Cryptococcus neoformans VNII as the Main Cause of Cryptococcosis in Domestic Cats from Rio de Janeiro, Brazil.

Cryptococcosis is a systemic fungal disease acquired from contaminated environments with propagules of the basidiomycetous yeasts of the Cryptococcus neoformans and C. gattii species complexes. The C. neoformans species complex classically comprises four major molecular types (VNI, VNII, VNIII, and VNIV), and the C. gattii species complex comprises another four (VGI, VGII, VGIII, and VGIV) and the newly identified molecular type VGV. These major molecular types differ in their epidemiological and ecological features, clinical presentations, and therapeutic outcomes. Generally, the most common isolated types are VNI, VGI, and VGII. The epidemiological profile of cryptococcosis in domestic cats is poorly studied and cats can be the sentinels for human infections. Therefore, the present study aimed to determine the molecular characterization of Cryptococcus spp. isolated from domestic cats and their dwellings in the metropolitan area of Rio de Janeiro, Brazil. A total of 36 Cryptococcus spp. strains, both clinical and environmental, from 19 cats were subtyped using multilocus sequence typing (MLST). The ploidy was identified using flow cytometry and the mating type was determined through amplification with specific pheromone primers. All strains were mating type alpha and 6/36 were diploid (all VNII). Most isolates (63.88%) were identified as VNII, a rare molecular type, leading to the consideration that this genotype is more likely related to skin lesions, since there was a high percentage (68.75%) of cats with skin lesions, which is also considered rare. Further studies regarding the molecular epidemiology of cryptococcosis in felines are still needed to clarify the reason for the large proportion of the rare molecular type VNII causing infections in cats.

Cryptococcosis due to Cryptococcus gattii VGII in southeast Brazil: The One Health approach revealing a possible role for domestic cats.

Two cats infected by C. gattii, presented lesions on the nasal region and respiratory signs. Strains were typed as molecular type VGII, mating type alpha, MLST subtypes ST442 and ST185. Since Rio de Janeiro is known as an endemic area for C. neoformans VNI, these cases might be a warning for a possible emergence of C. gattii VGII in southeast Brazil.

Cell-free antigens of Sporothrix brasiliensis: antigenic diversity and application in an immunoblot assay.

Sporotrichosis is a subcutaneous mycosis diagnosed by isolation of the fungus in culture. Serological tests for help in diagnosis in general do not use purified or recombinant antigens, because there is a paucity of described immunoreactive proteins, especially for the new described Sporothrix species, such as Sporothrix brasiliensis. This study aims to characterise antigens from S. brasiliensis and verify their application in serodiagnosis of sporotrichosis. An immunoblot assay allied with computer-based analysis was used to identify putative antigenic molecules in a cell-free extracts of both morphological phases of this fungus, and to delineate antigenic polymorphism among seven S. brasiliensis isolates and one S. schenckii Brazilian strain. The mycelial and yeast phase of the fungus originated 14 and 23 reactive bands, respectively, which were variable in intensity. An 85 kDa antigen, verified in the yeast phase of the fungus, was observed in all strains used and the immunodominant protein was identified. This protein, however, cross-react with serum samples from patients infected with other pathogens. The results show that the S. brasiliensis cell-free antigen extract is a single and inexpensive source of antigens, and can be applied on the sporotrichosis serodiagnosis.

Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil.

An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD) technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20% similarity to each primer and 60% when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8%) and the oral cavities (66%) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.

Molecular characterisation of Sporothrix schenckii isolates from humans and cats involved in the sporotrichosis epidemic in Rio de Janeiro, Brazil

An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD) technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20 percent similarity to each primer and 60 percent when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8 percent) and the oral cavities (66 percent) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.

[Importance of culture of urine in the diagnosis of AIDS associated cryptococcosis]. / Criptococose associada à Aids: a importância do cultivo da urina no seu diagnóstico.

In this retrospective study we analyzed 70 HIV patients with cryptococcosis over a 16-years period. The specimens with the best positivity were CSF (97.8%) followed by the culture of urine sediment (86.7%) and blood culture (58.8%). We conclude that the urine could be a useful tool for the diagnosis of cryptococcosis.

Criptococose associada à Aids: a importância do cultivo da urina no seu diagnóstico / Importance of culture of urine in the diagnosis of AIDS associated cryptococcosis

Neste estudo retrospectivo analisamos 70 pacientes HIV positivos com criptococose em um período de 16 anos. Os espécimes com melhor rendimento diagnóstico foram o LCR (97,8 por cento), seguido do cultivo do sedimento urinário (86,7 por cento) e sangue (58,8 por cento). Concluímos que a urina pode ser uma ferramenta útil para o diagnostico da criptococose.

