Antifungal activity of liriodenine on clinical strains of Cryptococcus neoformans and Cryptococcus gattii species complexes

Background: Cryptoccocal meningitis continues to present high incidence among AIDS patients. The treatment of choice is the synergistic combination of flucytosine (5-FC) with amphotericin B deoxycholate (AmBd) or its lipid formulations. However, 5-FC is unavailable in many countries and AmB demands hospitalization. The combination of AmB with the fungistatic fluconazole (FLC) or the use of high FLC daily doses alone became the choice. Nonetheless, sterilization of cerebrospinal fluid is delayed with FLC monotherapy, mainly with high fungal burden. These findings suggest the search for new antifungal compounds, such as liriodenine. Methods: Liriodenine antifungal activity was evaluated by three procedures: determining the minimum inhibitory concentration (MIC) on 30 strains of the Cryptococcus neoformans (C. neoformans) complex and 30 of the Cryptococcus gattii (C. gattii) complex, using EUCAST methodology and amphotericin B deoxycholate as control; performing the time-kill methodology in two strains of the C. neoformans complex and one of the C. gattii complex; and injury to cryptococcal cells, evaluated by transmission electron microscopy (TEM). Liriodenine absorption and safety at 0.75 and 1.50 mg.kg-1 doses were evaluated in BALB/c mice. Results: Liriodenine MICs ranged from 3.9 to 62.5 µg.mL-1 for both species complexes, with no differences between them. Time-kill methodology confirmed its concentration-dependent fungicidal effect, killing all the strains below the limit of detection (33 CFU.mL-1) at the highest liriodenine concentration (32-fold MIC), with predominant activity during the first 48 hours. Liriodenine induced severe Cryptococcus alterations ­ cytoplasm with intense rarefaction and/or degradation, injury of organelles, and presence of vacuoles. Liriodenine was better absorbed at lower doses, with no histopathological alterations on the digestive tract. Conclusion: The fungicidal activity confirmed by time-kill methodology, the intense Cryptococcus injury observed by TEM, the absorption after gavage administration, and the safety at the tested doses indicate that the liriodenine molecule is a promising drug lead for development of anticryptococcal agents.(AU)

Peritoneal cryptococcosis in a dog

Background: Cryptococcosis is an opportunistic, systemic mycotic disease caused by yeasts of genus Cryptococcus.Cryptococcus neoformans might cause serious public health problems, as they can be transmitted to humans, domesticand wild animals. In cats and dogs, the main site of infection is the upper lower respiratory tract, however, the infectioncan extend and affect other sites in the organism, however, the peritoneal manifestation of the disease is rare. Thus, thepresent work aims to report a case of cryptococcosis in a canine, mixed breed, female, adult, from the Metropolitan Regionof Belém, PA, Brazil.Case: A mixed breed dog, female, adult was admitted in a veterinary hospital at Belém, presenting abdominal discomfortand, after the clinical examination, emergency exploratory laparotomy was indicated, and it were observed disseminatedlesions in the abdominal cavity, with multiple nodules attached to the peritoneum and intestinal serosa and bladder, uterus,among other organs. Fragments from the surgical biopsy were fixed, and processed routinely according to the techniquesfor histological tissue processing. Histological examination revealed a predominance of granulomatous lesions in peritoneum and mesentery organs. It was observed a discrete inflammatory reaction of macrophages, epithelioid cells andgiant cells, many in phagocytosis of spherical or ovoid organisms, with a thick capsule and a clear perinuclear halo. Inthe PAS staining, the Cryptococcus conidia presented eosinophilic characteristics and some of them had the capsule welldemarcated, which was shown to be radiated. Also it was possible to see the budding yeast and in the Grocott stain, theCryptococcus stained in black.Discussion: Cryptococcosis with peritoneal involvement in animals is rarely reported in the literature. Cryptococcus is afungus that presents tropism by the...

