Sandro Antonio Pereira

Phylogenetic Analysis Reveals a High Prevalence of Sporothrix brasiliensis in Feline Sporotrichosis Outbreaks

Sporothrix schenckii, previously assumed to be the sole agent of human and animal sporotrichosis, is in fact a species complex. Recently recognized taxa include S. brasiliensis, S. globosa, S. mexicana, and S. luriei, in addition to S. schenckii sensu stricto. Over the last decades, large epidemics of sporotrichosis occurred in Brazil due to zoonotic transmission, and cats were pointed out as key susceptible hosts. In order to understand the eco-epidemiology of feline sporotrichosis and its role in human sporotrichosis a survey was conducted among symptomatic cats. Prevalence and phylogenetic relationships among feline Sporothrix species were investigated by reconstructing their phylogenetic origin using the calmodulin (CAL) and the translation elongation factor-1 alpha (EF1α) loci in strains originated from Rio de Janeiro (RJ, n = 15), Rio Grande do Sul (RS, n = 10), Paraná (PR, n = 4), São Paulo (SP, n = 3) and Minas Gerais (MG, n = 1). Our results showed that S. brasiliensis is highly prevalent among cats (96.9%) with sporotrichosis, while S. schenckii was identified only once. The genotype of Sporothrix from cats was found identical to S. brasiliensis from human sources confirming that the disease is transmitted by cats. Sporothrix brasiliensis presented low genetic diversity compared to its sister taxon S. schenckii. No evidence of recombination in S. brasiliensis was found by split decomposition or PHI-test analysis, suggesting that S. brasiliensis is a clonal species. Strains recovered in states SP, MG and PR share the genotype of the RJ outbreak, different from the RS clone. The occurrence of separate genotypes among strains indicated that the Brazilian S. brasiliensis epidemic has at least two distinct sources. We suggest that cats represent a major host and the main source of cat and human S. brasiliensis infections in Brazil.

Canine sporotrichosis in Rio de Janeiro, Brazil: clinical presentation, laboratory diagnosis and therapeutic response in 44 cases (1998 /2003)

A sporotichosis epidemic involving forty-four dogs in the Metropolitan area of Rio de Janeiro is described. Solitary skin lesions were noted in 18 dogs (40.9%), 2-4 such lesions were observed in 17 animals (38.6%), and nine (20.5%) animals had five or more lesions. Twenty-five (56.8%) animals had single ulcerated skin lesions on the nose and nine (20.5%) showed nasal mucosal involvement (three of which also has a skin lesion). Respiratory symptoms were observed in 17 (38.6%) dogs and were found to be the most common extracutaneous signs of infection. Anemia, leukocytosis with neutrophilia, hypoalbuminemia and hyperglobulinemia were the most frequent hematological abnormalities. Histopathological analysis of skin biopsies in most cases revealed granulomatous reactions characterized by histiocytic hyperplasia and neutrophil infiltration. Yeast-like cells were observed in seven (16.7%) of 42 dogs examined histologically. During the study, eight (18.2%) animals were lost to follow-up and three (6.8%) were submitted to euthanasia. Of the remaining 33 dogs, five (15.2%) presented spontaneous regression of the lesions, 26 (78.8%) were cured after treatment, and two (6%) continue to be treated. The present cases indicate that many dogs with sporotrichosis respond well to treatment and in a few dogs, the disease may be self-limiting.

Response to azolic antifungal agents for treating feline sporotrichosis

The effectiveness and safety of treatment with ketoconazole and itraconazole were compared in 773 sporotrichosis-infected cats over a four-year period (2002 to 2005). Five hundred and ninety-eight cats received oral ketoconazole and 175 received oral itraconazole. Treatment was successful in 238 (30.8 per cent) cats, of which 171 (28.6 per cent) of 598 received 13.5 to 27.0 mg/kg/day ketoconazole and 67 (38.3 per cent) of 175 received 8.3 to 27.7 mg/kg/day itraconazole. Adverse effects were reported in 306 (39.6 per cent) of the cats, 105 (13.6 per cent) died and 430 (55.6 per cent) dropped out of treatment or were still under treatment at the time of data analysis.

