Browsing by Author "Carvalho, Dinah"
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- Characterisation of respiratory disease during the 2010/2011 influenza winter season in Portugal: Contribute of the Laboratory Network for the Diagnosis of Influenza A(H1N1)2009 InfectionPublication . Pechirra, Pedro; Gonçalves, Paulo; Conde, Patrícia; Guiomar, Raquel; Duque, Vítor; Vaz, João; Ribeiro, Graça; Cabral, Rita; Mota Vieira, Luísa; Almeida Santos, Madalena; Silvestre, Maria José; Pimentel Couto, Ana Rita; Bruges Armas, Jacome; Castro, Ana Paula; Ramos, Maria Helena; Janeiro, André; Mimoso, Paula; Marcelino, Rute; Fernandes, Aida; Milho, Luís; Rego, João; Beleza, Álvaro; Barreto, Maria do Rosário; Carvalho, Dinah; Ribeiro, Carlos Manuel; Fernandes, Paula; Andrade, Graça; Sobrinho Simões, Joana; Costa, Maria do Rosário; Guimarães, João Tiago; Corte Real, Rita; Branquinho, Paula; Caldeira, Filomena; Maurílio, Manuel
- Eumicetoma causado por Parathyridaria percutanea num doente imunodeprimido - caso importadoPublication . Fraga, Marta; Carvalho, Dinah; Sabino, Raquel; Veríssimo, Cristina; Simões, Helena; Sapinho, Guilherme; Alho4, Ana; Espada, Eduardo; Moreno, Raul; Forjaz de Lacerda, J.; Lito, L. Marques; Cristino, J. MeloEumicetoma é uma infeção fúngica subcutânea crónica caracterizada por edema, fistulização e libertação de grânulos. Parathyridaria percutanea é uma causa rara de feohifomicose subcutânea, não havendo casos descritos de formação de fistulas ou drenagem de grãos devido a este fungo. Apresenta-se um caso clínico de eumicetoma causado por Parathyridaria percutanea
- Eumycetoma due to Parathyridaria percutânea in an immunosuppressed patient - an imported case in PortugalPublication . Fraga, Marta; Carvalho, Dinah; Sabino, Raquel; Veríssimo, Cristina; Simões, Helena; Sapinho, Guilherme; Alho, Ana; Espada, Eduardo; Moreno, Raul; Forjaz de Lacerda, J.; Marques Lito, L.; Melo Cristino, J.Objectives: Eumycetoma is a chronic subcutaneous fungal infection characterized by swelling, fistulization and discharge of fungal granules. These infections occur after traumatic inoculation and progress slowly over months or even years into a chronic form. Parathyridaria percutanea is a rare cause of subcutaneous phaeohyphomycoses and there are no published reports of discharging sinus or granuloma formation due to this fungus. We present a rare case of eumycetoma due to Parathyridaria percutanea in a 10-years-old patient from Angola, presenting after allogeneic hematopoietic stem cell transplant (alloHSCT) for severe aplastic anemia. Material and methods: A 10-year-old boy from Angola with severe aplastic anemia was transferred to Portugal and submitted to alloHSCT from a mismatched unrelated donor at a single HLA-locus (9/10). After secondary graft failure due to post-transplant hemophagocytic lymphohistiocytosis, he underwent a second alloHSCT, without engraftment. Twenty-three days after the 3rd alloHSCT from the same donor and seven days under prophylaxis with posaconazole, the patient presented with febrile neutropenia with no clear focus. He complained of painless swelling of the right knee, with slight fluctuation. An ultrasound was performed revealing a heterogeneous hypoechogenic area with imprecise contours, inside of which there was a nodular formation of approximately 10 mm. Two weeks later, a spontaneous drainage of a grossly spherical grain from the right knee occurred. It was characterized as extra-articular, subcutaneous with filamentous aspect. The material was sent to the laboratory where its macroscopic observation allowed the identification of a granule of possible mycotic etiology, consistent with the diagnosis of eumycetoma. This sample was promptly processed for bacteriology (aerobic and anaerobic) and mycology studies. Results: Direct microscopic examination of the grain revealed septate hyphae with irregular hyphal swellings. Cultures for bacteria (aerobic and anaerobic) were sterile. After 1 week, slow growing colonies appeared on Sabouraud dextrose agar (SDA) incubated at 25 °C. Those colonies were flat, spreading with sparse aerial hyphae, become creamy after 5 to 10 days and with orange reverse and then turned dark brown after 4 weeks. Lactophenol cotton blue mount revealed nonsporulating dematiceous hyphae with clamidospores. Phenotypic identification of the organism was uncertain but sequencing of the internal transcribed spacer (ITS) region of ribosomal DNA identified as Parathyridaria percutanea (100% homology, 99% coverage). The isolate showed in vitro resistance to itraconazole, fluconazole and anidulofungin and susceptibility to posaconazole, voriconazole and amphotericin B. As this pathogen is extremely rare, no antifungal guidelines have been proposed so far. The patient received dual coverage with posaconazole and liposomal amphotericin-B but died 2 weeks later of complications of the underlying disease. Conclusions: The presented case maybe the first described case of eumycetoma caused by Parathyridaria percutanea associated with fistulae presentation. Molecular identification of rare fungi is essential, especially when they are poorly sporulated, as in this case. Moreover, rare fungi can present with odd clinical signs, as in this case, emphasizing the relevance of a polyphasic approach to identify the etiological agent.
