RESUMEN
Canine distemper virus (CDV) causes systemic, respiratory, cutaneous, and neurological manifestations, in dogs and other mammalians. Canine adenovirus A (CAdV) is divided in two types, CAdV-1 and CAdV-2, and cause infectious canine hepatitis and infectious tracheobronchitis, respectively. Canineparvovirus type 2 (CPV-2)-induced enteritis is one of the most common infectious cause of diarrhea inpuppies. The aim of this study was to detect the urinary excretion of major viral pathogens that cause systemic infections in dogs. From December 2011 to December 2012, 41 urine samples were collected from dogs that presented systemic clinical signs of infectious diseases. The samples were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) for CDV and by PCR for CAdV-1, CAdV- 2, and CPV-2. RT-PCR amplified a partial fragment of the CDV N gene (287 bp) in 15 (36.6%) urine samples. PCR for E gene of CAdV-1 (508 bp) and CAdV-2 (1,030 bp) was positive in 4 (9.8%) and 1 (2.4%) sample, respectively; CPV-2 partial VP2 capsid protein gene (583 bp) was amplified in 6 (14.6%) samples. These results suggest that urine can be used as a clinical sample for the ante mortem diagnosis by molecular tools such as RT-PCR and PCR for CDV, CAdV-1 and 2, and CPV-2 systemic infections in dogs. Urinary excretion might be an important route for maintaining these viruses within the environment, and should be considered as a source of infection for healthy dogs.(AU)
O vírus da cinomose canina (CDV) é um Morbilivirus que causa manifestações clínicas sistêmicas, respiratórias, cutâneas e neurológicas em cães e outros mamíferos. O adenovírus canino A (CAdV) pode ser diferenciado em dois tipos, CAdV-1 e CAdV-2, e causam hepatite infecciosa canina etraqueobronquite infecciosa, respectivamente. Enterite pelo parvovírus canino tipo 2 (CPV-2) é uma das diarreias infecciosas mais comuns em cães, especialmente em filhotes. O objetivo deste estudo foi detectar a excreção urinária de alguns dos principais vírus que ocasionam infecções sistêmicas em cães. No período de dezembro de 2011 a dezembro de 2012 foram colhidas 41 amostras de urina de cães com sinais clínicos sistêmicos. As amostras foram avaliadas por reação em cadeia da polimerase precedida por transcrição reversa (RT-PCR) para o CDV e por PCR para CAdV-1, CAdV-2 e CPV-2. A RT-PCR amplificou um fragmento do gene N do CDV (287 pb) em 15 (36,6%) amostras. A PCR para o gene E do CAdV-1 (508 pb) e do CAdV-2 (1030 pb) foi positiva em 4 (9,8%) e 1 (2,4%) amostra, respectivamente; o gene VP2 da proteína do capsídeo do CPV-2 (583 pb) foi amplificado em 6 (14,6%) amostras. Estes resultados sugerem que a urina pode ser utilizada como amostra clínica para o diagnóstico ante mortem de infecções sistêmicas por CDV, CAdV-1 e 2 e CPV-2 por técnicas moleculares como RT-PCR e PCR. Adicionalmente, a excreção viral pela urina parece ser uma importante rota para a manutenção destes vírus no ambiente e deve ser considerada como fonte de infecção para cães saudáveis.(AU)
Asunto(s)
Animales , Perros , Moquillo/orina , Infecciones por Adenoviridae/veterinaria , Adenovirus Caninos , Parvovirus Canino , Infecciones por Parvoviridae/veterinaria , Perros , Reacción en Cadena de la PolimerasaRESUMEN
Canine distemper virus (CDV) causes systemic, respiratory, cutaneous, and neurological manifestations, in dogs and other mammalians. Canine adenovirus A (CAdV) is divided in two types, CAdV-1 and CAdV-2, and cause infectious canine hepatitis and infectious tracheobronchitis, respectively. Canineparvovirus type 2 (CPV-2)-induced enteritis is one of the most common infectious cause of diarrhea inpuppies. The aim of this study was to detect the urinary excretion of major viral pathogens that cause systemic infections in dogs. From December 2011 to December 2012, 41 urine samples were collected from dogs that presented systemic clinical signs of infectious diseases. The samples were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) for CDV and by PCR for CAdV-1, CAdV- 2, and CPV-2. RT-PCR amplified a partial fragment of the CDV N gene (287 bp) in 15 (36.6%) urine samples. PCR for E gene of CAdV-1 (508 bp) and CAdV-2 (1,030 bp) was positive in 4 (9.8%) and 1 (2.4%) sample, respectively; CPV-2 partial VP2 capsid protein gene (583 bp) was amplified in 6 (14.6%) samples. These results suggest that urine can be used as a clinical sample for the ante mortem diagnosis by molecular tools such as RT-PCR and PCR for CDV, CAdV-1 and 2, and CPV-2 systemic infections in dogs. Urinary excretion might be an important route for maintaining these viruses within the environment, and should be considered as a source of infection for healthy dogs.
