Cintia de Moraes Borba

Comparison of virulence of different Sporothrix schenckii clinical isolates using experimental murine model

The virulence of two strains of Sporothrix schenckii isolated from patients with lymphocutaneous or disseminated sporotrichosis were examined in BALB/c mice (Group 1 and 2, respectively). The mice were inoculated subcutaneously into the left hind footpad with 4 x 10(6) S. schenckii yeast cells in order to evaluate (i) the development of cutaneous lesions, (ii) signs of inactivity, (iii) weight loss, (iv) survival rates, (v) number of viable yeast cells in the lungs and spleen, (vi) splenic index, (vii) extent of organ lesions, and (viii) immunological responses. Comparison of the two groups showed more severe disease in Group 2 mice that developed significant weight and hair loss associated with inactivity and left hind footpad lesions that extended close to the testicular area. The histopathology and large number of viable microorganisms isolated from the spleen confirmed the higher invasive ability of this strain. Moreover, a decrease of an in vitro specific lymphoproliferative response and IFN-gamma production were observed over time in Group 2 mice. As a result, at the end of the experiment, the S. schenckii-antigen (Ss-Ag) response was considered negative with a stimulation index (SI) = 2. In contrast, Group 1 mice presented a positive response to Ss-Ag (SI = 14.1). These results confirm the existence of different virulence profiles in S. schenckii strains. In addition, the use of subcutaneous inoculation as a suitable route for verification of the pathogenicity of this fungus in the murine model was confirmed.

Viability and morphological alterations of Paracoccidioides brasiliensis strains preserved under mineral oil for long periods of time

We evaluated the survival and the morphological alteration of 70 strains of Paracoccidioides brasiliensis maintained in the Fungal Culture Collection of Institute Oswaldo Cruz and initially preserved by successive subculturing and later under mineral oil at room temperature from 1923 to 1992. Of the 70 strains preserved under mineral oil, 18 (26%) continued to be viable. The mycelia of the 18 viable strains and the mycelia of four representatives of the non‐viable strains presented transitional micromorphology under mineral oil. Macroscopy and microscopy of the first subcultures of five of the seven strains, taken as random samples from the 18 viable strains and plated onto peptone‐yeast extract‐glucose agar, glucose‐glycine‐yeast extract agar and brain‐heart infusion agar media at room temperature, revealed that all samples appeared to be in a transition phase (Y⇄M). These five strains were unable to grow and complete thermal dimorphism at a temperature of 37°C. Only two strains were able to complete the entire dimorphic process. The period of preservation of these strains under oil was relatively short, i.e. 9 and 10 years. The results demonstrate that less widely spaced subculturing and more appropriate culture and environmental conditions are needed to preserve P. brasiliensis strains under oil.

Neglected and emerging fungal infections: review of hyalohyphomycosis by Paecilomyces lilacinus focusing in disease burden, in vitro antifungal susceptibility and management

Paecilomyces lilacinus is an emerging pathogenic fungus that can cause different clinical manifestations ranging from cutaneous and sub-cutaneous infections to severe oculomycosis. This review discusses infections caused by P. lilacinus, as well as their symptoms and correlates of immune responses, morphological characteristics of the fungus, therapies, in vitro susceptibility tests, laboratory diagnosis and the experimental models available.

Long-time survival and morphological stability of preserved Sporothrix schenckii strains

Summary. The survival and stability of morphological traits of 33 Sporothrix schenckii strains were evaluated. Strain subcultures were maintained under mineral oil for different periods of time lasting as long as 41 years, or in sterile distilled water for as long as 23 years. Of the 33 strains preserved under mineral oil, 28 (85%) maintained viability and unchanged macro‐ and microscopic characteristics. All of the 4 strains maintained in distilled water also remained viable, with no morphological changes. The results confirm the efficiency of the methods used to preserve strains of the pathogenous and anamorphous fungus Sp. schenckii, especially in laboratories of limited resources.