American cutaneous leishmaniasis in two cats from Rio de Janeiro, Brazil: first report of natural infection with Leishmania (Viannia) braziliensis

We describe the isolation of Leishmania (Viannia) braziliensis from two female cats with American cutaneous leishmaniasis in Rio de Janeiro, Brazil. The isolates were identified as L. (V.) braziliensis by isoenzyme electrophoresis.

Feline sporotrichosis: epidemiological and clinical aspects.

Feline sporotrichosis, which is caused by species of the Sporothrix schenckii complex, is endemic to Rio de Janeiro, Brazil. More than 4000 cases of the disease were diagnosed at Fundação Oswaldo Cruz, Brazil, between 1998 and 2012. Sporotrichosis in cats has been reported in several countries, but nowhere has an outbreak of animal sporotrichosis been as large as that seen in Brazil. The clinical manifestations of the disease range from an isolated skin lesion that can progress to multiple skin lesions and even fatal systemic involvement. Nodules and ulcers are the most common types of lesions, and respiratory signs and mucosa involvement are frequent. The definitive diagnosis depends on isolation of the etiologic agent in culture. Cytology, histopathology, and serology are useful tools for preliminary diagnosis. Severe pyogranulomatous inflammatory infiltrate, high fungal load, and extension of lesions to mucosa, cartilage, and bone in the nose of cats are indicative of an agent of high virulence in this endemic region. Itraconazole is the drug of choice, while, in refractory cases, amphotericin B or potassium iodide might be alternative treatments; however, recurrence after discharge may occur. Sporotrichosis persists as a neglected disease in Rio de Janeiro, and the treatment of cats remains a challenging and long-term endeavor.

Zoonotic Epidemic of Sporotrichosis: Cat to Human Transmission.

Most of the 51 species embedded in the genus Sporothrix are nonpathogenic environmental fungi that are closely related to decaying wood, plants, and soil. However, members of the Sporothrix schenckii complex are highly successful mammal pathogens, including S. brasiliensis, S. schenckii sensu stricto (s. str.), S. globosa, and S. luriei, the causative agents of human and animal sporotrichosis [1]. Their key to success during mammal infection lies at least in part with their ability to change from a mycelial saprophytic lifestyle at 25°C in the environment to a parasitic yeast cell at an elevated temperature (35°C–37°C), such as those developed by warm-blooded hosts [2]. Typically, infection develops after traumatic inoculation of contaminated soil, plants, and organic matter into skin or mucosa. Alternatively, infection may occur during the animal transmission (cat–cat or cat–dog) and zoonotic transmission (cat–human), which has been mostly associated with scratches or bites from infected cats [2]. In Brazil, S. brasiliensis is repeatedly associated with feline infection and has consistently shown higher virulence during epizootics, as well as in murine models of sporotrichosis. A hallmark of S. brasiliensis infection is its tendency to escalate to outbreaks or epidemics among cats with high potential for zoonotic transmission. Sporotrichosis is an emergent disease and, over the past two decades, the incidence of zoonotic sporotrichosis has been on the rise, particularly in Brazil. Judging from the epizootic and zoonotic epidemics taking place in Rio de Janeiro, Brazil, tackling sporotrichosis requires the engagement of animal and human health policies to reduce the transmission chain of Sporothrix.

The epidemiological scenario of feline sporotrichosis in Rio de Janeiro, State of Rio de Janeiro, Brazil

INTRODUCTION Sporotrichosis is a mycosis affecting both humans and animals. Within the context of the ongoing sporotrichosis epidemic in the State of Rio de Janeiro, Brazil, sick cats plays an important role in the zoonotic transmission. The aim of this study was to update the number of feline cases diagnosed at the Fundação Oswaldo Cruz (2005-2011). METHODS The medical records of the cats followed were reviewed; the inclusion criterion was the isolation of Sporothrix spp. in culture. RESULTS In total, 2,301 feline cases were identified. CONCLUSIONS These results should alert sanitary authorities to the difficulties associated with sporotrichosis control.