Peritoneal cryptococcosis in a dog

Background: Cryptococcosis is an opportunistic, systemic mycotic disease caused by yeasts of genus Cryptococcus.Cryptococcus neoformans might cause serious public health problems, as they can be transmitted to humans, domesticand wild animals. In cats and dogs, the main site of infection is the upper lower respiratory tract, however, the infectioncan extend and affect other sites in the organism, however, the peritoneal manifestation of the disease is rare. Thus, thepresent work aims to report a case of cryptococcosis in a canine, mixed breed, female, adult, from the Metropolitan Regionof Belém, PA, Brazil.Case: A mixed breed dog, female, adult was admitted in a veterinary hospital at Belém, presenting abdominal discomfortand, after the clinical examination, emergency exploratory laparotomy was indicated, and it were observed disseminatedlesions in the abdominal cavity, with multiple nodules attached to the peritoneum and intestinal serosa and bladder, uterus,among other organs. Fragments from the surgical biopsy were fixed, and processed routinely according to the techniquesfor histological tissue processing. Histological examination revealed a predominance of granulomatous lesions in peritoneum and mesentery organs. It was observed a discrete inflammatory reaction of macrophages, epithelioid cells andgiant cells, many in phagocytosis of spherical or ovoid organisms, with a thick capsule and a clear perinuclear halo. Inthe PAS staining, the Cryptococcus conidia presented eosinophilic characteristics and some of them had the capsule welldemarcated, which was shown to be radiated. Also it was possible to see the budding yeast and in the Grocott stain, theCryptococcus stained in black.Discussion: Cryptococcosis with peritoneal involvement in animals is rarely reported in the literature. Cryptococcus is afungus that presents tropism by the...(AU)

Yeast microbiota of free-ranging amphibians and reptiles from Caatinga biome in Ceará State, Northeast Brazil: High pathogenic potential of Candida famata / Leveduras da microbiota de anfíbios e répteis do bioma Caatinga no Ceará, Nordeste do Brasil: alto potencial patogênico de Candida famata

Studies on the fungal microbiota of reptiles and amphibians are necessary to better understand of host-microbe interactions and the establishment of fungal disease in these animals. However, these studies are limited. The present researchidentified yeasts from free-ranging reptiles and amphibians from the Caatinga biome andevaluated the virulence factors production, the antifungal susceptibility in planktonic and biofilm growth and the pathogenicity of Candida famata isolates. Twenty-nine isolates of the genera Candida, Cryptococcus and Rhodotorula were identified by phenotypic and/or molecular methods and production of hydrolytic enzymes in vitro by these genera of fungi was evaluated. In addition, susceptibility of planktonic cells and biofilms to azoles and amphotericin B was evaluated. The pathogenicity of C. famata, the most prevalent yeast species isolated, was evaluated using Caenorhabditis elegans model. C. famata was the most prevalent yeast in amphibian and reptilian microbiota. Phospholipase and protease production was observed in 18/29 and 11/29 of the yeast isolates, respectively, while 100% formed biofilms. Itraconazole presented high minimal inhibitory concentrations against C. famata and C. tropicalis. Amphotericin B reduced the biomass and metabolic activity of biofilms. C. famata induced the mortality of C. elegans. In conclusion, reptiles and amphibians are colonized by yeasts capable of producing important virulence factors, especially by Candida spp. that present low susceptibility to azoles which may result from imbalances in ecosystem. Finally, C. famata isolated from these animals presented high pathogenicity, showing the importance of the study of reptile and amphibians fungal microbiota.(AU)

Estudos sobre a microbiota fúngica de répteis e anfíbios são necessários para melhor compreender as interações hospedeiro-microrganismo e o estabelecimento de doenças fúngicas nesses animais. No entanto, esses estudos são limitados. O objetivo da presente pesquisa foi identificar leveduras isoladas de répteis e anfíbios do bioma Caatinga e avaliar a produção de fatores de virulência, a sensibilidade a antifúngicos no crescimento planctônico e de biofilme e a patogenicidade de Candida famata. Vinte e nove isolados dos gêneros Candida, Cryptococcus e Rhodotorula foram identificados por métodos fenotípicos e/ou moleculares e a produção de enzimas hidrolíticas in vitro por esses gêneros de fungos foi avaliada. Além disso, foi avaliada a suscetibilidade de células planctônicas e biofilmes a azólicos e anfotericina B. A patogenicidade de C. famata, a espécie de levedura isolada mais prevalente, foi avaliada usando Caenorhabditis elegans. C. famata foi a levedura mais prevalente na microbiota de anfíbios e répteis. A produção de fosfolipase e protease foi observada em 18/29 e 11/29 dos isolados de levedura, respectivamente, enquanto 100% formaram biofilmes. O itraconazol apresentou altas concentrações inibitórias mínimas contra C. famata e C. tropicalis. A anfotericina B reduziu a biomassa e atividade metabólica dos biofilmes. C. famata induziu a mortalidade de C. elegans. Em conclusão, répteis e anfíbios são colonizados por leveduras capazes de produzir importantes fatores de virulência, especialmente por cepas de Candida spp. que apresentam baixa suscetibilidade a azólicos que podem resultar de desequilíbrio no ecossistema. Por fim, C. famata isolados desses animais apresentaram alta patogenicidade, mostrando a importância do estudo da microbiota fúngica de répteis e anfíbios.(AU)

Criptococose em felino - Relato de caso / Cryptococcosis in a feline - case report

This paper analyzes a case report of cryptococcosis in a 11 years old crossbreed domestic feline, that was attended at Dr. Vicente Borelli"  Centro Universitário Fundação de Ensino Octávio Bastos Veterinary Hospital. The animal presented increased nasal volume. After physical examination, it was collected material from the lesion in order to do fungal culture and histologic analysis. It was observed pyogranulomatous dermatitis and presence of Cryptococcus spp., an microorganism able to affect humans, domestics and wild animals, corroborating the diagnosis of Cryptococcosis.