Construção do conhecimento em biossegurança: uma revisão da produção acadêmica nacional na área de saúde (1989-2009)

Este artigo refere-se a uma revisao bibliografica realizada atraves do portal da Biblioteca Virtual de Saude com objetivo de identificar os artigos cientificos relacionados a biosseguranca, publicados em periodicos nacionais, visando a conhecer inicialmente de que forma esse tema vem sendo abordado no Brasil e como esta inserido no nexo trabalho e na educacao do profissional da area de saude. Foram identificados inicialmente 126 artigos publicados durante o periodo de 1989 a outubro de 2009, em periodicos nacionais na area de saude. Do total de artigos, 46 estavam relacionados diretamente a tematica educacao, mas apenas oito abordaram a pratica educacional em biosseguranca. Identificou-se tambem que a regiao Sudeste concentrou o maior numero de publicacao (77 artigos) acerca do tema biosseguranca. Apesar da abrangencia dos artigos analisados, ainda existe muito que pesquisar sobre o assunto, de forma a ampliar o debate sobre a educacao profissional para o setor saude, onde o trabalhador seja considerado como sujeito da aprendizagem, capaz de ter uma postura critica-reflexiva do seu ambiente ocupacional.Este artigo refere-se a uma revisao bibliografica realizada atraves do portal da Biblioteca Virtual de Saude com objetivo de identificar os artigos cientificos relacionados a biosseguranca, publicados em periodicos nacionais, visando a conhecer inicialmente de que forma esse tema vem sendo abordado no Brasil e como esta inserido no nexo trabalho e na educacao do profissional da area de saude. Foram identificados inicialmente 126 artigos publicados durante o periodo de 1989 a outubro de 2009, em periodicos nacionais na area de saude. Do total de artigos, 46 estavam relacionados diretamente a tematica educacao, mas apenas oito abordaram a pratica educacional em biosseguranca. Identificou-se tambem que a regiao Sudeste concentrou o maior numero de publicacao (77 artigos) acerca do tema biosseguranca. Apesar da abrangencia dos artigos analisados, ainda existe muito que pesquisar sobre o assunto, de forma a ampliar o debate sobre a educacao profissional para o setor saude, onde o trabalhador seja considerado como sujeito da aprendizagem, capaz de ter uma postura critica-reflexiva do seu ambiente ocupacional.

Inoculation experimental animals withParacoccidioides brasiliensis strains: An attempt to reestablish the dimorphic process and variation in pathogenicity as a function of time of preservation under mineral oil

In an attempt to reestablish the dimorphic process in strains ofParacoccidioides brasiliensis in the transition phase (Y ⇄ M) and to reisolate them, five strains in the transitional phase due to the long time of preservation under mineral oil and two strains in the yeast-like phase were inoculated into male albino rats. The animals were then studied for the presence of paracoccidioidomycotic granulomata. Of the seven strains inoculated, five caused granulomatous nodules in several organs of the animals and only two of these five strains, which had been preserved for the shortest period of time (9 years) were reisolated in culture. Two strains were unable to provoke infection, with no lesions detected in any organ. It is assumed that the long period of time during which the strains were left under oil favored the alteration of celt wall contents, leading to differences in pathogenicity.

Viability and molecular authentication of Coccidioides immitis strains from culture collection of the Instituto Oswaldo Cruz, Rio de Janeiro, Brazil

Twenty Coccidioides immitis strains were evaluated. Only 5 of the 20 strains kept under mineral oil maintained their viability while all 5 subcultures preserved in water remained viable and none of the 13 subcultures kept in soil were viable. A 519 bp PCR product from the csa gene confirmed the identity of the strains.

Variants of Sporothrix schenckii with attenuated virulence for mice

Strains of Sporothrix schenckii preserved under mineral oil were examined for virulence in BALB/c mice. The mice were inoculated with S. schenckii conidia and development of cutaneous lesions, signs of inactivity, weight loss, survival rates, number of viable yeast cells in lung and spleen, splenomegaly and organ lesions were evaluated. After intravenous injection of 7.5 x 10(6) conidia, two of five S. schenckii strains were unable to induce systemic disease and to kill the mice, only producing cord-like lesions on the tail that regressed with mouse maturation. Very small numbers of viable cells isolated from the spleen confirmed the lower invasive ability of these strains when compared with other strains studied here. These results suggest a relationship between the attenuation of virulence and the storage method under mineral oil after long periods of time.

