RESUMO
Mycoplasma hemofelis is the most pathogenic hemoplasma species that affect cats. M. hemofelis may cause an acute infection that leads to hemolytic anaemia. The objective of this study was to detect and to quantify the load of M. hemofelis in cats by conventional polymerase chain reaction (PCR) and by quantitative PCR (qPCR) and to describe the possible hematological changes. M. hemofelis DNA was detected in 28.6% of the randomly selected cats (42 of 147) attended at the Veterinary Hospital of the Federal University of Mato Grosso, Brazil. The agreement between conventional PCR and qPCR was substantive (k 0.6). Females were twice as likely to acquire infection as males (odds ratio, 2.31). There was no statistically significant association (P > .05) and little/no correlation between the hematological parameters and the average of bacterial load. The results indicate that M. hemofelis infection is not related to clinical signs and bacterial blood load in cats. The agreement between conventional and quantitative PCR made it possible to detect infection by M. hemofelis in a larger number of cats.
Assuntos
Doenças do Gato , Infecções por Mycoplasma , Mycoplasma , Animais , Carga Bacteriana/veterinária , Gatos , DNA Bacteriano , Feminino , Masculino , Mycoplasma/genética , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/veterinária , Reação em Cadeia da Polimerase/veterináriaRESUMO
Cryptococcosis is caused by fungi of the genus Cryptococcus. Owing to its importance, this study aimed to analyze the genetic diversity of C. gattii isolates from animals, humans, and the environment in Mato Grosso State (MT), Brazil, during November 2010-December 2017. All isolates of the C. gattii species complex were subjected to molecular genotyping via Restriction Fragment Length Polymorphism (PCR-RFLP) and Multi-locus Sequence Typing (MLST). PCR-RFLP analysis revealed that 21 isolates presented the genotype VGII, which is considered the most common and virulent genotype globally among. MLST analysis revealed the presence of 14 sequence types (STs), of which 5 are considered new genotypes. Clonal Complex (CC) CC182 (n = 5; 23,80%) and CC309 (n = 3; 14,28%) were the most frequent. CC distribution in relation to origin revealed that three CCs were found in animals with a predominance of CC182 (66,66%), while nine were found in humans, and two CCs were found in the environment. Extensive genetic variability was observed among the isolates in the State of Mato Grosso. STs belonging to the already described clonal complexes (CC) indicate the global expansion and adaptation of isolates in several other countries. Therefore, detection of clonal complexes and STs already described in other regions and the occurrence of new STs in the present study help further the current understanding of the geographic dispersion and genetic origin of the C. gattii species complex.
Assuntos
Criptococose/microbiologia , Criptococose/veterinária , Cryptococcus gattii/classificação , Cryptococcus gattii/genética , Microbiologia Ambiental , Variação Genética , Animais , Brasil/epidemiologia , Criptococose/epidemiologia , Cryptococcus gattii/isolamento & purificação , Genótipo , Humanos , Tipagem de Sequências Multilocus , Técnicas de Tipagem Micológica , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
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...
Assuntos
Animais , Bovinos , Fluoracetatos , Intoxicação por Plantas/imunologia , Intoxicação por Plantas/veterinária , Plantas Tóxicas/imunologia , Rubiaceae , Rúmen/microbiologiaRESUMO
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)
Assuntos
Animais , Bovinos , Rúmen/microbiologia , Fluoracetatos , Rubiaceae , Plantas Tóxicas/imunologia , Intoxicação por Plantas/imunologia , Intoxicação por Plantas/veterináriaRESUMO
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)
Assuntos
Animais , Gatos , Infecções por Mycoplasma/epidemiologia , Mycoplasma , Medidas de Ocorrência de Doenças , BrasilRESUMO
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.
Assuntos
Animais , Gatos , Infecções por Mycoplasma/epidemiologia , Medidas de Ocorrência de Doenças , Mycoplasma , BrasilRESUMO
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 Spearman's 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.
Assuntos
Doenças do Cão/sangue , Hemostasia , Leishmania infantum , Leishmaniose Visceral/veterinária , Carga Parasitária/veterinária , Animais , Cães , Leishmaniose Visceral/sangueRESUMO
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)
Assuntos
Animais , Cães , Cães/parasitologia , Leishmania infantum , Leishmaniose/veterinária , Coagulação SanguíneaRESUMO
Abstract 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 Spearman’s 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.
Resumo 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.
Assuntos
Animais , Cães , Leishmania infantum , Doenças do Cão/sangue , Carga Parasitária/veterinária , Hemostasia , Leishmaniose Visceral/veterinária , Leishmaniose Visceral/sangueRESUMO
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)
Assuntos
Animais , Cães , Doenças do Cão , Colite/diagnóstico , Colite/terapia , Colite/veterinária , Pythium/patogenicidadeRESUMO
ABSTRACT: 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.
RESUMO: 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.