Histoplasmosis in a Brazilian center: clinical forms and laboratory tests.

Histoplasmosis, caused by the dimorphic fungus Histoplasma capsulatum, is endemic in many regions of the Americas, Asia and Africa. It has a wide spectrum of clinical manifestations, from asymptomatic infection to severe disseminated disease. A retrospective study was carried out to describe the clinical forms and assess the clinical significance of the laboratory diagnostic tests of patients with histoplasmosis during the period of July 1987 to December 2003 at Instituto de Pesquisa Clínica Evandro Chagas/ FIOCRUZ, RJ, Brazil. Seventy-four patients were included. Forty-nine percent of the cases (n = 36) occurred in HIV positive patients who presented with disseminated disease. The remaining 38 cases were classified in different clinical forms. Histoplasma capsulatum was isolated from 69.5% of the clinical specimens sent to culture. Immunodiffusion and immunoblot were positive in 72.6% and 100% of the performed tests, respectively. Histopathologic findings suggestive of H. capsulatum were found in 63.2% of the performed exams. Serology had a lower proportion of positivity amongst AIDS patients, when compared with HIV negative patients (X2 = 6.65; p lower than 0.008). Statistical differences between AIDS and non-AIDS patients were not observed with culture and histopathology. The specific role of each test varies according to the clinical form. Physicians need to know the value and limitations of the available diagnostic tests, but before that, they have to think about histoplasmosis and consider this clinical entity in their differential diagnosis.

Primary conjunctival sporotrichosis: two cases from a zoonotic epidemic in Rio de Janeiro, Brazil.

PURPOSE: To describe Sporothrix schenckii conjunctivitis in 2 owners of cats with sporotrichosis. METHODS: Small case series and literature review. RESULTS: Two women had been caring for their pet cats with sporotrichosis for 2 months but did not recall any traumatic injury such as scratches or bites. Each presented a conjunctival granulomatous lesion measuring 4 to 5 mm accompanied by local hyperemia, secretion, and edema, in addition to painful facial subcutaneous nodes and regional lymph node enlargement. Pyogenic material was collected from the conjunctival sac and from cutaneous lesions on the cats. In both the patients and their respective cats, fungal colonies were isolated and identified as S. schenckii. Treatment with oral itraconazole 100 mg/d for 3 months resulted in complete healing of lesions in both patients. Patients remained clinically cured 15 months after end of treatment. CONCLUSION: Sporotrichosis is presently occurring as an emerging zoonosis in Rio de Janeiro, and some unusual clinical forms have been diagnosed in humans. The cases reported here suggest atraumatic exposure to cats infected by S. schenckii.

Positive Montenegro skin test among patients with sporotrichosis in Rio De Janeiro.

We studied 52 patients with sporotrichosis confirmed by isolation of Sporothrix schenckii and reactivity to the Montenegro skin test (MST) during an ongoing outbreak of this mycosis in Rio de Janeiro. The objective was to emphasize the importance of parasitological confirmation and the possibility of incorrect diagnosis based on the lesion's appearance, epidemiological information, and immunological tests. The antigen used for the MST was conserved in either thimerosal 1:10,000 (group 1) or 0.4% phenol (group 2). Nineteen patients (39%) in group 1 and seven (12%) in group 2 presented an induration>or=10 mm (p<0.001). Sera from three patients (6.7%) reacted to indirect immunofluorescence (IIF) for leishmaniasis, while sera from 10 patients (22.2%) reacted to enzyme-linked immunosorbent assay (ELISA). Fifteen patients (28.8%) presented up to two lesions, with a predominance of ulcers. Forty-four patients (84.6%) were treated with itraconazole. In the differential diagnosis between sporotrichosis and cutaneous leishmaniasis, the possibility of co-infection, allergy to the reagent diluent, and cross-reactions should be further investigated, especially in regions with limited laboratory facilities.

Utilidade do coágulo sangüíneo para o isolamento de Sporothrix schenckii de gatos naturalmente infectados / Sporothrix schenckii isolation from blood clot of naturally infected cats

O diagnóstico de esporotricose disseminada costuma ser obtido através da necrópsia e o isolamento de Sporothrix schenckii do sangue é raro. Fungemia foi demonstrada in vivo através do isolamento do S. schenckii do sangue periférico de 13 (n=38; 34,2%) gatos com esporotricose naturalmente adquirida. A coinfecção com FIV e com FeLV encontradas, respectivamente, em 6 (n=34; 17,6%) casos e 1 (n=34; 2,9%), aparentemente não alterou a freqüência do isolamento de S. schenckii do sangue periférico. Comparando estes resultados aos dos hemocultivos realizados simultaneamente houve concordância de 84,2%. Assim, propomos o cultivo do coágulo como um método alternativo prático, eficiente e econômico para o diagnóstico de esporotricose disseminada em gatos in vivo.

