Native valve Aspergillus fumigatus endocarditis with blood culture positive and negative for galactomannan antigen. Case report and literature review

Native valve endocarditis caused by Aspergillus spp. is an uncommon disease with a high mortality rate. Generally, Aspergillus is isolated from affected valve in post-mortem or biopsy specimens. However, its isolation from blood cultures is exceedingly rare. We report a case of fungal endocarditis in a native mitral valve with the isolation of Aspergillus fumigatus both in valve vegetation and in blood culture bottles. The patient underwent valve replacement and antifungal treatment with voriconazole and caspofungin, but he died on post-operative day 45 with disseminated aspergillosis confirmed by necropsy. Paradoxically, galactomannan antigen detection in serum was negative. This is the third case of Aspergillus endocarditis with positive blood culture reported in the literature.     

Renal abscess due to Aspergillus fumigatus as the only sign of disseminated aspergillosis in a patient with AIDS

BackgroundAspergillus fumigatus can cause a wide variety of clinical syndromes, especially in the three largest immunocompromised groups, such as HIV-infected patients. Primary renal aspergillosis is an extremely rare entity.AimsWe report an unusual case of renal abscess due to Aspergillus fumigatus in a patient with AIDS.MethodsWe review clinical and laboratory records, and provide follow up of the patient.ResultsA 38-year-old man, HIV seropositive, was admitted to our hospital with fever, lumbar pain and respiratory symptoms. Abdominal ultrasound and computerised tomography showed a single and large lesion consistent with an abscess located in the left kidney. Aspergillus fumigatus was isolated from clinical sample obtained by ultrasound-guided needle aspiration. Despite a correct treatment based on amphotericin B and drainage of the abscess, surgery was necessary and nephrectomy was carried out. Histopathological examination of the surgical specimen confirmed the diagnosis of renal aspergillosis. Systemic antifungal therapy based on intravenous and oral voriconazole and highly active antiretroviral therapy was started after surgery. The patient had a good response to the established treatment and he remains in a good clinical condition at one year of follow up.ConclusionsCombined medical and surgical treatment is the elective therapy for renal abscesses due to Aspergillus when percutaneous drainage and the administration of systemic antifungal drugs, such as amphotericin B and/or oral voriconazole or itraconazole, fail. This case emphasizes renal fungal infections should be included in the differential diagnosis of kidney abscesses in AIDS patients.     

Aerobiological,biochemical and immunological studies on some of the dominant <Emphasis Type="Italic">Aspergillus</Emphasis> species of South Assam (India)

Two years atmospheric survey of air-borne Aspergillus was carried out in the environmental conditions of South Assam. The survey revealed a total of 16 different species of Aspergillus with marked seasonal and annual variations. Aspergillus fumigatus was found to be the dominant atmospheric fungal species followed by Aspergillus flavus, Aspergillus niger, etc. Among the sample extracts tested, highest quantity of soluble protein was recorded in Aspergillus fumigatus (95.0 mg/g) whereas highest quantity of soluble carbohydrate (40.8 mg/g) and free amino acid (135.0 mg/g) was recorded in the sample extract of Aspergillus niger per gram of dry weight, respectively. The highest numbers of protein polypeptide bands were detected in the sample extract of Aspergillus fumigatus followed by Aspergillus flavus and lowest in Aspergillus niger. The maximum numbers of immunoglobulin E binding protein fractions were found in Aspergillus fumigatus, followed by Aspergillus flavus, Aspergillus clavatus, etc.     

Evolutionary Relationships Among <Emphasis Type="Italic">Aspergillus terreus</Emphasis> Isolates and their Relatives

