Prevalence of Aspergillus fumigatus and other fungal species in the sputum of adult patients with cystic fibrosis

Aspergillus fumigatus is often found in the respiratory tract secretions of patients with cystic fibrosis (CF), although the role of the fungus for progression of pulmonary disease remains unclear. This study aimed to investigate the frequency of A. fumigatus and other fungi in sputum of adult CF patients using different methods for culture and microscopy. Results from the analysis of 369 samples from 94 patients showed that A. fumigatus could be isolated in 45.7% of patients. Other moulds were rare, but the yeast Candida albicans was another frequent isolate, detected in 75.5% of patients. A comparison of different culture media showed no difference between a selective medium developed to specifically inhibit Pseudomonas aeruginosa and a standard fungal culture medium for growth of A. fumigatus, although both were more efficient for detection of fungi than other bacterial culture media. Fluorescent microscopy with calcofluor white was more sensitive for detection of fungal hyphae in undiluted sputum than standard methylene blue staining. This study shows that A. fumigatus and C. albicans have a high frequency in adult CF patients. Microbiological analysis should routinely include methods for specific identification of fungi to monitor for potential complications arising from fungal disease in these patients.     

免疫抑制宿主和健康宿主在鼠肺部烟曲霉感染时基因表达的差异

目的 观察免疫抑制宿主和健康宿主在遭遇条件致病真菌初期时基因表达的差异,筛选新的真菌免疫相关基因,以便对条件致病真菌导致系统性真菌病的发病机制有更新的认识。方法 免疫抑制和健康小鼠各6只,经鼻内接种烟曲霉孢子3d后处死,取肺组织,抽提总RNA,进行基因表达谱芯片研究。结果 在4096条基因中共筛选出66条差异表达基因。其中,免疫抑制组编码CD22、CD53、24p3以及与单核巨噬细胞吞噬消化有关的基因表达上调。编码免疫球蛋白入链、肺泡表面蛋白C以及与肺组织代谢有关的基因表达下调。编码IL18、CD8、CD28等的基因无差异表达。结论 免疫抑制组CD22基因上调可能是宿主受到免疫抑制的途径之一,而CD8则未发现差异。CD53可能间接参与对单核巨噬细胞的调控。24p3蛋白作为应激蛋白,可能保护宿主免受过度炎症反应。     

A novel `two-component' protein containing histidine kinase and response regulator domains required for sporulation in Aspergillus nidulans

We have characterised a novel Aspergillus nidulans gene encoding a `two-component' signalling protein (tcsA). tcsA encodes both a histidine kinase domain and a response regulator domain similar to those found in bacterial, lower eukaryotic and plant members of the two-component family of proteins, while two PAS domains in the amino-terminal region of the predicted tcsA product may monitor the signal which regulates a tcsA histidine kinase-response regulator phosphorelay. While tcsA is nonessential for vegetative growth, cells lacking the gene are unable to produce conidia on standard Aspergillus growth media. However, tcsA is not absolutely required for production since this defect is suppressed by growth on 1 M sorbitol. Received: 23 December 1999 / 1 March 2000     

Assessing risk factors for systemic fungal infections

The incidence of invasive fungal infection has increased in recent years. Most infections are caused by Candida albicans and Aspergillus spp. but the emergence of other fungal infections is changing the spectrum of disease. Immunosuppression and breakdown of anatomical barriers such as the skin are the major risk factors for fungal infections. Health care workers encounter at-risk patients in various settings, including AIDS clinics and intensive care, transplantation and oncology units. Patients with prolonged and deep neutropenia (haematological malignancy patients) are most at risk and are therefore most likely to receive prophylactic therapy. Practical measures can be taken to avoid exposing the patient to fungi (air filtration, regular hand washing, avoiding plants and flowers) and antifungal agents can be administered to prevent systemic fungal infection. Most fungal infections have non-specific symptoms; this makes recognition of the signs and symptoms of the disease important but also makes diagnosis difficult and empirical treatment necessary. Some antifungal agents have limitations but new formulations will improve therapy and play a key role in future antifungal strategies.     

Molecular approaches for the diagnosis and epidemiological investigation of Aspergillus infection

Molecular techniques have been applied to the diagnosis of invasive aspergillosis and to investigate the ecology and epidemiology of Aspergillus. Recent advances in diagnosis include the development of PCRs targeting either panfungal or Aspergillus-specific sequences, using whole blood or serum samples. When a sensitive PCR is used, invasive aspergillosis in bone marrow transplant patients can be detected several weeks before antigen tests become positive, and a positive PCR often pre-dates the institution of antifungal therapy. The role of PCR in monitoring response to therapy in immunocompromised patients is unclear. No prospective studies have yet demonstrated that management incorporating PCR alters the poor outcome of invasive aspergillosis in immunocompromised patients. Molecular typing of Aspergillus fumigatus has shown wide geographical dispersal of indistinguishable strains. This, combined with the observation that multiple strains may be isolated from individual colonised patients with cystic fibrosis and from immunocompromised patients with disseminated disease, makes the elucidation of the epidemiology of aspergillosis relatively complex.     

