Detection and characterization of bovine rumen microorganismsresistant to sodium fluoroacetate

Background: Poisoning of animals due to toxic plants is found in Brazil and other countries. One of the known toxic plantsin Brazil, with the active ingredient sodium fluoroacetate (SF), is Palicourea marcgravii. Dehalogenases that inactivatethe fluor-carbon bonds are enzymes found in microorganisms and may prevent intoxication. This study evaluated the occurrence of rumen microorganisms naturally resistant to SF.Materials, Methods & Results: Two samples of rumen fluid of cattle from the Experimental Farm of Federal University ofMato Grosso fed with Brachiaria sp. were obtained via fistula in flasks. An aliquot of 2 mL was placed in a microtube andcentrifuged at 9000 g for 1 min. Then, the sample was inoculated into 2 tubes, one containing 100 µL of clarified rumenfluid in 2 mL of modified liquid culture medium (0.1% ammonium sulfate, 0.1% potassium phosphate monobasic, 0.05%sodium phosphate dibasic, 0.01% magnesium sulfate, 0.01% yeast extract, pH 7.0) and 0.4% of SF and the other samplecontaining 2 mL of liquid culture medium and 100 µL of clarified rumen fluid. The 2 samples were incubated at 40°C for24 h. Dilutions were performed under the same conditions every 24 h until the attainment of microorganisms resistant toSF, and the finaldilution containing 50 µL of each sample was plated in the middle containing SF (0.4%) and incubated at40°C for 24 h for the isolation of bacteria. The bacterial colonies resistant to SF were identified by morphological methods, stained, and subjected to DNA extraction sequencing using the universal primers 27f and 1492r (16S rDNA) for theidentification of the bacterial genus using Blast DNA identity analysis. These bacteria were cultured with and without SF(0.4%), and the presence of fluoride ions was detected by an ion-selective electrode (fluoride) during incubation for 0, 30,60, 90, and 120 min. Two resistant microorganisms were isolated, one was a Gram-positive coccus and the other was aGram-positive rod...(AU)

Detection and characterization of bovine rumen microorganismsresistant to sodium fluoroacetate

Background: Poisoning of animals due to toxic plants is found in Brazil and other countries. One of the known toxic plantsin Brazil, with the active ingredient sodium fluoroacetate (SF), is Palicourea marcgravii. Dehalogenases that inactivatethe fluor-carbon bonds are enzymes found in microorganisms and may prevent intoxication. This study evaluated the occurrence of rumen microorganisms naturally resistant to SF.Materials, Methods & Results: Two samples of rumen fluid of cattle from the Experimental Farm of Federal University ofMato Grosso fed with Brachiaria sp. were obtained via fistula in flasks. An aliquot of 2 mL was placed in a microtube andcentrifuged at 9000 g for 1 min. Then, the sample was inoculated into 2 tubes, one containing 100 µL of clarified rumenfluid in 2 mL of modified liquid culture medium (0.1% ammonium sulfate, 0.1% potassium phosphate monobasic, 0.05%sodium phosphate dibasic, 0.01% magnesium sulfate, 0.01% yeast extract, pH 7.0) and 0.4% of SF and the other samplecontaining 2 mL of liquid culture medium and 100 µL of clarified rumen fluid. The 2 samples were incubated at 40°C for24 h. Dilutions were performed under the same conditions every 24 h until the attainment of microorganisms resistant toSF, and the finaldilution containing 50 µL of each sample was plated in the middle containing SF (0.4%) and incubated at40°C for 24 h for the isolation of bacteria. The bacterial colonies resistant to SF were identified by morphological methods, stained, and subjected to DNA extraction sequencing using the universal primers 27f and 1492r (16S rDNA) for theidentification of the bacterial genus using Blast DNA identity analysis. These bacteria were cultured with and without SF(0.4%), and the presence of fluoride ions was detected by an ion-selective electrode (fluoride) during incubation for 0, 30,60, 90, and 120 min. Two resistant microorganisms were isolated, one was a Gram-positive coccus and the other was aGram-positive rod...

Natural hemoplasma infection of cats in Cuiaba, Mato Grosso, Brazil / Hemoplasmas em felinos domésticos naturalmente infectados em Cuiabá, Mato Grosso, Brasil

The aim of this research was to investigate natural hemoplasma infection in cats treated at the Veterinary Hospital of the Federal University of Mato Grosso, and the factors associated with infection. Blood samples from 151 cats of different sexes, breeds, and ages were analyzed by PCR and blood count. The overall occurrence of hemoplasma was 25.8%. Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm), and Candidatus Mycoplasma turicensis (CMt) were observed in 15.2%, 14.6% and 2.6% of cats, respectively. In 6.6 % of cases, co-infection was observed. Male felines or mixed breed cats were associated with infection by CMhm (P = 0.02 and 0.04, respectively). The data obtained demonstrated an occurrence of 25.8% for hemoplasma infection in felines coming from clinical care in the city of Cuiabá, where males were at higher risk of acquiring the infection by these agents, in addition to a higher risk for CMhm in felines with no specific breed.