- Influenza severe cases in hospitals, between 2014 and 2016 in PortugalPublication . Guiomar, Raquel; Pechirra, Pedro; Cristóvão, Paula; Costa, Inês; Conde, Patrícia; Corte-Real, Rita; Branquinho, Paula; Silvestre, Maria José; Almeida Santos, Madalena; Fernandes, Isabel; Dias, Isabel; Rodrigues, Sónia; Sena, Nadir; Lazzara, Daniela; Sobrinho Simões, Joana; Costa, Maria do Rosário; Guimarães, João Tiago; Rodrigues, Fernando; Pereira-Vaz, João; Correia, Lurdes; Andrade, Graça; Freitas, Ludivina; Figueira, Neuza; Sanches, Raquel; Marques, Mónica; Barros, Margarida; Mota Vieira, Luísa; Cabral Veloso, Rita; Castelo Branco, Cláudia; Pimentel, Sílvia; Duarte, Joana; Pereirinha, Tânia; Bulhões, Sara; Moniz, Raquel; Brilhante, Maria José; Bruges Armas, Jácome; Pimentel Couto, Ana Rita; Santos, Margarida; Soares, Marta; Melo Cristino, José; Ribeiro, Carlos; Carvalho, Dinah; Barreto, Rosário; Ramos, Maria Helena; Castro, Ana Paula; Matos Santos, Ana Cláudia; Cunha, Mário; Martins, Luís; Almeida, Sofia; Peres, Maria João; Viseu, Regina; Inácio, Filipe; Mota, PaulaBackground: Since 2009, the Portuguese Laboratory Network (PLNID) for Influenza Diagnosis has integrated 15 Laboratories in mainland and Atlantic Islands of Azores and Madeira. This PLNID added an important contribute to the National Influenza Surveillance Program regarding severe and hospitalized influenza cases. The present study aims to describe influenza viruses detected in influenza like illness (ILI) cases: outpatients (Outp), hospitalized (Hosp), and intensive care units (ICU), between 2014 and 2016. Methods: The PLNID performs influenza virus diagnosis by biomolecular methodologies. Weekly reports to the National Influenza Reference Laboratory ILI cases tested for influenza. Reports include data on detecting viruses, hospital assistance, antiviral therapeutics, and information on death outcome. Were reported during two winter seasons 8059 ILI cases,being 3560 cases in 2014/15 (1024 in Outp, 1750 Hosp, and 606 in ICU) and 4499 cases in 2015/2016 (1933 in Outp, 1826 Hosp, and 740 in ICU). Results: The higher percentage of influenza positive cases were detected in Outp in both seasons, 18% during 2014/15 and 20% in 2015/16. In 2014/15,influenza cases were more frequent in individuals older than 65 years old and these required more hospitalizations,even in ICU. In 2015/16,the influenza cases were mainly detected in individuals between 15-64 years old. A higher proportion of influenza positive cases with hospitalization in ICU were observed in adults between 45-64 years old.During the study period,the predominant circulating influenza viruses were different in the two seasons: influenza B and A(H3) co-circulated in 2014/15,and influenza A(H1)pdm09 was predominant during 2015/16. Even when influenza A is notthe dominant virus, A(H3) and A(H1)pdm09 subtypes correlate with higher detection rate in hospitalized cases (Hosp and UCI), with higher frequencies in adults older than 45. Influenza B,detected in higher proportion in outpatients, was frequently relatedwith influenza cases in younger age groups: 0-4 and 5-14 years old. Conclusions: This study highlights the correlation of theinfluenza virus type/subtype that circulates in each season with the possible need for hospitalization and intensive care in special groups of the population. Circulation of influenza A subtypes can cause more frequentdisease in individuals older than 45, with need of hospitalization including intensive care. On the other hand, influenza B is more frequently associated with less severe cases and with infection in children and younger adults. Influenza B circulation might predict lower number of hospitalizations.The identification of influenza type in circulation,byPLNID ineach season, could guide action planning measures in population health care.
