Candidose sistêmica em cães associada à cinomose / Systemic candidosis in dogs associated with canine distemper virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidosis associated to canine distemper virus in dogs, highlighting the main epidemiological, clinical and pathological aspects. Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were...

Candidose sistêmica em cães associada à cinomose / Systemic candidosis in dogs associated with canine distemper virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidosis associated to canine distemper virus in dogs, highlighting the main epidemiological, clinical and pathological aspects. Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were...(AU)

Systemic infection by Spencermartinsiella sp. in a Nile crocodile (Crocodylus niloticus) / Infecção sistêmica por Spencermartinsiella sp. em um crocodilo do Nilo (Crocodylus niloticus)

A male adult crocodile (Crocodylus niloticus) was diagnosed with systemic yeast infection. Histologically, there were extensive areas of necrosis in the lung, which were associated with a diffuse severe lympho-plasmo-histiocytic inflammatory infiltrate, with numerous multinucleated giant cells, and myriads of intralesional pseudo-hyphae and yeast like organisms within distended foveolae. Necrotic foci were also observed in the mucosa of the digestive tract, trachea, tunica intima of arteries, liver, and heart, with a marked inflammatory lympho-histiocytic infiltrate, with large numbers of epithelioid macrophages and giant cells, and intralesional and intravascular pseudo-hyphae and yeast-like organisms. Oval yeast structures with 4 to 6 µm in diameter and 5 to 8 µm thick paralleled-wall pseudo-hyphae were observed in PAS or GMS stained sections. PCR with DNA template extracted from paraffin embedded tissues amplified the D1/D2 domains of the large subunit rRNA gene, which was sequenced and found to be identical to sequences of a new species, isolated from rotting wood in Brazil, of the genus Spencermartinsiella, which its closest relative is Spencermartinsiella cellulosicola.(AU)

Um crocodilo macho adulto (Crocodylus niloticus) foi diagnosticado com infecção fúngica sustêmica. Histologicamente, havia extensas áreas de necrose no pulmão, que estavam associadas com infiltrado inflamatório linfo-plasmo-histiocitário, com numerosas células gigantes multinucleadas e miríade de pseudo-hifas e organismos leveduriformes intralesionais, dentro de favéolas distendidas. Focos necróticos também foram observados na mucosa do trato digestório, traquéia, túnica íntima de artérias, fígado e coração, com acentuado infiltrado inflamatório linfo-histiocitário, com grande número de macrófagos epitelioides e células gigantes e hifas e organismos leveduriformes intralesionais e intravasculares. Cortes corados por PAS e GMS evidenciaram estruturas leveduriformes ovais com 4 a 6 µm de diâmetro e pseudo-hifas de paredes espessas e paralelas com 5 a 8 µm. PCR realizado com DNA extraído de material parafinizado amplificou os domínios D1/D2 da subunidade maior do gene rRNA, cuja sequencia foi idêntica a sequências de uma nova espécie, isolada no Brasil de madeira em decomposição, do gênero Spencermartinsiella, cuja espécie mais próxima é Spencermartinsiella cellulosicola.(AU)

Systemic infection by Spencermartinsiella sp in a Nile crocodile (Crocodylus niloticus) / Infecção sistêmica por Spencermartinsiella sp em um crocodilo do Nilo (Crocodylus niloticus)

A male adult crocodile (Crocodylus niloticus) was diagnosed with systemic yeast infection. Histologically, there were extensive areas of necrosis in the lung, which were associated with a diffuse severe lympho-plasmo-histiocytic inflammatory infiltrate, with numerous multinucleated giant cells, and myriads of intralesional pseudo-hyphae and yeast like organisms within distended foveolae. Necrotic foci were also observed in the mucosa of the digestive tract, trachea, tunica intima of arteries, liver, and heart, with a marked inflammatory lympho-histiocytic infiltrate, with large numbers of epithelioid macrophages and giant cells, and intralesional and intravascular pseudo-hyphae and yeast-like organisms. Oval yeast structures with 4 to 6 m in diameter and 5 to 8 m thick paralleled-wall pseudo-hyphae were observed in PAS or GMS stained sections. PCR with DNA template extracted from paraffin embedded tissues amplified the D1/D2 domains of the large subunit rRNA gene, which was sequenced and found to be identical to sequences of a new species, isolated from rotting wood in Brazil, of the genus Spencermartinsiella, which its closest relative is Spencermartinsiella cellulosicola.(AU)