O presente trabalho analisa um caso de criptococose em um felino doméstico, sem raça definida, com 11 anos de idade, atendido no Hospital Veterinário "Dr. Vicente Borelli" do Centro Universitário Fundação de Ensino Octávio Bastos, São João da Boa Vista - SP. O animal apresentava aumento de volume em região nasal. Após exame clínico, foi realizada a colheita de material para exame de cultura fúngica e histopatológico. Neste último, observou-se uma dermatite piogranulomatosa e presença do microrganismo Cryptococcus spp., capaz de acometer humanos e animais domésticos e silvestres, confirmando o diagnóstico de Criptococose.

Criptococose em felino - Relato de caso / Cryptococcosis in a feline - case report

This paper analyzes a case report of cryptococcosis in a 11 years old crossbreed domestic feline, that was attended at Dr. Vicente Borelli"  Centro Universitário Fundação de Ensino Octávio Bastos Veterinary Hospital. The animal presented increased nasal volume. After physical examination, it was collected material from the lesion in order to do fungal culture and histologic analysis. It was observed pyogranulomatous dermatitis and presence of Cryptococcus spp., an microorganism able to affect humans, domestics and wild animals, corroborating the diagnosis of Cryptococcosis.(AU)

O presente trabalho analisa um caso de criptococose em um felino doméstico, sem raça definida, com 11 anos de idade, atendido no Hospital Veterinário "Dr. Vicente Borelli" do Centro Universitário Fundação de Ensino Octávio Bastos, São João da Boa Vista - SP. O animal apresentava aumento de volume em região nasal. Após exame clínico, foi realizada a colheita de material para exame de cultura fúngica e histopatológico. Neste último, observou-se uma dermatite piogranulomatosa e presença do microrganismo Cryptococcus spp., capaz de acometer humanos e animais domésticos e silvestres, confirmando o diagnóstico de Criptococose.(AU)

Criptococose nasal causada por Cryptococcus gattii em ovino na Bahia, Nordeste do Brasil / Nasal cryptococcosis caused by Cryptococcus gattii in sheep in Bahia, northeastern Brazil

Background: Cryptococcosis is a potentially fatal systemic mycosis that can affect several animal species. The disease is caused by the yeast species Cryptococcus neoformans and C. gattii and is usually associated with immunosuppressive diseases or debilitating factors. In Brazil, cryptococcosis in ruminants has been described in several states, but there have been only two instances of infection in sheep. Until the present, cryptococcosis caused by C. gattii has only been reported in goats. The objective of this study was to describe, for the first time, a case of nasal cryptococcosis caused by C. gattii in a sheep in the state of Bahia. Case: The disease affected a 4-year-old Santa Inês sheep, created in extensive system in the municipality of Ribeira do Pombal, Bahia, Brazil. The animal presented weight loss, with an asymmetric increase of volume in the right nasal cavity, accompanied by dyspnea and bloody nasal discharge. Samples of the tumor mass were collected for cytological analysis of yeast structures compatible with Cryptococcus spp. After cytological diagnosis, the sheep was sent to the Livestock Development Center of the Federal University of Bahia. On physical examination, there was a large and firm irregular mass, strongly adhering to the right nasal cavity and causing facial asymmetry. Surgical removal was attempted, but a marked infiltrative character of the lesion was observed, with extensive tissue destruction. Owing to the unfavorable prognosis, euthanasia was carried out followed by necropsy, in which a large irregular mass was observed that almost completely obliterated the right nasal cavity. Resorption of the right nasal bone and the dorsal and middle nasal turbinates was also observed. Fragments of the nasal lesion were collected, fixed in 10% formalin, routinely processed for histopathology, and stained with hematoxylin and eosin (HE) and Mayer’s...

