologic demonstration of hyphae or growth in culture.

Adult↗

Influence of medium components and metabolic inhibitors on glucose oxidase production by Aspergillus niger mycelium.

The effect of some medium components and metabolic inhibitors on glucose oxidase (GOD) production by mutant Aspergillus niger was studied in shaken culture. Altering the composition of the basal medium, particularly substitution of (NH4)2HPO4 for NaNO3, and lack of Mg2+ ions caused an increase in GOD activity by 269.6%. The highest yield of extracellular GOD activity of A. niger mycelium was obtained in the presence of hematin (1 mM), choline (40 mM) and Tween 80 (0.1%), which improved the activity of this enzyme by about 31.4-53.9%. A significant increase (68.3%) in intracellular GOD activity was found in the presence of sodium orthovanadate (1 mM). The time course of growth and enzyme production by A. niger in the optimized medium is also reported.

Acid Phosphatase↗

Cloning and function determination of promoter region of glucoamylase gene from Aspergillus niger T21.

A 850 bp fragment of the 5' flanking region of the glucoamylase gene (glaA) was synthesized from A. niger T21 genome using PCR. The function detection vector was constructed by fusing this fragment to E. coli hygromycin B phosphotransferase gene (hph) and was used to transform A. niger. The high hygromycin resistance transformants thus obtained verified that the synthesized fragment functioned as a promoter in filamentous fungi. Southern blot analysis showed the hph gene has been integrated into genomic DNA of A. niger transformant.

Anti-Bacterial Agents↗

[Visceral leishmaniasis in Niger: six new parasitologically confirmed cases].

From January 1992 to January 1995, six cases of Kala-azar have been observed in young soldiers at Niamey, Niger. All the patients had spent some time at Tin-Galene, in Aïr mountains, Northern Niger where they had been apparently contaminated. One patient was also infected with Salmonella and an other with Mycobacterium, but none of the six was positive for HIV. The 6 cases have been confirmed by the presence of Leishmania in the sternum bone-marrow. Four patients recovered after a treatment with Glucantime; two died because the treatment was too late. In Niger, Kala-azar prevalence is probably much higher than estimated previously. So far all the cases described or suspected were in the Saharan mountains of Aïr. The strains have not been typed and it is not possible to state if it is L. infantum or L. donovani. The vector of the two species Ph. orientalis and Ph. alexandri are known to occur in the area.

Adult↗

[Eco-epidemiology of malaria in Niamey and in the river valley, the Republic of Niger, 1992-1995].

The Niger valley is an original ecosystem in the Sahelian belt. For more than 25 years it has been affected by dryness and rainfall decreased by more than 30% as compared to the period 1950-1960. Moreover the demography has sharply increased and the capital town Niamey grew by 10% a year. The Niger valley including Niamey has about 1 million inhabitants. Three sites have been selected to study the status of malaria and its evolution for the last 30 years: Niamey, Karma 40 km on the N.W. on the river and the nearby fossil valley of Fatay-Karma. In the sixties the malaria vectors were An. gambiae, An. arabiensis et An. funestus. The last species was no more harvested after 1970 because its breeding places have been destroyed on the combined action of dryness and human activities. In Niamey, parasite index was very low in the dry season but grew by 5 to 10 times during the rainy season to reach 50% in certain corners. The highest plasmodic index (PI) were recorded along the river banks where vectors are found all the year long. It decreases in the central part of the city and becomes very low in some peripheral suburban settlements. This situation is quite different of most of the cities of the area where prevalence decrease from outskirts to city centre. The serology confirms these points. In Karma, along the Niger and despite a perennial transmission, the PI removes low as well as the malaria antibodies, probably because of the self use of antimalarial drugs by the population. In Fatay-Karma the PI of 23.9% after the rainy season drops to 6% in the dry one. In data prior to 1970 the PI was over 60%, reaching 89% in young children of Niamey suburbs. Obviously it has strongly decreased. It is very likely that this is due partly to the disappearance of An. funestus after the drought.