- Invasive and Subcutaneous Infections Caused by Filamentous Fungi: Report from a Portuguese Multicentric Surveillance ProgramPublication . Veríssimo, Cristina; Toscano, Cristina; Ferreira, Teresa; Abreu, Gabriela; Simões, Helena; Diogo, José; Carvalho, Dinah; Santiago, Felicidade; Lima, Ana; Queirós, Ana Maria; Sabino, RaquelInvasive fungal infections (IFI) have significantly increased over the past years due to advances in medical care for the at-risk immunocompromised population. IFI are often difficult to diagnose and manage, and can be associated with substantial morbidity and mortality. This study aims to contribute to understanding the etiology of invasive and subcutaneous fungal infections, their associated risk factors, and to perceive the outcome of patients who developed invasive disease, raising awareness of these infections at a local level but also in a global context. A laboratory surveillance approach was conducted over a seven-year period and included: (i) cases of invasive and subcutaneous fungal infections caused by filamentous/dimorphic fungi, confirmed by either microscopy or positive culture from sterile samples, (ii) cases diagnosed as probable IFI according to the criteria established by EORTC/MSG when duly substantiated. Fourteen Portuguese laboratories were enrolled. Cases included in this study were classified according to the new consensus definitions of invasive fungal diseases (IFD) published in 2020 as follows: proven IFI (N = 31), subcutaneous fungal infection (N = 23). Those proven deep fungal infections (N = 54) totalized 71.1% of the total cases, whereas 28.9% were classified as probable IFI (N = 22). It was possible to identify the etiological fungal agent in 73 cases (96%). Aspergillus was the most frequent genera detected, but endemic dimorphic fungi represented 14.47% (N = 11) of the total cases. Despite the small number of cases, a high diversity of species were involved in deep fungal infections. This fact has implications for clinical and laboratory diagnosis, and on the therapeutic management of these infections, since different species, even within the same genus, can present diverse patterns of susceptibility to antifungals.