Um crocodilo macho adulto (Crocodylus niloticus) foi diagnosticado com infecção fúngica sustêmica. Histologicamente, havia extensas áreas de necrose no pulmão, que estavam associadas com infiltrado inflamatório linfo-plasmo-histiocitário, com numerosas células gigantes multinucleadas e miríade de pseudo-hifas e organismos leveduriformes intralesionais, dentro de favéolas distendidas. Focos necróticos também foram observados na mucosa do trato digestório, traquéia, túnica íntima de artérias, fígado e coração, com acentuado infiltrado inflamatório linfo-histiocitário, com grande número de macrófagos epitelioides e células gigantes e hifas e organismos leveduriformes intralesionais e intravasculares. Cortes corados por PAS e GMS evidenciaram estruturas leveduriformes ovais com 4 a 6 μm de diâmetro e pseudo-hifas de paredes espessas e paralelas com 5 a 8 μm. PCR realizado com DNA extraído de material parafinizado amplificou os domínios D1/D2 da subunidade maior do gene rRNA, cuja sequencia foi idêntica a sequências de uma nova espécie, isolada no Brasil de madeira em decomposição, do gênero Spencermartinsiella, cuja espécie mais próxima é Spencermartinsiella cellulosicola.(AU)

Systemic infection by Spencermartinsiella sp in a Nile crocodile (Crocodylus niloticus) / Infecção sistêmica por Spencermartinsiella sp em um crocodilo do Nilo (Crocodylus niloticus)

A male adult crocodile (Crocodylus niloticus) was diagnosed with systemic yeast infection. Histologically, there were extensive areas of necrosis in the lung, which were associated with a diffuse severe lympho-plasmo-histiocytic inflammatory infiltrate, with numerous multinucleated giant cells, and myriads of intralesional pseudo-hyphae and yeast like organisms within distended foveolae. Necrotic foci were also observed in the mucosa of the digestive tract, trachea, tunica intima of arteries, liver, and heart, with a marked inflammatory lympho-histiocytic infiltrate, with large numbers of epithelioid macrophages and giant cells, and intralesional and intravascular pseudo-hyphae and yeast-like organisms. Oval yeast structures with 4 to 6 m in diameter and 5 to 8 m thick paralleled-wall pseudo-hyphae were observed in PAS or GMS stained sections. PCR with DNA template extracted from paraffin embedded tissues amplified the D1/D2 domains of the large subunit rRNA gene, which was sequenced and found to be identical to sequences of a new species, isolated from rotting wood in Brazil, of the genus Spencermartinsiella, which its closest relative is Spencermartinsiella cellulosicola.

Um crocodilo macho adulto (Crocodylus niloticus) foi diagnosticado com infecção fúngica sustêmica. Histologicamente, havia extensas áreas de necrose no pulmão, que estavam associadas com infiltrado inflamatório linfo-plasmo-histiocitário, com numerosas células gigantes multinucleadas e miríade de pseudo-hifas e organismos leveduriformes intralesionais, dentro de favéolas distendidas. Focos necróticos também foram observados na mucosa do trato digestório, traquéia, túnica íntima de artérias, fígado e coração, com acentuado infiltrado inflamatório linfo-histiocitário, com grande número de macrófagos epitelioides e células gigantes e hifas e organismos leveduriformes intralesionais e intravasculares. Cortes corados por PAS e GMS evidenciaram estruturas leveduriformes ovais com 4 a 6 μm de diâmetro e pseudo-hifas de paredes espessas e paralelas com 5 a 8 μm. PCR realizado com DNA extraído de material parafinizado amplificou os domínios D1/D2 da subunidade maior do gene rRNA, cuja sequencia foi idêntica a sequências de uma nova espécie, isolada no Brasil de madeira em decomposição, do gênero Spencermartinsiella, cuja espécie mais próxima é Spencermartinsiella cellulosicola.

