C W Arns

Publications (12)19.23 Total impact

• Article: Molecular data of UL24 homolog gene (ORF37) from Brazilian isolates of equine herpesvirus type 1.

  R F Carvalho, F R Spilki, E M Cunha, R C Stocco, C W Arns
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  ABSTRACT: Equine herpesvirus type 1 (EHV-1) is associated with abortions, respiratory distress, and neurological disturbances in horses. The ORF37 of EHV-1 encodes a protein homolog to UL24 gene product of human herpesvirus that has been associated with neurovirulence. In the present work, ORF37 PCR fragments derived from two Brazilian EHV-1 isolates, a German isolate and an American reference strain were sequenced and characterized by molecular phylogenetic analysis. This genomic region is highly conserved an allowed to infer genetic distances between EHV-1 strains and other animal herpesvirus.
  Research in Veterinary Science 06/2011; 93(1):494-7. · 1.65 Impact Factor
• Article: Genetic diversity of avian infectious bronchitis virus isolated from domestic chicken flocks and coronaviruses from feral pigeons in Brazil between 2003 and 2009.

  P A N Felippe, L H A da Silva, M M A B Santos, F R Spilki, C W Arns
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  ABSTRACT: To detect the presence of infectious bronchitis virus or avian coronavirus, a nested reverse transcriptase PCR (RT-PCR) method was developed with the aim of amplifying a fragment of 530 bases, comprising the gene coding S1 protein. In the first step, all samples were submitted to RNA extraction, RT-PCR, and nested PCR. Next, only the positive nested-PCR samples were propagated in specific-pathogen-free (SPF) embryonated chicken eggs for virus isolation. Positive samples were then sequenced and analyzed using a molecular phylogeny approach. Tracheal swab samples were collected from 23 different domestic chickens distributed in three regions of Brazil, in the period between 2003 and 2009. Also analyzed were six swab samples (tracheal and cloacal) from asymptomatic pigeons (Columba livia), caught in an urbanized region in southeastern Brazil. The study revealed two major phylogenetic groups: one clustered with the Massachusetts vaccine serotype and another joined with the D207 strain. Interestingly, samples grouped with the Connecticut and Arkansas serotypes were also found. Pigeon isolates clustered with the Massachusetts serotype showed significant similarity (close to 100%) to those obtained from chickens. Only one pigeon isolate was seen to be grouped with the Connecticut serotype, and no correlation was observed between sample grouping and region origin. Understanding the diversity of genotypes and eco-epizootiology of the disease in different environments is expected to be helpful for vaccine production aimed at the main circulating variants. In this respect, one could also expect benefits in the management of other bird species that may act as avian coronavirus reservoirs.
  Avian Diseases 12/2010; 54(4):1191-6. · 1.46 Impact Factor
• Article: Infection and apparent invasion of vero cells by Paracoccidioides brasiliensis

  M.J.S. Mendes-Giannini, L.C. Ricci, M.A. Uemura, E. Toscano, C.W. Arns
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  ABSTRACT: Paracoccidioides brasiliensis probably uses many different mechanisms to establish itself in the host and cause disease. In this work, we assess an in vitro model system which uses cultured mammalian cells to investigate the virulence factors of P. brasiliensis. We were able to demonstrate an invasion process of the yeast form of this fungus in Vero cell cultures. We deduced that the overall invasive process involved three steps: adhesion, followed by invasion of individual epithelial cells and spread to adjacent cells.
  07/2009; 32(3):189-197.
• Source

  Available from: Fernando Rosado Spilki

  Article: Phylogenetic comparison of the carboxy-terminal region of glycoprotein C (gC) of bovine herpesviruses (BoHV) 1.1, 1.2 and 5 from South America (SA).

