Wastewater-Agar as a selection environment: A first step towards a fungal in-situ bioaugmentation strategy.

Viable and metabolically active fungi in toxic mixed liquors, treating landfill leachates and municipal wastewaters, were identified by culture depending methods. A selective culture medium consisting of wastewater and agar (WA) restrained fungi that could be randomly present (94% of the 51 taxa retrieved on WA were sample-specific), overcoming the problem of fast growing fungi or mycoparasite fungi. Moreover, WA allowed the isolation of fungi with a possible role in the degradation of pollutants typically present in the two wastewaters. Phoma medicaginis var. medicaginis, Chaetomium globosum, and Geotrichum candidum were mainly found in municipal wastewater, whereas Pseudallescheria boydii, Scedosporium apiospermum, Aspergillus pseudodeflectus, and Scopulariopsis brevicaulis were typical of landfill leachate.

Scleral Buckle Infection With Pseudallescheria boydii.

Pseudallescheria boydii is a ubiquitous fungus that infects soft tissues and is known to cause ocular disease, including keratitis and endophthalmitis, in rare cases. In immunocompromised hosts, P. boydii can disseminate to or from the eye and other organs with lethal consequences. Postoperative P. boydii infections have, in rare cases, complicated several types of ocular surgeries in immunocompetent patients, but never for a scleral buckle. The authors present the first case of an infected scleral buckle from P. boydii. [Ophthalmic Surg Lasers Imaging Retina. 2017;48:676-678.].

Towards proteomic species barcoding of fungi - An example using Scedosporium/Pseudallescheria complex isolates.

MALDI-ToF mass spectrometry offers fast and reliable species identification for bacteria and yeasts under clinical routine conditions. Here, we produced mass spectra for identification of clinically important species of the Pseudallescheria/Scedosporium complex using the recently suggested new nomenclature and use this example to discuss to what extent the principle of DNA barcoding might be transferred to mass spectrometry.

Structural analysis of glucosylceramides (GlcCer) from species of the Pseudallescheria/Scedosporium complex.

Glucosylceramides (GlcCer) are the main neutral glycosphingolipids expressed in fungal cells. In this work, glucosylceramides (GlcCer) were extracted from three strains of Scedosporium (Pseudallescheria) boydii, one strain of Pseudallescheria ellipsoidea and one strain of Pseudallescheria angusta and purified by several chromatographic steps. Using high-performance thin layer chromatography (HPTLC), we found a similarity between GlcCer obtained from all of the analysed strains. A detailed structural analysis of the P. ellipsoidea GlcCer was performed via electrospray ionization mass spectrometry (ESI-MS) and confirmed in 1- and 2-D heteronuclear NMR experiments ((1)H-(13) C HSQC). GlcCer species produced by mycelial forms of these strains displayed the same structure previously demonstrated by our group for P. boydii, Cryptococcus neoformans, Pseudallescheria minustipora, Fusarium solani, and Colletotrichum gloesporioides. A monoclonal antibody (mAb) against GlcCer was used for immunofluorescence experiments. Our results revealed that GlcCer is present on the surface of these fungi, and no difference was observed in the GlcCer structure of the present set of strains in terms of geographic or clinical origin, suggesting a conserved GlcCer structure similar to those previously described for Scedosporium apiospermum, Scedosporium aurantiacum, and P. minutispora. The surface distribution of GlcCer in these fungi is suggestive of the involvement of this molecule in fungal growth.

Pseudallescheria boydii with Aspergillus fumigatus and Aspergillus terreus in a Critically Ill Hematopoietic Stem Cell Recipient with ARDS.

Pseudallescheria boydii is a fungal organism known to affect immunocompromised patients. This organism is known to cause, in severe cases, invasive infection of various organs such as the central nervous, cardiovascular, and respiratory systems. We report an unusual case of pulmonary P. boydii pneumonia in an immunocompromised critically ill patient with a co-infection of Aspergillus fumigatus and Aspergillus terreus with ARDS. This case highlights the importance of a high index of suspicion for superimposed fungal infections in patients who are critically ill and immunocompromised. Uncommon fungal pathogens should be considered in the differential diagnosis of respiratory failure, especially if diagnostic markers such as galactomannan (from BAL and serum) or 1,3-beta-D-glucan are elevated. Further diagnostic interventions are warranted when insufficient clinical improvement is observed to prevent treatment failure and adverse outcomes.