O vírus da cinomose canina (CDV) é um Morbilivirus que causa manifestações clínicas sistêmicas, respiratórias, cutâneas e neurológicas em cães e outros mamíferos. O adenovírus canino A (CAdV) pode ser diferenciado em dois tipos, CAdV-1 e CAdV-2, e causam hepatite infecciosa canina etraqueobronquite infecciosa, respectivamente. Enterite pelo parvovírus canino tipo 2 (CPV-2) é uma das diarreias infecciosas mais comuns em cães, especialmente em filhotes. O objetivo deste estudo foi detectar a excreção urinária de alguns dos principais vírus que ocasionam infecções sistêmicas em cães. No período de dezembro de 2011 a dezembro de 2012 foram colhidas 41 amostras de urina de cães com sinais clínicos sistêmicos. As amostras foram avaliadas por reação em cadeia da polimerase precedida por transcrição reversa (RT-PCR) para o CDV e por PCR para CAdV-1, CAdV-2 e CPV-2. A RT-PCR amplificou um fragmento do gene N do CDV (287 pb) em 15 (36,6%) amostras. A PCR para o gene E do CAdV-1 (508 pb) e do CAdV-2 (1030 pb) foi positiva em 4 (9,8%) e 1 (2,4%) amostra, respectivamente; o gene VP2 da proteína do capsídeo do CPV-2 (583 pb) foi amplificado em 6 (14,6%) amostras. Estes resultados sugerem que a urina pode ser utilizada como amostra clínica para o diagnóstico ante mortem de infecções sistêmicas por CDV, CAdV-1 e 2 e CPV-2 por técnicas moleculares como RT-PCR e PCR. Adicionalmente, a excreção viral pela urina parece ser uma importante rota para a manutenção destes vírus no ambiente e deve ser considerada como fonte de infecção para cães saudáveis.
Asunto(s)
Animales , Perros , Adenovirus Caninos , Moquillo/orina , Infecciones por Adenoviridae/veterinaria , Infecciones por Parvoviridae/veterinaria , Parvovirus Canino , Perros , Reacción en Cadena de la PolimerasaRESUMEN
Listeriosis is a disease primarily of ruminants caused by the Gram-positive bacterium Listeria monocytogenes. Ruminants either demonstrate manifestations of the encephalitic, septicemic, or reproductive form of listeriosis. The pathological and molecular findings with encephalitic listeriosis in a 5.5-month-old, male, mixed-breed goat and a 3-year-old Texel-crossed sheep from northern Paraná, Brazil are described. Clinically, the kid demonstrated circling, lateral protrusion of the tongue, head tilt, and convulsions, the ewe presented ataxia, motor incoordination, and lateral decumbency. Brainstem dysfunctions were diagnosed clinically and listeriosis was suspected. Necropsy performed on both animals did not reveal remarkable gross lesions, significant histopathological alterations were restricted to the brainstem (medulla oblongata, rhombencephalitis) and were characterized as meningoencephalitis that consisted of extensive mononuclear perivascular cuffings, neutrophilic and macrophagic microabscesses, and neuroparenchymal necrosis. PCR assay and direct sequencing, using genomic bacterial DNA derived from the brainstem of both animals, amplified the desired 174 base pairs length amplicon of the listeriolysin O gene of L. monocytogenes. Phylogenetic analyses demonstrated that the strains associated with rhombencephalitis during this study clustered with known strains of L. monocytogenes lineage I from diverse geographical locations and from cattle of the state of Paraná with encephalitic listeriosis. Consequently, these strains should be classified as L. monocytogenes lineage I. These results confirm the active participation of lineage I strains of L. monocytogenes in the etiopathogenesis of the brainstem dysfunctions observed during this study, probably represent the first characterization of small ruminant listeriosis by molecular techniques in Latin America, and suggest that ruminants within the state of Paraná were infected by the strains of the same lineage of L. monocytogenes.(AU)
Asunto(s)
Listeriosis , Encefalitis , Rumiantes , Coloración y Etiquetado , FilogeniaRESUMEN
Listeriosis is a disease primarily of ruminants caused by the Gram-positive bacterium Listeria monocytogenes. Ruminants either demonstrate manifestations of the encephalitic, septicemic, or reproductive form of listeriosis. The pathological and molecular findings with encephalitic listeriosis in a 5.5-month-old, male, mixed-breed goat and a 3-year-old Texel-crossed sheep from northern Paraná, Brazil are described. Clinically, the kid demonstrated circling, lateral protrusion of the tongue, head tilt, and convulsions; the ewe presented ataxia, motor incoordination, and lateral decumbency. Brainstem dysfunctions were diagnosed clinically and listeriosis was suspected. Necropsy performed on both animals did not reveal remarkable gross lesions; significant histopathological alterations were restricted to the brainstem (medulla oblongata; rhombencephalitis) and were characterized as meningoencephalitis that consisted of extensive mononuclear perivascular cuffings, neutrophilic and macrophagic microabscesses, and neuroparenchymal necrosis. PCR assay and direct sequencing, using genomic bacterial DNA derived from the brainstem of both animals, amplified the desired 174 base pairs length amplicon of the listeriolysin O gene of L. monocytogenes. Phylogenetic analyses demonstrated that the strains associated with rhombencephalitis during this study clustered with known strains of L. monocytogenes lineage I from diverse geographical locations and from cattle of the state of Paraná with encephalitic listeriosis. Consequently, these strains should be classified as L. monocytogenes lineage I. These results confirm the active participation of lineage I strains of L. monocytogenes in the etiopathogenesis of the brainstem dysfunctions observed during this study, probably represent the first characterization of small ruminant listeriosis by molecular techniques in Latin America, and suggest that ruminants within the state of Paraná were infected by the strains of the same lineage of L. monocytogenes.