  P A Esteves, O A Dellagostin, L S Pinto, A D Silva, F R Spilki, J R Ciacci-Zanella, S O Hübner, R Puentes, J Maisonnave, A C Franco, F A M Rijsewijk, H B C R Batista, T F Teixeira, D Dezen, A P Oliveira, C David, C W Arns, P M Roehe
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  ABSTRACT: Different types and subtypes of bovine herpesvirus 1 and 5 (BoHV-1 and BoHV-5) have been associated to different clinical conditions of cattle, in such a way that type/subtype differentiation has become an essential tool for understanding the pathogenesis and epidemiology of BoHV infections. In search for a genomic region that would allow a clear distinction between BoHV-1 and BoHV-5, the carboxy-terminal portion of glycoprotein C (gC), corresponding to residues 321-450 (BoHV-1) and 301-429 (BoHV-5) of 23 South American (SA) isolates (Brazil mostly) was amplified and sequenced. The nucleotide sequence alignments revealed levels of genomic similarity ranging from 98.7 to 99.8% among BoHV-1 isolates, 88.3 to 92% between BoHV-1/BoHV-5 and 96 to 99.7% among BoHV-5 isolates. At the amino acid level, sequence similarity varied ranging from 97.5 to 99.5% among BoHV-1, 77.5 to 84.4% between BoHV-1/BoHV-5 and 92.1 to 99.5% (BoHV-5/BoHV-5). The isolates could be clearly separated into BoHV-1.1, BoHV-1.2 and BoHV-5 after phylogenetic analysis. The results suggest that the phylogenetic analysis performed here can be used as a potential molecular epidemiological tool for herpesviruses.
  Virus Research 02/2008; 131(1):16-22. · 2.94 Impact Factor
• Article: Circulation of bovine respiratory syncytial virus in Brazil.

  R S Almeida, H G Domingues, F R Spilki, L E Larsen, S Hägglund, S Belák, C W Arns
  The Veterinary record 06/2006; 158(18):632-4. · 1.25 Impact Factor
• Article: Characterization of bovine respiratory syncytial virus isolated in Brazil.

  C W Arns, J Campalans, S C B Costa, H G Domingues, R C F D'Arce, R S Almeida, L T Coswig
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  ABSTRACT: This paper presents the first isolation of bovine respiratory syncytial virus in Brazil and its physicochemical, morphological and molecular characterization. The virus was isolated from 33 samples of nasotracheal secretions, successively inoculated into a Madin-Darby bovine kidney cell culture, which was characterized by physicochemical tests and morphological observation by electron microscopy. The Brazilian sample is an RNA pleomorphic, enveloped, thermolabile and non-hemagglutinating spicular virus. Reverse transcription, followed by nested polymerase chain reaction (nRT-PCR) assay was carried out using oligonucleotides B1, B2A, B3 and B4 for the fusion proteins (F) and B5A, B6A, B7A and B8 for the attachment protein (G). The nRT-PCR-F amplified a fragment of 481 bp corresponding to part of the gene that codes for protein F, whereas nRT-PCR-G amplified a fragment of 371 bp, in agreement with part of the G gene. The virus isolated from Brazilian samples in this study corresponded to the bovine respiratory syncytial virus, and RT-PCR proved to be useful for the diagnosis of bovine clinical samples.
  Brazilian Journal of Medical and Biological Research 03/2003; 36(2):213-8. · 1.13 Impact Factor
• Source

  Available from: Fernando Rosado Spilki

  Article: Restriction endonuclease and monoclonal antibody analysis of Brazilian isolates of bovine herpesviruses types 1 and 5.

  R C F D'Arce, R S Almeida, T C Silva, A C Franco, F Spilki, P M Roehe, C W Arns
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  ABSTRACT: Twelve Brazilian isolates and three reference strains of bovine herpesviruses (BHVs) were subjected to restriction endonuclease analysis (REA) and monoclonal antibody (MAb) analysis. Viral DNA was cleaved with BamHI, BstEII, EcoRI, HindIII and PstI. The monoclonal antibody panel allowed the differentiation between types 1 and 5 viruses, while REA with BstEII and HindIII showed the distinction between BHV-1 and -5 subtypes. Typical 1.1 and 1.2a patterns were observed with two isolates from respiratory disease. An isolate from semen of a clinically healthy bull displayed 1.2b profile, whereas another displayed a clear 5a pattern, which was never reported before. Seven out of nine Brazilian type 5 (BHV-5) isolates displayed REA patterns similar to the Australian BHV-5 strain N569 (BHV-5a), and differing from the Argentinean A663 strain (BHV-5b) virus. Another two BHV-5 isolates, which displayed an unusual MAb pattern of reactivity, showed a BstEII profile different from both reference strains of BHV-5. These two viruses were considered BHV-5 "non-a/non-b" subtype.
  Veterinary Microbiology 10/2002; 88(4):315-24. · 3.33 Impact Factor
• Source

  Available from: Tereza Cristina Cardoso

  Article: Replication of classical infectious bursal disease virus in the chicken embryo related cell line.