RESUMO
The dengue virus (DENV), which is frequently involved in large epidemics, and the yellow fever virus (YFV), which is responsible for sporadic sylvatic outbreaks, are considered the most important flaviviruses circulating in Brazil. Because of that, laboratorial diagnosis of acute undifferentiated febrile illness during epidemic periods is frequently directed towards these viruses, which may eventually hinder the detection of other circulating flaviviruses, including the Saint Louis encephalitis virus (SLEV), which is widely dispersed across the Americas. The aim of this study was to conduct a molecular investigation of 11 flaviviruses using 604 serum samples obtained from patients during a large dengue fever outbreak in the state of Mato Grosso (MT) between 2011 and 2012. Simultaneously, 3,433 female Culex spp. collected with Nasci aspirators in the city of Cuiabá, MT, in 2013, and allocated to 409 pools containing 1-10 mosquitoes, were also tested by multiplex semi-nested reverse transcription PCR for the same flaviviruses. SLEV was detected in three patients co-infected with DENV-4 from the cities of Cuiabá and Várzea Grande. One of them was a triple co-infection with DENV-1. None of them mentioned recent travel or access to sylvatic/rural regions, indicating that transmission might have occurred within the metropolitan area. Regarding mosquito samples, one pool containing one Culex quinquefasciatus female was positive for SLEV, with a minimum infection rate (MIR) of 0.29 per 1000 specimens of this species. Phylogenetic analysis indicates both human and mosquito SLEV cluster, with isolates from genotype V-A obtained from animals in the Amazon region, in the state of Pará. This is the first report of SLEV molecular identification in MT.
Assuntos
Culex/virologia , Dengue/epidemiologia , Vírus da Encefalite de St. Louis/genética , Encefalite de St. Louis/epidemiologia , RNA Viral/genética , Animais , Brasil/epidemiologia , Estudos Transversais , Vírus da Encefalite de St. Louis/isolamento & purificação , Feminino , Genótipo , Humanos , Filogenia , Análise de Sequência de DNARESUMO
The dengue virus (DENV), which is frequently involved in large epidemics, and the yellow fever virus (YFV), which is responsible for sporadic sylvatic outbreaks, are considered the most important flaviviruses circulating in Brazil. Because of that, laboratorial diagnosis of acute undifferentiated febrile illness during epidemic periods is frequently directed towards these viruses, which may eventually hinder the detection of other circulating flaviviruses, including the Saint Louis encephalitis virus (SLEV), which is widely dispersed across the Americas. The aim of this study was to conduct a molecular investigation of 11 flaviviruses using 604 serum samples obtained from patients during a large dengue fever outbreak in the state of Mato Grosso (MT) between 2011 and 2012. Simultaneously, 3,433 female Culex spp. collected with Nasci aspirators in the city of Cuiabá, MT, in 2013, and allocated to 409 pools containing 1-10 mosquitoes, were also tested by multiplex semi-nested reverse transcription PCR for the same flaviviruses. SLEV was detected in three patients co-infected with DENV-4 from the cities of Cuiabá and Várzea Grande. One of them was a triple co-infection with DENV-1. None of them mentioned recent travel or access to sylvatic/rural regions, indicating that transmission might have occurred within the metropolitan area. Regarding mosquito samples, one pool containing one Culex quinquefasciatus female was positive for SLEV, with a minimum infection rate (MIR) of 0.29 per 1000 specimens of this species. Phylogenetic analysis indicates both human and mosquito SLEV cluster, with isolates from genotype V-A obtained from animals in the Amazon region, in the state of Pará. This is the first report of SLEV molecular identification in MT.
O vírus da dengue (DENV), frequentemente envolvido em epidemias de grande proporção, e o vírus da febre amarela (YFV), responsável por surtos silvestres esporádicos, são considerados os flavivírus circulantes mais importantes no Brasil. Por este motivo, o diagnóstico laboratorial de doença febril aguda indiferenciada durante períodos epidêmicos é frequentemente direcionado para dengue e febre amarela no país, dificultando a detecção de outros arbovírus possivelmente circulantes, incluindo o vírus da encefalite de Saint Louis (SLEV), que é amplamente disperso nas Américas. O objetivo deste estudo foi investigar molecularmente a presença de 11 flavivírus no soro de 604 pacientes durante grande epidemia de dengue no estado de Mato Grosso (MT), Centro-Oeste do Brasil, entre 2011- 2012. Concomitantemente, 3.433 fêmeas de Culex spp. capturadas com aspirador de Nasci na cidade de Cuiabá, MT e alocadas em 409 pools com 1-10 mosquitos em 2013 foram testadas por multiplex seminested RT-PCR para os mesmos flavivírus. O SLEV foi detectado em três pacientes co-infectados com o DENV-4 das cidades de Cuiabá e Várzea Grande, MT. Um dos pacientes apresentava tripla co-infecção com DENV-1. Nenhum paciente referiu histórico recente de viagem ou acesso a áreas rurais/silvestres. Um pool contendo uma fêmea de Culex quinquefasciatus foi positivo para o SLEV, apresentando taxa de infecção mínima (MIR) de 0,29 por 1000 espécimes desta espécie. A análise filogenética indica que ambas as amostras formam um cluster com isolados do genótipo V-A do SLEV obtidos de animais na região amazônica do estado do Pará. Este é o primeiro relato de identificação molecular do SLEV no MT.
