Journal of Basic and Applied Pharmaceutical Sciencies, 2015
O calicivirus felino (FCV) e um importante patogeno de gatos que causa lesoes ulcerativas orais e... more O calicivirus felino (FCV) e um importante patogeno de gatos que causa lesoes ulcerativas orais e infeccoes respiratorias. O virus tem sido utilizado como modelo experimental para avaliacao de agente antivirais contra norovirus (NoVs). Nesse estudo, investigou-se a acao dos oleos essenciais de alecrim (Rosmarinus officinalis L.), oregano mexicano (Lippia graveolens HBK.) e tomilho (Thymus vulgaris L.) frente ao FCV, in vitro. A toxicidade celular foi testada pelo metodo de MTT e os ensaios antivirais pelo teste de reducao de placas. Tres protocolos foram aplicados: a) diferentes concentracoes nao toxicas dos oleos essenciais (CNTOE) foram incubadas com o virus por 1 hora antes da inoculacao (ensaio virucida); b) CNTOE foram adicionadas as celulas CRFK e incubadas por 1 hora antes da adsorcao viral (ensaio de pre-tratamento); c) CNTOE foram adicionadas as celulas apos a inoculacao do FCV e mantidas por 18 horas (ensaio de pos-tratamento). A CC50 para os oleos de alecrim, oregano mexi...
Feline calicivirus (FCV) and felid herpesvirus type-1 (FeHV-1) are the main infectious agents of ... more Feline calicivirus (FCV) and felid herpesvirus type-1 (FeHV-1) are the main infectious agents of domestic and wild felines worldwide. The FCV and FeHV-1 viruses were isolated in Brazil in 1988 and 2012, respectively. Serology surveys were performed among domestic feline in the State of Rio Grande do Sul and among wild felines in central Brazilian States. Felines with acute or chronic infections may become carriers for both viruses and, viral transmission occurs mainly by ocular and nasal secretions. In addition, FCV may be transmitted by oropharyngeal secretion and fomites. The clinical signs commonly observed in cats are fever, sneezing, coughing and nasal and ocular discharge; however, oral lesions are restricted to FCV infection. A systemic syndrome showing hemorrhagic lesions, alopecia, facial edema and jaundice has been associated with FCV. Attenuated as well as inactivated vaccines against FCV and FeHV-1 were developed in the middle 1970s, and they are effective at reducing th...
Feline calicivirus (FCV) is an important pathogen that affects domestic cats, inducing acute oral... more Feline calicivirus (FCV) is an important pathogen that affects domestic cats, inducing acute oral and upper respiratory tract clinical signs. The aim of this study was to analyze the variability of the capsid protein in different FCV isolates from southern Brazil. The sequencing analyses of thirteen Brazilian FCV samples, phylogenetic analyses and assessments of ten previously published sequences were conducted by examining the open reading frame 2 (ORF2, regions B-F). Comparisons of the predicted amino acid sequences of the ORF2 in Brazilian FCV isolates with those of the FCV-F9 strain indicated that the main differences are located within the regions C and hypervariable E (HVR_E). Epitopes that were mapped to the regions D, 5'HVR_E and conserved E also presented with some variability when compared to the strain F9. This is the first study describing sequence analyses and the phylogenetic relationships among FCV isolates from Brazil. The results presented here may expand upon c...
Esse artigo relata a avaliação da resposta sorológica e proteção fetal conferida por uma vacina e... more Esse artigo relata a avaliação da resposta sorológica e proteção fetal conferida por uma vacina experimental contendo duas amostras atenuadas do vírus da diarréia viral bovina tipos 1 (BVDV-1) e 2 (BVDV-2). Vacas foram imunizadas com a vacina experimental (n=19) e juntamente com controles não-vacinadas (n=18) foram colocadas em cobertura e desafiadas, entre os dias 60 e 90 de gestação, pela inoculação intranasal de quatro amostras heterólogas de BVDV-1 e BVDV-2. A resposta sorológica foi avaliada por testes de soro-neutralização realizados a diferentes intervalos após a vacinação (dias 34, 78 e 138 pós-vacinação [pv]). A proteção fetal foi monitorada por exames ultra-sonográficos e clínicos realizados durante o restante da gestação; e pela pesquisa de vírus e anticorpos no sangue pré-colostral coletado dos fetos abortados e/ou dos bezerros recém nascidos. No dia do desafio (dia 138 pv), todas as vacas vacinadas apresentavam anticorpos neutralizantes em títulos altos contra o BVDV-1 ...
Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum recognized in hors... more Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum recognized in horses, dogs, cats, cattle and humans. In dogs, the disease involves the gastrointestinal tract and skin. A 2-year-old male Labrador Retriever was referred to the Veterinary Hospital at the Federal University of Santa Maria (HV-UFSM), because of vomiting, diarrhea and weight loss. On clinical examination, there was a palpable mass in the abdominal cavity, confirmed by exploratory laparotomy. Because of the poor prognosis the dog was euthanatized. On histological examination, thick hyphae (4-7 μm) could be easily seen in sections stained with GMS. The diagnosis was confirmed through isolation of the agent, and the species P. insidiosum was confirmed by nested polymerase chain reaction (PCR)-based assay.
BLUETONGUE is an arthropod-borne disease of ruminants caused by the bluetongue virus (BTV). The d... more BLUETONGUE is an arthropod-borne disease of ruminants caused by the bluetongue virus (BTV). The distribution of BTV depends on the distribution of Culicoides species insect populations and appropriate climatic conditions. Except for some sheep breeds, most BTV infections in domestic ruminants are
Many aspects of the biology of orf virus (ORFV) infection remain poorly understood and attempts t... more Many aspects of the biology of orf virus (ORFV) infection remain poorly understood and attempts to establish animal models have yielded conflicting and non-reproducible results. We herein describe the characterization of ORFV infection and disease in rabbits and mice. A protocol of intradermal inoculation was employed to inoculate 10 8.5 TCID 50 /mL of ORFV strain IA-82 in the skin of ears, of the back and labial commissures. All inoculated rabbits presented a clinical course characterized by erythema, macules, papules/vesicles or pustules that eventually dried originating scabs. Local signs started around days 3 and 4 post-inoculation (pi) and lasted 3e10 days. Virus was recovered from lesions between days 2 and 14pi. Histological examination of lesions revealed focal proliferative dermatitis with ballooning degeneration and eosinophilic intracytoplasmic inclusion bodies in keratinocytes, histological hallmarks of contagious ecthyma in sheep. A similar, albeit milder clinical course occurred in 5/10 inoculated mice; virus was recovered from lesions from three animals. Inoculated lambs e used as controls e developed severe lesions of contagious ecthyma. VN tests performed at day 28pi failed to detect neutralizing antibodies in all inoculated animals. In contrast, convalescent rabbit sera were positive by ELISA at dilutions from 100 to 400. These results show that rabbits are susceptible to ORFV infection and thus may be used to study selected aspects of ORFV biology.
Feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1) are the two primary causes of uppe... more Feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1) are the two primary causes of upper respiratory tract disease in cats. The aim of this study was to demonstrate the distribution of FCV and FHV-1 among the feline population of several counties in Rio Grande do Sul State, Brazil. To this end, conjunctival and nasal swabs were collected from 302 cats from different locations, including households, breeding catteries, veterinary clinics, animal hospitals and experimental research facilities. The samples were collected between July 2006 to June 2009. The virus isolation was performed in CRFK cells and, subsequently, the identification was confirmed by PCR. FCV, FHV-1, or both were isolated from 55 cats from 28 different locations. FCV alone was isolated from 52.7% (29/55) of the animals that tested positively, FHV-1 alone was isolated from 38.2% (21/55) of the animals that tested positively, and coinfection were detected in 9.1% (5/55) of the animals that tested positively. Virus detection was more prevalent in cats that were less than 1 year old, among animals that shared a living space with other cats, and females. FCV and FHV-1 were isolated from vaccinated cats. In addition, both viruses were isolated from cats that showed no signs of disease. The results suggest that a carrier state is common for both viruses in the evaluated population. A search for other causes of respiratory disease in that population is necessary; and further studies relating to the molecular characterization of viruses and vaccine efficacy are also necessary.
