Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro

Abstract The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.516 g/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

Resumo O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,516 g/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios semelhantes àqueles induzidos pelo vírus em células não tratadas, indicando danos citotóxicos e deletérios. Nossos resultados corroboram achados relatados anteriormente de que as NETs contribuem para a imunopatologia desenvolvida por pacientes infectados com hRSV.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro / Efeitos das redes extracelulares dos neutrófilos durante a infecção in vitro pelo vírus sincicial respiratório

The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5–16 μg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,5–16 μg/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios [...].

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro / Efeitos das redes extracelulares dos neutrófilos durante a infecção in vitro pelo vírus sincicial respiratório

The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5–16 μg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.(AU)

O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,5–16 μg/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios [...].(AU)

Granulomatous rhinitis by Neoconidiobolus lamprauges in a mule / Rinite granulomatosa por Neoconidiobolus lamprauges em uma mula

ABSTRACT: Conidiobolomycosis has a wide distribution, predominantly in humid tropical regions, affecting several species with significant mortality rates. The genus Conidiobolus is now divided into four genera: Capillidium, Conidiobolus, Microconidiobolus, and Neoconidiobolus. There are no confirmed reports of infection by these fungi in Equidae in Brazil. We present a rhinofacial rhinitis caused by Neoconidiobolus lamprauges in a mule from Rio de Janeiro, Brazil. The mule presented bilateral semi-occlusion of the nostrils, difficulty breathing, and weight loss. The histological examination of the nostril biopsied mass revealed multifocal necrotizing areas with nonstained images of fungal hyphae in the Splendore-Hoeppli reaction and surrounded by macrophages, eosinophils, neutrophils, and multinucleated giant cells. The Grocott methenamine silver staining revealed thin-walled, rarely septated, irregular branching hyphae, with a varying diameter of 12 μm (± 3.63 μm), and terminal ballooning dilations. The determining etiology of this rhinitis was based on the hyphae staining by immunohistochemistry and by amplifying the DNA fragment of N. lamprauges by polymerase chain reaction. Conidiobolomycosis should be included in the differential diagnosis of the causes of rhinitis in equids, mainly in tropical regions.

RESUMO: A conidiobolomicose apresenta ampla distribuição geográfica, com predominância em regiões tropicais úmidas, afeta várias espécies e apresenta taxa de mortalidade significativa. O gênero Conidiobolus está agora dividido em quatro gêneros: Capillidium, Conidiobolus, Microconidiobolus e Neoconidiobolus. Não há relatos confirmados de infecção por esses fungos em equídeos no Brasil. Relata-se o caso de uma mula com rinite rinofacial causada por Neoconidiobolus lamprauges no Rio de Janeiro, Brasil. A mula apresentava uma massa que semiocluía as narinas bilateralmente, dificuldade respiratória e emagrecimento. O exame histológico de biópsia da massa da narina revelou áreas de necrose multifocais com imagens negativas de hifas fúngicas em meio à reação de Splendore-Hoeppli, circundadas por macrófagos, eosinófilos, neutrófilos e células gigantes multinucleadas. O exame histoquímico metenamina prata de Grocott revelou hifas de parede fina, raramente septadas, com ramificação irregular, grau variável de paralelismo, e diâmetro médio de 12 μm (± 3,63) e dilatações balonosas terminais. O diagnóstico etiológico foi realizado pela associação da imuno-histoquímica e da amplificação do fragmento de DNA de N. lamprauges pela reação em cadeia da polimerase. A conidiobolomicose deve ser incluída no diagnóstico diferencial das causas de rinite em equídeos, principalmente em regiões tropicais.

Diarrhea in cats infected with Cystoisospora spp. - evaluation of the neutrophil-to-lymphocyte ratio and monocyte-to-lymphocyte ratio

Background: Cystoisospora spp. is considered a potential pathogen to cause diarrhoea in cats, and the disease occurs clinically, especially in young, weak and immunocompromised cats. The present study aimed to evaluate the clinical utility and diagnostic roles of neutrophil-to-lymphocyte ratio (NLR) and monocyte-to-lymphocyte (MLR) ratios in diarrheal cats infected with Cystoisospora spp. Materials, Methods & Results: Twenty-five cats with diarrhea due to infection caused by Cystoisospora spp. and 15 healthy cats were included in this study. The anamnesis, clinical and laboratory findings of all cats were recorded. A clinical severity score was generated for diarrheal cats, which included activity, appetite, presence of vomit, and faecal consistency. Also, cats infected with Cystoisospora spp. were classified as systemic inflammatory response syndrome (SIRS) positive (+) and SIRS (-) group according to the presence of at least 3 of the SIRS criteria. The Cystoisospora spp. oocysts were diagnosed by direct smear and simple faecal flotation. Cats found positive for other parasitic and protozoal agents by faeces examination were excluded from the study. Complete blood counts (CBC) were performed with an automated blood cell counter for all cats. Blood smears were prepared (with May-Grunwald Giemsa) from samples, and the CBC results were confirmed microscopically (obj.100x). White blood cell (WBC), neutrophil, lymphocyte and monocyte counts were recorded directly from the CBC. The neutrophil-to-lymphocyte ratio was calculated as the ratio of the absolute neutrophil count to the absolute lymphocyte count. The monocyte-to-lymphocyte was calculated as the absolute monocyte count ratio to the absolute lymphocyte count. White blood cell (P = 0.015), neutrophil (P = 0.001), monocyte (P = 0.033), NLR (P = 0.000) and MLR (P = 0.042) of diarrheal cats infected with Cystoisospora spp. were significantly higher than in the healthy group. There was no significant difference between groups in the lymphocyte counts (P > 0.05). SIRS (+) group had statistically significantly higher WBC (P = 0.014), neutrophil counts (P = 0.000), NLR (P = 0.000) and MLR (P = 0.037) than the healthy group. Also, neutrophil counts and NLR of the SIRS (+) group were statistically significantly higher than the SIRS (-) group (P = 0.032; P = 0.001, respectively). However, there was no significant difference SIRS (+), SIRS (-), and healthy groups regarding lymphocyte and monocyte counts (P > 0.05). The clinical severity of the disease positively correlated with NLR and MLR. Also, the best cut-off value of NLR to predict SIRS was > 1.67, with 92.86% sensitivity and 84.62% specificity. MLR was not a useful predictor for SIRS. Discussion: NLR and MLR are parameters derived from complete blood count. In recent years, they have been used in human and veterinary medicine as a biomarker in the diagnosis, prognosis and treatment follow-up of various diseases and provide valuable information about inflammation. In cats, there are limited studies on NLR, and to our knowledge, there are no studies on MLR. The current study suggests that NLR and MLR may be used to determine the severity of the inflammatory condition in diarrheal cats infected with Cystoisospora spp., and NLR is a useful marker for predicting SIRS status in these cats. In addition, this study also will contribute to larger-scale studies in the future.