Cutaneous Pseudallescheria boydii/Scedosporium apiospermum Complex (Molecular type: Scedosporium apiospermum [Clade 4]) Infection: A Case Report and Literature Review of Cases from Japan.

We report a case of subcutaneous Pseudallescheria boydii/Scedosporium apiospermum complex infection occurring in a 77-year-old Japanese female farmer suffering from interstitial pneumonia. Seven months prior to the current presentation, she noticed nodes on her right forearm after pulling up weeds, and the nodes grew larger. Two soft dome-shaped, protruded nodes ( 15 mm and 30 mm in size ) had fused together on the extensor surface of the right forearm. Yellowish-white, rice-grain-sized pustules clustered on the surface. Histopathological examination of the skin specimen showed large and small abscesses surrounded by epithelioid granuloma; separate branching hyphae within the granulation tissue were stained with PAS. No grains were observed. Fungal culture yielded fast-growing, grayish-white, fluffy colonies which were identified as Scedosporium apiospermum (Clade 4) using sequence analysis of the ß-tubulin gene. We also reviewed 28 previously reported Japanese cases of P. boydii or S. apiospermum infection presenting with skin manifestations.

In vitro antifungal activity of antipsychotic drugs and their combinations with conventional antifungals against Scedosporium and Pseudallescheria isolates.

In the present study, in vitro antifungal activities of five antipsychotic drugs (i.e., chlorpromazine hydrochloride, CPZ; trifluoperazine hydrochloride, TPZ; amantadine hydrochloride; R-(-)-deprenyl hydrochloride, and valproic acid sodium salt) and five conventional antifungal drugs (i.e., amphotericin B, AMB; caspofungin, CSP; itraconazole; terbinafine, TRB and voriconazole, VRC) were investigated in broth microdilution tests against four clinical and five environmental Scedosporium and Pseudallescheria isolates. When used alone, phenothiazines CPZ and TPZ exerted remarkable antifungal effects. Thus, their in vitro combinations with AMB, CSP, VRC, and TRB were also examined against the clinical isolates. In combination with antifungal agents, CPZ was able to act synergistically with AMB and TRB in cases of one and two isolates, respectively. In all other cases, indifferent interactions were revealed. Antagonism was not observed between the tested agents. These combinations may establish a more effective and less toxic therapy after further in vitro and in vivo studies for Scedosporium and Pseudallescheria infections.

Human-impacted areas of France are environmental reservoirs of the Pseudallescheria boydii/Scedosporium apiospermum species complex.

Species of the Pseudallescheria boydii/Scedosporium apiospermum complex (PSC) are emerging fungal pathogens able to chronically colonize the airways of patients with cystic fibrosis (CF). As P. boydii was found more frequently colonizing the lungs of CF patients in France than in other European countries in a previous report, the present study was conducted in order to clarify distribution of PSC species in France and to characterize their natural habitat. The highest densities of PSC isolates were found in human-impacted areas, i.e. agricultural areas, fluids obtained from wastewater treatment plants, playgrounds and industrial areas. PSC was not detected from soil samples collected in forests. Most PSC culture-positive soil samples exhibited a pH range of 6-8. Scedosporium dehoogii, the most abundant species, was detected in all human-impacted area types except vineyards, whereas Scedosporium aurantiacum was mostly found in agricultural areas. Pseudallescheria boydii and S. apiospermum were predominantly isolated from seashores and playgrounds respectively. Pseudallescheria minutispora was found only once from a playground. This study highlights potential sources of contamination of the patients, especially in the CF context.

[Invasive fungal disease due to Scedosporium, Fusarium and mucorales]. / Enfermedad fúngica invasora por Scedosporium, Fusarium y Mucor.