Adolescent↗

Enhanced retinal longwave sensitivity using a chlorophyll-derived photosensitiser in Malacosteus niger, a deep-sea dragon fish with far red bioluminescence.

Through partial bleaching of both visual pigment extracts and cell suspensions we show that the deep-sea stomiid Malacosteus niger, which produces far red bioluminescence, has two visual pigments within its retina which form a rhodopsin/porphyropsin pigment pair with lambda max values around 520 and 540 nm, but lacks the very longwave sensitive visual pigments (lambda max > 550 nm) observed in two other red light producing stomiids. The presence of only a single opsin gene in the M. niger genome was confirmed by molecular and cladistic analysis. To compensate for its apparently reduced longwave sensitivity compared to related species, the outer segments of M. niger contain additional pigments, which we identify as a mixture of defarnesylated and demetallated derivatives of bacteriochlorophylls c and d, that are used as a photosensitiser to enhance its sensitivity to longwave radiation.

Animals↗

Molecular basis of glucoamylase overproduction by a mutagenised industrial strain of Aspergillus niger.

We have compared a mutagenized strain of Aspergillus niger (S1), used industrially for glucoamylase production, and a related low glucoamylase-producing strain (S2) with a laboratory strain of A. niger (AB4.1). Our aim was to assess the properties of S1 in relation to the laboratory strain and to account at the molecular level for the basis of its glucoamylase overproduction. Both S1 and S2 have similar multiple copies of the glucoamylase-encoding gene (glaA) but only S1 has enhanced glaA transcript and glucoamylase levels compared to AB4.1 that has a single copy of the glaA gene. Glucoamylase production by S1 and AB4.1 was repressed by xylose and induced by starch but, in S2, remained unaffected by carbon source. S1 also secreted elevated levels of alpha-amylase relative to both S2 and AB4.1 but the production of alpha-glucosidase was low in all three strains. The gene encoding aspergillopepsin (pepA), an abundant secreted aspartyl protease, was present as a single copy in all strains but no aspergillopepsin could be detected by Western blotting in either S1 or S2 culture supernatants. We conclude that A. niger strain improvement by mutagenesis and screening for glucoamylase overproduction has led to glaA gene multiplication and an expression defect in the pepA gene.

Journal Article↗

GlaA promoter controlled production of a mutant green fluorescent protein (S65T) by recombinant aspergillus niger during growth on defined medium in batch and fed-batch cultures

The first successful expression of the Aequorea victoria green fluorescent protein (GFP) gene in Aspergillus niger is described. When the wild-type GFP gene was expressed in A. niger, neither the fluorescence nor the full translation product of the wtGFP gene was detectable. However, the expression of a mutant form of the green fluorescent protein (S65TGFP) gene resulted in the formation of a functional fluorescent polypeptide. The synthesis of S65TGFP was used to study glaA promoter controlled heterologous gene expression by recombinant A. niger in batch and fed-batch cultures using a defined growth medium. Cells were grown on xylose as noninducing carbon source, and the production of S65TGFP was accomplished by the addition of maltose. The recombinant protein accumulated up to 10 or 25 mg of S65TGFP g-1 cell dry weight using either a maltose pulse for induction or continuous addition of the inducing carbon source, respectively. Irrespective of the induction protocol, the recombinant protein started to accumulate 2 h after addition of the inducing carbon source and reached its maximum specific concentration 10 h after induction. Bright green fluorescing fungal pellets were first detectable by fluorescence microscopy 4-5 h after the onset of maltose addition.

Journal Article↗

Polyprenol phosphate as an acceptor of mannose from guanosine diphosphate mannose in Aspergillus niger.