Criptococose nasal causada por Cryptococcus gattii em ovino na Bahia, Nordeste do Brasil / Nasal cryptococcosis caused by Cryptococcus gattii in sheep in Bahia, northeastern Brazil

Background: Cryptococcosis is a potentially fatal systemic mycosis that can affect several animal species. The disease is caused by the yeast species Cryptococcus neoformans and C. gattii and is usually associated with immunosuppressive diseases or debilitating factors. In Brazil, cryptococcosis in ruminants has been described in several states, but there have been only two instances of infection in sheep. Until the present, cryptococcosis caused by C. gattii has only been reported in goats. The objective of this study was to describe, for the first time, a case of nasal cryptococcosis caused by C. gattii in a sheep in the state of Bahia. Case: The disease affected a 4-year-old Santa Inês sheep, created in extensive system in the municipality of Ribeira do Pombal, Bahia, Brazil. The animal presented weight loss, with an asymmetric increase of volume in the right nasal cavity, accompanied by dyspnea and bloody nasal discharge. Samples of the tumor mass were collected for cytological analysis of yeast structures compatible with Cryptococcus spp. After cytological diagnosis, the sheep was sent to the Livestock Development Center of the Federal University of Bahia. On physical examination, there was a large and firm irregular mass, strongly adhering to the right nasal cavity and causing facial asymmetry. Surgical removal was attempted, but a marked infiltrative character of the lesion was observed, with extensive tissue destruction. Owing to the unfavorable prognosis, euthanasia was carried out followed by necropsy, in which a large irregular mass was observed that almost completely obliterated the right nasal cavity. Resorption of the right nasal bone and the dorsal and middle nasal turbinates was also observed. Fragments of the nasal lesion were collected, fixed in 10% formalin, routinely processed for histopathology, and stained with hematoxylin and eosin (HE) and Mayers...(AU)

Antifungal activity of liriodenine on agents of systemic mycoses, with emphasis on the genus Paracoccidioides

Endemic systemic mycoses remain a health challenge, since these opportunistic diseases are increasingly infecting immunosuppressed patients. The simultaneous use of antifungal compounds and other drugs to treat infectious or non-infectious diseases has led to several interactions and undesirable effects. Thus, new antifungal compounds should be investigated. The present study aimed to evaluate the activity of liriodenine extracted from Annona macroprophyllata on agents of systemic mycoses, with emphasis on the genus Paracoccidioides. Methods: The minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined by the microdilution method. The cellular alterations caused by liriodenine on a standard P. brasiliensis (Pb18) strain were evaluated by transmission and scanning electron microscopy. Results: Liriodenine was effective only in 3 of the 8 strains of the genus Paracoccidioides and in the Histoplasma capsulatum strain, in a very low concentration (MIC of 1.95 µg.mL-1); on yeasts of Candida spp. (MIC of 125 to 250 µg.mL-1), including C. krusei (250 µg.mL-1), which has intrinsic resistance to fluconazole; and in Cryptococcus neoformans and Cryptococcus gattii (MIC of 62.5 µg.mL-1). However, liriodenine was not effective against Aspergillus fumigatus at the studied concentrations. Liriodenine exhibited fungicidal activity against all standard strains and clinical isolates that showed to be susceptible by in vitro tests. Electron microscopy revealed cytoplasmic alterations and damage to the cell wall of P. brasiliensis (Pb18). Conclusion: Our results indicate that liriodenine is a promising fungicidal compound that should undergo further investigation with some chemical modifications.(AU)

Doenças infecciosas e não infecciosas nos peixes-boi do Brasil / Infectious and non-infectious diseases in manatees in Brazil

Background: Antillean manatee (Trichechus manatus) and Amazonian manatee (Trichechus inunguis) are the two species of manatees that occur in Brazil, and both are endangered. Studies on manatee disease in Brazil were previously based mainly on Florida manatees. In recent years, these studies have increased in Brazil. Thus, the objective was to carry out a survey of the publications of infectious and non-infectious diseases in the two species of manatees that occur in the country, to assist in the identification of threats to the species and in the health assessment of manatees in the country. Review: Regarding infectious and contagious diseases, parasites have affected both species of manatees in Brazil, such as Pulmonicola cochleotrema in wild T. manatus, Toxoplasma gondii in both species, and Cryptosporidium spp. also in both species, this being an important agent of zoonotic character. Giardia sp., another zoonotic parasite, has also been reported in both species of manatees in Brazil. Mycotic lesions caused by B. hawaiiensis have been described in calves of T. manatus in captivity. Meanwhile, Candida sp., Trichosporon sp., Rhodotorula sp., and Cryptococcus sp. have been identified in T. inunguis. Studies on viruses in manatees are still not widespread compared to other species; however, it has been verified that rotavirus is present in T. manatus in captivity. In studies carried out to identify bacteria in T. manatus, the following bacteria were isolated: Arizona sp., Bacillus sp., Corynebacterium equi, Corynebacterium pseudodiphteriticum, Corinebacterium sp., Edwasiella tarda, Enterobacter aerogenes, Enterobacter agglomerans, Enterobacter sakasakii, Escherichia coli, Leptospira spp., Proteus sp., Proteus vulgaris, Providencia rettgeri...

