Anti-Brucella canis antibodies in dogs naturally infected with Leishmania infantum and associated histological alterations in the genital tract.

In canine visceral leishmaniasis, coinfections can aggravate the disease. Our aim was to investigate Brucella canis in dogs infected with Leishmania infantum. One hundred and six L. infantum-seropositive dogs were submitted to serology for B. canis, PCR for B. canis and L. infantum, and histopathological analysis of the genital tract. Anti-B. canis antibodies were detected in seven dogs whose clinical signs, L. infantum load and histological alterations were similar to those of seronegative animals. The circulation of anti-B. canis antibodies was low but demonstrates the exposure of dogs to this bacterium in a visceral leishmaniasis-endemic area.

Geo-epidemiology and socioeconomic aspects of human sporotrichosis in the municipality of Duque de Caxias, Rio de Janeiro, Brazil, between 2007 and 2016.

BACKGROUND: Sporotrichosis is usually caused by the traumatic inoculation of pathogenic species of fungi of the genus Sporothrix. The most prevalent species in Brazil is Sporothrix brasiliensis, which is generally associated with transmission involving infected cats. Sporotrichosis is hyperendemic in the state of Rio de Janeiro and Duque de Caxias is one of the most affected municipalities. METHODS: This was a cross-sectional, geo-epidemiological and socioeconomic study of human sporotrichosis in the municipality of Duque de Caxias using geoprocessing information for the construction of thematic maps. RESULTS: Eight hundred and twenty-seven cases of sporotrichosis from Duque de Caxias were reported between 2007 and 2016, most of them in women from 25-59 years. The most affected areas had low per capita income and scarce supply of treated water. Human sporotrichosis expanded throughout the territory of the municipality over time. CONCLUSIONS: An increase in both the number of reported cases and their spatial distribution occurred throughout the studied decade. The concentration of the disease was more intense in areas with greater vulnerability of the population, expressed by low per capita income and deficient provision of basic sanitation services. Sporotrichosis requires measures to better control the disease in Duque de Caxias and in the state of Rio de Janeiro.

Synthesis and Identification of Pentathiepin-Based Inhibitors of Sporothrix brasiliensis.

Sporothrix brasiliensis is the causative agent of zoonotic sporotrichosis in Brazil and is currently referred to as the most virulent species among those of clinical importance within the genus. Sporotrichosis is an emergent disease that has come to the forefront over two decades with a recent hot spot of sporotrichosis infection emerging in the state of Rio de Janeiro. The source of these infections is now at epidemic proportions with more than 4000 cases reported in Rio de Janeiro, Brazil, alone since 1998. We developed a focused library of a rare pentathiepin ring system and identified a potent substitution pattern that yielded compounds 21 and 22. These compounds were more potent than itraconazole which is the current standard of care for sporotrichosis.

Association of itraconazole and potassium iodide in the treatment of feline sporotrichosis: a prospective study.

Feline sporotrichosis is an endemic disease in Rio de Janeiro, Brazil, where zoonotic transmission of Sporothrix spp. has been reported since 1998. Itraconazole (ITZ) remains the first choice for treating this disease in cats. However, there have been reports of therapeutic failure and a long-term endeavor. Potassium iodide (KI), considered in the past as a drug with variable effectiveness in cats with sporotrichosis, arises as an important option in the treatment of cats from the endemic area of Rio de Janeiro. In order to evaluate the effectiveness of the association of ITZ and KI in naive cats with sporotrichosis, a prospective cohort study was conducted on 30 cats receiving ITZ 100 mg/day and KI 2.5 mg-20 mg/kg/day. Clinical and laboratory adverse effects were assessed once a month according to the standard care protocol. The cure rate was 96.15% within a median of 14 weeks of treatment. Adverse effects were observed in 50% of cats and were managed with a temporary drug suspension and/or a hepatoprotective therapy. The association of ITZ and KI emerges as an effective option for the treatment of feline sporotrichosis.

Severe feline sporotrichosis associated with an increased population of CD8low cells and a decrease in CD4⁺ cells.