Aspergillus terreus is a ubiquitous fungus in our environment. It is an opportunistic human pathogen and economically important as the main producer of lovastatin, a cholesterol lowering drug. Our aim was to examine the genetic variability of A. terreus and closely related species using molecular and analytical techniques. Lovastatin production was examined by HPLC. Lovastatin was produced by seven isolates belonging to the species A. terreus. RAPD analyses were carried out using 25 different random primers. Neighbor-joining analysis of RAPD data (120 characters) resulted in clustering of the A. terreus isolates into distinct groups. Some correlation was observed between lovastatin producing abilities of the isolates and their position on the dendrogram based on RAPD profiles. The internal transcribed spacer region and the 5.8S rRNA gene of A. terreus and related isolates was also sequenced. Phylogenetic analysis of sequence data let us classify the isolates into different clades which mostly correspond to the species Aspergillus terreus, Aspergillus flavipes, Aspergillus niveus, Aspergillus carneus and Aspergillus janus/A. janus var. brevis. Aspergillus allahabadii, A. terreus var. aureus and A. niveus var. indicus belonged to the A. niveus clade, while an Aspergillus isolate previously classified as A. niveus was most closely related to A. flavipes isolates. Aspergillus anthodesmis formed a distinct branch on the tree. Although it was previously suggested based on 28S rDNA sequence data that Aspergillus section Terrei should include A. carneus and A. niveus isolates, phylogenetic analysis of ITS sequences indicate that A. flavipes isolates are more closely related to A. terreus than A. carneus isolates. Our data suggest that sections Terrei and Flavipedes should be merged. However, further loci should be analysed to draw more definite conclusions.     

Biosorption of an Azo Dye by <Emphasis Type="Italic">Aspergillus niger</Emphasis> and <Emphasis Type="Italic">Trichoderma</Emphasis> sp. Fungal Biomasses

Biosorption is an eco-friendly and cost-effective method for treating the dye house effluents. Aspergillus niger and Trichoderma sp. were cultivated in bulk and biomasses used as biosorbents for the biosorption of an azo dye Orange G. Batch biosorption studies were performed for the removal of Orange G from aqueous solutions by varying the parameters like initial aqueous phase pH, biomass dosage, and initial dye concentration. It was found that the maximum biosorption was occurred at pH 2. Experimental data were analyzed by model equations such as Langmuir and Freundlich isotherms, and it was found that both the isotherm models best fitted the adsorption data. The monolayer saturation capacity was 0.48 mg/g for Aspergillus niger and 0.45 mg/g for Trichoderma sp. biomasses. The biosorption kinetic data were tested with pseudo first-order and pseudo second-order rate equations, and it was found that the pseudo second-order model fitted the data well for both the biomasses. The rate constant for the pseudo second-order model was found to be 10–0.8 (g/mg min⁻¹) for Aspergillus niger and 8–0.4 (g/mg min⁻¹) for Trichoderma sp. by varying the initial dye concentrations from 5 to 25 mg/l. It was found that the biomass obtained from Aspergillus niger was a better biosorbent for the biosorption of Orange G dye when compared to Trichoderma sp.     

Aspergillosis of Biliary Tract After Liver Transplantation: A Case Report

We reported a case of aspergillosis presented as cholangitis in a patient after liver transplantation, even with prophylactic use of fluconazole. The patient had multiple predisposing factors, such as leukocytopenia, immunosuppressive drug therapy. He died 2 days after an exploratory laparotomy was carried out. The histopathologic finding of biopsy specimen from biliary tract was positive for Aspergillus. And the pathogen was identified as Aspergillus flavus by mycological culture and PCR. The patient was confirmed as a case of Aspergillosis of biliary tract that was responded poorly to fluconazole. This indicated that azole should have been switched to more effective antifungal agents at earlier stage when the patients responded poorly to the original treatment.     

The solubilization and biological activities of Aspergillus beta-(1 --> 3)-D-glucan

We have recently demonstrated that the cell wall beta-glucan of Candida albicans could be solubilized by sodium hypochlorite, followed by dimethylsulfoxide-extraction (NaClO-DMSO method). In this study, applying this method to Aspergillus spp., we prepared mycelial cell wall beta-glucan and examined its physical properties and immunotoxicological activity. The acetone-dried mycelia of Aspergillus spp. were oxidized by the NaClO-DMSO method. An analysis of (13)C NMR spectra revealed the preparations to be composed of alpha-(1 --> 3) and beta-(1 --> 3)-D-glucan. Also, the proportion of alpha-(1 --> 3) and beta-(1 --> 3)-D-glucan varied. Furthermore, a solubilized Aspergillus beta-glucan (ASBG) was prepared from OX-Asp by urea-autoclave treatment. ASBG showed limulus activity similar to Candida solubilized beta-glucan (CSBG), and there was little difference in the activity of ASBG between various Aspergillus spp. ASBG affected the production of IL-8 by human peripheral blood mononuclear cells (PBMC). ASBG should be useful for analyzing the clinical role of beta-glucan.     