Sensitivity of cytopathological examination in the diagnosis of feline sporotrichosis.

Sporotrichosis is caused by Sporothrix schenckii. The cat is the animal species most affected by this mycosis and plays an important role in the zoonotic transmission of this disease. The definitive diagnosis is made by isolation of the fungus in culture; however, cytopathological examination is frequently used in cats. Medical records from cats treated at Instituto de Pesquisa Clínica Evandro Chagas/Fiocruz, Rio de Janeiro, Brazil, between 2004 and 2006 were reviewed. Criteria for inclusion were a diagnosis by isolation of S schenckii in culture and cytopathological examination of the same lesion as that used for collection of the culture material. Eight hundred and six cats were included in the study. Yeast-like structures suggestive of S schenckii were observed in 636 cases. The sensitivity of the method was 78.9%. Its practicality, low cost and sensitivity validate cytopathology as a safe technique for the presumptive diagnosis of sporotrichosis in cats.

Serodiagnosis of sporotrichosis infection in cats by enzyme-linked immunosorbent assay using a specific antigen, SsCBF, and crude exoantigens.

Abstract The main objective of this study is to standardize an ELISA for the diagnosis of feline sporotrichosis. Sporothrix schenckii is the etiological agent of human and animal sporotrichosis. Cats may act as reservoirs for S. schenckii and can transmit the infection to humans by a bite or scratch. There are few methods for the serological diagnosis of fungal diseases in animals. In this paper, an ELISA test for the diagnosis of cat sporotrichosis is proposed, which detects S. schenckii-specific antibodies in feline sera. Two different kinds of antigens were used: “SsCBF”, a specific molecule from S. schenckii that consists of a Con A-binding fraction derived from a peptido-rhamnomannan component of the cell wall, and a S. schenckii crude exoantigen preparation. The ELISA was developed, optimized, and evaluated using sera from 30 cats with proven sporotrichosis (by culture isolation); 22 sera from healthy feral cats from a zoonosis center were used as negative controls. SsCBF showed 90% sensitivity and 96% specificity in ELISA; while crude exoantigens demonstrated 96% sensitivity and 98% specificity. The ELISA assay described here would be a valuable screening tool for the detection of specific S. schenckii antibodies in cats with sporotrichosis. The assay is inexpensive, quick to perform, easy to interpret, and permits the diagnosis of feline sporotrichosis.

Is Leishmania (Viannia) braziliensis preferentially restricted to the cutaneous lesions of naturally infected dogs

Nineteen dogs naturally infected with Leishmania (Viannia) braziliensis were studied in order to determine the presence of the parasite outside cutaneous lesions. Eleven (57.9%) animals showed single cutaneous or mucosal lesions and eight (42.1%) presented two or three lesions. Twenty-eight active lesions were biopsied. Isolation in culture and characterization by enzyme electrophoresis were possible in 100% of cases and amastigote forms were visualized upon histopathological examination in three samples (n=25, 12%). Isolation of the parasite in culture from peripheral blood and intact skin fragments obtained from the scapular region was negative in all animals, as was the histopathological analysis of skin from this region. Serological reactivity determined by an immunofluorescent antibody test and/or enzyme-linked immunosorbent assay was demonstrated in 15 animals. The results obtained suggest that L. braziliensis preferentially remains at the site of lesion, in contrast to the systemic distribution of parasites observed in dogs infected with L. (Leishmania) chagasi. A better understanding of this aspect may help direct diagnostic and control strategies applicable to areas characterized by the simultaneous occurrence of the cutaneous and visceral forms of leishmaniasis, as is the case for the Municipality of Rio de Janeiro, Brazil.