Granulomatous rhinitis by Neoconidiobolus lamprauges in a mule / Rinite granulomatosa por Neoconidiobolus lamprauges em uma mula

Conidiobolomycosis has a wide distribution, predominantly in humid tropical regions, affecting several species with significant mortality rates. The genus Conidiobolus is now divided into four genera: Capillidium, Conidiobolus, Microconidiobolus, and Neoconidiobolus. There are no confirmed reports of infection by these fungi in Equidae in Brazil. We present a rhinofacial rhinitis caused by Neoconidiobolus lamprauges in a mule from Rio de Janeiro, Brazil. The mule presented bilateral semi-occlusion of the nostrils, difficulty breathing, and weight loss. The histological examination of the nostril biopsied mass revealed multifocal necrotizing areas with nonstained images of fungal hyphae in the Splendore-Hoeppli reaction and surrounded by macrophages, eosinophils, neutrophils, and multinucleated giant cells. The Grocott methenamine silver staining revealed thin-walled, rarely septated, irregular branching hyphae, with a varying diameter of 12 µm (± 3.63 µm), and terminal ballooning dilations. The determining etiology of this rhinitis was based on the hyphae staining by immunohistochemistry and by amplifying the DNA fragment of N. lamprauges by polymerase chain reaction. Conidiobolomycosis should be included in the differential diagnosis of the causes of rhinitis in equids, mainly in tropical regions.

A conidiobolomicose apresenta ampla distribuição geográfica, com predominância em regiões tropicais úmidas, afeta várias espécies e apresenta taxa de mortalidade significativa. O gênero Conidiobolus está agora dividido em quatro gêneros: Capillidium, Conidiobolus, Microconidiobolus e Neoconidiobolus. Não há relatos confirmados de infecção por esses fungos em equídeos no Brasil. Relata-se o caso de uma mula com rinite rinofacial causada por Neoconidiobolus lamprauges no Rio de Janeiro, Brasil. A mula apresentava uma massa que semiocluía as narinas bilateralmente, dificuldade respiratória e emagrecimento. O exame histológico de biópsia da massa da narina revelou áreas de necrose multifocais com imagens negativas de hifas fúngicas em meio à reação de Splendore-Hoeppli, circundadas por macrófagos, eosinófilos, neutrófilos e células gigantes multinucleadas. O exame histoquímico metenamina prata de Grocott revelou hifas de parede fina, raramente septadas, com ramificação irregular, grau variável de paralelismo, e diâmetro médio de 12 µm (± 3,63) e dilatações balonosas terminais. O diagnóstico etiológico foi realizado pela associação da imuno-histoquímica e da amplificação do fragmento de DNA de N. lamprauges pela reação em cadeia da polimerase. A conidiobolomicose deve ser incluída no diagnóstico diferencial das causas de rinite em equídeos, principalmente em regiões tropicais.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro / Efeitos das redes extracelulares dos neutrófilos durante a infecção in vitro pelo vírus sincicial respiratório

Abstract The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.5-16 μg/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

Resumo O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,5-16 μg/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios semelhantes àqueles induzidos pelo vírus em células não tratadas, indicando danos citotóxicos e deletérios. Nossos resultados corroboram achados relatados anteriormente de que as NETs contribuem para a imunopatologia desenvolvida por pacientes infectados com hRSV.

Outbreak of leishmaniasis caused by Leishmania enriettii in guinea pigs (Cavia porcellus) / Surto de leishmaniose causada por Leishmania enriettii em cobaias (Cavia porcellus)

We describe an outbreak of leishmaniasis in seven guinea pigs (Cavia porcellus) in which nodular ulcerated skin lesions of varying sizes were observed in the nasal cavity, upper lip, pinnae, vulva, and periarticular region of the limbs. Cytologic exam of collected samples of the lesions in the auricle of one of the animals revealed macrophages containing parasitophorous vacuoles of approximately 4.0μm in diameter in their cytoplasm with morphology suggestive of Leishmania sp. Although skin lesions spontaneously regressed in two of the Guinea pigs, only one survived. All six animals that died were necropsied. Grossly, all animals showed bloody nodular cutaneous lesions with crusts. One of the guinea pigs had distended dark red and firm lungs. Histopathology of the skin lesions revealed histiocytic interstitial acanthotic dermatitis associated with a myriad of Leishmania organisms within macrophages cytoplasm. In the lung, the lesions were characteristic of broncho-interstitial pneumonia with focal infiltrates of neutrophils, epithelioid macrophages, and multinucleated giant cells containing 2µm basophilic amastigotes with morphology compatible with Leishmania spp. A focal granulomatous lesion ,associated with the causal agent in the lung is a novel description of leishmaniasis in guinea pigs caused by L. enriettii. The polymerase chain reaction (PCR) technique with mini-exon primer performed in samples of lesions from two affected guinea pigs was positive and equal to the reference strain, identifying Leishmania enriettii. The cytological, macroscopic, and histological lesions associated with the PCR technique allowed the diagnosis of leishmaniasis and the identification of the specie L. enriettii.

Descrevemos um surto de leishmaniose em sete cobaias (Cavia porcellus), com lesões cutâneas nodulares ulceradas de tamanhos variados observadas na cavidade nasal, lábio superior, pavilhões auriculares, vulva e região periarticular dos membros. No exame citológico foram encontrados macrófagos contendo vacúolos parasitóforos no citoplasma de aproximadamente 4.0μm em diâmetro com morfologia sugestiva de Leishmania sp. Apesar de regressão espontânea das lesões cutâneas terem ocorrido em duas das sete cobaias, apenas um sobreviveu. Seis dos sete animais afetados morreram e foram necropsiados. Macroscopicamente, todos os animais apresentaram lesões cutâneas nodulares, crostosas e sanguinolentas. Uma das cobaias tinha pulmões vermelho-escuros, distendidos e firmes. A histopatologia das lesões cutâneas revelou dermatite acantótica intersticial histiocítica associada a miríades de organismos de Leishmania no citoplasma de macrófagos. Nos pulmões as lesões eram características de pneumonia bronco-intersticial com infiltrado focal de neutrófilos, eosinófilos, macrófagos epitelioides e células gigantes multinucleadas contendo amastigotas basofílicos de 2µm com morfologia compatível com Leishmania spp. Lesões granulomatosas focais associadas ao agente no pulmão são um achado inédito na leishmaniose causada por L. enriettii em cobaias. A técnica de reação em cadeia da polimerase (PCR) com primer mini-exon realizada em amostras de lesões de duas cobaias afetadas foi positiva, identificando Leishmania enriettii. Os aspectos macroscópicos, citológicos, e histológicos associados à técnica da (PCR), permitiram o diagnóstico da leishmaniose e a identificação da espécie L. enriettii.