Reciprocal oxylipin-mediated cross-talk in the Aspergillus–seed pathosystem

In Aspergilli, mycotoxin production and sporulation are governed, in part, by endogenous oxylipins (oxygenated, polyunsaturated fatty acids and metabolites derived therefrom). In Aspergillus nidulans , oxylipins are synthesized by the dioxygenase enzymes PpoA, PpoB and PpoC. Structurally similar oxylipins are synthesized in seeds via the action of lipoxygenase (LOX) enzymes. Previous reports have shown that exogenous application of seed oxylipins to Aspergillus cultures alters sporulation and mycotoxin production. Herein, we explored whether a plant oxylipin biosynthetic gene ( ZmLOX3 ) could substitute functionally for A. nidulans ppo genes. We engineered ZmLOX3 into wild-type A. nidulans , and into a Δ ppoAC strain that was reduced in production of oxylipins, conidia and the mycotoxin sterigmatocystin. ZmLOX3 expression increased production of conidia and sterigmatocystin in both backgrounds. We additionally explored whether A. nidulans oxylipins affect seed LOX gene expression during Aspergillus colonization. We observed that peanut seed pnlox2–3 expression was decreased when infected by A. nidulans Δ ppo mutants compared with infection by wild type. This result provides genetic evidence that fungal oxylipins are involved in plant LOX gene expression changes, leading to possible alterations in the fungal/host interaction. This report provides the first genetic evidence for reciprocal oxylipin cross-talk in the Aspergillus –seed pathosystem.     

Endogenous <Emphasis Type="Italic">Aspergillus versicolor</Emphasis> Endophthalmitis in an Immuno-competent HIV-positive Patient

A 25-year-old white, HIV-positive, immuno-competent man was referred to us because of a progressive blurred vision in his right eye. Clinical characteristics were suggestive for an unilateral fungal endophthalmitis, and thereby fluconazole firstly, followed by conventional amphotericin B were intravenously administered, without any significant improvement. Thus, a pars plana vitrectomy was performed. Aspergillus versicolor was isolated from the cultures of the vitreous sample and intravenous liposomal amphotericin B was administered. An increase of visual acuity together with a reduction of vitreous inflammation occurred. This case of ours represents the first report describing an endogenous endophthalmitis induced by Aspergillus versicolor.     

Role of Respiration in the Germination Process of the Pathogenic Mold <Emphasis Type="Italic">Aspergillus fumigatus</Emphasis>

Inhalation of resting conidia is usually the first step of a systemic infection caused by the opportunistic fungal pathogen Aspergillus fumigatus. In the lung, the inhaled spores encounter an environment that permits germination. However, the relative importance of certain environmental conditions for conidial activation and subsequent hyphae formation has so far not been analyzed in detail. In this study, we studied the role of oxygen during germination. We found that inhibitors of the respiratory chain were nearly as efficient in blocking germination as cycloheximide, an inhibitor of protein synthesis, which is already known to prevent germination of Aspergillus nidulans. We also found that A. fumigatus is unable to grow or germinate under anaerobic conditions, and using the fluorescent mitotracker dye we detected active mitochondria already at the stage of swollen conidia, which indicates that respiration is an early event during germination. In line with these data, we found that significant oxygen consumption was detectable early during germination, whereas no oxygen consumption was measurable in suspensions of resting conidia. In summary, the present study provides evidence that respiration is absolutely required for the germination of A. fumigatus conidia. Anela Taubitz and Bettina Bauer contributed equally.     

Long term trends in outdoor <Emphasis Type="Italic">Aspergillus/Penicillium</Emphasis> spore concentrations in Derby,UK from 1970 to 2003 and a comparative study in 1994 and 1996 with the indoor air of two local houses

Aspergillus/Penicillium spore concentrations have been monitored in Derby since 1970 using a volumetric spore trap, with full year data from 1991. In addition a short comparative study with the indoor air was undertaken at two local houses in 1994 and 1996. Aspergillus/Penicillium spores were present in the Derby air throughout the year and often reached maximum monthly cumulative concentrations in the autumn, although they were occasionally the dominant spores in the winter when total spore concentrations were low. Very high daily concentrations could occur at any time of year with a count of over 5000 recorded. Peak days in the autumn and winter of 2002–2003 were examined on a two hourly basis showing higher concentrations in the middle of the day. There was a positive correlation of cumulative monthly Aspergillus/Penicillium totals with maximum temperature. Indoor data from the two houses was examined on a daily basis and compared with simultaneously sampled outdoor daily spore concentrations. The elevated Aspergillus/Penicillium spore levels found in the older of the two houses occurred on all of the days sampled. Compared to the modern house, the Aspergillus/Penicillium spore concentrations in the old house represented a much higher percentage of the total spore count than in the modern one. The correlation between outdoor Aspergillus/Penicilliumspore concentrations and the indoor air of the old house was 0.62, whereas in the modern house it was 0.31. Peak hourly samples of Aspergillus/Penicillium spore counts occurred at times of greatest activity.     