The number of emerging organisms causing invasive fungal infections has increased in the last decades. These etiological agents include Scedosporium, Fusarium and mucorales. All of them can cause disseminated, virulent, and difficult-to treat infections in immunosuppressed patients, the most affected, due to their resistance to most available antifungal agents. Current trends in transplantation including the use of new immunosuppressive treatments, the common prescription of antifungal agents for prophylaxis, and new ecological niches could explain the emergence of these fungal pathogens. These pathogens can also affect immunocompetent individuals, especially after natural disasters (earthquakes, floods, tsunamis), combat wounds or near drowning. All the invasive infections caused by Scedosporium, Fusarium, and mucorales are potentially lethal and a favourable outcome is associated with rapid diagnosis by direct microscopic examination of the involved tissue, wide debridement of infected material, early use of antifungal agents including combination therapy, and an improvement in host defenses, especially neutropenia.

[A case of filamentous fungal infection showing tumor-like expansion in the cisterns].

A 67-year-old woman was admitted for headache. The initial MRI showed a gadolinium-enhanced lesion in the prepontine area. Initial and repeated CSF examinations were negative for the fungal infection. Since the enhanced lesion expanded in the cisterns, and showed tumor-like appearance, brain biopsy was performed at 3 months from her first admission. Histological studies revealed filamentous fungal infection probably caused by the pseudallescheria boydii. Intrathecal miconazole injection through the Ommaya reservoir successfully ameliorated patient's symptoms and the MRI findings. Primary cisternal fungal infection showing tumor-like expansion typically fails to demonstrate its supportive findings by the CSF examination. Therefore, histopathological assessment after brain biopsy should be considered in cases that are not conclusive by means of conventional laboratory examinations.

Multilocus sequence typing of Scedosporium apiospermum and Pseudallescheria boydii isolates from cystic fibrosis patients.

BACKGROUND: Scedosporium and Pseudallescheria species are the second most common lung-colonising fungi in cystic fibrosis (CF) patients. For epidemiological reasons it is important to trace sources of infection, routes of transmission and to determine whether these fungi are transient or permanent colonisers of the respiratory tract. Molecular typing methods like multilocus sequence typing (MLST) help provide this data. METHODS: Clinical isolates of the P. boydii complex (including S. apiospermum and P. boydii) from CF patients in different regions of Germany were studied using MLST. Five gene loci, ACT, CAL, RPB2, BT2 and SOD2, were analysed. RESULTS: The S. apiospermum isolates from 34 patients were assigned to 32 sequence types (STs), and the P. boydii isolates from 14 patients to 8 STs. The results revealed that patients can be colonised by individual strains for years. CONCLUSIONS: The MLST scheme developed for S. apiospermum and P. boydii is a highly effective tool for epidemiologic studies worldwide. The MLST data are accessible at http://mlst.mycologylab.org/.

[Isolation of Scedosporium apiospermum (teleomorph: Pseudallescheria apiosperma) from an acute myeloid leukemia patient]. / Bir akut miyeloid lösemi hastasindan Scedosporium apiospermum (teleomorf: Pseudallescheria apiosperma) izolasyonu.

Scedosporium apiospermum is an emerging opportunistic pathogen that may lead to life-threatening infections especially in immunosuppressive individuals. In this report, S.apiospermum infection in a 62 year old male patient with acute myeloid leukemia was presented. During remission-induction chemotherapy, piperacillin-tazobactam therapy was started for febrile neutropenia. Since fever had continued, treatment was switched to imipenem and also amphotericin B deoxycholate was added to the treatment protocol. Because of allergic reaction to amphotericin B, caspofungin was started at the fifth day of neutropenic fever. Following imaging studies with high resolution computerized thorasic tomography, antifungal therapy was changed to voriconazole due to findings suggestive of invasive aspergillosis. Since galactomannan antigen was found negative at the first day of voriconazole therapy, bronchoalveolar lavage material from apical segment of the left lower lobe was cultured onto various microbiologic media. S.apiospermum (Teleomorph: Pseudallescheria apiosperma) was isolated on the fourth day of cultivation. According to CLSI M38-A2 microdilution procedure, minimum inhibitory concentrations (MIC) of voriconazole, caspofungin, amphotericin B and posaconazole were found as 0.06, 2, 8 and 4 µg/ml, respectively. Since neutropenia was resolved, the patient was discharged with continued voriconazole therapy. It was concluded that antifungal susceptibility tests should be performed for Scedosporium species and the results should be compared to the clinical response. The determination of MIC breakpoints may provide useful information for the recommendation and use of optimal choices for the treatment of Scedosporium infections.