Antifungal activity of liriodenine on agents of systemic mycoses, with emphasis on the genus Paracoccidioides

Endemic systemic mycoses remain a health challenge, since these opportunistic diseases are increasingly infecting immunosuppressed patients. The simultaneous use of antifungal compounds and other drugs to treat infectious or non-infectious diseases has led to several interactions and undesirable effects. Thus, new antifungal compounds should be investigated. The present study aimed to evaluate the activity of liriodenine extracted from Annona macroprophyllata on agents of systemic mycoses, with emphasis on the genus Paracoccidioides. Methods: The minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined by the microdilution method. The cellular alterations caused by liriodenine on a standard P. brasiliensis (Pb18) strain were evaluated by transmission and scanning electron microscopy. Results: Liriodenine was effective only in 3 of the 8 strains of the genus Paracoccidioides and in the Histoplasma capsulatum strain, in a very low concentration (MIC of 1.95 µg.mL-1); on yeasts of Candida spp. (MIC of 125 to 250 µg.mL-1), including C. krusei (250 µg.mL-1), which has intrinsic resistance to fluconazole; and in Cryptococcus neoformans and Cryptococcus gattii (MIC of 62.5 µg.mL-1). However, liriodenine was not effective against Aspergillus fumigatus at the studied concentrations. Liriodenine exhibited fungicidal activity against all standard strains and clinical isolates that showed to be susceptible by in vitro tests. Electron microscopy revealed cytoplasmic alterations and damage to the cell wall of P. brasiliensis (Pb18). Conclusion: Our results indicate that liriodenine is a promising fungicidal compound that should undergo further investigation with some chemical modifications.(AU)

Cryptococcosis in a cat

Background: Cryptococcosis is a mycosis that primarily affects domestic cats and is caused by fungi of the genus Cryptococcus, resulting in cutaneous, ocular, respiratory, and neurological manifestations. Diagnosis is based on the microscopicevaluation of the lesions and isolation of the causative agent. The aim of this study was to describe a case of feline cryptococcosis diagnosed by cytology and treated at the Veterinary Hospital of the State University of Santa Cruz (HV-UESC).Case: A 5-year-old uncastrated male cat was taken to the HV-UESC small animal clinic, owing to a gradual increase inthe volume of the nasal plane spanning over a year, accompanied by respiratory distress. Upon physical examination, theanimal was found to have regular nutritional status, dehydration, hypocorous mucosae, and enlarged popliteal lymph nodes.In addition, multifocal areas of ulceration on the body as well as increased volume in the nasal plane (clown nose) wereobserved. Peripheral blood samples were collected for complete blood count and biochemical analysis. Due to clinicalsuspicion of fungal lesions, fine needle aspiration of the nasal lesion was performed and cytological slides were sent tothe HV-UESC histopathology laboratory. Probable diagnoses included rhinosporidiosis, cryptococcosis, and histoplasmosis. The animal was subjected to thoracic radiography in the imaging division of the HV-UESC. Blood counts revealedthrombocytopenia (130,000/mm3) and biochemical tests presented hypoproteinemia (3.2 g/dL), with hypoalbuminemia(1.52 g/dL); however, chest radiography did not show pulmonary alterations. Results of cytological analysis indicatedpyogranulomatous inflammation associated with intracellular organisms, demonstrating a pathogen morphology similarto that of Cryptococcus spp. Before the diagnosis, itraconazole1 5 mg/kg (1 capsule, twice a day for 30 days) and fipronil2(1 ampoule 0.5 mL, on the back) were prescribed...