Asunto(s)
Animales , Femenino , Masculino , Toxinas Bacterianas/genética , Enfermedades de las Cabras/patología , Proteínas de Choque Térmico/genética , Proteínas Hemolisinas/genética , Listeriosis/veterinaria , Meningoencefalitis/veterinaria , Enfermedades de las Ovejas/patología , Brasil , Tronco Encefálico/patología , Análisis por Conglomerados , Genotipo , Cabras , Enfermedades de las Cabras/microbiología , Histocitoquímica , Listeria monocytogenes/genética , Listeriosis/microbiología , Listeriosis/patología , Meningoencefalitis/microbiología , Meningoencefalitis/patología , Filogenia , Reacción en Cadena de la Polimerasa , Homología de Secuencia , Ovinos , Enfermedades de las Ovejas/microbiologíaRESUMEN
The concomitant infections of Canine distemper virus (CDV), Canine adenovirus A types 1 (CAdV-1) and 2 (CAdV-2), Canine parvovirus type 2 (CPV-2), and Toxoplasma gondii are described in a 43-day-old mixed-breed puppy. Clinically, there were convulsions and blindness with spontaneous death; 14 siblings of this puppy, born to a 10-month-old dam, which was seropositive (titer: 1,024) for T. gondii, also died. Necropsy revealed unilateral corneal edema (blue eye), depletion of intestinal lymphoid tissue, non-collapsible lungs, congestion of meningeal vessels, and a pale area in the myocardium. Histopathology demonstrated necrotizing myocarditis associated with intralesional apicomplexan protozoa; necrotizing and chronic hepatitis associated with rare intranuclear inclusion bodies within hepatocytes; necrotizing bronchitis and bronchiolitis; interstitial pneumonia associated with eosinophilic intracytoplasmic inclusion bodies within epithelial cells; atrophy and fusion of intestinal villi with cryptal necrosis; and white matter demyelination of the cerebrum and cerebellum associated with intranuclear inclusion bodies within astrocytes. Polymerase chain reaction (PCR) amplified the partial fragments (bp) of the CDV N gene (290 bp), CPV-2c VP2 capsid protein gene (583 bp), and CAdV-1 (508 bp) and CAdV-2 (1,030 bp) E gene from urine and tissue samples. The PCR assays demonstrated that the apicomplexan protozoa observed within several organs contained DNA specific for T. gondii; genotyping revealed T. gondii type III. The findings support the characterization of concomitant infections of CDV, CAdV-1, CAdV-2, CPV-2, and T. gondii in this puppy. Further, seroreactivity to T. gondii of the dam in association with the systemic disease observed in the puppy described herein is suggestive of congenital toxoplasmosis.