Monitoring periodontal lesions and their effects during pregnancy: microbiological aspects of the oral cavity and amniotic fluid in pregnant ewes / Monitoramento de lesões periodontais e seus efeitos na gestação: aspectos microbiológicos da cavidade bucal e líquido amniótico de ovelhas gestantes

Periodontitis affects the teeth supporting tissues, leading to tooth loss and damage to animal health. Evidence in humans suggests that oral microorganisms spread systemically, increasing the risk of pregnancy disorders such as miscarriage, prematurity, and low birth weight. This study aimed to verify whether periodontopathogenic microorganisms reach the transplacental unit, culminating in problems in pregnant ewes. After analyzing the oral cavity, 10 clinically healthy pregnant ewes (OGCH group) and 10 pregnant ewes with periodontitis (OGP group) were selected. The subgingival biofilm was collected for the polymerase chain reaction (PCR) test and amniotic fluid for both the PCR and interleukin (IL) analysis. Peripheral blood was collected for complete blood count, and analyses of IL-6, IL1-ß, and tumor necrosis factor-α were performed. Placental fragments were collected to assess the inflammatory changes using optical microscopy. After giving birth, both the ewes and their lambs were weighed. On clinical examination, a positive correlation between bleeding and suppuration (correlation index - CI=0.54), suppuration and marginal gingivitis (CI=0.34), and marginal gingivitis and edema (CI=0.54) was observed. The weights of the ewes (p=0.013) and their respective lambs (p=0.04) in the OGP group were lower than those of their OGCH group counterparts. The hematological analysis revealed that the OGP group ewes showed a slight increase in the mean corpuscular volume (p=0.2447), segmented cells (p=0.3375), and eosinophils (p=0.3823) when compared with the OGCH group ewes, without a statistical difference. Regarding the microorganisms detected in the oral cavity, there was a significant difference between the occurrence of periodontal pockets and the presence of Fusobacterium necrophorum (p=0.0328), Porphyromonas asaccharolytica (p=0.0392), and the Mollicutes class (p=0.0352). Staphylococcus genus (p=0.9107) and Archaea domain (p=0.7245) were detected in the amniotic samples of both groups, without a significant difference, whereas P. asaccharolytica (p=0.2685) was only detected in one sample in the OGCH group. The expression of cytokine IL-6 in the OGP group differed significantly between the prepartum and postpartum periods (p=0.0039); moreover, it differed significantly in the postpartum period between the OGCH and OGP groups (p=0.0198). Histological examination showed a higher percentage of placental changes in the OGP group (70%) than in the OGCH group, such as the presence of macrophages, neutrophils, plasma cells, and multifocal areas of calcification. These results do not corroborate the hypothesis of dissemination of oral microorganisms to the placental unit, suggesting that it constitutes placental isolation in sheep.

A periodontite afeta os tecidos de suporte dos dentes levando à perda dentária e danos à saúde do animal. Evidências em humanos sugerem que os microrganismos orais se espalham sistemicamente, aumentando o risco de distúrbios da gravidez, como aborto espontâneo, prematuridade e baixo peso ao nascer. Este estudo teve como objetivo verificar se microrganismos periodontopatogênicos atingem a unidade transplacentária, culminando em problemas em ovelhas gestantes. Após análise da cavidade oral, foram selecionadas 10 ovelhas gestantes clinicamente saudáveis (grupo OGCH) e 10 ovelhas gestantes com periodontite (grupo OGP). O biofilme subgengival foi coletado para o teste de reação em cadeia da polimerase (PCR) e o líquido amniótico para teste de PCR e análise de interleucina (IL). Sangue periférico foi coletado para hemograma completo e análises de IL-6, IL1-ß e fator de necrose tumoral-α foram realizadas. Fragmentos de placenta foram coletados para avaliação das alterações inflamatórias por meio de microscopia óptica. Após o parto, as ovelhas e seus cordeiros foram pesados. Ao exame clínico, observou-se correlação positiva entre sangramento e supuração (índice de correlação - IC=0,54), supuração e gengivite marginal (IC=0,34) e gengivite marginal e edema (IC=0,54). Os pesos das ovelhas (p=0,013) e de seus respectivos cordeiros (p=0,04) do grupo OGP foram inferiores aos do grupo OGCH. A análise hematológica revelou que as ovelhas do grupo OGP apresentaram discreto aumento no volume corpuscular médio (p=0,2447), células segmentadas (p=0,3375) e eosinófilos (p=0,3823) quando comparadas com as ovelhas do grupo OGCH, sem diferença estatística diferença. Em relação aos microrganismos detectados na cavidade oral, houve diferença significativa entre a ocorrência de bolsas periodontais e a presença de Fusobacterium necrophorum (p=0,0328), Porphyromonas asaccharolytica (p=0,0392) e da classe Mollicutes (p=0,0352). O gênero Staphylococcus (p=0,9107) e o domínio Archaea (p=0,7245) foram detectados nas amostras amnióticas de ambos os grupos, sem diferença significativa, enquanto P. asaccharolytica (p=0,2685) foi detectado apenas em uma amostra do grupo OGCH. A expressão da citocina IL-6 no grupo OGP diferiu significativamente entre os períodos pré e pós-parto (p=0,0039); além disso, diferiu significativamente no período pós-parto entre os grupos OGCH e OGP (p=0,0198). O exame histológico mostrou maior porcentagem de alterações placentárias no grupo OGP (70%) do que no grupo OGCH, como a presença de macrófagos, neutrófilos, plasmócitos e áreas multifocais de calcificação. Esses resultados não corroboram a hipótese de disseminação de microrganismos orais para a unidade placentária, sugerindo que se trata de um isolamento placentário em ovinos.