Sporotrichosis is a subcutaneous mycosis with worldwide distribution, especially in tropical and subtropical areas. Zoonotic transmission is described with cats being the main animal species involved. The occurrence of severe feline sporotrichosis with high fungal levels demonstrates the susceptibility of cats to this disease and the importance of studying its pathogenesis. This study describes the leukocytes profile in blood of cats with sporotrichosis by flow cytometry and its correlation with histopathology and fungal load. The cats with sporotrichosis were separated into groups L1, L2, and L3 (lesions at one, two, and three or more noncontiguous skin locations, respectively) and were classified as good, fair, or poor general conditions. The highest percentage of CD4+ cells was associated to L1 (P = .04) and to good general condition (P = .03). The percentage of CD8+ cells was greater in L2 and L3 (P = .01). CD8(low) expression occurred in 20 animals with sporotrichosis, mainly in L3 (P = .01) and was not observed in healthy controls. This expression was related to macrophage granulomas (P = .01) and predominated in cases with high fungal load. Altogether, the results indicated that control over feline sporotrichosis, with maintenance of a good general condition, fixed lesions, well-organized response and lower fungal load, is associated with increased CD4+ cells percentages. In contrast, a poor general condition, disseminated lesions and high fungal load were related to increased CD8+ cell percentages and increased expression of CD8(low). As conclusion these results point to an important role of the CD4:CD8 balance in determining the clinical outcome in feline sporotrichosis.

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.

Detection of serum antibodies against Bartonella species in cats with sporotrichosis from Rio de Janeiro, Brazil.

Cat scratch disease is a zoonosis caused by Bartonella species, transmitted to humans through scratches or bites from infected cats and via direct contact with infected feces. Sporotrichosis, caused by the fungal complex Sporothrix, is transmitted by traumatic inoculation of the fungus. Cats are important in zoonotic transmission. Serum samples from 112 domestic cats with sporotrichosis and 77 samples from healthy cats were analyzed by indirect immunofluorescence assay (IFA), using the commercial kit Bartonella henselae IFA IgG (Bion). The presence of antibodies against feline leukemia virus (FeLV) and of feline immunodeficiency virus (FIV) core antigens was detected using the commercial kit Snap Combo FIV-FeLV (Idexx). The group of animals with sporotrichosis contained 93 males with a median age of 22 months, eight (7.1%) of which were positive for FIV and 15 (13.4%) for FeLV. The group of animals without sporotrichosis contained 36 males with a median age 48 months, 10 (13.0%) of which were positive for FIV and eight (10.4%) for FeLV. Of the 112 cats with sporotrichosis and 77 cats without mycosis, 72 (64.3%) and 35 (45.5%), respectively, were IFA reactive. No association was found between age, sex, FIV/FeLV and the presence of antibodies to Bartonella species. The results suggest that the study population can be considered a potential source of zoonotic infection for both diseases.

Feline sporotrichosis: histopathological profile of cutaneous lesions and their correlation with clinical presentation.

Cutaneous lesions of feline sporotrichosis show high fungal load and are associated with severe disease and elevated zoonotic potential. The present study describes the histopathology and fungal load of the lesions in different clinical presentations of feline sporotrichosis. Cats with sporotrichosis were separated into groups L1, L2 and L3 (lesions in one, two and three or more locations, respectively) and subjected to skin biopsies for histopathology. Eighty-six cats were included in the study. Lesions were suppurative granulomatous in 84 cases and poorly formed granulomas were predominant. The well-formed granulomas were associated with group L1. The high fungal load was predominant in group L3 and in poorly formed granuloma cases and did not occur in well-formed granulomas cases. The good general condition was associated with low fungal load. These findings suggest that the fungal load control in animals with more localized lesions and well-organized response is linked with the improvement in the outcome of infected cats.

Potassium iodide capsule treatment of feline sporotrichosis.

Sporotrichosis is a mycosis caused by Sporothrix schenckii. The most affected animal is the cat; it has played an important role in the zoonotic transmission of this disease, especially in Rio de Janeiro, Brazil, since 1998. In order to evaluate the treatment of feline sporotrichosis with potassium iodide, an observational cohort was conducted in 48 cats with sporotrichosis at Instituto de Pesquisa Clínica Evandro Chagas, Fiocruz. All cats received potassium iodide capsules, 2.5 mg/kg to 20 mg/kg q24h. The cure rate was 47.9%, treatment failure was 37.5%, treatment abandonment was 10.4% and death was 4.2%. Clinical adverse effects were observed in 52.1% of the cases. Thirteen cats had a mild increase in hepatic transaminase levels during the treatment, six of them presented clinical signs suggestive of hepatotoxicity. Compared to previous studies with itraconazole and iodide in saturated solution, potassium iodide capsules are an alternative for feline sporotrichosis treatment.