Asunto(s)
Infecciones por Adenoviridae/veterinaria , Moquillo/parasitología , Infecciones por Parvoviridae/veterinaria , Toxoplasmosis Animal/virología , Infecciones por Adenoviridae/parasitología , Infecciones por Adenoviridae/virología , Adenovirus Caninos/genética , Adenovirus Caninos/aislamiento & purificación , Animales , ADN Viral/química , ADN Viral/genética , Moquillo/virología , Virus del Moquillo Canino/genética , Virus del Moquillo Canino/aislamiento & purificación , Perros , Resultado Fatal , Histocitoquímica , Masculino , Infecciones por Parvoviridae/parasitología , Infecciones por Parvoviridae/virología , Parvovirus Canino/genética , Parvovirus Canino/aislamiento & purificación , Reacción en Cadena de la Polimerasa/veterinaria , Toxoplasma/genética , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/parasitologíaRESUMEN
Listeriosis is a disease primarily of ruminants caused by the Gram-positive bacterium Listeria monocytogenes. Ruminants either demonstrate manifestations of the encephalitic, septicemic, or reproductive form of listeriosis. The pathological and molecular findings with encephalitic listeriosis in a 5.5-month-old, male, mixed-breed goat and a 3-year-old Texel-crossed sheep from northern Paraná, Brazil are described. Clinically, the kid demonstrated circling, lateral protrusion of the tongue, head tilt, and convulsions; the ewe presented ataxia, motor incoordination, and lateral decumbency. Brainstem dysfunctions were diagnosed clinically and listeriosis was suspected. Necropsy performed on both animals did not reveal remarkable gross lesions; significant histopathological alterations were restricted to the brainstem (medulla oblongata; rhombencephalitis) and were characterized as meningoencephalitis that consisted of extensive mononuclear perivascular cuffings, neutrophilic and macrophagic microabscesses, and neuroparenchymal necrosis. PCR assay and direct sequencing, using genomic bacterial DNA derived from the brainstem of both animals, amplified the desired 174 base pairs length amplicon of the listeriolysin O gene of L. monocytogenes. Phylogenetic analyses demonstrated that the strains associated with rhombencephalitis during this study clustered with known strains of L. monocytogenes lineage I from diverse geographical locations and from cattle of the state of Paraná with encephalitic listeriosis. Consequently, these strains should be classified as L. monocytogenes lineage I. These results confirm the active participation of lineage I strains of L. monocytogenes in the etiopathogenesis of the brainstem dysfunctions observed during this study, probably represent the first characterization of small ruminant listeriosis by molecular techniques in Latin America, and suggest that ruminants within the state of Paraná were infected by the strains of the same lineage of L. monocytogenes.
Asunto(s)
Toxinas Bacterianas/genética , Enfermedades de las Cabras/patología , Proteínas de Choque Térmico/genética , Proteínas Hemolisinas/genética , Listeriosis/veterinaria , Meningoencefalitis/veterinaria , Enfermedades de las Ovejas/patología , Animales , Tronco Encefálico/patología , Brasil , Análisis por Conglomerados , Femenino , Genotipo , Enfermedades de las Cabras/microbiología , Cabras , Histocitoquímica , Listeria monocytogenes/genética , Listeriosis/microbiología , Listeriosis/patología , Masculino , Meningoencefalitis/microbiología , Meningoencefalitis/patología , Filogenia , Reacción en Cadena de la Polimerasa , Homología de Secuencia , Ovinos , Enfermedades de las Ovejas/microbiologíaRESUMEN
Bovine coronavirus (BCoV) may cause acute diarrhea in newborn calves, leading to significant economic losses for cattle farmers. There are several diagnostic techniques used to detect BCoV in calf fecal samples, but virus isolation still has advantages for antigenic and genomic characterization. This study describes the isolation in HRT-18 cells and molecular characterization of Brazilian BCoV wild-type strains. Three fecal samples from diarrheic 30 day-old calves were inoculated in HRT-18 cell monolayers, which were then evaluated for HA titers and tested using semi-nested PCR followed by RFLP and sequencing. Two samples were successfully isolated and presented HA titers of 16 and 32 units per 25 mL. The results were confirmed using semi-nested PCR and RFLP. Molecular analyses identified a cell culture-adapted strain and a wild-type strain that were genetically similar (99 percent) to each other, but more distinct than BCoV strains circulating in other countries, even in the conserved N gene.
O coronavírus bovino (BCoV) pode causar diarreia aguda em bezerros recém-nascidos, ocasionando consideráveis perdas econômicas para a pecuária bovina. Várias técnicas de diagnóstico podem ser empregadas na detecção do BCoV a partir de amostras fecais de bezerros. Porém, o isolamento do BCoV em cultivo celular apresenta a vantagem de possibilitar a caracterização antigênica e molecular da estirpe viral. O presente estudo descreve o isolamento em células HRT-18, e a caracterização molecular de estirpes brasileiras do BCoV. Três amostras de fezes diarreicas de bezerros com 30 dias de idade foram inoculadas em culturas de células HRT-18. Os isolados foram avaliados por hemaglutinação (HA) e por uma semi-nested PCR seguida de RFLP e sequenciamento. Duas amostras foram isoladas e a confirmação foi verificada na semi-nested PCR e também RFLP. Na HA os títulos foram de 16 e 32 unidades por 25 mL. Análises moleculares identificam a estirpe adaptada em cultura celular e uma estirpe selvagem, como estirpes de BCoV semelhantes (99 por cento) entre si, mas distintas das circulantes em outros países, mesmo em um gene de uma proteína conservada (gene N).