- Isolation of Rasamsonia argillacea species complex in a Cystic Fibrosis Adult Patient - First Case in PortugalPublication . Carvalho, Dinah; Sabino, Raquel; Veríssimo, Cristina; Simões, Helena; Azevedo, Pilar; Marques Lito, Luis; Melo Cristino, JoséObjectives: Cystic fibrosis (CF) is the most common monogenetic autosomal reces sive disease in the human population. An important fungal biota has been described in respiratory secretions of patients suffering from CF being Aspergillus fumigatus and Candida albicans the most common fungi found. We report the isolation, for the first time in Portugal, of the emerging fungal pathogen Rasamsonia argillacea species complex, from a respiratory sample of an adult patient with CF. Material & Methods: A 51-year-old male patient with heterozygous CF due to muatations ΔF508/P205S, is being followed in Cystic Fibrosis Reference Center for about 10 years. In the last years, he has been consistently colonized with Methicilin-susceptible Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus section Fumigati whereby is under chronic suppression therapy with two inhaled antibiotics. Recently, there has been a progressive clinical respiratory functional deterioration. In a periodic evaluation, a miccrobiology control sputum was requested. Sample was cultured, in parallel, for bacteriol ogy and mycology evaluation. Results: Apart from detection of S. aureus and Raoultella ornithinolytica, after 3-5 days of incubation at 37ºC, the cultures showed several cream-coloured colonies, powdery to velvety. Microscopic examination showed hyaline septate hyphae, Penicillium-like conid iophores with rough wall, ovoid to cylindrical phyalides with a narrow neck and cylin drical unicellular smooth-walled microconidia, arranged in unbranched basipetal chains arising from phialides. The isolate was identified as Rasamsonia argillacea species complex based on its mor phology and confirmed by MALDI-TOF mass spectrometry. As no septate hyphae were seen on direct examination, a new sample was requested to exclude extrinsic contamina tion. The second sample was inoculated as previously, confirming the persistent presence of Rasamsonia argillacea species complex in the sputum of this patient. The identification of this isolate was further confirmed by sequencing the internal transcribed spacer (ITS) region of ribosomal DNA, showing 100% homology with sequences deposited on data bases. Antifungal susceptibility testing showed high minimal inhibitory concentrations (MIC) to almost all tested antifungals (posaconazole, voriconazole, amphotericin B) and low MIC to anidulafungin. The patient had no great exacerbation of his respiratory prob lems and the isolated fungus was interpreted as colonization being the patient under more frequent surveillance. Conclusion: Although colonization of the upper respiratory tract in CF patients by R. argillacea species complex has been described as an emerging situation, the role of these fungi in clinical or functional deterioration of the disease remains controversial. Indeed, data about its real prevalence in the CF population are lacking. However, taking into ac count the ability of this species to predominantly affect the lungs, to induce pneumonia and to disseminate to adjacent organs or even to the central nervous system (CNS) in imunocompromised patients, it is essential to promote its accurate identification that is often misidentified as Penicillium spp. or Paecilomyces species. Antifungal susceptibility testing should be performed for epidemiological purposes and to guide therapy, as Rasamsonia spp. usually presents a marked antifungal resistance profile
- Rede Portuguesa de Laboratórios para o Diagnóstico da Gripe: inverno 2013/2014Publication . Guiomar, Raquel; Pechirra, Pedro; Conde, Patrícia; Cristóvão, Paula; Maia, Ana Carina; Silvestre, Maria José; Almeida Santos, Madalena; Sobrinho Simões, Joana; Costa, Maria do Rosário; Pinto, Rita; Guimarães, João Tiago; Ribeiro, Graça; Pereira-Vaz, João; Correia, Lurdes; Fernandes, Paula Luísa; Andrade, Graça; Mota Vieira, Luísa; Cabral Veloso, Rita; Moniz, Raquel; Pereirinha, Tânia; Bruges Armas, Jácome; Pimentel Couto, Ana Rita; Soares, Marta; Melo Cristino, José; Ribeiro, Carlos; Carvalho, Dinah; Barreto, Raquel; Côrte-Real, Rita; Branquinho, Paula; Ramos, Maria Helena; Castro, Ana Paula; Cunha, Mário; Martins, Luís; Almeida, Sofia; Peres, Maria João; Viseu, Regina; Inácio, FilipeA Rede Portuguesa de Laboratórios para o Diagnóstico da Gripe (RPLDG) integra, atualmente, 15 laboratórios maioritariamente hospitalares e é coordenada pelo Laboratório