MUCORMICOSE E CANDIDÍASE EM CÃES

Ultimamente, as doenças fúngicas tem sido responsáveis por elevada mortalidade na espécie canina. A zigomicose e a candidíase estão inseridas entre as principais micoses que eventualmente podem provocar infecções sistêmicas e fatais em indivíduos imunossuprmidos. Esta Dissertação, foi elaborada em dois capítulos, composta por dois artigos originais. O primeiro submetido à Journal of Comparative Pathology relata dois casos de infecções por fungos da ordem Mucorales com envolvimento sistêmico e hepatogástrico em cães, descrevendo os aspectos epidemiológicos, clínicos e patológicos. A doença ocorreu em cães Poodle, machos com idade de um e dez anos. As alterações clínicas incluíam sinais respiratórios, neurológicos e gastrointestinais que progrediram evoluindo para morte em três a dezenove dias. Macroscopicamente, no caso 1, havia áreas irregulares amareladas ou avermelhadas e elevadas que aprofundavam-se ao corte nos pulmões, coração e encéfalo. No caso 2, as alterações consistiam em ruptura de estômago com bordos e mucosa gástrica recobertos por espesso material brancacento e friável, além de múltiplas áreas extensas nodulares e cavitações profundas amarelo-brancacentas na superfície capsular do fígado. Histologicamente, foram observados piogranulomas em todos os órgãos afetados, associados a hifas fúngicas, vasculite e trombose. O diagnóstico de mucormicose foi realizado através das lesões microscópicas associadas às características morfotintoriais das hifas. As hifas foram fortemente imunomarcadas pelo anticorpo monoclonal anti-Rhizopus arrhizus. O segundo artigo, foi submetido à revista Arquivo Brasileiro de Medicina Veterinária e Zootecnia, descreve os aspectos epidemiológicos, clínicos e patológicos de três casos de candidíase sistêmica associada a cinomose em cães. Os cães acometidos tinham idade de quatro meses a cinco anos. Desses, dois eram cães sem raça definida e um Poodle. As manifestações clínicas eram predominantemente neurológicas, seguidas de alterações inespecíficas com evolução de cinco a trinta dias. Na necropsia havia áreas branco-amareladas circundadas por bordas avermelhadas nos rins, coração, fígado, linfonodo submandibular e pulmão. Adicionalmente, no caso 2, havia material grumoso brancacento aderido à superfície epicárdica, artéria aorta, saco pericárdico e baço. Além de infarto no testículo direito, hidroureter, pielonefrite e hidronefrose. No caso 1, havia aumento da articulação carpo-metacárpica direita com conteúdo avermelhado e turvo. No encéfalo, as lesões caracterizavam-se por áreas enegrecidas e/ou avermelhadas e friáveis. Microscopicamente, as lesões consistiam em áreas de necrose e hemorragia, circundadas por neutrófilos associada a vasculite, congestão, trombose, infarto e fungos com vários padrões morfológicos que incluíam leveduras, pseudo-hifas e hifas. Os três cães estavam imunossuprimidos decorrente da infecção concomitante pelo vírus da cinomose canina. Os casos 2 e 3 foram diagnosticados ainda com dermatofitose generalizada e linfoma, respectivamente. O diagnóstico de candidíase sistêmica foi realizado através das características morfotintoriais do agente e imunomarcação pelo anticorpo policlonal anti-Candida albicans. Com a realização desses trabalhos foi possível constatar que a mucormicose e a candidíase são doenças esporadicamente diagnosticadas na rotina do Laboratório de Patologia Animal do Hospital Veterinário Universitário Prof. Dr. Ivon Macêdo Tabosa da UFCG. Embora pouco frequentes, essas micoses constituem importantes causas de morte em cães.

Lately, fungal diseases have been responsible for high mortality in the canine species. The zygomycosis, and candidiasis are inserted between the main mycoses which eventually may cause systemic and fatal infections in immunosuppressed individuals. This Dissertation, was elaborated in two chapters, composed by two original articles. The first one submitted to the Journal of Comparative Pathology, describe two cases of fungal infection of the order Mucorales with systemic and hepatogastric involvement in dogs. The disease occurred in Poodle dogs, males aged one and ten years. The clinical signs included respiratory, neurological and gastrointestinal symptoms that progressed to death in three and nineteen days. Macroscopically, in case 1, there were irregular yellowish or reddish raised areas that deepened the cut in the lungs, heart and brain. In case 2, the alterations consisted of rupture of the stomach with borders and gastric mucosa covered by a thick and whitish, but also friable material, in addition to multiple extensive nodular areas and deep white-yellowish cavitations on the capsular surface of the liver. Histologically, piogranulomas were observed in all affected organs, associated with fungal hyphae, vasculitis and thrombosis. The diagnosis of mucormycosis was based on the microscopic lesions associated with the morphotinctorial features of the hyphae. The hyphae were strongly immunomarked by the anti-Rhizopus arrhizus monoclonal antibody. The second article, was submitted to the journal Arquivo Brasileiro de Medicina Veterinária e Zootecnia, describes the epidemiological, clinical and pathological aspects of three cases of systemic candidiasis associated with distemper in dogs. The affected dogs were between four months and five years. Of these, two were without defined breed dogs and one Poodle. The clinical manifestations were predominantly neurological, followed by nonspecific changes with evolution from five to thirty days. At necropsy there were yellowish-white areas surrounded by reddish edges in the kidneys, heart, liver, submandibular lymph node and lung. In addition, in case 2, there were lumpy materials whitish adhered to the epicardial surface, aortic artery, pericardial sac and spleen. In addition to infarction in the right testicle, hydroureter, pyelonephritis and hydronephrosis. In case 1, there was an increase in the right carpal-metacarpal joint, that when cutting reddish and cloudy content flowed. In the encephalon, the lesions were characterized by blackened and/or reddish and friable areas. Microscopically, the lesions consisted of areas of necrosis and hemorrhage, surrounded by neutrophils associated with vasculitis, congestion, thrombosis, infarction, and fungi with various morphological patterns including yeasts, pseudo-hyphae and hyphae. The three dogs were immunosuppressed due to the concomitant infection with the distemper virus. Cases 2 and 3 were diagnosed still with generalized dermatophytosis and lymphoma, respectively. The diagnosis of systemic candidiasis was made through the morphotinctorial characteristics of the agent and immunostaining by the polyclonal anti-Candida albicans antibody. With the accomplishment of these works it was possible to verify that mucormycosis and candidiasis are diseases sporadically diagnosed in the routine of the Laboratório de Patologia Animal of the Hospital Veterinário Universitário Prof. Dr. Ivon Macêdo Tabosa of the UFCG. Although uncommon, these mycoses are important causes of death in dogs.