Cat-transmitted sporotrichosis epidemic in Rio de Janeiro, Brazil: description of a series of cases.

Sporotrichosis is the most common subcutaneous mycosis in South America. Classic infection is associated with traumatic inoculation of soil, vegetables, and organic matter contaminated with Sporothrix schenckii. Zoonotic transmission has been described in isolated cases or in small outbreaks. Since 1998, we have been observing an increasing number of cases of sporotrichosis in persons from the city of Rio de Janeiro, Brazil, and surroundings. From 1998 to 2001, 178 cases of culture-proven sporotrichosis had been diagnosed. Female patients predominated, and the median age was 39 years. The most frequent clinical presentation was lymphocutaneous disease. Of the 178 patients, 156 reported domiciliary or professional contact with cats with sporotrichosis, and 97 of these patients had a history of receipt of cat scratch or bite. The patients received itraconazole as first-line treatment. This study suggests that feline transmission of sporotrichosis was associated with a large and long-lasting outbreak of the disease in Rio de Janeiro.

Sporotrichosis with widespread cutaneous lesions: report of 24 cases related to transmission by domestic cats in Rio de Janeiro, Brazil.

BACKGROUND: Sporotrichosis most commonly presents as a localized lymphocutaneous infection following traumatic inoculation of soil, vegetables or organic substrates contaminated with the dimorphic fungus Sporothrix schenckii. Cases of widespread cutaneous lesions are rare. There have been isolated reports of household outbreaks of sporotrichosis involving cats and humans. METHODS: We report 24 cases of culture-proven sporotrichosis presenting with widespread cutaneous lesions. RESULTS: These 24 cases are part of an epidemic currently occurring in Rio de Janeiro. All patients reported contact with cats with sporotrichosis and 17 reported a history of a scratch or bite. Clinical manifestations included fixed lesions at multiple anatomical sites, and fixed lesions associated with the lymphocutaneous, bilateral lymphocutaneous and mucosal forms of the disease. Two patients were alcoholics and one patient was diabetic, while the remaining patients did not present any immunosuppressing condition. All patients responded to treatment with itraconazole. CONCLUSIONS: The domestic cat has played an important role in the transmission of sporotrichosis in Rio de Janeiro and seems to have contributed to this unusual clinical manifestation.

Sporothrix schenckii isolated from domestic cats with and without sporotrichosis in Rio de Janeiro, Brazil.

A total of 148 cats with a clinical and mycologic diagnosis of sporotrichosis and 84 apparently healthy cats with domiciliary contact with the affected animals were studied. Sporothrix schenckii was isolated from 148 (n = 148; 100%) clinical samples of cutaneous lesion (biopsy, swab or aspiration of purulent secretion), 47 (n = 71; 66.2%) nasal cavities, 33 (n = 79; 41.8%) oral cavities, and 15 (n = 38; 39.5%) nails of cats with sporotrichosis. Histopathological examination revealed yeast-like structures in 50 (n = 70; 71.4%) of the biopsies studied. S. schenckii was isolated from the blood culture of one cat (n = 5, 20%) with the disseminated cutaneous form of the disease. On another occasion, the fungus was isolated from the testis of one (n = 7; 14.3%) of the animals submitted to sterilization. In the group of cats with domiciliary contacts, 3 (n = 84; 3.57%) oral swabs showed positive cultures. Isolation of S. schenckii from different clinical specimens during both the clinical and preclinical phase reinforces the zoonotic potential of feline sporotrichosis.

Sporotrichosis: an emergent zoonosis in Rio de Janeiro

During the period from 1987 to 1998, 13 cases of human sporotrichosis were recorded at the Research Center Evandro Chagas Hospital (CPqHEC) in Rio de Janeiro. Two of these patients related scratch by a sick cat. During the subsequent period from July 1998 to July 2000, 66 human, 117 cats and 7 dogs with sporotrichosis were diagnosed at the CPqHEC. Fifty-two humans (78.8 percent) reported contact with cats with sporotrichosis, and 31 (47 percent) of them reporting a history of a scratch or bite. This epidemic, unprecedented in the literature, involving cats, dogs and human beings may have started insidiously before 1998