  T C Cardoso, P Rahal, D Pilz, M C Teixeira, C W Arns
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  ABSTRACT: Infectious bursal disease (IBD) is an acute, highly contagious viral disease. The diagnosis of IBD depends on time-consuming and costly procedures, like virus isolation on chick embryos and histopathological examination. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), immunoperoxidase and reverse transcription polymerase chain reaction (RT-PCR) were applied in this study to detect classical IBD virus (IBDV) after three blind passages of the Lukert strain on chicken embryo related (CER) cell monolayer after different periods of infection: 6, 12, 24 and 48h. Cytophatic effects were most evident 12 h post-infection (p.i.) but were observed at 6 h p.i. The maximum discrimination between IBDV-infected and uninfected cell suspensions obtained by the use of DAS-ELISA for virus detection corresponded to 0.597 +/-0.02 and 0.010 +/-0.01 after 12 h p.i., respectively. The RT-PCR was performed using the set of primers A3.1 and A3.2 to amplify the VP2 region of the IBDV genome. This molecular technique demonstrated that from 6h p.i., it was possible to detect the viral RNA. The results show that the CER cell line can be used for classical IBDV propagation, confirmed by the DAS-ELISA, immunoperoxidase and RT-PCR assay.
  Avian Pathology 07/2000; 29(3):213-7. · 1.71 Impact Factor
• Article: Molecular characterization of Brazilian avian pneumovirus isolates: comparison between immunochemiluminescent Southern blot and nested PCR.

  M A Dani, E L Durigon, C W Arns
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  ABSTRACT: Avian pneumovirus (APV) causes acute respiratory tract infection both in turkeys (turkey rhinotracheitis) and chickens (swollen head syndrome (SHS)) with sudden onset and rapid spread through the flocks. In this study, an immunochemiluminescent Southern blot RT-PCR assay was employed to detect a F gene transcript of the APV in two European turkey isolates and two Brazilian chicken isolates. Limiting dilution PCR was carried out to compare the sensitivity of immunochemiluminescent Southern blot assay and nested PCR assay (nPCR). The sensitivity and specificity of immunochemiluminescent Southern blot RT-PCR assay were comparable to that of nPCR, and at least 100 fold more sensitive than a single PCR amplification. Sequence analysis of the 175 bp product of the F gene revealed 100% identity with APV sequences described earlier.
  Journal of Virological Methods 06/1999; 79(2):237-41. · 2.01 Impact Factor
• Article: Virulence factors of avian Escherichia coli associated with swollen head syndrome.

  V R Parreira, C W Arns, T Yano
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  ABSTRACT: Virulence characteristics of 50 strains of Escherichia coli isolated from chickens with swollen head syndrome were examined. The results were the following: in the absence of D-mannose, 74% of strains agglutinated guinea pig erythrocytes, but in the presence of D-mannose 32% agglutinated guinea-pig erythrocytes and 62% agglutinated human erythrocytes. When slide agglutination assays were carried out with antisera to adhesin of bovine and swine origin (K88, K99, F41, F42 987P and 2134P), only 14% of strains agglutinated with antiserum to F41. Colicin V was produced by 78% of the E. coli strains and 80% produced aerobactin. In the serum resistance test, 36 (72%) of strains showed resistance to normal chicken serum. Only seven (14%) strains expressed K1 capsular antigen, while motility was found in 62% of the strains.
  Avian Pathology 05/1998; 27(2):148-54. · 1.71 Impact Factor
• Article: An agar-overlay method for detection of toxins produced by Escherichia coli.

  V R Parreira, C W Arns, T Yano
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  ABSTRACT: An agar overlay method, with Vero and HeLa cells, was used for detection of heat-labile enterotoxin and verotoxin from Escherichia coli. The method is more sensitive than the conventional cell culture assay, is rapid, easy to perform, and is suitable for epidemiological studies.
  FEMS Microbiology Letters 08/1994; 120(3):303-6. · 2.04 Impact Factor
• Article: Infection and apparent invasion of Vero cells by Paracoccidioides brasiliensis.

  M J Mendes-Giannini, L C Ricci, M A Uemura, E Toscano, C W Arns
  [show abstract] [hide abstract]
  ABSTRACT: Paracoccidioides brasiliensis probably uses many different mechanisms to establish itself in the host and cause disease. In this work, we assess an in vitro model system which uses cultured mammalian cells to investigate the virulence factors of P. brasiliensis. We were able to demonstrate an invasion process of the yeast form of this fungus in Vero cell cultures. We deduced that the overall invasive process involved three steps: adhesion, followed by invasion of individual epithelial cells and spread to adjacent cells.
  Journal of medical and veterinary mycology: bi-monthly publication of the International Society for Human and Animal Mycology 02/1994; 32(3):189-97.