Enteritis associated with Clostridium difficile and opportunistic candidiasis in a foal

A twelve-day-old foal was admitted to the Veterinary School of UFMG with profuse diarrhea. The animal was treated with several medications, including antibiotics. Despite the clinical care, the clinical condition did not improve. Due to poor prognosis, the foal was euthanized. Gross and histopathological examinations, showed pseudomembranous and/or membranous glossitis, esophagitis and gastritis with intense amount of pseudohyphae and blastoconidia compatible with Candida spp Also, fibrinous and necrotizing typhlocolitis was observed. Clostridium difficile was isolated from intestinal contents which were also positive for toxins A/B by ELISA. Using a multiplex-PCR assay, genes encoding toxin A (tcdA) and toxin B (tcdB) were detected. The severe lesions caused by Candida in this foal probably occurred due to changes in the microbiota induced by the treatment with antibiotics. However, the possibility of an acquired immunodeficiency cannot be excluded because information about quantity and quality of the colostrum ingested by this foal was not obtained.(AU)

Mixed mycotic rhinitis and pneumonia in wild boars

Background: Wild boar population is present worldwide. Contact between wild boars and domestic pigs may occur occasionally, and several diseases, as well as the occurrence of opportunistic infections are observed in both species. Mycotic rhinitis and pneumonia were reported before in pig herds, mainly associated with immunosuppression caused by viral infection. This study reports the occurrence of mycotic rhinitis in two wild boars due to Aspergillus fumigatus, A. fl avus and Candida albicans, together with Pneumocystis sp. in the lungs, originating from a herd infected with PCV2. Cases: In a commercial wild boar herd, poor body condition, sneezing and diarrhea were observed. Three animals were euthanized and, in two of them, yellow and green plaque-like masses of fungal growth in the mucosal and in cartilage surface and accentuated atrophy of nasal turbinates were observed. Additionally, multifocal subcutaneous abscesses in the maxillary area and bilateral reddening of the ocular mucosa with muco-purulent discharge were noted. Microscopically, in fragments from the nasal cavity of the two affected wild pigs, massive ulceration of the mucosal surface and presence of hyphae with septations and dichotomous branching and pseudohyphae were observed. Multifocal moderated interstitial pneumonia and alveolar edema were the main histological lesions founded in the lungs of 3 animals. In the lymph nodes multifocal moderated lymphoid depletion and lymphohistiocytic infi ltrated was the main microscopical lesion. Aspergillus fumigatus, A. fl avus and Candida albicans were isolated in nasal cavity. Pseudomonas aeruginosa was isolated from the subcutaneous abscesses and Staphylococcus hyicus and Streptococcus equisimilis from ocular swab. Pneumocystis was detected in lungs from the three wild boars by nested PCR, Grocott´s staining and immunohistochemistry (IHC). Porcine circovirus 2 (PCV2) was detected in lungs by PCR. Virus detection by IHC was only confi rmed in one wild boar. Discussion: Diagnostic of mycotic rhinitis and pneumonia was based on macroscopical and microscopical fi ndings, as well as mycological analysis, IHC and Groccott ´s methenamine staining. Pneumocystis carinii, Aspergillus spp. and Candida spp. are considered as opportunistic fungal pathogens commonly associated with immunosuppression in animals and humans and have been found in lungs and in muco-cutaneous tissue of PMWS affected pigs. Clinically, immunodeficiency is usually associated with illness caused by organisms of low pathogenicity or well-know secondary pathogens, among other factors. Besides immunodefi ciency, prolonged antimicrobial therapy is another predisposing factor to the development of mycotic infections, well described in animals. In the present report, antimicrobial therapy was performed when respiratory signs were noted in therapeutic doses, suggesting that massive antibiotic use was not the trigger of mycotic rhinitis. PCV2 IHC result positive only in one wild pig, although all the samples were positive by PCR. This fi nding could indicate a subclinical infection or a recovery phase of the disease in the IHC negative cases, as previously suggested for domestic and wild pigs using in situ hybridization. PCV2 load in wild boar was lower when compared with domestic pigs. A viral load higher than 108 PCV2 genomes per 500 ng DNA was required to give a visible IHC staining in swine. Although quantitative PCR it was not used in order to detect PCV2 in the present report, the viral load could be another possible explanation for the IHC negative cases observed. The role of PCV2 as a cause of immunosupression, facilitating the infection with secondary agents as Aspergillus, Candida and Pneumocystis cannot be ruled out.(AU)