Carcinoma de células escamosas em gato - tratamento com quimioterapia metronômica / Squamous cell carcinoma in a cat - treatment with metronomic chemotherapy

Background: Squamous cell carcinoma (SCC) is a malignant neoplasm associated with prolonged exposure to ultraviolet light and lack of skin pigment in white-coated animals. The disease diagnosis is based on the animal's history, clinical signs and histopathology of the samples collected. The therapeutic alternatives are diverse. Among them, one of the most used is the metronomic chemotherapy which is based on the use of cytostatics drugs, applied in low and frequent doses for a longer period of time. One of the mechanisms of action of this type of therapy is the decrease in angiogenesis. Non-steroidal anti-inflammatory drugs (NSAIDs) are simultaneously used to maximize the angiogenic effect. The objective of this study is to report the results of a treatment accomplised with metronomic chemotherapy (cyclophosphamide with meloxicam) in a domestic feline which presented a squamous cell tumor. Case: A 7-year-old female cat, mixed breed, not neutered, weighing 3.5 kg, was assisted at the Veterinary Specialties Teaching Clinic (CEEV) of the Technical University of Machala (UTMACH), presenting bilateral ulcerative lesion in the nasal plane for 5 months. Chronic exposure to sunlight is referred to in the patient's anamnesis. Aspiration cytology performed in the patient revealed an inflammatory process with a predominance of neutrophils and macrophages, and showed absence of neoplastic cells. A treatment based on antibiotics and corticosteroids was adopted. As there were no favorable results, an excisional biopsy procedure was chosen to collect the affected tissues. The result of the histopathological analysis was a level 2 squamous cell carcinoma (SCC). The initial treatment consisted of surgical removal of the tumor and later the use of metronomic chemotherapy based on Cyclophosphamide, Meloxicam, DHA (docosahexaenoic acid) and Sucralfate for 5 months was adopted. At the beginning of the second phase of treatment, the patient developed a macula that progressively diminished. Currently, the patient does not have any recurrence of the diagnosed neoplasm. Discussion: The diagnosis of squamous cell carcinoma was based on clinical and histopathological findings. The macroscopic lesions were crusted ulcers that do not heal in the nasal plane and auricular pavilion, as reported in the literature. SCC is characterized by the infiltration of squamous cells in the dermal layer of the skin, forming epithelial islands with keratin pearls. The histopathological findings of this case present the same characteristics. The climatic conditions of Ecuador, high temperatures with greater incidence of sunlight, hypopigmented areas of the animal's body and chronic exposure to solar radiation are risk factors present in this case that foster the development of this malignant neoplasm. There are multiple treatments to be introduced in this sort of pathology. In this case, surgical removal of the tumor was adopted, followed by metronomic chemotherapy based on cyclophosphamide at 10 mg/m2 and meloxicam at a 0.05 mg/kg per day for a period of time of five months, obtaining favorable results and absence of recurrence. Despite metronomic chemotherapy being easy to administer, with fewer side effects, which includes a better acceptance of care takers who are reluctant to administer conventional chemotherapy, further studies are still necessary in what concerns certain variables such as age, carcinoma level, appropriate dosages and drugs according to each neoplasm.

Extensive emphysematous pyelonephritis in a nondiabetic female cat - treatment with unilateral nephroureterectomy

Background: Emphysematous pyelonephritis (EPN) is an acute, severe necrotizing infection of the renal parenchyma and surrounding tissues that results in gas formation in the kidney, collecting system, or surroundings. EPN is a rare condition in veterinary medicine and occurs most frequently in dogs with diabetes mellitus. Although the prognosis of medical management in animals is poor, the standardized treatment protocol according to EPN severity is unclear. This report describes the first case of a nondiabetic female cat with extensive EPN and good prognosis following direct nephroureterectomy (NU). Case: A 10-year-old spayed female cat presented with the chief complaint of an acute loss of weight within 1 week, vomiting, and disorientation including stumbling, discoordination, circling, wobbling, head tilting, and difficulties in standing. At presentation, the patient had a body condition score of 1/9 and weighed 2.6 kg. Blood examination revealed leukocytosis, anemia, and hypoproteinemia. Abdominal radiography revealed severely decreased serosal details. A massive gas silhouette observed in the peritoneal and retroperitoneal cavities, was diagnosed as abdominal free gas. Abdominal ultrasound showed an accumulation of moderately anechoic fluid mixed with gas and cyst-like capsules around the left kidney. Left partial ureteral obstruction and dilation were also observed. Computed tomography (CT) was performed without sedatives or anesthetic drugs. The findings showed severe inflammatory changes in the peritoneum and a loss of the normal inner structure in the left kidney. A pyelogram of the left kidney was not observed after injection of the contrast material. Diffuse fat stranding and free gas observed in the mesentery of the entire abdominal cavity and around the left kidney were considered septic peritonitis. Urinalysis revealed proteinuria and hematuria. Numerous neutrophils with rod-type bacteria were observed in the ascites. Following diagnostic examinations, the patient was diagnosed with extensive left EPN, including inflammatory ascites and abdominal free gas. Therefore, emergency NU of the nonfunctional left kidney and ruptured ureter and thorough abdominal lavage were conducted. Diffuse inflammation and a nephrolith were observed in the section of the harvested kidney. The nephrolith was composed of 100% calcium oxalate monohydrate. The realtime polymerase chain reaction (RT-PCR) test for feline infectious peritonitis (FIP) was negative. Escherichia coli was detected in the ascites, and antibiotic therapy was administered following the antibiotic sensitivity test. The histological findings from the left kidney and ureter included marked chronic inflammation and fibrosis. The patient was discharged 4 days after surgery. During the 8-month follow-up period, the patient's condition improved. Discussion: This was a unique case of EPN in a nondiabetic cat and the first reported case of EPN with a ruptured ureter, including abdominal free gas, inflammatory ascites, and peritonitis. This patient had a bacterial urinary tract infection with E. coli, which is the most frequently isolated pathogen in humans. This gas-forming bacteria produced a massive amount of gas and inflammation that were considered to have ruptured the urinary tract, so that the gas was released into the abdomen. This case corresponded to class 3B, with two risk factors according to the human EPN classification system. Direct NU and abdominal lavage were performed as emergency surgeries. The patient stabilized gradually and showed a good prognosis. Immediate surgical intervention is recommended in animal patients showing the extensive EPN stage.