In-vitro study of the host–parasite interactions between mouse macrophages and the opportunistic fungus Paecilomyces lilacinus

Paecilomyces lilacinus (Thom) Samson 1974 is a cosmopolitan, filamentous, asexual and hyaline fungus. Although detected mainly as a saprophyte, in soil and decaying vegetables, it has also been found parasitising insects and nematodes — sparking interest in its potential use as a biocontrol agent (Garbino et al., 2002) — and, increasingly, infecting humans (Motswaledi et al., 2009). In culture at 25uC, Pae. lilacinus grows fast, producing a colony, consisting of a basal felt with a floccose over-growth of aerial mycelium, that is initially white but becoming vinaceous as the fungus sporulates (Samson, 1974). The vegetative hyphae, which are smooth-walled, hyaline and with conidiophores, occasionally form loose, thick-walled yellow or purple synnemata (Samson, 1974). Each synnema consists of verticillate branches with whorls of two, three or four phialides. Each phialide has a swollen basal part that tapers into a thin distinct neck, which is about 1 mm wide. Conidia, which form divergent, sometimes tangled chains, are ellipsoidal to fusiform, smooth-walled to slightly roughened, hyaline and purple in mass, differing from those of the genus Penicillium; chlamydospores are absent (Samson, 1974). Because a hyaline mycelium develops in the host tissue, human infections with Paecilomyces spp. (like those with species in several other genera, including Acremonium, Fusarium, Scopulariopsis and Beauveria) are classed as cases of hialohyphomycosis (Vartivarian et al., 1993; Matsumoto et al., 1994). Although most human infections produce cutaneous or subcutaneous disease or oculomycosis, onychomycosis, vaginitis, osteomyelitis, generalized infection, fungaemia, and respiratory-tract infections have also been described (Pastor and Guarro, 2006). Localized infections of the skin and subcutaneous nodules tend to develop in immunocompetent individuals whereas disseminated lesions occur in the immunocompromised (Walsh et al., 2004). The main routes of fungal entrance are the respiratory tract, the skin and surgical implants, such as prosthetic devices. Although there are very few reports of human infection with Pae. lilacinus in Brazil, this may simply reflect a lack of general awareness of the problem posed by the fungus to human health (Itin et al., 1998; Hofling-Lima et al., 2005). Currently, no effective treatment has been established for hialohyphomycosis caused by Pae. lilacinus, and the amphotericin B, flucytosine, fluconazole, miconazole or itraconazole that have been used, in some cases of skin, subcutaneous or ocular infection, have often given unsatisfactory results. Encouragingly, however, some of the socalled ‘new azoles’, such as posaconazole, ravuconazol and voriconazole, have recently demonstrated good activity against Pae. lilacinus in vitro (Pastor and Guarro, 2006; Yaun et al., 2008, Rodriguez et al., 2010). Constrained skin lesions may be treated by debridement and, whenever possible, by the removal of any foreign bodies that might be the source of the infection. Previous studies on hialohyphomycosis caused by Pae. lilacinus have focused on clinical manifestations, treatment, prognosis Annals of Tropical Medicine & Parasitology, Vol. 104, No. 6, 529–534 (2010)

Fusarium species in sandy soil from Ipanema Beach, Rio de Janeiro, Brazil

The presence of the genus Fusarium was investigated in sandy soil from Ipanema Beach, Rio de Janeiro, RJ, Brazil during a period of one year in order to determine the diversity, and seasonal distribution of species. A total of 123 strains were isolated, for a total of 34 different species, with higher incidence in spring and winter. The species detected in greater numbers during the various seasons of the year were, in decreasing order, F. solani var. solani, F. oxysporum var. oxysporum and F. sacchari var. subglutinans, described in the literature (GUARRO and GENÉ 1992) as infectious agents for humans and animals. Few species were detected in summer and fall, probably due to the higher rainfull occurring during these seasons.