O objetivo desta pesquisa foi investigar a ocorrência de Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm) e Candidatus Mycoplasma turicensis (CMt) em felinos atendidos no Hospital Veterinário da Universidade Federal de Mato Grosso e os fatores associados à positividade. Amostras de sangue de 151 felinos, dos distintos gêneros, raças e idades foram analisadas pela PCR e hemograma. A ocorrência geral de hemoplasmas foi de 25,8% e positividade para Mhf, CMhm e CMt foi observada em 15,2%, 14,6% e 2,6% felinos, respectivamente. Em 6.6% dos casos foi observado co-positividade. Apenas a variável sexo, macho (p = 0,02), foi associada à positividade para hemoplasmas. Felinos sem raça definida foram associados à positividade para CMhm (p = 0,04). Os dados obtidos demonstram prevalência de 25,8% para hemoplasmas em felinos provenientes de atendimento clínico do Hospital Veterinário da UFMT, onde os machos apresentaram maior risco de adquirir a infecção por estes agentes, além de maior risco para CMhm nos felinos sem definição racial.

Natural hemoplasma infection of cats in Cuiaba, Mato Grosso, Brazil / Hemoplasmas em felinos domésticos naturalmente infectados em Cuiabá, Mato Grosso, Brasil

The aim of this research was to investigate natural hemoplasma infection in cats treated at the Veterinary Hospital of the Federal University of Mato Grosso, and the factors associated with infection. Blood samples from 151 cats of different sexes, breeds, and ages were analyzed by PCR and blood count. The overall occurrence of hemoplasma was 25.8%. Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm), and Candidatus Mycoplasma turicensis (CMt) were observed in 15.2%, 14.6% and 2.6% of cats, respectively. In 6.6 % of cases, co-infection was observed. Male felines or mixed breed cats were associated with infection by CMhm (P = 0.02 and 0.04, respectively). The data obtained demonstrated an occurrence of 25.8% for hemoplasma infection in felines coming from clinical care in the city of Cuiabá, where males were at higher risk of acquiring the infection by these agents, in addition to a higher risk for CMhm in felines with no specific breed.(AU)

O objetivo desta pesquisa foi investigar a ocorrência de Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm) e Candidatus Mycoplasma turicensis (CMt) em felinos atendidos no Hospital Veterinário da Universidade Federal de Mato Grosso e os fatores associados à positividade. Amostras de sangue de 151 felinos, dos distintos gêneros, raças e idades foram analisadas pela PCR e hemograma. A ocorrência geral de hemoplasmas foi de 25,8% e positividade para Mhf, CMhm e CMt foi observada em 15,2%, 14,6% e 2,6% felinos, respectivamente. Em 6.6% dos casos foi observado co-positividade. Apenas a variável sexo, macho (p = 0,02), foi associada à positividade para hemoplasmas. Felinos sem raça definida foram associados à positividade para CMhm (p = 0,04). Os dados obtidos demonstram prevalência de 25,8% para hemoplasmas em felinos provenientes de atendimento clínico do Hospital Veterinário da UFMT, onde os machos apresentaram maior risco de adquirir a infecção por estes agentes, além de maior risco para CMhm nos felinos sem definição racial.(AU)

Hemostatic assessment of dogs associated with hepatic parasite load of Leishmania infantum chagasi / Avaliação hemostática de cães associada com a carga parasitária hepática de Leishmania infantum chagasi

Leishmania infantum chagasi liver parasite load was compared to hemostatic abnormalities, as well as to clinical, laboratorial, and histopathological findings in dogs with visceral leishmaniasis. The liver parasite load of 30 dogs L. infantum chagasi naturally-infected was evaluated by quantitative real- time PCR and the results were compared with serum biochemistry and primary and secondary hemostasis findings. Moreover, hepatic histological lesions were described in these dogs. Prolonged bleeding time, prothrombin time (PT), and activated partial thromboplastin time (APTT), were observed in the group with visceral leishmaniasis. Eleven dogs presented inflammatory liver lesions, with predominance of mild multifocal mononuclear periportal hepatitis. No association between the presence of parasites and abnormalities in screening tests was observed by Spearmans rank correlation coefficient. The clinical progression in leishmaniasis is associated with the occurrence of hemorrhagic diathesis, which depends not only on the presence of the parasite but also the inflammatory process, compromised immunological response, hepatic and renal failure in symptomatic dogs.(AU)