Caudal vena cava thrombosis in cows

Background: Caudal vena cava thrombosis (CVCT) is a serious disease that affects cattle. Due to being commonly a fatal pathology, it causes economic losses for producers and national livestock. Thus, the present study describes the epidemiological, clinical, ultrasonographic imaging and pathological findings in 4 cattle with CVCT attended at the Veterinary Hospital (HV) of the Institute of Veterinary Medicine (IMV) of the Federal University of Pará (UFPA). Cases: The animals were crossbreds of the Gir x Holstein and Jersey x Holstein breeds, aged between 4 and 8 years old, raised in semi-extensive and intensive systems. The main clinical signs were pale mucous membranes, reluctance to move, markedly positive venous pulse, engorged jugular with positive stasis test, and serous to mucopurulent nasal exudation. The auscultation of the lung fields revealed tachypnea, silent areas, wheezing, and pleural friction, in addition to coughing, expiratory dyspnea, mouth breathing, and expiratory grunts. One animal had severe hemoptysis. The ultrasound examination performed on a bovine revealed a circular and dilated caudal vena cava in cross-section. Laboratory tests in 3 cattle revealed anemia, leukocytosis with neutrophil left shift, and increased liver enzymes. At necropsy, all cattle had thrombi in the hepatic segment of the caudal vena cava. In the lung, multiple abscesses and areas of parenchymal consolidation, crateriform areas, as well as thrombi in the arteries were observed. Pleural effusion and ascites were seen in all cattle. Clotted blood was seen in the trachea, bronchi, and on rumen contents of an animal. Histopathological alterations seen in the liver were centrilobular hepatocytes with frequent intensely eosinophilic cytoplasm, and pyknotic, karyorrhexic, or absent nuclei and cell borders barely distinguishable. In the lung were nodular and random formations, with a thick wall of mature connective tissue and a central area full of cellular debris, necrotic cells, and intact and degenerated neutrophils (abscesses). Discussion: The set of diagnostic tools that include epidemiology, clinical signs and clinical examinations, ultrasound, necropsy, and histopathology were efficient in the diagnosis of CVCT. The possible causes that led the animals to develop CVCT were diffuse septic pododermatitis in the medial nail of the right pelvic limb associated with traumatic reticuloabomasitis and liver abscesses. In 1 cow, it was not possible to establish the probable cause of CVCT, but for the other cattle in the present study, the probable causes are in agreement with studies that have shown that this disease can occur as a sequel to several septic conditions such as jugular phlebitis, mastitis, hoof rot, enteritis, pneumonia, traumatic reticulopericarditis, acidosis and rumen laminitis, as well as omphalophlebitis in calves. The tachypnea, serous to purulent nasal exudation, pulmonary wheezing, pleural friction, coughing, and expiratory dyspnea, usually with open mouth breathing and expiratory grunts evidenced in the animals of this study, occurred as a result of embolic abscess pneumonia. The presence of multiple lung abscesses, areas of parenchymal consolidation, crateriform foci, and thrombi in the pulmonary arteries and chronic suppurative pneumonia, found at necropsy of the animals in the present study, are related to the development of a thrombus in the caudal vena cava that detaches and embolizes and lodges in the pulmonary arteries. The histopathological findings in 1 cow are compatible with lesions found at necropsy and draw attention to embolic pneumonia and liver lesions, which, are related to thrombi in pulmonary arteries and abscesses formed from CVCT, as well as venous stasis exerted in the return circulation.

Mansoa alliacea extract improves the growth performance and innate immune response of Arapaima gigas challenged with Aeromonas hydrophila and handling stress / Extrato de Mansoa alliacea melhora o desempenho de crescimento e resposta imunológica de Arapaima gigas desafiado com Aeromonas hydrophila e estresse de manejo

This study investigated the effects of dietary supplementation with Mansoa alliacea hydroalcoholic extracts on growth, blood and immune parameters of Arapaima gigas. Fish were fed for 30 days with diets enriched with 0, 4, 8, and 12 g kg-1 of M. alliacea hydroalcoholic extract and subjected to infection with Aeromonas hydrophila and handling stress. Fish fed with 8 g kg-1 of extract showed significant increase in final weight, specific growth rate and feed efficiency when compared to the other groups. Glucose, triglycerides, total proteins, and globulins increased significantly in fish fed with 8 g kg-1 of extract, whereas albumin decreased. The number of thrombocytes increased significantly with the dietary supplementation of 8 and 12 g kg-1 of extract. After the challenge with A. hydrophila and handling stress, fish fed with 8 g kg-1 of extract had significantly higher levels of glucose, globulins, and albumins, and fish fed with 8 and 12 g kg-1 of extract showed an increment of respiratory burst. Triglyceride levels dropped significantly in fish fed with 4, 8, and 12 g kg-1 of extract, whereas the number of neutrophils increased, and total thrombocytes, leukocytes and lymphocytes were higher in fish fed with 12 g kg-1 of extract. Dietary supplementation with M. alliacea extract at 8 g kg-1 was efficient in improving the growth and innate immunity of A. gigas, being potentially useful in fish farming to control the development of A. hydrophila infections.(AU)

Investigou-se os efeitos da suplementação com extrato hidroalcólico de Mansoa alliacea sobre o crescimento e parâmetros sanguíneos e imunológicos de Arapaima gigas. Os peixes foram alimentados por 30 dias com dietas enriquecidas com 0, 4, 8 e 12 g kg-1 de extrato hidroalcoólico de M. alliacea e submetidos à infecção por Aeromonas hydrophila e estresse de manejo. Os peixes alimentados com 8 g kg-1 de extrato apresentaram aumento significativo no peso final, taxa de crescimento específico e eficiência alimentar quando comparados aos demais grupos. Glicose, triglicerídeos, proteínas totais e globulinas aumentaram significativamente nos peixes alimentados com 8 g kg-1 de extrato, enquanto a albumina diminuiu. O número de trombócitos aumentou significativamente com a suplementação dietética de 8 e 12 g kg-1 de extrato. Após infecção com A. hydrophila e estresse de manejo, os peixes alimentados com 8 g kg-1 de extrato apresentaram níveis significativamente mais altos de glicose, globulinas e albuminas, e os peixes alimentados com 8 e 12 g kg-1 de extrato apresentaram incremento de explosão respiratória. Os níveis de triglicerídeos decresceram nos peixes alimentados com 4, 8 e 12 g kg-1 de extrato, enquanto o número de neutrófilos aumentou, e o número total de trombócitos, leucócitos e linfócitos foi maior nos peixes alimentados com 12 g kg-1 de extrato. A suplementação com a 8 g kg-1 de extrato de M. alliacea foi eficiente em melhorar o crescimento e a imunidade inata de A. gigas, sendo potencialmente útil na piscicultura para controlar o desenvolvimento de infecções por A. hydrophila.(AU)