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.

Intralesional amphotericin B in a cat with refractory localised sporotrichosis.

The present report concerns a case of sporotrichosis in a 3-year-old male crossbred cat. After 9 months of oral itraconazole (20mg/kg) therapy, all skin lesions had resolved with the exception of a single nodular lesion located on the bridge of the nose. Therefore, a combined therapy that included intralesional (IL) amphotericin B (1mg/kg) was started. The lesion resolved completely after three weekly administrations of IL amphotericin B, given in concert with oral itraconazole. The cat remains well 1 year after discontinuing therapy, with no signs of recurrence.

Sporotrichosis: the main differential diagnosis with tegumentary leishmaniosis in dogs from Rio de Janeiro, Brazil.

Seventy-four dogs from the State of Rio de Janeiro with ulcerated cutaneous lesions were submitted to clinical, dermatological, parasitological, mycological, histopathological and cytopathological exams, a leishmanin skin test, an indirect immunofluorescence (IIF) test for leishmaniosis, and nonspecific laboratory tests such as blood count and serum biochemistry. Sporothrix schenckii was isolated from 41 dogs and Leishmania (Viannia) braziliensis was isolated from 33 animals. Most dogs with sporotrichosis were from the municipality of Rio de Janeiro (53.7%) and presented ulcerated cutaneous lesions on the head (68.3%). Laboratory alterations in these animals included anemia (58.5%), hypoalbuminemia (83%) and hyperglobulinemia (75.6%). Histopathology revealed the predominance of a chronic granulomatous inflammatory infiltrate (70.7%), and yeast-like structures were detected in 17% of the histopathological exams and in 32% of the cytological exams. Three of 41 dogs with sporotrichosis were seropositive by IIF for leishmaniosis and 2 of 20 animals tested within this group had a positive leishmanin skin test. Similarly, most of the 33 dogs with leishmaniosis were from the municipality of Rio de Janeiro (69.7%) and had ulcerated cutaneous lesions on the head (84.8%). Laboratory alterations in these animals included anemia (66.7%), hypoalbuminemia (100%) and hyperglobulinemia (91%). Histopathology showed the predominance of a chronic granulomatous inflammatory infiltrate (63.6%) and amastigote forms were detected in 30.3% of the histopathological exams and in 31.8% of the 22 cytological exams performed. About 72.7% of the dogs were seropositive by IIF and five of seven animals had a positive skin test. Due to the clinical similarities, histopathological and nonspecific laboratory results similarities, the serological and skin tests for leishmaniosis positive in dogs with sporotrichosis, and the overlapping endemic areas in Rio de Janeiro, the differential diagnosis between the two diseases requires the demonstration of their respective etiological agents.

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.

Post mortem parasitological evaluation of dogs seroreactive for Leishmania from Rio de Janeiro, Brazil.

A parasitological study was conducted on 66 dogs seroreactive for Leishmania captured as a control measure of visceral leishmaniasis in the State of Rio de Janeiro, Brazil. Biological samples from different anatomical sites were collected during autopsy of the animals and cultured on biphasic medium (NNN/Schneider). The Leishmania isolates were characterized by isoenzyme electrophoresis. Leishmania was isolated from 80.3% of the animals: 12 animals with Leishmania (Viannia) braziliensis isolated exclusively from cutaneous lesions, 39 with L. (L.) chagasi isolated from different sites in the same animal, and 2 with simultaneous isolation of L. (V.) braziliensis from cutaneous lesions and L. (L.) chagasi from different sites. Isolation in culture revealed the absence of Leishmania parasites in 13 animals. The results obtained confirm the existence of mixed infections in dogs in Rio de Janeiro and indicate the need to complement the investigation of seroreactive dogs using methods for the parasitological diagnosis and identification of Leishmania species.

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.