Production and characterization of monoclonal antibodies to serine proteinase allergens in Penicillium and Aspergillus species.

BACKGROUND: Alkaline and/or vacuolar serine proteinases are major allergens in prevalent airborne Penicillium and Aspergillus species. OBJECTIVE: The object of this study is to generate and characterize monoclonal antibodies against these serine proteinase allergens. METHODS: BALB/c mice were immunized individually with the Penicillium citrinum culture medium or the crude extract and culture medium preparations of Aspergillus fumigatus. Hybridoma cells that secrete monoclonal antibodies against serine proteinase allergens were selected by immunoblotting. Antigens in three different Penicillium (P. citrinum, P. notatum and P. oxalicum) and two different Aspergillus species (A. fumigatus, and A. flavus) recognized by these monoclonal antibodies were analysed by sodium dodecyl sulphate and two-dimensional polyacrylamide gel electrophoresis immunoblotting and N-terminal amino acid sequence analysis. RESULTS: Four (PCM8, PCM10, PCM16 and PCM39) and one (FUM20) monoclonal antibodies against serine proteinase allergens were generated after fusion of NS-1 cells with spleen cells obtained from BALB/c mice immunized with antigens from P. citrinum and A. fumigatus, respectively. Immunoblotting results showed that PCM8 reacted with an alkaline serine proteinase allergen in P. citrinum and P. notatum. PCM10 and PCM39 reacted with the alkaline serine proteinase in two Penicillium (P. citrinum, P. notatum) and two Aspergillus species (A. fumigatus, and A. flavus) tested. PCM16 reacted with the alkaline serine proteinase allergen in P. citrinum, A. fumigatus and A. flavus but not with that in P. notatum. MoAb FUM20 reacted with the alkaline serine proteinase allergen in two Aspergillus species (A. fumigatus and A. flavus) but not with that in two different Penicillium species (P. citrinum, P. notatum) tested. Among these five monoclonal antibodies generated, only PCM39 and FUM20 can react with the vacuolar serine proteinase allergen in P. notatum, P. oxalicum and in A. fumigatus. The 35 kDa P. citrinum component that reacted with FUM20 has an N-terminal amino acid sequence of DSPSVEKNAP. CONCLUSION: Five monoclonal antibodies against different epitopes of the serine proteinase major allergens in prevalent Penicillium and Aspergillus species were generated in the present study. Antibodies obtained may be useful in the characterization and standardization of serine proteinase allergens in crude fungal extracts.     

Pulmonary hydatid disease with coexistent aspergillosis: An incidental finding

There are only a few case reports in the literature on the coexistence of aspergillosis and echinococcosis. We report a case of a 45-year-old immunocompetent patient who presented with a history of intermittent fever and cough with haemoptysis. Chest x-ray and CECT showed a large cystic lesion in right lower lobe with multiple floating membranes. Histopathological examination of cyst wall revealed the laminated membrane of hydatid cyst along with infiltration of its wall with septate fungal hyphae with acute angle branching suggestive of aspergillosis.     

Mycotic Dermatitis in Domestic Fowl. A Case Report./Pilzdermatitis am Haushuhn. Ein Fallbericht

Summary: Two cases of mycotic dermatitis, probably caused by Aspergillus spec, are presented.
Zusammenfassung: Es werden zwei Fälle von Pilzdermatitis, wahrscheinlich hervorgerufen durch Aspergillus, vorgestellt     

菊芋原料同步糖化发酵生产丁二酸

对菊芋原料发酵生产丁二酸进行了研究,用 Actinobacillus succinogenes 和 Aspergillus niger 同步糖化发酵,发现同步糖化发酵效果优于糖化后再发酵,在同步糖化发酵过程中还原糖质量浓度始终保持在10～40 g/L,可以避免高浓度的还原糖对 A.succinogenes 的抑制.5 L搅拌罐中同步糖化补料分批发酵96 h产丁二酸98.2 g/L,对消耗糖产率95.4%,生产强度1.02 g/(L·h) .     

A strategy for managing fungal infections in haematopoietic stem cell transplantation

As more indications continue to be found for allogeneic haematopoietic transplantation, more patients are at risk for invasive fungal infectious diseases (IFID), particularly candidiasis and aspergillosis. Risk factors for disease are becoming better defined and diagnostic methods have improved considerably. In addition, there is now international agreement that three elements form the basis for defining IFID (host factors, clinical evidence, and mycological results), that imaging is acceptable for diagnosing disease, and that indirect tests such as antigen detection are also adequate mycological proof of cause. There are also more drugs available and still more to come, offering the potential for selective prophylaxis as well as preemptive and specific therapy, as well as for flexible administration. Hence, all the elements are in place for designing and testing an effective and economically sound strategy for dealing with IFID.