Growth of Aspergillus niger in the presence of [2-(14)C]mevalonate and (32)P(i) led to the formation of a lipid, containing (14)C (0.14% of dose) and (32)P (0.009% of dose), that had chromatographic properties identical with those of exo-methylene-hexahydropolyprenol phosphate. When a particulate enzyme preparation from the thallus of A. niger was incubated with GDP-[(14)C]mannose, the main radioactive products were mannose 1-phosphate (57% of products) and mannose (18%). In addition radioactive mannan (8%) and a mannolipid (2%) were formed. The latter was identified as exo-methylene-hexahydropolyprenol phosphate mannose on the basis of its chromatographic properties, acid lability and on the increase in formation of the mannolipid when the phosphate of exo-methylene-hexahydropolyprenols was added to the incubation mixture. The phosphates of ficaprenols and cetyl alcohol caused no corresponding increase. These observations are interpreted as evidence that the thallus of A. niger contains a mannose transferase that uses the phosphate of exo-methylene-hexahydropolyprenols as an acceptor. This situation is discussed in the light of the analogous involvement of a prenol phosphate mannose as an intermediate in the biosynthesis of bacterial wall mannan.

Alcohols↗

Responses of the ant Lasius niger to various compounds perceived as sweet in humans: a structure-activity relationship study.

A behavioural study on the ant Lasius niger was performed by observing its feeding responses to 85 compounds presented in a two-choice situation (tested compound versus water control or sucrose solution). Among these compounds, only 21 were phagostimulating: six monosaccharides (D-glucose, 6-deoxy-D-glucose, L-galactose, L-fucose, D-fructose, L-sorbose), four derivatives of D-glucose (methyl alpha-D-glucoside, D-gluconolactone and 6-chloro- and 6-fluoro-deoxy-D-glucose), five disaccharides (sucrose, maltose, palatinose, turanose and isomaltose), one polyol glycoside (maltitol), three trisaccharides (melezitose, raffinose and maltotriose) and two polyols (sorbitol and L-iditol). None of the 16 non-carbohydrate non-polyol compounds tested, although perceived as sweet in humans, was found to be active in ants. The molar order of effectiveness of the major naturally occuring compounds (melezitose > sucrose = raffinose > D-glucose > D-fructose = maltose = sorbitol) is basically different from the molar order of their sweetness potency in humans (sucrose > D-fructose > melezitose > maltose > D-glucose = raffinose = sorbitol). On a molar basis melezitose is in L. niger about twice as effective as sucrose or raffinose, while D-glucose and D-fructose are three and four times less effective, respectively, than sucrose or raffinose. From a structure-activity relationship study it was inferred that the active monosaccharides and polyols should interact with the ant receptor through only one type of receptor, through the same binding pocket and the same binding residues, via a six-point interaction. The high effectiveness of melezitose in L. niger mirrors the feeding habits of these ants, which attend homopterans and are heavy feeders on their honeydew, which is very rich in this carbohydrate.

Animals↗

Citric Acid Fermentation by Aspergillus niger on Low Sugar Concentrations and Cotton Waste.

The possible use of cotton waste as a carbohydrate source of citric acid production by Aspergillus niger was examined. No citric acid was produced when A. niger was grown on cotton waste as a sole carbon source. In two-stage fermentations, however, mycelium obtained from surface cultures in cotton waste medium yielded more citric acid when transferred to sucrose-containing media than when directly inoculated to sucrose-containing media. It is concluded that cotton waste can be used for saving sucrose and for increasing yields of citric acid fermentation by A. niger.

Journal Article↗

Secretion and properties of a hybrid Kluyveromyces lactis-Aspergillus niger beta-galactosidase.

