Trends in the molecular epidemiology and population genetics of emerging Sporothrix species.

Sporothrix (Ophiostomatales) comprises species that are pathogenic to humans and other mammals as well as environmental fungi. Developments in molecular phylogeny have changed our perceptions about the epidemiology, host-association, and virulence of Sporothrix. The classical agent of sporotrichosis, Sporothrix schenckii, now comprises several species nested in a clinical clade with S. brasiliensis, S. globosa, and S. luriei. To gain a more precise view of outbreaks dynamics, structure, and origin of genetic variation within and among populations of Sporothrix, we applied three sets of discriminatory AFLP markers (#3 EcoRI-GA/MseI-TT, #5 EcoRI-GA/MseI-AG, and #6 EcoRI-TA/MseI-AA) and mating-type analysis to a large collection of human, animal and environmental isolates spanning the major endemic areas. A total of 451 polymorphic loci were amplified in vitro from 188 samples, and revealed high polymorphism information content (PIC = 0.1765-0.2253), marker index (MI = 0.0001-0.0002), effective multiplex ratio (E = 15.1720-23.5591), resolving power (Rp = 26.1075-40.2795), discriminating power (D = 0.9766-0.9879), expected heterozygosity (H = 0.1957-0.2588), and mean heterozygosity (Havp  = 0.000007-0.000009), demonstrating the effectiveness of AFLP markers to speciate Sporothrix. Analysis using the program structure indicated three genetic clusters matching S. brasiliensis (population 1), S. schenckii (population 2), and S. globosa (population 3), with the presence of patterns of admixture amongst all populations. AMOVA revealed highly structured clusters (PhiPT = 0.458-0.484, P < 0.0001), with roughly equivalent genetic variability within (46-48 %) and between (52-54 %) populations. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for S. schenckii (χ2 = 2.522; P = 0.1122), supporting random mating. In contrast, skewed distributions were found for S. globosa (χ2 = 9.529; P = 0.0020) with a predominance of MAT1-1 isolates, and regional differences were highlighted for S. brasiliensis with the overwhelming occurrence of MAT1-2 in Rio de Janeiro (χ2 = 14.222; P = 0.0002) and Pernambuco (χ2 = 7.364; P = 0.0067), in comparison to a higher prevalence of MAT1-1 in the Rio Grande do Sul (χ2 = 7.364; P = 0.0067). Epidemiological trends reveal the geographic expansion of cat-transmitted sporotrichosis due to S. brasiliensis via founder effect. These data support Rio de Janeiro as the centre of origin that has led to the spread of this disease to other regions in Brazil. Our ability to reconstruct the source, spread, and evolution of the ongoing outbreaks from molecular data provides high-quality information for decision-making aimed at mitigating the progression of the disease. Other uses include surveillance, rapid diagnosis, case connectivity, and guiding access to appropriate antifungal treatment.

Comparison of the Sensitivity of Three Methods for the Early Diagnosis of Sporotrichosis in Cats.

Sporotrichosis is caused by species of fungi within the Sporothrix schenckii complex that infect man and animals. In Rio de Janeiro, Brazil, an epidemic has been observed since 1998, with most of the cases being related to transmission from infected cats. Although the definitive diagnosis of feline sporotrichosis is made by fungal culture, cytopathological and histopathological examinations are used routinely, because the long culture period may delay treatment onset. However, alternative methods are desirable in cases of low fungal burden. Immunohistochemistry (IHC) has been described as a sensitive method for diagnosing human and canine sporotrichosis, but there are no reports of its application to cats. The aim of this study was to analyse the sensitivity of cytopathological examination (Quick Panoptic method), histopathology (Grocott silver stain) and anti-Sporothrix IHC by blinded comparisons, using fungal culture as the reference standard. Samples were collected from 184 cats with sporotrichosis that exhibited skin ulcers. The sensitivities of Grocott silver stain, cytopathological examination and IHC were 91.3%, 87.0% and 88.6%, respectively. Grocott silver stain showed the best performance. IHC showed high sensitivity, as did cytopathological examination and these may be considered as alternative methodologies. When the three methods were combined, the diagnosis was established in 180 (97.8%) out of 184 cases. Taken together, these findings indicate the need to implement these methods as routine tools for the early diagnosis of sporotrichosis in cats, notably when fungal culture is not available.