Cryptococcosis in a cat

Background: Cryptococcosis is a mycosis that primarily affects domestic cats and is caused by fungi of the genus Cryptococcus, resulting in cutaneous, ocular, respiratory, and neurological manifestations. Diagnosis is based on the microscopicevaluation of the lesions and isolation of the causative agent. The aim of this study was to describe a case of feline cryptococcosis diagnosed by cytology and treated at the Veterinary Hospital of the State University of Santa Cruz (HV-UESC).Case: A 5-year-old uncastrated male cat was taken to the HV-UESC small animal clinic, owing to a gradual increase inthe volume of the nasal plane spanning over a year, accompanied by respiratory distress. Upon physical examination, theanimal was found to have regular nutritional status, dehydration, hypocorous mucosae, and enlarged popliteal lymph nodes.In addition, multifocal areas of ulceration on the body as well as increased volume in the nasal plane (clown nose) wereobserved. Peripheral blood samples were collected for complete blood count and biochemical analysis. Due to clinicalsuspicion of fungal lesions, fine needle aspiration of the nasal lesion was performed and cytological slides were sent tothe HV-UESC histopathology laboratory. Probable diagnoses included rhinosporidiosis, cryptococcosis, and histoplasmosis. The animal was subjected to thoracic radiography in the imaging division of the HV-UESC. Blood counts revealedthrombocytopenia (130,000/mm3) and biochemical tests presented hypoproteinemia (3.2 g/dL), with hypoalbuminemia(1.52 g/dL); however, chest radiography did not show pulmonary alterations. Results of cytological analysis indicatedpyogranulomatous inflammation associated with intracellular organisms, demonstrating a pathogen morphology similarto that of Cryptococcus spp. Before the diagnosis, itraconazole1 5 mg/kg (1 capsule, twice a day for 30 days) and fipronil2(1 ampoule 0.5 mL, on the back) were prescribed...(AU)

Pneumonia e leptomeningite criptocócica em felino: relato de caso / Cryptococcal pneumonia and leptomeningitis in feline: a case report

A criptococose é uma doença fúngica infecciosa, tegumentar e/ou sistêmica, crônica, de distribuição global, causada por levedura sa-próbia do gênero Cryptococcus que pode infectar humanos, animais silvestres e mamíferos domésticos, especialmente felinos domésticos. Duas espécies estão mais diretamente envolvidas com a doença: o Cryptococcus neoformans e o Cryptococus gattii, porém pacientes imunocomprometidos têm, como principal causador, o Cryptococcus neoformans. Todavia, O Cryptococcus gattii, além de afetar hospedei-ros imunossuprimidos, também pode causar a doença em indivíduos imunocompetentes. Este relato apresenta os achados clínicos e o diagnóstico firmado com o exame histopatológico de um caso de pneumonia e leptomeningite criptocócica em um felino, macho, sem raça definida, de cinco anos de idade, atendido no Hospital Veterinário “Dr. Vicente Borelli” – do Centro Universitário da Fundação de Ensino Octávio Bastos (Unifeob).

Cryptococcosis is a chronic, global, infectious, integumentary and/or systemic fungal disease caused by yeast from the genus Cryptococcus that can infect humans, wild animals and domestic mammals, especially domestic cats. Two species are most directly involved with the disease Cryptococcus neoformansand Cryptococcus gattii, but immunocompromised patients have, as the main cause, Cryptococcus neoformans. However, Cryptococcus gattii, despite affecting immunosuppressed hosts, can also cause disease in immunocompetent individuals. The present report presents the clinical findings and the diagnosis confirmed through the histopathological examination of a cryptococcal pneumonia and leptomeningitis case observed in a male feline, undefined breed, five-year-old, treated at the Veterinary Hospital “Dr. Vicente Borreli”– do Centro Universitário da Fundação de Ensino Octávio Bastos (Unifeob).

Pneumonia e leptomeningite criptocócica em felino: relato de caso / Cryptococcal pneumonia and leptomeningitis in feline: a case report

A criptococose é uma doença fúngica infecciosa, tegumentar e/ou sistêmica, crônica, de distribuição global, causada por levedura sa-próbia do gênero Cryptococcus que pode infectar humanos, animais silvestres e mamíferos domésticos, especialmente felinos domésticos. Duas espécies estão mais diretamente envolvidas com a doença: o Cryptococcus neoformans e o Cryptococus gattii, porém pacientes imunocomprometidos têm, como principal causador, o Cryptococcus neoformans. Todavia, O Cryptococcus gattii, além de afetar hospedei-ros imunossuprimidos, também pode causar a doença em indivíduos imunocompetentes. Este relato apresenta os achados clínicos e o diagnóstico firmado com o exame histopatológico de um caso de pneumonia e leptomeningite criptocócica em um felino, macho, sem raça definida, de cinco anos de idade, atendido no Hospital Veterinário “Dr. Vicente Borelli” do Centro Universitário da Fundação de Ensino Octávio Bastos (Unifeob). (AU)

Cryptococcosis is a chronic, global, infectious, integumentary and/or systemic fungal disease caused by yeast from the genus Cryptococcus that can infect humans, wild animals and domestic mammals, especially domestic cats. Two species are most directly involved with the disease Cryptococcus neoformansand Cryptococcus gattii, but immunocompromised patients have, as the main cause, Cryptococcus neoformans. However, Cryptococcus gattii, despite affecting immunosuppressed hosts, can also cause disease in immunocompetent individuals. The present report presents the clinical findings and the diagnosis confirmed through the histopathological examination of a cryptococcal pneumonia and leptomeningitis case observed in a male feline, undefined breed, five-year-old, treated at the Veterinary Hospital “Dr. Vicente Borreli”do Centro Universitário da Fundação de Ensino Octávio Bastos (Unifeob).(AU)

