Date: 26 November 2013
cultures grown from BAL fluid showing formation of sclerotia.
Copyright:
Kindly donated by Dr Claudia Venturelli and Dr Giorgia Bertazzoni, Laboratory of Microbiology – Policlinico of Modena-Italy. © Fungal Research Trust
Notes:
These colonies were isolated from a BAL, (also with bacterial qrowth of S.aureus and S.maltophilia) from a patient with a VAP (undergoing corticosteroid treatment). The growth medium used is sabouraud dextrose agar , incubated at 37° C The identification is made by microscopic/macroscopic observation criteria.
Colonies on CYA 60-70 mm diam, plane, sparse to moderately dense, velutinous in marginal areas at least, often floccose centrally, sometimes deeply so; mycelium only conspicuous in floccose areas, white; conidial heads usually borne uniformly over the whole colony, but sparse or absent in areas of floccose growth or sclerotial production, characteristically Greyish Green to Olive Yellow (1-2B-E5-7), but sometimes pure Yellow (2-3A7-8), becoming greenish in age; sclerotia produced by about 50% of isolates, at first white, becoming deep reddish brown, density varying from inconspicuous to dominating colony appearance and almost entirely suppressing conidial production; exudate sometimes produced, clear, or reddish brown near sclerotia; reverse uncoloured or brown to reddish brown beneath sclerotia. Colonies on MEA 50-70 mm diam, similar to those on CYA although usually less dense. Colonies on G25N 25-40 mm diam, similar to those on CYA or more deeply floccose and with little conidial production, reverse pale to orange or salmon. No growth at 5°C. At 37°C, colonies usually 55-65 mm diam, similar to those on CYA at 25°C, but more velutinous, with olive conidia, and sometimes with more abundant sclerotia.
Sclerotia produced by some isolates, at first white, rapidly becoming hard and reddish brown to black, spherical, usually 400- 800 µm diam. Teleomorph not known. Conidiophores borne from subsurface or surface hyphae, stipes 400 µm to 1 mm or more long, colourless or pale brown, rough walled; vesicles spherical, 20-45 µm diam, fertile over three quarters of the surface, typically bearing both metulae and phialides, but in some isolates a proportion or even a majority of heads with phialides alone; metulae and phialides of similar size, 7-10 µm long; conidia spherical to subspheroidal, usually 3.5-5.0 µm diam, with relatively thin walls, finely roughened or, rarely, smooth.
Distinctive features
Aspergillus flavus is distinguished by rapid growth at both 25°C and 37°C, and a bright yellow green (or less commonly yellow) conidial colour. A. flavus produces conidia which are rather variable in shape and size, have relatively thin walls, and range from smooth to moderately rough, the majority being finely rough.
Images library
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A. Necrotic mass prolapsing in and out of the distal right intermediate bronchus obscuring both the basal stem and basal division
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History : A 20 yr old female with cystic fibrosis complicated by CF-related Diabetes mellitus (diagnosed at age 12 years) & ABPA (diagnosed at
15 years ). She was fairly stable until last 9 months, when she started to experience frequent pulmonary exacerbations, which have prompted intensive therapies.
Her serum IgE at the time of diagnosis was 5060 IU/L, skin prick test for aspergillus was positive, and serum was positive for precipitating antibodies to Aspergillus. She was treated with oral prednisone (1 mg/kg/day) for first two weeks followed by prednisone at 0.5 mg/kg every other day for at least 6 months with some clinical and serologic improvement. Over the following 5 years, she presented with a pattern of repeated episodic exacerbations with wheezing and crackles, increases in IgE and need to increase prednisone dosage. In an attempt to control her frequent ABPA relapses, itraconazole was added at 200mg twice a day, with some clinical & serologic improvement.
2-3 weeks prior to the scans and X rays, she developed severe protracted coughing spells associated with minor hemoptysis, low grade intermittent fever, and weight loss. Her FEV1 declined in a 3 months period from 56% to 33%.
A chest-x ray (Fig 3) did not reveal any new changes when compared to the one obtained almost a year before. A CT scan of the chest Figs 1 and 2- lateral view), however shows an ovoid soft tissue density within an ectatic bronchi in the anterior basal segment of the LLL, felt to be an aspergilloma.She was started on oral voriconazole (3 months after the above scans and X-rays) which is better absorbed in CF patients than itraconazole, at 200mg twice a day. Her prednisone was gradually reduced to 5 mgs/day and her FEV1 increased to 46% of predicted (03/31/10), her IgE level was 1167 kU/L (previously in January 2009, her IgE level was 3053 kU/L)and her weight has plateaued.
Since she was not fit enough for surgery at that time, removing the aspergilloma by flexible bronchoscopy was unsuccessful. The aspergilloma can be seen at bronchoscopy in Fig 4.,
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Widespread mucoid impaction also seen, particularly in left side, suggestive of allergic bronchopulmonary aspergillosis.
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Microscopy of fungal hyphae isolated from fungal culture (lactophenol blue)
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A CT scan performed at this time showed an enhancing lesion with ill defined margins involving the pterigopalatine fossa, the maxillary antrum , orbital apex and middle cranial fossa. There was also destruction of boney architecture locally.
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The patient underwent a pneumonectomy because of the severity of her disease process, and uncertainty about the diagnosis, prior to serology results being obtained.
Serology showed an IgE of 2600, with a strongly positive Aspergillus RAST test and weakly positive Aspergillus precipitins. Material removed at bronchoscopy showed eosinophilia. These features confirm a diagnosis of allergic bronchopulmonary aspergillosis (ABPA).
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