This study aims to expand the knowledge about fecal contamination by humans and animals using Ade... more This study aims to expand the knowledge about fecal contamination by humans and animals using Adenovirus (AdV) as bioindicators in different water sources from rural areas, to evaluate the viral infectivity, and to compare the different techniques used to detect the Human mastadenovirus (HAdV). For that, 124 samples were collected (86 from groundwater and 38 from surface water) along the Rio dos Sinos Basin. Escherichia coli count was carried out, and the samples were submitted for the detection and characterization tests of AdV by different methods (qPCR, multiplex qPCR, and nested PCR). In addition, the viral infectivity was realized by integrated cell culture quantitative PCR (ICC-qPCR). E. coli was detected in 63% of groundwater samples (geometric mean of 16.7 MPN/100 mL) and 68% (geometric mean: 5.08×102 MPN/100 mL) in surface waters. Among the viral indicator in the groundwater, the HAdV was detected in 49% of the samples, followed by Canine mastadenovirus (CAV, 20%), Bovine mastadenovirus (BAdV, 17%), Aviadenovirus (AvAdV, 15%), and Porcine mastadenovirus (PAdV, 03%). In surface water, HAdV was detected in 45%, followed by CAV (42%), BAdV (29%), and PAdV and AvAdV (13%). The quantification of genomic copies per liter ranged from 9.40×104 to 5.54×1010 gc/L. In groundwater samples, it was possible to observe infectious adenovirus in 12% of the samples, as well as in surface water for 18%. The results showed an increase in the sensitivity of positive samples when a combined set of techniques were used for HAdV detection.
Journal of Basic and Applied Pharmaceutical Sciencies, 2015
O calicivirus felino (FCV) e um importante patogeno de gatos que causa lesoes ulcerativas orais e... more O calicivirus felino (FCV) e um importante patogeno de gatos que causa lesoes ulcerativas orais e infeccoes respiratorias. O virus tem sido utilizado como modelo experimental para avaliacao de agente antivirais contra norovirus (NoVs). Nesse estudo, investigou-se a acao dos oleos essenciais de alecrim (Rosmarinus officinalis L.), oregano mexicano (Lippia graveolens HBK.) e tomilho (Thymus vulgaris L.) frente ao FCV, in vitro. A toxicidade celular foi testada pelo metodo de MTT e os ensaios antivirais pelo teste de reducao de placas. Tres protocolos foram aplicados: a) diferentes concentracoes nao toxicas dos oleos essenciais (CNTOE) foram incubadas com o virus por 1 hora antes da inoculacao (ensaio virucida); b) CNTOE foram adicionadas as celulas CRFK e incubadas por 1 hora antes da adsorcao viral (ensaio de pre-tratamento); c) CNTOE foram adicionadas as celulas apos a inoculacao do FCV e mantidas por 18 horas (ensaio de pos-tratamento). A CC50 para os oleos de alecrim, oregano mexi...
Feline calicivirus (FCV) and felid herpesvirus type-1 (FeHV-1) are the main infectious agents of ... more Feline calicivirus (FCV) and felid herpesvirus type-1 (FeHV-1) are the main infectious agents of domestic and wild felines worldwide. The FCV and FeHV-1 viruses were isolated in Brazil in 1988 and 2012, respectively. Serology surveys were performed among domestic feline in the State of Rio Grande do Sul and among wild felines in central Brazilian States. Felines with acute or chronic infections may become carriers for both viruses and, viral transmission occurs mainly by ocular and nasal secretions. In addition, FCV may be transmitted by oropharyngeal secretion and fomites. The clinical signs commonly observed in cats are fever, sneezing, coughing and nasal and ocular discharge; however, oral lesions are restricted to FCV infection. A systemic syndrome showing hemorrhagic lesions, alopecia, facial edema and jaundice has been associated with FCV. Attenuated as well as inactivated vaccines against FCV and FeHV-1 were developed in the middle 1970s, and they are effective at reducing th...