Nacional de Referência para o Vírus da Gripe (LNRVG) do Departamento de Doenças Infecciosas do Instituto Nacional de Saúde Doutor Ricardo Jorge, I.P. A RPLDG realiza o diagnóstico laboratorial do vírus da gripe assim como de outros vírus respiratórios, permitindo um conhecimento mais preciso da etiologia das infeções respiratórias, particularmente em casos hospitalizados de infeção respiratória aguda grave, constituindo um complemento valioso para o PNVG. Os casos de SG provenientes de emergências hospitalares e casos de Infecção Respiratória Aguda Grave, incluindo casos com internamento em unidade de cuidados intensivos, foram notificados pelos laboratórios da Rede ao LNRVG. Dos 15 laboratórios da Rede, 13 notificaram casos de doença respiratória durante a época de 2013/2014. Os dados recolhidos foram inseridos em suporte informático tendo as bases de dados sido agregadas numa base de dados comum submetida a um processo de validação de congruência de dados. Os dados analisados correspondem ao período que decorreu entre a semana 38 de 2013 e a semana 21 de 2014. Foram notificados pelos Laboratórios da Rede um total de 3790 casos de infeção respiratória. O maior número de notificações foi observado no mês de janeiro e fevereiro (semanas 2/2014 a 8/2014), com um pico de ocorrência na semana 4/2014 com a notificação de 454 casos de infeção respiratória. O vírus da gripe foi detetado em 822 casos de infeção respiratória. O vírus influenza A foi identificado em 807 (98,2%) dos casos positivos, destes 403 (49,0%) pertencem ao subtipo A(H1)pdm09, 98 (12,0%) ao subtipo A(H3) e 306 (37,0%) vírus influenza A não foram subtipados. O vírus influenza B foi detetado em 14 (2,0%) casos. Foi identificada 1 infecção mista por vírus influenza A(H1)pdm09 e A(H3) (0,1%). A maior percentagem de casos de gripe foi observada em indivíduos entre os 15 e os 64 anos sendo o vírus influenza A(H1)pdm09 o predominantemente detetado. Nas crianças com menos de 4 anos o vírus influenza foi detetado numa proporção reduzida, apenas em 8,8% dos casos analisados laboratorialmente, sendo o agente mais detetado neste grupo etário, o vírus sincicial respiratório (dados não mostrados). A Rede Portuguesa de Laboratórios para o Diagnóstico da Gripe permitiu a deteção dos vírus da gripe em meio hospitalar, incluindo doentes em internamento e UCI. Os vírus influenza A foram predominantes e detetados em maior percentagem nos jovens e adultos.
- Severe acute respiratory infections in the 2012/2013 season studied by the Portuguese Laboratory Network for Influenza DiagnosisPublication . Guiomar, Raquel; Pechirra, Pedro; Conde, Patrícia; Cristóvão, Paula; Silvestre, Maria José; Almeida Santos, Madalena; Sobrinho Simões, Joana; Costa, Maria do Rosário; Amaral, Susana; Guimarães, João Tiago; Ribeiro, Graça; Correia, Lurdes; Fernandes, Aida; Milho, Luís; Fernandes, Paula Luísa; Andrade, Graça; Mota Vieira, Luísa; Cabral, Rita; Moniz, Raquel; Pereirinha, Tania; Bruges Armas, Jacome; Pimentel Couto, Ana Rita; Soares, Marta; Melo Cristino, José; Carvalho, Dinah; Ribeiro, Carlos; Barreto, Rosário; Côrte-Real, Rita; Branquinho, Paula; Ramos, Maria Helena; Castro, Ana Paula; Caldeira, Filomena; Maurílio, Manuel; Cunha, Mário; Ornelas, Carmo; Almeida, SofiaDuring the 2009/10 influenza pandemic, a network of 14 laboratories located in the main reference hospitals from Portugal mainland, Madeira and Azores was established for the diagnosis of the new influenza A(H1N1)2009 pandemic strain. Since then, the network performs laboratory diagnosis of influenza as well as other respiratory pathogens, thus contributing to the laboratory diagnosis of respiratory disease in Portugal. This network is a valuable complement of the National Influenza Surveillance Programme (mainly based on primary healthcare units), enabling a more accurate knowledge of the aetiology of the severe respiratory infections, especially in hospitalized cases. The present study describes the severe acute respiratory infections, in the 2012/2013 season, diagnosed by the laboratory network. From the 14 laboratories, 11 reported cases of respiratory disease during 2012/2013 season. The laboratory network performs diagnosis of influenza A and B viruses and other respiratory agents by PCR based methods, also enabling the detection of mixed infections. All 14 laboratories perform the detection of influenza A(H1)pdm09, 4 perform the influenza A(H1) seasonal and A(H3) subtyping, and 10 participants also detect influenza B. Eight laboratories implemented methodologies for the detection of other infectious agents associated with respiratory