A carga parasitária de Leishmania infantum chagasi do fígado foi comparada às anormalidades hemostáticas, bem como aos achados clínicos, laboratoriais e histopatológicos em cães com leishmaniose visceral. A carga parasitária do fígado de 30 cães naturalmente infectados por L. infantum chagasi foi avaliada por PCR quantitativo em tempo real e os resultados foram comparados com bioquímica sérica e achados de hemostasia primária e secundária. Além disso, foram descritas as lesões hepáticas nestes cães. Prolongado tempo de sangramento, tempo de protrombina (TP) e tempo de tromboplastina parcial ativada (TTPA) foram observados no grupo com leishmaniose visceral. Onze cães apresentaram lesões inflamatórias no fígado, predominando hepatite periportal mononuclear multifocal. Não foi observada associação entre a presença de parasitos e as anormalidades nos testes laboratoriais por correlação de Spearman. A progressão clínica na leishmaniose está associada com a ocorrência de diátese hemorrágica, que depende não só da presença do parasito, mas também do processo inflamatório, do comprometimento da resposta imunológica e da falência renal e hepática em cães sintomáticos.(AU)

Pythium insidiosum colitis in a dog: treatment and clinical outcome / Colite por Pythium insidiosum em um cão: tratamento e evolução clínica

The aim of this report is to describe the clinical, pathological and imaging findings and treatment of colitis caused by Pythium insidiosum in a canine presenting haematochezia and progressive weight loss. Through imaging, a thickening of the transverse and descending colon was observed. Histopathological analysis of the large intestine fragment revealed the presence of hyphae, confirmed by immunohistochemistry and PCR as P. insidiosum. Antifungal treatment with itraconazole implemented after partial surgical resection, resulted in control of the disease.(AU)

O objetivo deste relato é descrever os achados clínico-patológicos, de imagem e o tratamento de colite por Pythium insidiosum em canino apresentando hematoquezia e emagrecimento progressivo. Nos exames de imagem, foram observados espessamento do cólon transverso e descendente. Análise histopatológica de fragmento do intestino grosso revelou a presença de hifas, confirmado pela imuno-histoquímica e PCR como P. insidiosum. Terapia antifúngica com itraconazol foi instituída pós-ressecção cirúrgica parcial, obtendo-se controle da doença.(AU)

Identification of genes expressed by Cryptococcus gattii during iron deprivation

Cryptococcus neoformans and C. gattii are pathogenic yeasts that cause life-threatening diseases in humans and animals. Iron is an essential nutrient for virtually every organism as it functions as a cofactor in numerous essential enzymatic reactions. In the literature, the competition for iron between microbes and mammalian hosts during infection is well documented. In this study, we used representational difference analysis (RDA) in order to gain a better understanding of how C. gattii responds to iron starvation. A total of 15 and 29 genes were identified as having altered expression levels due to iron depletion after 3 h and 12 h, respectively. Of these, eight genes were identified in both libraries. The transcripts were related to many biological processes, such as cell cycle, ergosterol metabolism, cell wall organization, transportation, translation, cell respiration and the stress response. These data suggest a remodeling of C. gattii metabolism during conditions of iron deprivation.

Expressão diferencial de proteínas do fungo Conidiobolus lamprauges cultivado em diferentes temperaturas / Differential expression of protein of the fungus Conidiobolus lamprauges cultivated at different temperatures

Para sobreviverem na temperatura corpórea de seu hospedeiro, os fungos patogênicos têm desenvolvido mecanismos moleculares importantes, como a expressão de proteínas relacionadas ao crescimento em altas temperaturas. Assim, o objetivo deste trabalho foi analisar o crescimento in vitro de Conidiobolus lamprauges em diferentes temperaturas e comparar o perfil de proteínas expressas através de eletroforese bidimensional (2D), em duas temperaturas distintas, sendo uma considerada baixa (28°C) e alta (37°C). Para análise do crescimento em diferentes temperaturas, cinco isolados de C. lamprauges, oriundos de ovinos doentes, foram incubados a 20, 25, 30, 35 e 40°C e o crescimento radial foi medido a cada 24 horas. Para análise da expressão diferencial, realizou-se a extração de proteínas do fungo cultivado a 28°C e a 37°C por 48 horas. A média de crescimento radial dos isolados foi diferente nas temperaturas analisadas, sendo 35°C a melhor temperatura para crescimento em todas as amostras. A temperatura ótima ajustada variou entre 33,3°C a 34,8°C. Os limites inferior e superior de inibição de crescimento foram 18°C e 42°C, respectivamente. Na análise da expressão diferencial, foram encontrados 16 spots diferencialmente expressos, sete (7/16) estavam com expressão diminuída e nove (9/16) com expressão aumentada a 37°C, quando comparado a 28°C. Além disso, oito spots estavam presentes apenas a 28°C e seis a 37°C. Sugere-se que C. lamprauges produza um perfil de proteínas relacionadas à termorregulação desencadeado pela alta temperatura do hospedeiro.(AU)