Feline calicivirus (FCV) is an important pathogen that affects domestic cats, inducing acute oral... more Feline calicivirus (FCV) is an important pathogen that affects domestic cats, inducing acute oral and upper respiratory tract clinical signs. The aim of this study was to analyze the variability of the capsid protein in different FCV isolates from southern Brazil. The sequencing analyses of thirteen Brazilian FCV samples, phylogenetic analyses and assessments of ten previously published sequences were conducted by examining the open reading frame 2 (ORF2, regions B-F). Comparisons of the predicted amino acid sequences of the ORF2 in Brazilian FCV isolates with those of the FCV-F9 strain indicated that the main differences are located within the regions C and hypervariable E (HVR_E). Epitopes that were mapped to the regions D, 5'HVR_E and conserved E also presented with some variability when compared to the strain F9. This is the first study describing sequence analyses and the phylogenetic relationships among FCV isolates from Brazil. The results presented here may expand upon c...
Esse artigo relata a avaliação da resposta sorológica e proteção fetal conferida por uma vacina e... more Esse artigo relata a avaliação da resposta sorológica e proteção fetal conferida por uma vacina experimental contendo duas amostras atenuadas do vírus da diarréia viral bovina tipos 1 (BVDV-1) e 2 (BVDV-2). Vacas foram imunizadas com a vacina experimental (n=19) e juntamente com controles não-vacinadas (n=18) foram colocadas em cobertura e desafiadas, entre os dias 60 e 90 de gestação, pela inoculação intranasal de quatro amostras heterólogas de BVDV-1 e BVDV-2. A resposta sorológica foi avaliada por testes de soro-neutralização realizados a diferentes intervalos após a vacinação (dias 34, 78 e 138 pós-vacinação [pv]). A proteção fetal foi monitorada por exames ultra-sonográficos e clínicos realizados durante o restante da gestação; e pela pesquisa de vírus e anticorpos no sangue pré-colostral coletado dos fetos abortados e/ou dos bezerros recém nascidos. No dia do desafio (dia 138 pv), todas as vacas vacinadas apresentavam anticorpos neutralizantes em títulos altos contra o BVDV-1 ...
Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum recognized in hors... more Pythiosis is a granulomatous disease caused by the oomycete Pythium insidiosum recognized in horses, dogs, cats, cattle and humans. In dogs, the disease involves the gastrointestinal tract and skin. A 2-year-old male Labrador Retriever was referred to the Veterinary Hospital at the Federal University of Santa Maria (HV-UFSM), because of vomiting, diarrhea and weight loss. On clinical examination, there was a palpable mass in the abdominal cavity, confirmed by exploratory laparotomy. Because of the poor prognosis the dog was euthanatized. On histological examination, thick hyphae (4-7 μm) could be easily seen in sections stained with GMS. The diagnosis was confirmed through isolation of the agent, and the species P. insidiosum was confirmed by nested polymerase chain reaction (PCR)-based assay.