Pelger-Huёt anomaly in a bitch Basenji

Background: Pelger-Huët anomaly (PHA) is characterized by morphological changes in all granulocytes, being more evident in neutrophils. Granulocytic function in these animals remains unchanged. Hereditary form of PHA should be differentiated from pseudo-PHA,a condition acquired from infections and/or inflammation conditions. Recognition of PHA is important to avoid misleading leukogram interpretation, since hyposegmentation of neutrophils can be confused with left shift, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The objective of this case report was to demonstrate the importance of laboratory diagnosis in PHA. Case: A 11-month-old bitch Basenji, was presented to perform preoperative evaluation for elective ovariosalpingohisterectomy at Veterinary Hospital (HUVET) of the Universidade Federal Fluminense (UFF). Tutor reported that animal was healthy, vaccination status was current, had deworming protocol applied and had not made use of medications recently. Animal presented normophagy, normodipsia, normuria and normochezia. Upon physical examination, animal was alert consciousness level, with adequate hydration, hyperemic oral mucosa, a less than 2 s capillary perfusion time, normal lymph nodes (submandibular, pre-scapular, inguinal and popliteal), rectal temperature of 39.2°C, heart rate of 160 beats per minute and respiratory rate of 60 movements per minute, possibly due to the animal's agitated state. Abdominal palpation showed no changes. Physical examination presented no alterations. Preoperative exams included complete blood count (CBC) and biochemistry profile (ALT, AP, glucose, creatinine, urea, total proteins and fractions). Samples were sent to Hospital's Veterinary Clinical Pathology Laboratory (LABHUVET/UFF) for analysis. CBC was performed using automatic method. Blood smears were stained with hematological stain and then a cytomorphological evaluation was performed. The first CBC revealed 23% of neutrophils with nuclear hyposegmentation and 44% of neutrophils were bands. A follow up was performed after 9 months, and a Complete Blood Count was performed again in which 12% of neutrophils showed nuclear hyposegmentation with mature chromatin pattern, 40% of neutrophils were bands, 1% of meta-myelotcytes neutrophils, 1% of myelocytes neutrophils and, also, eosinophils with nuclear hyposegmentation. Animal was healthy, and had no alterations on physical examination suggesting a diagnosis of PHA. Discussion: Recognition of PHA is important to avoid misleading leukogram interpretation, since neutrophils hyposegmentation can be confused with left shift, which is considered severe with poor prognosis, making it necessary to carry out diagnostic tests and treatment for the disease that is generating the deviation. The diagnosis of PHA was considered by the shape of the leukocytes nuclei, without evidence of inflammatory disease, during the patient follow up. Therefore, this anomaly should be considered as a differential diagnosis of left shift, thus avoiding unnecessary clinical and therapeutic procedures. Guidance in face of this hereditary hematological syndrome is important. The responsible guardian of the animal must not allow it to act as a breeder in order to interrupt possible transmission of this anomaly to offspring, because there is a fatal form when it comes to homozygotes.

Biochemical energy and hematological profile of sheep supplemented with soy molasses

and proteins. Despite the great variety of by-products and being potentially beneficial, it is not yet fully understood how the interaction between the nutrients in this product and the physiological changes in sheep occurs. Thus, the objective was to evaluate the effects of supplementation levels with soy molasses under the energy metabolites and hematological parameters of sheep. The treatments consisted of 0, 3, 6, 9, and 12% of inclusion of soy molasses in the dry matter for a basal diet of corn silage, which met the nutritional needs of the sheep in maintenance, distributed in a 5 × Latin square design. 5. The energy metabolites were evaluated: cholesterol, triglycerides, very-low-density lipoproteins (VLDL), and fructosamine. The hematological parameters were red blood cells, hemoglobin, hematocrits, and platelets, in addition to leukocytes, rods, neutrophils, eosinophils, monocytes, and lymphocytes. Supplementation with soy molasses did not significantly alter (P> 0.05) the serum concentrations of cholesterol, triglycerides, VLDL, and fructosamine. There was no significant difference in the blood cell, hemoglobin, hematocrit, and platelet concentrations of sheep with soy molasses supplementation (P> 0.05), which may indicate that the level of inclusion was sufficient to meet the nutritional needs of the animals, without damage. Leukocyte, rod, neutrophil, e

Soy molasses is a by-product of the soy industry, with low commercial cost and rich in carbohydrates and proteins. Despite the great variety of by-products and being potentially beneÞ cial, it is not fully understood how the interaction between the nutrients in this product and the physiological changes in sheep occurs. Thus, the objective was to evaluate the effects of supplementation levels with soy molasses on the energy metabolites and hematological parameters of sheep. The treatments consisted of 0, 3, 6, 9, and 12% of inclusion of soy molasses in dry matter basis for a basal diet of corn silage, which met the nutritional needs of the sheep in maintenance, distributed in a 5 × 5 Latin square design. The energy metabolites evaluated were: cholesterol, triglycerides, very-low-density lipoproteins (VLDL), and fructosamine. The hematological parameters were: red blood cells, hemoglobin, hematocrits, and platelets, in addition to leukocytes, rods, neutrophils, eosinophils, monocytes, and lymphocytes. Supplementation with soy molasses did not signiÞ cantly alter (P> 0.05) the serum concentrations of cholesterol, triglycerides, VLDL, and fructosamine. There was no signiÞ cant difference in the blood cell, hemoglobin, hematocrit, and platelet concentrations in sheep receiving soy molasses supplementation (P> 0.05), which may indicate that the level of inclusion was

Refractory feline sporotrichosis: a comparative analysis on the clinical, histopathological, and cytopathological aspects