To survive at the body temperature of their hosts, pathogenic fungi have developed important molecular mechanisms, such as protein expression associated with growth at high temperatures. Thus, the aim of this study was to analyze the in vitro growth of Conidiobolus lamprauges at different temperatures and compare proteins expressed by two-dimensional electrophoresis (2D), for the pathogen cultivated at low (28°C) and high (37°C) temperatures. For the analysis of growth temperatures, five isolates of C. lamprauges from sick sheep were incubated at 20, 25, 30, 35 and 40°C and radial growth was measured every 24 hours. For the analysis of differential expression, protein extraction and two-dimensional polyacrylamide gel electrophoresis were performed with C. lamprauges cultivated at 28°C and 37°C for 48 hours. The average radial growth was different at the temperatures tested, and 35°C was found to be the best growth temperature for all isolates. The optimum adjusted temperature ranged between 33.3°C and 34.8°C. The upper and lower limits of growth inhibition were 18°C and 42°C, respectively. Upon expression analysis, a total of 16 spots were differentially expressed, seven (7/16) proteins were downregulated and nine (9/16) were over-expressed at 37ºC compared to 28°C. In addition, eight spots were present only at 28ºC and six were present only at 37ºC. It is suggested that C. lamprauges produces a profile of proteins that is related to thermoregulation triggered by the high temperature of the host.(AU)

Visceral leishmaniasis and infectious hepatitis in bush dog from captivity in Brazil - Report of case / Leishmaniose visceral e hepatite infecciosa em cachorro-vinagre mantido em cativeiro no Brasil - Relato de Caso

Among mammals, the canids are most often infected by Leishmania sp. This paper reports the occurrence of Leishmania (Leishmania) infantum chagasi as well as the presence of intranuclear inclusions resembling viral particles of canine adenovirus-1 in a bush dog (Speothos venaticus) kept in captivity at the zoo of Federal University of Mato Grosso, central area of Brazil.  The report demonstrates the importance of wild canids as reservoirs of agents of veterinary and public health importance, and it emphasizes the necessity of further studies with these animals concerning their habits, behavior and sanitary management.

Dentre os mamíferos, os canídeos são os mais frequentemente infectados por Leishmania sp. O presente artigo relata a ocorrência de Leishmania (Leishmania) infantum chagasi e inclusões intranucleares sugestivas de adenovírus canino 1, em cachorro vinagre (Speothos venaticus) mantido no zoológico da Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso. O relato demonstra a importância dos canídeos silvestres como reservatórios de agentes de importância veterinária e de saúde pública, reforçando a necessidade de estudos com esses animais acerca de seus hábitos, comportamento e manejo sanitário.

Visceral leishmaniasis and infectious hepatitis in bush dog from captivity in Brazil - Report of case / Leishmaniose visceral e hepatite infecciosa em cachorro-vinagre mantido em cativeiro no Brasil - Relato de Caso

Among mammals, the canids are most often infected by Leishmania sp. This paper reports the occurrence of Leishmania (Leishmania) infantum chagasi as well as the presence of intranuclear inclusions resembling viral particles of canine adenovirus-1 in a bush dog (Speothos venaticus) kept in captivity at the zoo of Federal University of Mato Grosso, central area of Brazil.  The report demonstrates the importance of wild canids as reservoirs of agents of veterinary and public health importance, and it emphasizes the necessity of further studies with these animals concerning their habits, behavior and sanitary management. 

Dentre os mamíferos, os canídeos são os mais frequentemente infectados por Leishmania sp. O presente artigo relata a ocorrência de Leishmania (Leishmania) infantum chagasi e inclusões intranucleares sugestivas de adenovírus canino 1, em cachorro vinagre (Speothos venaticus) mantido no zoológico da Universidade Federal de Mato Grosso, Cuiabá, Mato Grosso. O relato demonstra a importância dos canídeos silvestres como reservatórios de agentes de importância veterinária e de saúde pública, reforçando a necessidade de estudos com esses animais acerca de seus hábitos, comportamento e manejo sanitário.