BACKGROUND: The beta-galactosidase from Kluyveromyces lactis is a protein of outstanding biotechnological interest in the food industry and milk whey reutilization. However, due to its intracellular nature, its industrial production is limited by the high cost associated to extraction and downstream processing. The yeast-system is an attractive method for producing many heterologous proteins. The addition of a secretory signal in the recombinant protein is the method of choice to sort it out of the cell, although biotechnological success is not guaranteed. The cell wall acting as a molecular sieve to large molecules, culture conditions and structural determinants present in the protein, all have a decisive role in the overall process. Protein engineering, combining domains of related proteins, is an alternative to take into account when the task is difficult. In this work, we have constructed and analyzed two hybrid proteins from the beta-galactosidase of K. lactis, intracellular, and its Aspergillus niger homologue that is extracellular. In both, a heterologous signal peptide for secretion was also included at the N-terminus of the recombinant proteins. One of the hybrid proteins obtained has interesting properties for its biotechnological utilization. RESULTS: The highest levels of intracellular and extracellular beta-galactosidase were obtained when the segment corresponding to the five domain of K. lactis beta-galactosidase was replaced by the corresponding five domain of the A. niger beta-galactosidase. Taking into account that this replacement may affect other parameters related to the activity or the stability of the hybrid protein, a thoroughly study was performed. Both pH (6.5) and temperature (40 degrees C) for optimum activity differ from values obtained with the native proteins. The stability was higher than the corresponding to the beta-galactosidase of K. lactis and, unlike this, the activity of the hybrid protein was increased by the presence of Ni2+. The affinity for synthetic (ONPG) or natural (lactose) substrates was higher in the hybrid than in the native K. lactis beta-galactosidase. Finally, a structural-model of the hybrid protein was obtained by homology modelling and the experimentally determined properties of the protein were discussed in relation to it. CONCLUSION: A hybrid protein between K. lactis and A. niger beta-galactosidases was constructed that increases the yield of the protein released to the growth medium. Modifications introduced in the construction, besides to improve secretion, conferred to the protein biochemical characteristics of biotechnological interest.

Journal Article↗

Cloning and expression of Aspergillus niger icdA gene encoding mitochondrial NADP+-specific isocitrate dehydrogenase.

The complementary DNA (cDNA) and chromosomal DNA (icdA) encoding the NADP+-specific isocitrate dehydrogenase (EC 1.1.1.42) of Aspergillus niger WU-2223L, a citric acid-producing strain, were cloned. Two cDNA clones (cDNA-1, 2.0 kb; cDNA-2, 1.5 kb) were obtained and sequenced, and an ORF of 1494 base pairs (bp) encoding a protein of 498 amino acids (aa) was identified in cDNA-1. The predicted amino acid sequence showed 73% and 67% sequence identities with those of the mitochondrial NADP+-ICDHs from Saccharomyces cerevisiae and pig, respectively. The sequence analysis of cDNA-1 and -2 revealed that the cDNA-2 lacks a 500-bp fragment from cDNA-1 which contains a mitochondrial targeting motif. A peroxisomal targeting motif at the C-terminus was found on the aa sequences of cDNA-1 and cDNA-2, but the cDNA-2 product seemed to be localized in the cytoplasm since the peroxisomes were not found in the mycelia of WU-2223L cultivated under the conditions of citric acid production. The expression of both cDNAs in Escherichia coli DEK2004, an isocitrate dehydrogenase-deficient mutant, revealed that both cDNAs complemented the glutamate-requiring phenotype, and that the transformants retained NADP(+)-ICDH activities. Therefore, it was clarified that both of the cDNA-1 and -2 products are fully functional. The chromosomal DNA, icdA, was cloned to correspond to cDNA-1, and its nucleotide sequence revealed that it contains seven introns. Southern hybridization using cDNA-1 and cDNA-2 indicated that there is only one copy of icdA on the chromosomes of A. niger WU-2223L. Northern hybridization analysis as for total RNA of WU-2223L revealed that two mRNAs of different sizes, 2.0 kb and 1.5 kb, were hybridized to the ORF of cDNA-1 used as a probe. Therefore, it was found that approximately 1500-nt and 2000-nt mRNAs were transcribed from only one icdA chromosomal gene in A. niger. Such a transcription has not been observed for ICDH, which is one of the key regulatory enzymes in TCA cycle, in any other organisms.

Journal Article↗

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↗

[A new morphological character for the identification of Stomoxys calcitrans and S. niger (Diptera: Muscidae). Comparison of La Réunion island populations].