Proven Invasive Pulmonary Aspergillosis in Stem Cell Transplant Recipient Due to <Emphasis Type="Italic">Aspergillus sublatus</Emphasis>, a Cryptic Species of <Emphasis Type="Italic">A. nidulans</Emphasis>

Invasive fungal disease represents one of the severe complications in haematopoietic stem cell transplant recipients. We describe a case of a patient treated for relapse of chronic lymphoblastic leukaemia 6 years after HSCT. The patient was treated for invasive pulmonary aspergillosis but died 3 months later from multiple organ failures consisting of haemorrhagic necrotizing fungal pneumonia, refractory chronic hepatic graft versus host disease and cytomegalovirus hepatitis. Autopsy samples revealed histopathological evidence of fungal hyphae and an unusual Aspergillus nidulans-like species was isolated in pure culture. More precise identification was achieved by using scanning electron microscopy of ascospores and sequencing of calmodulin gene, and the isolate was subsequently re-identified as A. sublatus (section Nidulantes) and showed good in vitro susceptibility against all classes of antifungals. Commonly used ITS rDNA region and β-tubulin gene fail to discriminate A. sublatus from related pathogenic species, especially A. quadrilineatus and A. nidulans. Although this is the first case of proven IPA attributed to A. sublatus, we demonstrated that at least some previously reported infections due to A. quadrilineatus were probably caused by this cryptic species.     

Endo‐ and exo‐inulinases: Enzyme‐substrate interaction and rational immobilization

Three‐dimensional models of exoinulinase from Bacillus stearothermophilus and endoinulinase from Aspergillus niger were built up by means of homology modeling. The crystal structure of exoinulinase from Aspergillus awamori was used as a template, which is the sole structure of inulinase resolved so far. Docking and molecular dynamics simulations were performed to investigate the differences between the two inulinases in terms of substrate selectivity. The analysis of the structural differences between the two inulinases provided the basis for the explanation of their different regio‐selectivity and for the understanding of enzyme‐substrate interactions. Surface analysis was performed to point out structural features that can affect the efficiency of enzymes also after immobilization. The computational analysis of the three‐dimensional models proved to be an effective tool for acquiring information and allowed to formulate an optimal immobilized biocatalyst even more active that the native one, thus enabling the full exploitation of the catalytic potential of these enzymes. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

Production of Proteinase with Plasmin-Like and Prekallikrein Activating Activity by the Micromycete <Emphasis Type="Italic">Aspergillus terreus</Emphasis>

The effect of nitrogen sources in the fermentation medium and the cultivation conditions on the production of proteinases with plasmin-like and prekallikrein activation activity by micromycete Aspergillus terreus 2 was investigated. The highest secretion of proteinases was achieved when the micromycete was cultivated on a medium containing both amine- and mineral nitrogen sources at an initial pH of 5.5 and at 28°С. It was established that the extracellular micromycete proteinases are equally capable of hydrolyzing fibrin and fibrinogen.     

<Emphasis Type="Italic">Aspergillus vadensis</Emphasis>, a new species of the group of black Aspergilli

A strain from the group of black Aspergilli was analysed in detail to determine the species to which it belongs. A detailed analysis of morphology, RFLP patterns and metabolite profiles was carried out. In addition, a phylogenetic tree was constructed for the black Aspergilli using the ITS and the -tubulin sequences of the individual strains. The new species differs by its poor growth on glycerol and galacturonate and its unique extrolite profile consisting of aurasperone B, nigragillin, asperazine and kotanins. RFLP analysis using three genes as probes also resulted in a unique pattern. These data indicate that the strain was closely related but not identical to Aspergillus foetidus, Aspergillus niger and Aspergillus tubingensis. It was therefore designated as a novel species and named Aspergillus vadensis.     