Mixed mycotic rhinitis and pneumonia in wild boars

Background: Wild boar population is present worldwide. Contact between wild boars and domestic pigs may occur occasionally, and several diseases, as well as the occurrence of opportunistic infections are observed in both species. Mycotic rhinitis and pneumonia were reported before in pig herds, mainly associated with immunosuppression caused by viral infection. This study reports the occurrence of mycotic rhinitis in two wild boars due to Aspergillus fumigatus, A. fl avus and Candida albicans, together with Pneumocystis sp. in the lungs, originating from a herd infected with PCV2. Cases: In a commercial wild boar herd, poor body condition, sneezing and diarrhea were observed. Three animals were euthanized and, in two of them, yellow and green plaque-like masses of fungal growth in the mucosal and in cartilage surface and accentuated atrophy of nasal turbinates were observed. Additionally, multifocal subcutaneous abscesses in the maxillary area and bilateral reddening of the ocular mucosa with muco-purulent discharge were noted. Microscopically, in fragments from the nasal cavity of the two affected wild pigs, massive ulceration of the mucosal surface and presence of hyphae with septations and dichotomous branching and pseudohyphae were observed. Multifocal moderated interstitial pneumonia and alveolar edema were the main histological lesions founded in the lungs of 3 animals. In the lymph nodes multifocal moderated lymphoid depletion and lymphohistiocytic infi ltrated was the main microscopical lesion. Aspergillus fumigatus, A. fl avus and Candida albicans were isolated in nasal cavity. Pseudomonas aeruginosa was isolated from the subcutaneous abscesses and Staphylococcus hyicus and Streptococcus equisimilis from ocular swab. Pneumocystis was detected in lungs from the three wild boars by nested PCR, Grocott´s staining and immunohistochemistry (IHC). Porcine circovirus 2 (PCV2) was detected in lungs by PCR. Virus detection by IHC was only confi rmed in one wild boar. Discussion: Diagnostic of mycotic rhinitis and pneumonia was based on macroscopical and microscopical fi ndings, as well as mycological analysis, IHC and Groccott ´s methenamine staining. Pneumocystis carinii, Aspergillus spp. and Candida spp. are considered as opportunistic fungal pathogens commonly associated with immunosuppression in animals and humans and have been found in lungs and in muco-cutaneous tissue of PMWS affected pigs. Clinically, immunodeficiency is usually associated with illness caused by organisms of low pathogenicity or well-know secondary pathogens, among other factors. Besides immunodefi ciency, prolonged antimicrobial therapy is another predisposing factor to the development of mycotic infections, well described in animals. In the present report, antimicrobial therapy was performed when respiratory signs were noted in therapeutic doses, suggesting that massive antibiotic use was not the trigger of mycotic rhinitis. PCV2 IHC result positive only in one wild pig, although all the samples were positive by PCR. This fi nding could indicate a subclinical infection or a recovery phase of the disease in the IHC negative cases, as previously suggested for domestic and wild pigs using in situ hybridization. PCV2 load in wild boar was lower when compared with domestic pigs. A viral load higher than 108 PCV2 genomes per 500 ng DNA was required to give a visible IHC staining in swine. Although quantitative PCR it was not used in order to detect PCV2 in the present report, the viral load could be another possible explanation for the IHC negative cases observed. The role of PCV2 as a cause of immunosupression, facilitating the infection with secondary agents as Aspergillus, Candida and Pneumocystis cannot be ruled out.

Avaliação clínica, histológica, histoquímica e histomorfométrica do processo de cicatrização da pitiose cutânea em equinos tratados com acetonida de triancinolona