Distribution of the different species of the Pseudallescheria boydii/Scedosporium apiospermum complex in French patients with cystic fibrosis.

As various new sibling species within the Pseudallescheria boydii/Scedosporium apiospermum complex have been described recently with differences in their susceptibility to antifungals, this study was conducted in order to determine their respective frequency in cystic fibrosis. Results indicated that P. boydii largely predominated (62%), followed by S. apiospermum (24%), Scedosporium aurantiacum (10%) and Pseudallescheria minutispora (4%). Scedosporium dehoogii was not recovered in this study. The multiple correspondence factor analysis highlighted geographical discrepancies within species distribution: P. boydii was rarely encountered in Northern France, while S. apiospermum was less represented in the west of the country. Additionally, we demonstrated that all species encountered in the cystic fibrosis context were capable to chronically colonize the respiratory tract of patients. Molecular typing of a large set of environmental and clinical isolates should be conducted to delineate the epidemiology of each sibling species in the complex.

Arterial reconstruction for atypical mycotic aneurysms.

Our case involves a 67-year-old man with a  post lung transplantation status for idiopathic pulmonary fibrosis, who was initially referred to our institution for treatment of left hip septic arthritis due to Pseudallescheria boydii infection. One month following total hip arthroplasty, he noticed left upper thigh swelling with duplex imaging consistent with a 7-cm mid-superficial femoral artery aneurysm with popliteal vein compression and associated deep venous thrombosis. During operative resection, exploration of his contralateral saphenous vein showed inadequate conduit for use. Therefore, we proceeded with proximal and distal clamp control with intraoperative arterial shunting followed by cryoarterial reconstruction after complete aneurysm sac resection and debridement. Intraoperative arterial wall cultures remained negative for bacterial pathogens; however, final operative fungal cultures once again grew Pseudallescheria boydii. Our case highlights the complexity of adequate source control with effective arterial reconstruction in these immunosuppressed patient populations, particularly with fungal organisms that historically have poor therapeutic response to medical therapy alone.

El complejo Pseudallescheria/Scedosporium y taxas afines en la rutina del laboratorio micológico clínico / The Pseudallescheria/Scedoporium complex and related taxa in the routine of clinic mycological laboratory

El complejo Pseudallescheria/Scedosporium y taxas relacionados, pertenecientes a un grupo de microhongos con conidios viscosos e integrantes del orden Microascales, se presentan en diversos ambientes comunes asociados a las actividades humanas, ya sea en el suelo como en aguas contaminadas. Actualmente se consideran entre los mayores grupos de hongos filamentosos oportunistas causante de infecciones cutáneas y profundas en el hombre y otros mamíferos. El objetivo principal de este trabajo, consiste en reunir los datos primarios morfofisiológicos más relevantes, útiles para el micólogo médico en el laboratorio, con el aporte adicional de algunos aspectos, biológicos, ecológicos, taxonómicos y moleculares complementarios descritos en la literatura moderna.

The Pseudallescheria/Scedosporium complex and their related taxa are a group of fungus that conidia are presents in viscous mass and belong to the order Microascales. They are in several common enviroment related to human activities either in soil as in contaminated water. Nowadays they are considerated one of the most opportunistic group of filamentous fungus that may cause superficial and deep skin mycoses infections in man and other mammalian. The aim of this work is to gather the primary relevant morphophysiological aspects, usefull to the medical mycologist in the laboratory, plus the contribution of some of biological, ecological, taxonomical and moleculars complementary aspects that are describe in modern literature.