Histoplasmose nasocutânea em gato / Nasocutaneous histoplasmosis in cat

Histoplasmosis is a systemic fungal disease of an opportunistic nature that affects humans and animals. The objective of this work were to report a clinical case of histoplasmosis in a cat. In the Veterinary Hospital Professor Sylvio Barbosa Cardoso (HVSBC) a cat, no defined breed, female, of four years of age. No cytopathological examination was obtained through histoplasma spp. without cytoplasm of macrophages. Itraconazole in suspension (40 mg / mL / SID) was prescribed for the patient. On the return of the animal, after 21 days of treatment, it was not printed on a readiness scale, then the same drug was prescribed with dose adjustment (50 mg / capsule / SID for 90 days) and pharmaceutical form, and after 38 days was remission of the nasal region.(AU)

Histoplasmose nasocutânea em gato / Nasocutaneous histoplasmosis in cat

Histoplasmosis is a systemic fungal disease of an opportunistic nature that affects humans and animals. The objective of this work were to report a clinical case of histoplasmosis in a cat. In the Veterinary Hospital Professor Sylvio Barbosa Cardoso (HVSBC) a cat, no defined breed, female, of four years of age. No cytopathological examination was obtained through histoplasma spp. without cytoplasm of macrophages. Itraconazole in suspension (40 mg / mL / SID) was prescribed for the patient. On the return of the animal, after 21 days of treatment, it was not printed on a readiness scale, then the same drug was prescribed with dose adjustment (50 mg / capsule / SID for 90 days) and pharmaceutical form, and after 38 days was remission of the nasal region.

MELANOGÊNESE E SENSIBILIDADE DE CEPAS DO COMPLEXO DE ESPÉCIES Cryptococcus neoformans/Cryptococcus gattii IN VITRO: UMA MODULACÃO COM INIBIDORES DA BOMBA DE PRÓTONS, CATECOLAMINAS E PARAQUAT

Cryptococcus spp. são agentes etiológicos da criptococose. Para estabelecer a infecção, essas leveduras expressam fatores de virulência como a melanina e o biofilme. Esta tese foi dividida em três objetivos: 1. Avaliar os efeitos dos inibidores da bomba de prótons (IBPs) sobre o crescimento e a produção de melanina de Cryptococcus spp.; 2. Investigar a produção de melanina pelo complexo de espécies C. neoformans/C. gattii na presença de diferentes precursores da melanogênese, avaliando o efeito da melanização na sensibilidade antifúngica; 3. Avaliar a atividade de paraquat (PQ) contra Cryptococcus spp. nas formas planctônica e biofilme, bem como o efeito protetor de agentes antioxidantes sobre essa atividade. Buscou-se avaliar ainda a cinética da produção de melanina diante o PQ. As concentrações inibitórias mínimas (CIMs) de IBPs foram determinadas por microdiluição. O efeito dos IBPs na produção de melanina foi avaliado na presença de sulfato de cobre ou glutationa. Foram utilizados como substratos para a melanogênese, a epinefrina, norepinefrina, dopamina, ácido cafeico e L-dopa, sendo em seguida avaliada a sensibilidade in vitro. A atividade da anfotericina B em cepas melanizadas foi investigada no ensaio de time kill. A sensibilidade ao PQ foi avaliada e, em seguida, as cepas expostas ao PQ foram cultivadas em meio com ou sem ácido ascórbico (AA) e glutationa (GSH). Ademais, a produção de melanina foi avaliada na presença de L-dopa + PQ. Os IBPs mostraram CIMs variando de 125-1000 g/mL e redução da melanização em células de Cryptococcus. O sulfato de cobre e a glutationa restauraram a melanogênese das cepas na presença de IBPs. Adicionalmente, a presença de IBPs diminuiu a toxicidade do sulfato de cobre (1 mM). As cepas produziram melanina, em escalas diferenciada, de acordo com o substrato testado. As células melanizadas foram mais tolerantes à anfotericina B. No tocante ao efeito do PQ, observou-se CIM variando de 8-256 g/mL e uma inibição na formação do biofilme. O efeito inibitório planctônico foi restaurado por AA e GSH. Além disso, L-dopa + PQ alterou a cinética da produção de melanina. Em conclusão, observou-se que IBPs inibem o crescimento planctônico e a melanogênese em Cryptococcus spp., sendo este último efeito dependente da quelação do cobre ou inibição do transporte de cobre ATPase. Ademais, os dados em conjunto, evidenciaram que a melanina contribui para a maior resiliência de Cryptococcus spp. a drogas antifúngicas, como a anfotericina B, e agentes agressores ambientais, como o paraquat.