On La Reunion Island (France), two morphological closely related species of stable flies (Diptera: Muscidae), living in the same environment, Stomoxys calcitrans and S. niger, are involved in the transmission of blood parasites to the livestock. To facilitate a rapid identification of both species in the field conditions, we highlighted a diagnostic morphological character not yet described: the length of the maxillary palpus. The study of three populations of S. calcitrans and two populations of S. niger, collected at various elevations, showed that the maxillary palpi of S. niger were significantly longer than in S. calcitrans, independent of sex. This character, easily visible in the field with a simple magnifying glass, has been confirmed on individuals of both species from West Africa.

Animals↗

Nuclear configuration and neuronal types of the nucleus niger in the brain of the human adult.

The pigmentoarchitectonic analysis of the human nucleus niger reveals three main territories: Pars compacta, pars diffusa and pars reticulata. Seven subnuclei are recognized within the pars compacta. The nerve cell types forming the nucleus niger were investigated using a Golgi de-impregnation technique in combination with counterstaining of intraneuronally deposited pigment granules. Three principal types of neurons were defined: Type I was a medium-sized to large neuron, mainly encountered in the pars compacta, giving off a few thick and sparsely branching dendrites. These cells were richly endowed with elongated patches of Nissl material that were mainly found in the peripheral portions of the dendrites. One pole of the cell body contained tightly packed neuromelanin granules. Type II neurons were mainly found in the pars reticulata. They were variable in size and shape and generated, similar to type I neurons, extended and sparsely branching dendrites. Type II neurons were devoid of neuromelanin. A considerable number of these cells were lacking in lipofuscin deposits as well. Type III neurons occurred in all portions of the nuclear complex. The small cell body gave rise to a few thin and spineless dendrites. The axon and filiform processes of the dendrites showed small varicosities irregularly spaced apart. The pale cytoplasm contained small and intensely stained lipofuscin granules, which did not tend to agglomerate. Intraneuronally deposited neuromelanin and lipofuscin pigment can be considered a natural marker of the neuronal type in the nucleus niger of the human adult. The technique and the data provide a basis for investigations of the aged and the diseased human brain.

Adult↗

Order No. 60/MEN/FP/ALPHA relating to the creation of a national committee to eliminate illiteracy in the Niger, 3 May 1988.

This Order creates a National Committee to Eliminate Illiteracy in the Niger consisting of members appointed by the Minister of National Education and Professional Training. The objectives of the Committee are: 1) to lend its support to national education structures in pursuing the battle against illiteracy; 2) to collaborate in the definition of a national policy with respect to literacy and adult education; and 3) to formulate and execute a program making the public aware and giving them information about the problem of illiteracy and its impact on the development of the Niger.

Africa↗

Elevated temperature effects on the oxidant/antioxidant balance in submerged batch cultures of the filamentous fungus Aspergillus niger B1-D.

In the present study the relationship between oxidative stress and elevated culture temperature was examined in an industrially relevant fungal culture, Aspergillus niger B1-D. For the first time, both the intracellular levels of the main stressor species (superoxide radical [O(2) (.-)]) and activities of cellular defensive enzymes (superoxide dismutase [SOD], catalase [CAT], and glutathione peroxide [GPx]) were quantified at varying temperature (25, 30, 35, 40 degrees C) to more fully characterize culture response in different growth phases. Elevated culture temperature led to increased O(2) (.-) levels in various culture phases. In the exponential phase this was due to an enhanced generation of O(2) (.-), whereas in stationary phase a decreased dismutation rate may also have contributed. CAT activities generally increased with culture temperature, whereas GPx activity changed little as temperature rose, indicating that GPx played only a minor role in destroying H(2)O(2) in this A. niger. The combination of elevated temperature (35 degrees C) and increased O(2) supply (50% enrichment) led to decreased levels of O(2) (.-) compared to the cultivation at 35 degrees C gassed with air, probably due to enhanced activity of the alternative fungal respiratory pathway. Our findings indicate that while elevated cultivation temperature does clearly induce oxidative stress events, mechanistically, it does so by a rather more complex route than previous studies indicate. Elevated temperature caused a marked disparity in the activities of SOD and CAT, very distinct from the integrated increase in activity of these enzymes in response to oxidative stress.

Air↗