disease. The antigenic characterization of 8 isolated viruses [3 A(H1)pdm09 and 5 B/Yamagata] was performed at the National Influenza Reference Laboratory. The genetic analysis of the HA1 subunit of the haemagglutinin gene was performed in 17 viruses [7 A(H1)pdm09, 1 A(H3) and 9 B/Yamagata]. Twenty nine A(H1)pdm09 and 5 B/Yamagata were tested for antiviral susceptibility [PCR(NA)-H275Y and/or MUNANA phenotypic assays for oseltamivir and zanamivir]. The 11 laboratories reported a total of 1470 respiratory disease cases, from week 39/2012 to 21/2013 [peak of 205 (13.9%) cases during week 10/2013]. Influenza was identified in 504 cases. Influenza A was detected in 352 (70.0%) cases: 297 (59.0%) cases were A(H1)pdm09, 48 (10.0%) cases were not subtyped, and 7 (1.0%) cases were A(H3). Influenza B was identified in 152 (30%) of the influenza cases. During the 2012/2013 season, 311 (21.2%) reported cases were hospitalized in intensive care units (ICU), the majority of them had between 50-54 years (34; 10.9%), followed by the age groups 45-49 and 55-59 years old (28; 9.0% each). The causal agent was identified in 160 (51.4%) ICU cases. Influenza was identified in 120 (38.5%) patients, other respiratory agents were detected in 40 (12.8%), within these, multiple infections were present in 18 (5.7%). Bacteria were identified in 31 (10.0%) cases mainly associated with RSV and hRV. Among ICU influenza cases, the most detected virus was A(H1)pdm09 (82; 62.0%). However, cases of A(H3) (3; 2.0%), A unsubtyped (8; 7.0%) and B (27; 23.0%) were also detected. As expected, the highest number of ICU influenza positive cases was detected in week 10/2013 (18; 15.0%), coincident with the highest number of influenza cases during all season. ICU flu cases were detected predominantly in individuals between 50-54 years (18; 15.0%). From the ICU reported cases, 6 (1.9%) died. The influenza A(H1)pdm09 virus was detected in 2 man between 50-59 years old from these 6 fatal outcomes. The isolated influenza A viruses were similar to the 2012/2013 vaccine strains. The influenza B/Yamagata viruses showed a greater antigenic and genetic variability. The Portuguese Laboratory Network for Influenza Diagnosis plays a major role in the diagnosis of acute respiratory infections in Portugal, providing a more accurate knowledge of the respiratory agents involved. During the 2012/2013 season, the influenza A(H1)pdm09 virus predominated in co-circulation with influenza B virus. The A(H1)pdm09 virus was the responsible for the majority of the flu cases admitted in the ICU and may have been the cause of death in two cases. Bacterial and other viral agents have been identified in some of the severe cases reported. The majority of the characterized influenza viruses were similar to the vaccine strains and none of the virus showed reduced susceptibility to oseltamivir or zanamivir.
- Subcutaneous phaeohyphomycosis due to Phaeoacremonium parasiticum in a patient with Chronic Granulomatous DiseasePublication . Carvalho, Dinah; Sabino, Raquel; Veríssimo, Cristina; Simões, Helena; Lopes Silva, Susana; Marques, Tiago; Janeiro, João; Marques Lito, Luis; Melo Cristino, JoséObjectives: Phaeoacremonium parasiticum is a ubiquitous dematiaceous mold that rarely causes infection in humans. Its spectrum of disease ranges from subcutaneous infections to disseminated disease. The majority of those reported few cases involve immunocompromised patients. Chronic granulomatous disease (CGD) is an inherited disorder affecting nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. CGD patients are susceptible to a broad spectrum of opportunistic infections, being fungal infections a major determinant of survival. We report a case of Subcutaneous phaeohyphomycosis due to P. parasiticum in a young adult with CGD. Material and methods: A 28-year-old male patient with autossomic recessive CGD, due to mutations in CYBA, is followed in Primary Immunodeficiency Center since childhood. He has been under prophylaxis with itraconazole, cotrimoxazole and IFNg and tapering oral steroids, started for granulomatous colitits 2 years earlier. In a regular visit to the clinical center, he complained of persistent pain on his left leg with no history of recent injury in the affected area and without external inflammatory signs. An ultrasound was performed after 2 weeks revealing a heterogeneous liquid of slightly irregular contour in the sinus of the left anterior tibial muscle with heterogeneity of the adjacent muscle. This piomiositis collection was punctured, under ultrasound control, with drainage of about 7cm3 of