Cryptococcus spp. are etiological agents of cryptococcosis. To establish infection, these yeasts produce virulence factors such as melanin and biofilm. This thesis was divided into three aims: 1. Evaluate the effects of proton pump inhibitors (PPIs) on growth and melanin production by Cryptococcus spp.; 2. Investigate the production of melanin by the C. neoformans/C. gattii species complex in the presence of different precursors of melanogenesis, evaluating the effect of melanization on antifungal susceptibility. 3. Evaluate the activity of paraquat (PQ) against Cryptococcus spp. in planktonic and biofilm forms, as well as the protective role of antioxidant agents against PQ. We also sought to evaluate the kinetics of melanin production in the presence of PQ. Minimum inhibitory concentrations (MICs) of PPIs were determined by microdilution. The effect of PPIs on melanin production was evaluated in the presence of copper sulfate or glutathione. As substrates for melanogenesis, epinephrine, norepinephrine, dopamine, caffeic acid and L-dopa were used, then, in vitro susceptibility was evaluated. Amphotericin B activity against melanized strains was investigated by the time kill assay. The susceptibility to PQ was evaluated and the strains exposed to the PQ were cultivated in medium with or without ascorbic acid (AA) and glutathione (GSH). Furthermore, melanin production was evaluated in the presence of L-dopa + PQ. PPIs showed MICs ranging from 125-1000 g/mL and reduced melanization of Cryptococcus cells. Copper sulfate and glutathione restored the melanogenesis of the strains in the presence of PPIs. Additionally, the presence of PPIs decreased copper sulphate toxicity (1 mM). The strains produced melanin, at different levels, according to the tested substrate. Melanized cells were more tolerant to amphotericin B. Regarding the effect of PQ, MICs ranged from 8-256 g/mL and biofilm formation was inhibited. The inhibitory effect of PQ was hampered by AA and GSH. Furthermore, L-dopa + PQ altered the kinetics of melanin production. In conclusion, it was observed that PPIs inhibit planktonic growth and melanogenesis of Cryptococcus spp., with the latter being dependent on copper chelation or inhibition of copper ATPase transport. Furthermore, altogether, the data showed that melanin contributes to reduced susceptibility of Cryptococcus spp. to the antifungal drug amphotericin B, and the environmental aggressor paraquat.

Granulomatous leptomeningitis in a goat associated with infection by Cryptococcus neoformans

A case of granulomatous leptomeningitis caused by Cryptococcus neoformans is described in a 2-year-old mixed-breed, female goat which presented acute neurological signs including ataxia, nystagmus, bilateral blindness, opisthotonus, hyperesthesia, and spastic paresis of forelimbs. Granulomatous inflammation was detected in the pia arachnoid covering the frontal, parietal and occipital lobes of the telencephalon; cerebellum, thalamus, mesencephalon, pons, medulla, and cervical and thoracolumbar segments of the spinal cord. Yeast-like organisms with morphology compatible with Cryptococcus spp. were observed. A 350 base pair sequence was amplified from DNA extracted from the formalin fixed paraffin embedded (FFPE) tissue. The amplified sequence matched 100% Cryptococcus neoformans. It is suggested that cryptococcal meningitis should be included in the differential diagnosis list of goat diseases with neurological signs such as ataxia, opisthotonus, nystagmus, hyperesthesia and spastic paresis.

Granulomatous leptomeningitis in a goat associated with infection by Cryptococcus neoformans

A case of granulomatous leptomeningitis caused by Cryptococcus neoformans is described in a 2-year-old mixed-breed, female goat which presented acute neurological signs including ataxia, nystagmus, bilateral blindness, opisthotonus, hyperesthesia, and spastic paresis of forelimbs. Granulomatous inflammation was detected in the pia arachnoid covering the frontal, parietal and occipital lobes of the telencephalon; cerebellum, thalamus, mesencephalon, pons, medulla, and cervical and thoracolumbar segments of the spinal cord. Yeast-like organisms with morphology compatible with Cryptococcus spp. were observed. A 350 base pair sequence was amplified from DNA extracted from the formalin fixed paraffin embedded (FFPE) tissue. The amplified sequence matched 100% Cryptococcus neoformans. It is suggested that cryptococcal meningitis should be included in the differential diagnosis list of goat diseases with neurological signs such as ataxia, opisthotonus, nystagmus, hyperesthesia and spastic paresis.(AU)