ABSTRACT: Sporotrichosis is a chronic fungal infection caused by Sporothrix species. The occurrence of cases that are resistant to long-term treatment, especially in the nasal planum of cats, emphasizes the importance of studying its pathogenesis. The purpose of this study was to analyze and compare the inflammatory process of cutaneous lesions of feline refractory sporotrichosis to clinical aspects through cytopathological and histopathological examination. Moreover, the study included 13 cats with cutaneous lesions that had been resistant to itraconazole treatment for more than a year. Cutaneous lesions samples were collected for cytopathological, histopathological, and fungal culture analyses. Tissue fragments were processed and stained with hematoxylin-eosin (HE) and Grocott methenamine silver (GMS). Further, two clinical presentations had the highest occurrence: the localized cutaneous form in animals with good general condition and stable disease (n=9, 69.2%) and the disseminated cutaneous form in cats with poor general condition (n=4, 30.8%). In cats with refractory sporotrichosis, the nasal planum (84.6%) was the most common location of lesions. In the cytopathological study, cats with fewer than two lesions and in good general condition (n=9, 69.2%) showed absence or mild yeast intensity (up to 5 yeasts per field), lower intensity of macrophages and neutrophils, and higher intensity of epithelioid cells, lymphocytes, plasma cells, and eosinophils. On the other hand, (n=4, 30.8%) of the cats with disseminated sporotrichosis and a poor general condition had a marked intensity of yeasts, which were mostly phagocytosed by an increased number of macrophages and neutrophils. Of those animals with good general condition, the majority (n=6, 66.7%) had higher eosinophil intensity. In histopathology, malformed suppurative granuloma was the predominant type (n=9, 69.2%) in feline sporotrichosis lesions, followed by well-formed granulomas (n=4, 30.8%). Malformed granulomas showed mild to moderate fungal intensity (55.6%) in animals with good general condition and localized lesions while marked fungal intensity (44.4%) in cats with the disseminated form of the disease and poor general condition. Well-formed granulomas (n=4, 30.7%) had mild to moderate intensity of fungal load, and 75% of the animals with this type of granuloma had only one lesion and were in good general condition. Long-term itraconazole treatment in these cats with refractory sporotrichosis can keep the infection under control and localized lesions stable; however, fungus reactivation can occur, resulting in an exuberant and inefficient immune response.

RESUMO: A esporotricose é uma infecção fúngica crônica causada por espécies do gênero Sporothrix. A ocorrência de casos refratários ao tratamento de longo prazo, especialmente na região nasal de gatos, alerta para a importância do estudo de sua patogênese. Este estudo teve como objetivo analisar o processo inflamatório das lesões cutâneas de esporotricose felina refratária ao tratamento, por meio de exames citopatológico e histopalógico, e comparar com aspectos clínicos. Treze gatos com lesões cutâneas refratárias ao tratamento com Itraconazol por mais de um ano foram incluídos no estudo. Amostras de lesões cutâneas foram coletadas para análises citopatológicas, histopatológicas e cultura fúngica. Fragmentos de tecidos foram processados e corados pela hematoxilina-eosina (HE) e Prata Metenamina de Grocott (GMS). Duas apresentações clínicas tiveram maior ocorrência: a forma localizada cutânea em animais com bom estado geral e doença estável (n=9, 69,2%); e a forma disseminada cutânea em gatos com estado geral ruim (n=4, 30,8%). A região nasal (84,6%) foi a localização mais frequente das lesões nos gatos com esporotricose refratária ao tratamento. No estudo citopatológico, felinos com menos de duas lesões, e em bom estado geral, (n=9, 69,2%) revelaram ausência ou leve intensidade de leveduras (até 5 leveduras por campo), menor intensidade de macrófagos e neutrófilos, e maior intensidade de células epitelióides, linfócitos, plasmócitos e eosinófilos. Enquanto, (n=4, 30,8%) dos felinos que apresentavam a forma disseminada da esporotricose associada a um estado geral ruim, revelaram acentuada intensidade de leveduras em sua maioria fagocitadas por acentuado número de macrófagos e neutrófilos. Dos animais com bom estado geral, a maioria (n=6, 66,7%) apresentava maior intensidade de eosinófilos. Na histopatologia, o granuloma supurativo mal formado foi o tipo predominante (n=9, 69,2%) nas lesões de esporotricose felina, e (n=4, 30,8%) foram de granulomas bem formados. Granulomas mal formados apresentaram leve a moderada intensidade fúngica (55,6%) nos animais com bom estado geral e lesões localizadas, e acentuada intensidade fúngica (44,4%) nos gatos com a forma disseminada da doença e estado geral ruim. Granulomas bem formados (n=4, 30,7%) apresentaram leve a moderada intensidade de carga fúngica, sendo 75% dos animais com esse tipo de granuloma com apenas uma lesão e bom estadogeral. O tratamento de longo prazo com itraconazol, nesses gatos com esporotricose refratária, pode manter a infecção controlada e lesões localizadas estáveis de forma temporária, contudo a reativação do fungo pode ocorrer levando a uma resposta imunológica exuberante e ineficiente.

The use of occlusive dressings: influence on excisional wound healing in animal model

Purpose: To analyze the influence of occlusive dressing on the healing of excisional skin wounds in mice. Methods: Pre-clinical, comparative, and translational study. Mice were divided into three experimental groups: wounds occluded with hydrocolloid (HD) dressings, transparent polyurethane film (TF) dressings, and without occlusion (WO), monitored at three, six and 14 days, with eight animals each. Closure rate, infiltration of neutrophils and macrophages, measurement of tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) and, histologically, angiogenesis were evaluated. Results: Wound closure was accelerated in the occlusive groups. There was a decrease in TNF-α levels in the HD group when compared to the WO and TF groups. Neutrophils accumulation decreased in the HD group. Increased dosages of macrophages were evidenced in the HD group, compared to the WO and TF groups. Levels of VEGF were increased in the TF and HD groups. Conclusions: It is suggested that the occlusion of wounds modulates the inflammatory response.

Hematology and blood biochemistry profile of the freshwater stingray Potamotrygon magdalenae as a tool for population assessment in artificial environments

Abstract Hematological and blood biochemical reference information is important to establish physiological status of freshwater stingray populations and improve care and management protocols in artificial environments. Here, we used a commercial freshwater stingray with high mortality rates in the market (Potamotrygon magdalenae), as an example to understand how artificial environments and handling protocols influence physiological status of captive freshwater stingrays. To this purpose, blood from five adult males and six adult females was collected to perform complete blood counts and blood chemistry analyses. All sampled animals showed good body condition with no differences between sexes. Differences between sexes were only found for the differential count of lymphocytes. Red blood results were consistent with previously studied potamotrygonids while white blood results showed higher values of leukocytes, thrombocytes, heterophils and lymphocytes in P. magdalenae compared to other Potamotrygonids. All types of leukocytes described for elasmobranchs were found except neutrophils and basophils. Blood metabolites showed an influence of ex situ diet in total protein, triglycerides and cholesterol. Glucose results were consistent while urea showed lower levels than those recorded for other freshwater stingrays. These results highlight the importance of physical, physiological and health analysis in freshwater stingrays as a part of welfare assessment to improve monitoring protocols and survival rates in public or private aquaria.