O estudo teve como objetivo avaliar clínica, histopatológica e histomorfometricamente o processo de recuperação tecidual do granuloma cutâneo da pitiose cutânea em equinos tratados com acetonida de triancinolona. Realizou-se um estudo de tipo descritivo, não probabilístico de conveniência em equinos infectados naturalmente com pitiose cutânea. Foram utilizados 24 equinos com pitiose cutânea, divididos em dois grupos de pesquisa de 12 animais cada. A um dos grupos foi aplicado 50 mg da acetonida de triancinolona intramuscular (GT) enquanto que o outro grupo não foi aplicado tratamento (GC). A todos os animais (GT e GC) se lês realizo caracterização clínica e macroscópica da lesão a cada 4 dias até a recuperação completa do granuloma no GT. Nessa avaliação foi verificada a diminuição do exsudato fibrino sanguinolento, prurido, presença de trajetos fistulosos e saída de kunkers, do mesmo modo, foram realizadas projeções radiológicas para avaliar o grau de compromisso ósseo e fibrose tecidual. Em todos os animais do estudo foram coletadas biópsias de tecido, que foram posteriormente processadas, laminadas e coradas com hematoxilina eosina (H&E), tricrômico de Gómori (TG), picrosirius red/polarization (PR/P) e prata metanamina de Grocott (GMS Grocott´s methenamine silver). Foram selecionados e fotomicrografados 10 campos histológicos e analisadas com ajuda do software BioEstat 5.0, obtendo-se as quantidades de células tissulares como eosinófilos, neutrófilos, macrófagos, fibroblastos e colágeno através da avaliação planimétrica por contagem de pontos. Na avaliação macroscópica da lesão, encontrou-se que todos os animais do GT apresentaram diminuição progressiva nas características clínicas da doença aos 16±1,4 dias após a aplicação do tratamento. Com respeito à contração da ferida, observou-se que todos os granulomas do GT se recuperaram num 100% aos 60±3,4 dias após aplicação do tratamento, indicando diferencia altamente significativa (P<0,0001). No seguimento radiológico das lesões cutâneas crónicas, encontrou-se invasão aos tecidos subjacentes comprometendo estruturas ósseas, caracterizando-se por fibrose tecidual, exostoses moderada, assim como osteólise e osteomielite. Nas colorações de H&E, TG, PR/P e GMS, foram observadas que no GT o processo inflamatório foi diminuindo progressivamente, evidenciado na diminuição da camada fibrino leucocitária, PMN e dos fenómenos de Splendore Hoepli, assim como maior angiogênese, epiteliogênese e aumento da quantidade de fibroblastos e fibras colágenas gerais, assim mesmo na troca progressiva de colágeno tipo III a colágeno tipo I ao final do processo, além de que a presença de hifas intralesionais de Pythium insidiosum diminui-o na segunda semana. Na histomorfometria, observou-se na coloração de GSM, diminuição da presença de hifas intralesionais de P. insidiosum aos 16 dias, sendo estatisticamente significativa (p<0.05). Na coloração de H&E, observou-se diminuição do processo inflamatório, evidenciado nos PMN e os eosinófilos e na regulação dos neutrófilos e macrófagos, a análise estatística demostrou diferencia significativa nas atividades celulares aos 4, 8 e 12 dias apos aplicação da AT, principalmente nos eosinófilos (p=0,0001), neutrófilos (p=0,0001) e macrófagos (p=0,00001). Nas colorações de TG e PR/P, foram observadas aumento da quantidade de fibroblastos e fibras colágenas, assim mesmo na troca progressiva de colágeno tipo III a colágeno tipo I durante o processo de cicatrização, o aumento de fibroblastos mostrou diferencias altamente significativas (p=0,0001) desde o dia 16 e até o dia 40, a expressão de colágeno foi altamente significativo (p=0,0001) a partir do dia 16 e até o final do estudo, especialmente nas fibras tipo I. Estabeleceu-se correlação estatística significativa entre os neutrófilos e macrófagos (p=0,00018), colágeno e eosinófilo (p=0,03) e colágeno e fibroblastos (p=0,02). Nenhuns dos animais do GC apresentaram melhoria nas características clínicas, histológicas, histoquímicas e histomorfométricas da pitiose e se mantiveram iguais ao primeiro dia durante todo o estudo. É possível concluir que a acetonida de triancinolona produz 100% de recuperação clínica nos equinos com pitiose, demostrado pela constatação da histopatologia e da histomorfométria celular e tecidual.

The objective of the present study was to clinical, histopathological and histomorphometrical of the healing process with cutaneous pythiosis in horses treated with triamcinolone acetonide. We conducted a descriptive study, not probabilistic for convenience in live animals and equine patients naturally infected with cutaneous pythiosis. 24 horses were used for skin pythiosis divided into two research groups of 12 animals each. The one group was administered 50 mg of intramuscular triamcinolone acetonide (TG) while the other group was not applied treatment (GC). All animals (TG and CG) if you read realize clinical evaluation and macroscopic characterization of injury every four days until full recovery of the granuloma in GT. In this evaluation was a decreasing fibrin bloody exudate, pruritus, presence of fistulae and output kunkers, likewise radiographic projections were performed to assess the degree of compromise bone and tissue fibrosis. All animals in the study were collected tissue biopsies, further processed, laminated and stained with eosin hematoxylin (H&E), Gómori's trichrome (GT), picrosirius red/polarization (PR/P) and Grocott´s methenamine silver (GMS). They were selected and photomicrographed, 10 histologic fields and analyzed with the help of software BioEstat 5.0, obtaining quantities of tissue cells such as eosinophils, neutrophils, macrophages, fibroblasts and collagen by planimetric evaluation by count points. The macroscopic examination of the injury, it was found that all animals in the TG showed progressive decrease in the clinical characteristics of the disease at 16±1,4 days after treatment. With respect to wound contraction, it was observed that all the TG granulomas recovered in 100% at 60±3,4 days after treatment, indicating differentiates highly significant (P<0.0001). Radiological monitoring of chronic skin lesions, met invasion to underlying tissues compromising bone structures, characterized by tissue fibrosis, moderate exostosis, bake as osteolysis and osteomyelitis. In staining HE, GT, PR/P and GMS, if I note that in TG inflammation was gradually decreasing, as evidenced in decreased fibrin layer leukocyte, PMN and phenomena Splendore Hoepli, also in increased angiogenesis, epiteliogénesis, and increasing the overall amount of fibroblasts and collagen fibers, anyway in the progressive replacement of collagen type III to type I collagen at the end of the process, and that the presence of intralesional pseudo-hyphae of Pythium insidiosum reduces it to the second week. In GSM staining was observed decrease in the presence of intralesional hyphae of P. insidiosum to 16 days and statistically significant (p<0.05). Staining H & E, we observed a decrease of the inflammatory process, shown in PMN and eosinophils and in the regulation of neutrophils and macrophages, the statistical analysis it demonstrated differs significantly in the cell activities after 4, 8 and 12 days after application of the TA, especially in eosinophils (p=0.0001), neutrophils (p=0.0001), and macrophage (p=0.00001). In the staining of GT and PR/P increase the amount of fibroblasts and collagen fibers were observed, also the gradual exchange of type III collagen to type I collagen during the healing process, increased fibroblast show highly significant differences (p=0.0001) and from day 16 until day 40, the expression of collagen was highly significant (p=0.0001) from day 16 until the end of the study, particularly in the fiber type I. It was established statistically significant correlation between neutrophils and macrophages (p=0.00018), collagen and eosinophil (p=0.03) and fibroblasts and collagen (p=0.02). None of the animals in the control group showed improvement in histological and histochemical characteristics of pythiosis and maintained equal to the first day throughout the study. It is possible to conclude that triamcinolone acetonide produces 100% clinical recovery in horses with pythiosis, demonstrated by histomorphometric cellular and tissue observation.