我国首例肌曲霉病及其试验研究

目的报道国内首例肌曲霉病,对所分离的黄曲霉进行形态、药敏、rDNA序列分析研究。方法对患者病变肌组织行B超和病理检查;将分离菌株接种于察氏培养基观察形态学特征;微量法测定MIC值;酚氯仿法提取DNA,扩增ITS间区序列并进行多序列分析。结果B超示左腓肠肌不规则回声,病理示肌细胞变性、坏死,肌组织内见分枝分隔菌丝。表型及基因型特征为黄曲霉;分离菌株对多种抗真菌药物耐药;多基因序列分析显示所分离菌株与标准株及自然和临床分离株具有100%的相似性。给予特比萘芬治疗3个月后痊愈。结论黄曲霉可致肌肉感染;药敏试验对临床治疗具有指导意义;环境中普遍存在的真菌可引起免疫低下患者致死性感染。     

Cytotoxicity of Aspergillus fungi isolated from hospital environment

The majority of mycotoxins produced by Aspergillus fungi are immunosuppressive agents, and their cytotoxicity may impair defense mechanisms in humans. The objective of the study was evaluation of the cytotoxicity of fungi isolated from an environment where inpatients with impaired immunity were present. The materials comprised 57 fungal strains: Aspergillus fumigatus, Aspergillus niger: Aspergillus ochraceus, Aspergillus flavus, Aspergillus versicolor and Aspergillus ustus isolated from hospital rooms in Cracow. The cytotoxicity of all the strains was evaluated using the MTT test (3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide). To emphasize the differences in cytotoxicity among the particular strains, variance analysis (ANOVA) and Tukey's difference test were used. Out of 57 Aspergillus strains tested, 48 (84%) turned out to be cytotoxic. The cytyotoxicity was high (+++) in 21 strains, mainly in A. fumigatus. The least cytotoxic were A. niger fungi, this being statistically significant (p<0,05). To protect a patient from the adverse effects of mycotoxins, not only his or her immunity status should be evaluated but also the presence of fungi in hospital environment and their cytotoxicity should be monitored (possible exposure).     

Fatal-Mixed Cutaneous Zygomycosis–Aspergillosis: A Case Report

Mucorales and Aspergillus are molds responsible for infections in immunocompromised patients. In this report, we describe a case of a rare extensively mixed cutaneous infection caused by Lichtheimia ramosa, Aspergillus fumigatus and Aspergillus terreus in a neutropenic patient suffering from an acute leukemia. The fatal outcome of this patient can be attributed to its hematologic malignancy, the extensive nature of the lesions and the resistance of the strains to antifungals.     

拮抗链霉菌S24 发酵培养基的优化及其对黄曲霉的抑菌作用

拮抗链霉菌S24对黄曲霉、赭曲霉、黑曲霉等粮食和饲料中常见的曲霉菌具有广谱抗性。优化了该菌株抗菌物质高产发酵培养基配方,并分析该菌株产生的抗真菌物质对黄曲霉的抑菌作用。通过单次单因子试验、正交试验,对S24菌株抗菌物质的发酵培养基进行了优化,利用显微镜观察,研究了S24菌株产生的抗菌物质对黄曲霉菌丝及孢子的抑制作用。获得适合S24菌株抗菌物质产生的最佳培养基配方为：淀粉10 g、葡萄糖40 g、黄豆饼粉24 g、酵母浸膏粉8 g、KH2PO4 4 g、CaCO3 0.8 g、水 1 L。优化后的发酵培养基     

New application of Aspergillus versicolor in promoting plant growth after suppressing sterigmatocystin production via genome mining and engineering

Aspergillus genus is a key component in fermentation and food processing. However, sterigmatocystin (STE)—a mycotoxin produced by several species of Aspergillus—limits the use of some Aspergillus species (such as Aspergillus versicolor, Aspergillus inflatus, and Aspergillus parasiticus) because of its toxicity and carcinogenicity. Here, we engineered an STE-free Aspergillus versicolor strain based on genome mining techniques. We sequenced and assembled the Aspergillus versicolor D5 genome (34.52 Mb), in which we identified 16 scaffolds and 54 biosynthetic gene clusters (BGCs). We silenced cytochrome P450 coding genes STC17 and STC27 by insertional inactivation. The production of STE in the Δstc17 mutant strain was increased by 282% but no STE was detected in the Δstc27 mutant. Metabolites of Δstc27 mutant exhibited growth-promoting effect on plants. Our study makes significant progress in improving the application of some Aspergillus strains by restricting their production of toxic and carcinogenic compounds.