BLUETONGUE is an arthropod-borne disease of ruminants caused by the bluetongue virus (BTV). The d... more BLUETONGUE is an arthropod-borne disease of ruminants caused by the bluetongue virus (BTV). The distribution of BTV depends on the distribution of Culicoides species insect populations and appropriate climatic conditions. Except for some sheep breeds, most BTV infections in domestic ruminants are
Many aspects of the biology of orf virus (ORFV) infection remain poorly understood and attempts t... more Many aspects of the biology of orf virus (ORFV) infection remain poorly understood and attempts to establish animal models have yielded conflicting and non-reproducible results. We herein describe the characterization of ORFV infection and disease in rabbits and mice. A protocol of intradermal inoculation was employed to inoculate 10 8.5 TCID 50 /mL of ORFV strain IA-82 in the skin of ears, of the back and labial commissures. All inoculated rabbits presented a clinical course characterized by erythema, macules, papules/vesicles or pustules that eventually dried originating scabs. Local signs started around days 3 and 4 post-inoculation (pi) and lasted 3e10 days. Virus was recovered from lesions between days 2 and 14pi. Histological examination of lesions revealed focal proliferative dermatitis with ballooning degeneration and eosinophilic intracytoplasmic inclusion bodies in keratinocytes, histological hallmarks of contagious ecthyma in sheep. A similar, albeit milder clinical course occurred in 5/10 inoculated mice; virus was recovered from lesions from three animals. Inoculated lambs e used as controls e developed severe lesions of contagious ecthyma. VN tests performed at day 28pi failed to detect neutralizing antibodies in all inoculated animals. In contrast, convalescent rabbit sera were positive by ELISA at dilutions from 100 to 400. These results show that rabbits are susceptible to ORFV infection and thus may be used to study selected aspects of ORFV biology.
Feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1) are the two primary causes of uppe... more Feline calicivirus (FCV) and feline herpesvirus type 1 (FHV-1) are the two primary causes of upper respiratory tract disease in cats. The aim of this study was to demonstrate the distribution of FCV and FHV-1 among the feline population of several counties in Rio Grande do Sul State, Brazil. To this end, conjunctival and nasal swabs were collected from 302 cats from different locations, including households, breeding catteries, veterinary clinics, animal hospitals and experimental research facilities. The samples were collected between July 2006 to June 2009. The virus isolation was performed in CRFK cells and, subsequently, the identification was confirmed by PCR. FCV, FHV-1, or both were isolated from 55 cats from 28 different locations. FCV alone was isolated from 52.7% (29/55) of the animals that tested positively, FHV-1 alone was isolated from 38.2% (21/55) of the animals that tested positively, and coinfection were detected in 9.1% (5/55) of the animals that tested positively. Virus detection was more prevalent in cats that were less than 1 year old, among animals that shared a living space with other cats, and females. FCV and FHV-1 were isolated from vaccinated cats. In addition, both viruses were isolated from cats that showed no signs of disease. The results suggest that a carrier state is common for both viruses in the evaluated population. A search for other causes of respiratory disease in that population is necessary; and further studies relating to the molecular characterization of viruses and vaccine efficacy are also necessary.
This study aims to expand the knowledge about fecal contamination by humans and animals using Ade... more This study aims to expand the knowledge about fecal contamination by humans and animals using Adenovirus (AdV) as bioindicators in different water sources from rural areas, to evaluate the viral infectivity, and to compare the different techniques used to detect the Human mastadenovirus (HAdV). For that, 124 samples were collected (86 from groundwater and 38 from surface water) along the Rio dos Sinos Basin. Escherichia coli count was carried out, and the samples were submitted for the detection and characterization tests of AdV by different methods (qPCR, multiplex qPCR, and nested PCR). In addition, the viral infectivity was realized by integrated cell culture quantitative PCR (ICC-qPCR). E. coli was detected in 63% of groundwater samples (geometric mean of 16.7 MPN/100 mL) and 68% (geometric mean: 5.08×102 MPN/100 mL) in surface waters. Among the viral indicator in the groundwater, the HAdV was detected in 49% of the samples, followed by Canine mastadenovirus (CAV, 20%), Bovine mastadenovirus (BAdV, 17%), Aviadenovirus (AvAdV, 15%), and Porcine mastadenovirus (PAdV, 03%). In surface water, HAdV was detected in 45%, followed by CAV (42%), BAdV (29%), and PAdV and AvAdV (13%). The quantification of genomic copies per liter ranged from 9.40×104 to 5.54×1010 gc/L. In groundwater samples, it was possible to observe infectious adenovirus in 12% of the samples, as well as in surface water for 18%. The results showed an increase in the sensitivity of positive samples when a combined set of techniques were used for HAdV detection.
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