Filamentous growth in Saccharomyces cerevisiae

Fungal dimorphism is a complex phenomenon triggered by a large variety of environmental factors and consists of a reversible alternating pattern of growth between different elliptical and filamentous forms of cells. Understanding the mechanisms that regulate these events is of major interest because of their implications in fungal pathogenesis, cell differentiation and industry. Diploid cells of Saccharomyces cerevisiae transform from budding yeast to pseudohyphae when starved for nitrogen, giving the cells an advantage in food foraging, which is sensed by at least two signal transduction pathways: the MAP kinase (MAPK) and the PKA (cAMP-dependent protein kinase A) pathways. The output of these signalling pathways is the expression of pseudohypha-specific genes, whose expression profiles change and is accompanied by a G2 delay in the cell cycle and a prolonged period of polarized growth. Haploid yeast strains show a similar growth type after prolonged incubation on rich medium plates. The cells form chains and invade the agar on the edge of the colony, but they do not become elongated. This growth type is referred to as haploid invasive growth. Alcohols can also induce filamentous growth in S. cerevisiae, promoting aberrant and elongated morphology. The three forms of filamentous growth are revised in this article and also the pathways involved in sensing, signaling and signal transduction during filamentous growth.

O dimorfismo em fungos é um fenômeno complexo acionado por um grande número de fatores ambientais e consiste num padrão alternante e reversível de crescimento, oscilando entre formas elípticas e filamentosas de células. É de grande importância o entendimento dos mecanismos que regulam esses eventos devido as suas implicações na patogenicidade, diferenciação celular e indústria. Células diplóides de Saccharomyces cerevisiae mudam de células brotantes para pseudohifas quando em condições limitantes de nitrogênio, o que confere às células uma vantagem na procura por alimento. A deficiência de nitrogênio é 'percebida' por pelo menos dois caminhos sinalizadores: 'MAP kinase' (MAPK) e 'PKA' (cAMP-dependent protein kinase A). O resultado dessa sinalização é a expressão de genes específicos para filamentação, cujos perfis de expressão mudam e são acompanhados por um retardo da fase G2 do ciclo celular e um período prolongado de crescimento polarizado. Células haplóides mostram um tipo de crescimento similar após prolongada incubação em meio rico. As células formam cadeias e invadem o ágar na borda da colônia, mas não se tornam alongadas. Esse tipo de crescimento é conhecido como crescimento invasivo haplóide. Os álcoois podem também induzir crescimento filamentoso em S. cerevisiae, ocasionando uma morfologia alongada e aberrante. Nesse artigo revisamos as três formas de crescimento filamentoso incluindo os caminhos envolvidos na percepção, sinalização e transdução do sinal durante o crescimento filamentoso.

Hyphal-like extension and pseudohyphal formation in industrial strains of yeasts induced by isoamyl alcohol

Yeasts can produce pseudohyphae and hyphal-like extensions under certain growth conditions like isoamyl alcohol (IAA) induction, a chief constituent of fusel oil, which is a subproduct from the ethanolic fermentation. The morphology switch from yeast to a filamentous form can be troublesome to the process. In this work it was studied the influence of fusel alcohols, nitrogen sources (ammonium sulphate and leucine) and glifosate (a chemical maturator for sugar cane) added to a complex medium on some industrial strains of yeasts isolated from the fermentative process. Two industrial strains showed transition to hyphal-like extensions or pseudohyphae (clusters of cells) upon addition of IAA from 0.3 to 0.9% /v. The alterations were reversible when the yeasts were reinoculated in YEPD without IAA. Although pseudohyphae are a result of nitrogen-limited medium, we observed them as a result of IAA addition. No influence of the nitrogen source or isopropilic alcohol or glifosate was detected for any strain studied in the concentrations used.