Resumo A informação de referência hematológica e bioquímica do sangue é importante para estabelecer o estado fisiológico das populações de arraias de água doce e melhorar os protocolos de cuidado e manejo em ambientes artificiais. Aqui, usamos uma espécie comercial de arraia de água doce com elevadas taxas de mortalidade no mercado (Potamotrygon magdalenae) como espécie exemplo para compreender de que modo os ambientes artificiais e os protocolos de manipulação influenciam o estado fisiológico das arraias de água doce em cativeiro. Para este fim, foi coletado sangue de cinco machos adultos e seis fêmeas adultas para realizar contagens completas de células sanguíneas e análises bioquímicas de sangue. Todos os animais amostrados mostraram boa condição corpórea, sem diferenças entre os sexos. Diferenças entre os sexos foram encontradas só na contagem diferencial de linfócitos. Os resultados de células sanguíneas vermelhas foram consistentes com potamotrigonídeos previamente estudados, enquanto o leucograma revelou valores mais elevados de leucócitos, trombócitos, heterófilos e linfócitos em P. magdalenae, em comparação com outros potamotrigonídeos. Todos os tipos de leucócitos descritos para elasmobrânquios foram encontrados, exceto para neutrófilos e basófilos. Todos os tipos de leucócitos descritos para elasmobrânquios foram encontrados, exceto para neutrófilos e basófilos. Os metabólitos do sangue mostraram influência da dieta ex situ nas proteínas totais, triglicerídeos e colesterol. Os resultados da glicose foram consistentes, enquanto a ureia mostrou níveis mais baixos do que os registrados para outras espécies de arraias de água doce. Os resultados da glicose foram consistentes, enquanto a ureia mostrou níveis mais baixos em P. magdalenae. Estes resultados enfatizam a importância da análise física, fisiológica e de saúde em arraias de água doce como parte da avaliação do bem-estar para melhorar os protocolos de monitoramento e as taxas de sobrevivência em aquários públicos ou privados.

Development and characterization of poultry collagen-based hybrid hydrogels for bone regeneration

Purpose: Poultry by-products can contribute as an innovative natural source for the development of composites based on polymers and minerals aiming at bone regeneration. The objective of this study was the physicochemical and biological characterization of collagen-based hydrogels crosslinked with ultraviolet (UV)-riboflavin. Methods: Pure hydrogels of 100% collagen (G1) or hybrid hydrogels, 90% collagen:10% apatite (G2), 90% collagen:10% nanokeratin (G3), and 90% collagen:5% apatite:5% nanokeratin (G4) were characterized by scanning electron microscope, Fourier-transform infrared spectroscopy, differential scanning calorimetry, swelling degree and quali-quantitative histological analysis. Ectopic implantation in subcutaneous tissue in mice at one, three and nine weeks allowed to assess the inflammation (neutrophils, lymphocytes, macrophages, and giant cells) and repair (neovascularization, and connective tissue) to determine biocompatibility and the integrity of biomaterials to score their biodegradability. Histomorphometry on critical size defects in rat calvaria at one and three months evaluated the percentage of bone, connective tissue, and biomaterials in all groups. Results: The hydrogels presented porous microstructure, water absorption and physicochemical characteristics compatible with their polymeric and/or mineral composition. All materials exhibited biocompatibility, biodegradability, and low osteoconductivity. G2 showed greater density of new bone and biomaterial than the G1, G3 and G4. Conclusions: The collagen-apatite group formulation suggests potential for development as osteopromoting membrane.

Hematology and blood biochemistry profile of the freshwater stingray Potamotrygon magdalenae as a tool for population assessment in artificial environments / Perfil hematológico e bioquímico do sangue da arraia de água doce Potamotrygon magdalenae como ferramenta para avaliação da população em ambientes artificiais

Hematological and blood biochemical reference information is important to establish physiological status of freshwater stingray populations and improve care and management protocols in artificial environments. Here, we used a commercial freshwater stingray with high mortality rates in the market (Potamotrygon magdalenae), as an example to understand how artificial environments and handling protocols influence physiological status of captive freshwater stingrays. To this purpose, blood from five adult males and six adult females was collected to perform complete blood counts and blood chemistry analyses. All sampled animals showed good body condition with no differences between sexes. Differences between sexes were only found for the differential count of lymphocytes. Red blood results were consistent with previously studied potamotrygonids while white blood results showed higher values of leukocytes, thrombocytes, heterophils and lymphocytes in P. magdalenae compared to other Potamotrygonids. All types of leukocytes described for elasmobranchs were found except neutrophils and basophils. Blood metabolites showed an influence of ex situ diet in total protein, triglycerides and cholesterol. Glucose results were consistent while urea showed lower levels than those recorded for other freshwater stingrays. These results highlight the importance of physical, physiological and health analysis in freshwater stingrays as a part of welfare assessment to improve monitoring protocols and survival rates in public or private aquaria.

A informação de referência hematológica e bioquímica do sangue é importante para estabelecer o estado fisiológico das populações de arraias de água doce e melhorar os protocolos de cuidado e manejo em ambientes artificiais. Aqui, usamos uma espécie comercial de arraia de água doce com elevadas taxas de mortalidade no mercado (Potamotrygon magdalenae) como espécie exemplo para compreender de que modo os ambientes artificiais e os protocolos de manipulação influenciam o estado fisiológico das arraias de água doce em cativeiro. Para este fim, foi coletado sangue de cinco machos adultos e seis fêmeas adultas para realizar contagens completas de células sanguíneas e análises bioquímicas de sangue. Todos os animais amostrados mostraram boa condição corpórea, sem diferenças entre os sexos. Diferenças entre os sexos foram encontradas só na contagem diferencial de linfócitos. Os resultados de células sanguíneas vermelhas foram consistentes com potamotrigonídeos previamente estudados, enquanto o leucograma revelou valores mais elevados de leucócitos, trombócitos, heterófilos e linfócitos em P. magdalenae, em comparação com outros potamotrigonídeos. Todos os tipos de leucócitos descritos para elasmobrânquios foram encontrados, exceto para neutrófilos e basófilos. Todos os tipos de leucócitos descritos para elasmobrânquios foram encontrados, exceto para neutrófilos e basófilos. Os metabólitos do sangue mostraram influência da dieta ex situ nas proteínas totais, triglicerídeos e colesterol. Os resultados da glicose foram consistentes, enquanto a ureia mostrou níveis mais baixos do que os registrados para outras espécies de arraias de água doce. Os resultados da glicose foram consistentes, enquanto a ureia mostrou níveis mais baixos em P. magdalenae. Estes resultados enfatizam a importância da análise física, fisiológica e de saúde em arraias de água doce como parte da avaliação do bem-estar para melhorar os protocolos de monitoramento e as taxas de sobrevivência em aquários públicos ou privados.