Treatment abandonment in feline sporotrichosis - study of 147 cases.

This study describes the epidemiological, clinical and mycological aspects of feline sporotrichosis cases attending the Laboratory of Clinical Research on Dermatozoonosis in Domestic Animals - Evandro Chagas Clinical Research Institute (LAPCLIN-DERMZOO/IPEC/FIOCRUZ), from 1998 to 2005. It was possible to get in contact with 147 (19.2%) cat owners. One hundred and thirteen (76.9%) cats were male, 117 (79.6%) had no defined race and 87 (59.2%) were sexually intact. The age ranged from 72 to 216 months (median = 108 months). Nineteen cats were reassessed: eleven (57.8%) were male, thirteen (36.8%) were breed and fifteen (47.3%) castrated. Fourteen (52.6%) animals lived at home and did not roamed the streets. Seven (36.8%) had normal clinical findings and negative mycological examination. Twelve (63.1%) cats had skin lesions compatible with sporotrichosis. Thirty-one (21%, n = 147) cats disappeared after abandoning treatment, 36 (24.5%, n = 147) were alive and 80 (54.4%, n = 147) had died. Causes of death informed by the owners were: sporotrichosis in 35 (43.7%, n = 80), accidental death in 27 (33.7%, n = 80) and other diseases in 18 (22.5%, n = 80). Withdrawal of treatment occurred mainly at the time of clinical improvement and may represent a serious obstacle to the control of sporotrichosis.

Comparison of acetate tape impression with squeezing versus skin scraping for the diagnosis of canine demodicosis.

This study compared the sensitivity of acetate tape impression and skin squeezing with that of deep skin scraping for the diagnosis of demodicosis in dogs. Demodex canis was detected in 100% of acetate tape impressions obtained after skin squeezing and in 90% of deep skin scrapings. There was a significant difference (P < 0.001) between the techniques in the total number of mites detected. Acetate tape impression with skin squeezing was found to be more sensitive than deep skin scraping and is an alternative diagnostic method for canine demodicosis.

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.

Histopathology of canine sporotrichosis: a morphological study of 86 cases from Rio de Janeiro (2001-2007).

The present study reports the histopathological findings of 86 skin lesions of dogs with sporotrichosis from Rio de Janeiro. Suppurative granulomatous inflammation was the predominant finding and was observed in 76 (88.37%) cases. Plasma cells surrounding the suppurative granulomas were detected in 68 (89.5%) cases and an inflammatory infiltrate at the periphery of these granulomatous lesions was observed in 63 (82.9%). Fungus-specific staining revealed yeast cells compatible with Sporothrix schenckii in 36 cases. These fungal elements were only detected in lesions characterized by suppurative granulomatous inflammation. Thus, specific staining of serial sections is recommended in the case of dogs with skin lesions whose histopathological presentation is consistent with sporotrichosis. However, due to the generally small number of yeast cells in lesions, the hypothesis of sporotrichosis should not be ruled out even if the result is negative, especially in epidemic areas where correlation with epidemiological data is particularly useful.

First report of natural infection of a bush dog (Speothos venaticus) with Leishmania (Leishmania) chagasi in Brazil.

We report here the first known case of natural infection of a bush dog with Leishmania (Leishmania) chagasi in Brazil. The specimen was captured in the wild in the State of Mato Grosso and is currently being held in captivity at Fundação Jardim Zoológico, Rio de Janeiro, Brazil. The leishmaniasis was diagnosed by culture of promastigote forms in intact skin fragments and their characterization by isoenzyme electrophoresis. This report calls attention to the parasitological and etiological control of certain zoonoses, such as leishmaniasis, in wild animals kept in captivity, especially when animals are exchanged between zoos in Brazil.