As leveduras podem produzir pseudohifas e extensões semelhantes a hifas sob certas condições de crescimento, como a adição de álcool isoamílico. Esse álcool é o constituinte principal do óleo fusel, um subproduto da fermentação etanólica. A mudança de morfologia da levedura para a forma filamentosa pode causar problemas ao processo fermentativo. Neste trabalho, foi estudada a influência dos álcoois fusel (isoamílico e isopropílico), fontes de nitrogênio (sulfato de amônio e leucina) e glifosate (um maturador químico para cana-de-açúcar) adicionados a um meio complexo sobre linhagens industriais de leveduras isoladas do processo fermentativo. Duas linhagens industriais mostraram transição para pseudohifas (grupos de células) ou extensões semelhantes a hifas após adição de álcool isoamílico, de 0,3% a 0,9% v/v. Todas as alterações foram reversíveis quando as leveduras foram reinoculadas em YEPD sem adição de álcool. Apesar das pseudohifas serem comumente resultantes de meios com limitação de nitrogênio, observou-se que esta morfologia resultou da adição de álcool isoamílico. Não foi detectada nenhuma influência da fonte de nitrogênio ou álcool isopropílico ou glifosate para quaisquer linhagens estudadas.

Enteritis associated with Clostridium difficile and opportunistic candidiasis in a foal

A twelve-day-old foal was admitted to the Veterinary School of UFMG with profuse diarrhea. The animal was treated with several medications, including antibiotics. Despite the clinical care, the clinical condition did not improve. Due to poor prognosis, the foal was euthanized. Gross and histopathological examinations, showed pseudomembranous and/or membranous glossitis, esophagitis and gastritis with intense amount of pseudohyphae and blastoconidia compatible with Candida spp Also, fibrinous and necrotizing typhlocolitis was observed. Clostridium difficile was isolated from intestinal contents which were also positive for toxins A/B by ELISA. Using a multiplex-PCR assay, genes encoding toxin A (tcdA) and toxin B (tcdB) were detected. The severe lesions caused by Candida in this foal probably occurred due to changes in the microbiota induced by the treatment with antibiotics. However, the possibility of an acquired immunodeficiency cannot be excluded because information about quantity and quality of the colostrum ingested by this foal was not obtained.

Atypical Cryptococcus neoformans cells isolated from cerebrospinal fluid samples / Células atípicas de Cryptococcus neoformans var. neoformans observadas em líquido cefalorraquidiano

Most pathogenic fungi show dual morphology features in their saprophytic and parasitic stage. Occasionally, yeast cells produce short and aberrant hyphae, or large cells which can be seen on granulomas histological analysis, and otherwise as giant forms in fresh pus from abscesses, or also as highly encapsulated - serotype B in tissue specimens. The present study reports a HIV positive 39 years old single male patient, who presented oral candidiasis, pulmonary tuberculosis, cardiac insufficiency, Corynebacterium sp meningitis, and neurocryptococcosis due to C. neoformans var. neoformans. The patient was treated with amphotericin B for 45 days, who received approximately a total of 1,960 mg of drug. Atypical C. neoformans cells in diverse sizes and blastoconidia forms were isolated from cerebrospinal fluid (CSF). CSF stained with a China ink preparation revealed the presence of ovoid, branched, and pseudohyphae forms surrounded by a thick capsule. The isolate variety was identified as C. neoformans var. neoformans. Antifungal susceptibility testing performed by means of EUCAST5 technique indicated that the strain was sensitive to amphotericin B (MIC = 0.12g/mL), fluconazole (2g/mL), and itraconazole (0.06 g/mL). The previous reports on atypical yeast cells form described the occurrence of multiple and irregular blastoconidia in human clinical samples.

Muitos fungos patogênicos apresentam morfologias diferentes nas formas parasíticas e saprofíticas. Essas leveduras podem apresentar morfologias diversas como hifas curtas e aberrantes, formas gigantes em pus fresco, abscesso cerebral, cortes histológicos e presença de cápsula espessa geralmente no sorotipo B em tecidos. No presente relato é descrito o caso de um homem de 39 anos de idade, solteiro e HIV positivo, com os seguintes sintomas clínicos: candidíase oral, tuberculose pulmonar, insuficiência cardíaca, meningite por Corynebacterium sp e neurocriptococose por C. neoformans var. neoformans. O paciente foi tratado com anfotericina B durante 45 dias, com dose total de 1960mg. Foram observadas e isoladas do líquido cefalorraquidiano (LCR) células atípicas de C. neoformans de diferentes tamanhos e formas de blastoconídios. O exame direto do LCR em tinta da China apresentou células ovóides, ramificadas, com pseudo-hifas envolvidas por uma fina cápsula. A variedade foi identificada como C. neoformans var. neoformans. O teste de suscetibilidade aos antifúngicos realizados segundo a técnica de EUCAST indicou que a cepa isolada era sensível a: anfotericina B (Concentração Inibitória Mínima - MIC = 0,12g/mL), Fluconazol (2g/mL) e itraconazol (0,06g/mL). Os relatos anteriores sobre leveduras atípicas descrevem a ocorrência de blastoconídios múltiplos e irregulares em materiais bi