Assuntos
Animais , Feminino , Humanos , Culex/virologia , Dengue/epidemiologia , Vírus da Encefalite de St. Louis/genética , Encefalite de St. Louis/epidemiologia , RNA Viral/genética , Brasil/epidemiologia , Estudos Transversais , Vírus da Encefalite de St. Louis/isolamento & purificação , Genótipo , Filogenia , Análise de Sequência de DNARESUMO
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.
Assuntos
Cryptococcus gattii/genética , Cryptococcus gattii/metabolismo , Perfilação da Expressão Gênica , Ferro/metabolismo , Estresse Fisiológico , Cryptococcus gattii/fisiologia , Genes FúngicosRESUMO
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.
Assuntos
Cryptococcus gattii/genética , Cryptococcus gattii/metabolismo , Perfilação da Expressão Gênica , Ferro/metabolismo , Estresse Fisiológico , Cryptococcus gattii/fisiologia , Genes FúngicosRESUMO
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.
Assuntos
Cryptococcus gattii/genética , Cryptococcus gattii/metabolismo , Perfilação da Expressão Gênica , Ferro/metabolismo , Estresse Fisiológico , Cryptococcus gattii/fisiologia , Genes FúngicosRESUMO
INTRODUCTION: Cryptococcosis is a systemic fungal infection that affects humans and animals, mainly due to Cryptococcus neoformans and Cryptococcus gattii. Following the epidemic of acquired immunodeficiency syndrome (AIDS), fungal infections by C. neoformans have become more common among immunocompromised patients. Cryptococcus gattii has primarily been isolated as a primary pathogen in healthy hosts and occurs endemically in northern and northeastern Brazil. We to perform genotypic characterization and determine the in vitro susceptibility profile to antifungal drugs of the Cryptococcus species complex isolated from HIV-positive and HIV-negative patients attended at university hospitals in Cuiabá, MT, in the Midwestern region of Brazil. METHODOLOGY: Micromorphological features, chemotyping with canavanine-glycine-bromothymol blue (CGB) agar and genotyping by URA5-RFLP were used to identify the species. The antifungal drugs tested were amphotericin B, fluconazole, flucytosine, itraconazole and voriconazole. Minimum inhibitory concentrations (MICs) were determined according to the CLSI methodology M27-A3. RESULTS: Analysis of samples yelded C. neoformans AFLP1/VNI (17/27, 63.0%) and C. gattii AFLP6/VGII (10/27, 37.0%). The MICs ranges for the antifungal drugs were: amphotericin B (0.5-1 mg/L), fluconazole (1-16 mg/L), flucytosine (1-16 mg/L), itraconazole (0.25-0.12 mg/L) and voriconazole (0.06-0.5 mg/L). Isolates of C. neoformans AFLP1/VNI were predominant in patients with HIV/AIDS, and C. gattii VGII in HIV-negative patients. The genotypes identified were susceptible to the antifungal drugs tested. CONCLUSION: It is worth emphasizing that AFLP6/VGII is a predominant genotype affecting HIV-negative individuals in Cuiabá. These findings serve as a guide concerning the molecular epidemiology of C. neoformans and C. gattii in the State of Mato Grosso.
Assuntos
Antifúngicos/farmacologia , Criptococose/microbiologia , Cryptococcus/classificação , Cryptococcus/efeitos dos fármacos , Tipagem Molecular , Técnicas de Tipagem Micológica , Adulto , Idoso , Animais , Brasil/epidemiologia , Criança , Criptococose/epidemiologia , Cryptococcus/genética , Cryptococcus/isolamento & purificação , DNA Fúngico/genética , Feminino , Genótipo , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Epidemiologia Molecular , Polimorfismo de Fragmento de Restrição , Adulto JovemRESUMO
Little is known about the ecology of agents of cryptococcosis in Mato Grosso, without any data regarding to the sources of both agents in the environment. This study aimed to investigate Cryptococcus gattii and Cryptococcus neoformans associated with decay in tree hollows within the urban area of three different cities of Mato Grosso. Seventy-two environmental samples collected from 72 living trees in the cities of Cuiabá, Várzea Grande and Chapada dos Guimarães were sampled and analysed. One tree (Plathymenia reticulata, Leguminosae) in the city of Cuiabá yielded 19 colonies identified as C. gattii molecular type VGII. The isolation of C. gattii VGII in the downtown city of Cuiabá is important because it fits in the Northern Macroregion, suggesting expanding and urbanisation of this genotype in different Brazilian cities.
Assuntos
Criptococose/microbiologia , Cryptococcus gattii/isolamento & purificação , Fabaceae/microbiologia , Doenças das Plantas/microbiologia , Árvores/microbiologia , Brasil , Cryptococcus gattii/classificação , Cryptococcus gattii/genética , Cryptococcus neoformans/classificação , Cryptococcus neoformans/genética , Cryptococcus neoformans/isolamento & purificação , GenótipoRESUMO
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)
Assuntos
Conidiobolus/crescimento & desenvolvimento , Conidiobolus/isolamento & purificação , Proteínas , Regulação da Temperatura Corporal , Temperatura Baixa , Temperatura AltaRESUMO
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.
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.