hemato-purulent exsudate that was promptly processed for bacteriology (aerobic and anaerobic) and mycology studies. Results: Cultures for bacteria were sterile. Culture on Sabouraud was positive after 5 days of incubation, showing slow growing and initially white colonies. Microscopic examination showed hyaline, septate mycelium with long, thin conidiophores and curved, aseptate conidia.The fungus was initially identified as an Acremonium sp. and was sent to the Mycology Reference Laboratory for molecular identification and antifungal susceptibility testing. Its identification was performed by sequencing the internal transcribed spacer (ITS) region of ribosomal DNA, being the isolate identified as Phaeoacremonium parasiticum (100% homology). After 3 weeks incubation, coloration of the colonies emerged, becoming greyish black upon subculture, with velvety texture and black reverse. Microscopically, pigmented hyphae with tapering, funnel-shaped phialides were observed, and conidia were hyaline and oblong, forming clusters at the tip of the phialides. At this point, macroscopic and microscopic morphology was consistent with Phaeoacremonium species. Susceptibility pattern showed low minimal inhibitory concentrations (MIC) to posaconazole and voriconazole, and higher MIC values to anidulafungin and amphotericin B. The patient has improved under voriconazol therapy (200 mg; bid) Conclusions: Phaeoacremonium parasiticum is an uncommon infection and its appropriate identification is often difficult because morphologically, the genus Phaeoacremonium show morphological features resembling both Acremonium and Phialophora genera. Molecular identification is determinant to confirm morphology, as many species have indistinguishable characteristics that may lead to incorrect antifungal options. Also, susceptibility testing should be done for these so rare fungi as optimal treatment has not yet been clearly defined.
- Trends on Aspergillus Epidemiology-Perspectives from a National Reference Laboratory Surveillance ProgramPublication . Sabino, Raquel; Gonçalves, Paulo; Martins Melo, Aryse; Simões, Daniela; Oliveira, Mariana; Francisco, Mariana; Viegas, Carla; Carvalho, Dinah; Martins, Carlos; Ferreira, Teresa; Toscano, Cristina; Simões, Helena; Veríssimo, CristinaIdentification of Aspergillus to species level is important since sibling species may display variable susceptibilities to multiple antifungal drugs and also because correct identification contributes to improve the knowledge of epidemiological studies. Two retrospective laboratory studies were conducted on Aspergillus surveillance at the Portuguese National Mycology Reference Laboratory. The first, covering the period 2017-2018, aimed to study the molecular epidemiology of 256 Aspergillus isolates obtained from patients with respiratory, subcutaneous, or systemic infections and from environmental samples. The second, using our entire collection of clinical and environmental A. fumigatus isolates (N = 337), collected between 2012 and 2019, aimed to determine the frequency of azole-resistant A. fumigatus isolates. Aspergillus fumigatus sensu stricto was the most frequent species in both clinical and environmental samples. Overall, and considering all Aspergillus sections identified, a high frequency of cryptic species was detected, based on beta-tubulin or calmodulin sequencing (37% in clinical and 51% in environmental isolates). Regarding all Fumigati isolates recovered from 2012-2019, the frequency of cryptic species was 5.3% (18/337), with the identification of A. felis (complex), A. lentulus, A. udagawae, A. hiratsukae, and A. oerlinghauensis. To determine the frequency of azole resistance of A. fumigatus, isolates were screened for azole resistance using azole-agars, and 53 possible resistant isolates were tested by the CLSI microdilution reference method. Nine A. fumigatus sensu stricto and six Fumigati cryptic isolates showed high minimal inhibitory concentrations to itraconazole, voriconazole, and/or posaconazole. Real-time PCR to detect cyp51A mutations and sequencing of cyp51A gene and its promoter were performed. The overall frequency of resistance to azoles in A. fumigatus sensu stricto was 3.0%. With this retrospective analysis, we were able to detect one azole-resistant G54R mutant A. fumigatus environmental isolate, collected in 2015. The TR34/L98H mutation, linked to environmental transmission route of azole resistance, was the most frequently detected mutation (N = 4; 1.4%). Our findings underline the demand for correct identification and susceptibility testing of Aspergillus isolates.