Comparison of Serological and Molecular Tests for Diagnosis of Caprine Arthritis Encephalitis and Clinical Evaluation of Mammary Glands of Infected Dairy Goats

Authors

  • Dalva Alana Aragão de Azevedo Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Raymundo Rizaldo Pinheiro Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Vanderlan Warlington Souza de Santos Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Edgar Marques Damasceno Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Ana Lídia Madeira de Sousa Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Juscilânia Furtado Araújo Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Alice Andrioli Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Lúcia Helena Sider Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Renato Mesquita Peixoto Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.
  • Maria Fátima da Silva Teixeira Programa de Pós-graduação em Ciências Veterinárias (PPGCV), Universidade Estadual do Ceará (UECE), Fortaleza, CE, Brazil.

DOI:

https://doi.org/10.22456/1679-9216.92281

Abstract

Background: Caprine Arthritis Encephalitis (CAE) is a disease that causes productive losses in dairy goat flocks due to the reduction in milk production, followed by lesions in joints and mammary glands. An early diagnosis is essential, considering that there is frequent occurrence of asymptomatic animals. Hence, this study aimed to perform a comparison of immunological and molecular based diagnostic tests, represented by Agar Gel Immunodiffusion (AGID), Western Blot (WB) and nested Polymerase Chain Reaction (nPCR). In addition, the mammary glands (MG) of dairy goats were clinically evaluated. 

Material, Methods & Results: Blood collection and clinical examination were performed in 1191 dairy goats of 12 farms located in Northeastern and Southeastern regions of Brazil. Serological (AGID, WB) and molecular (nPCR) test results were compared and the data, along with MG alterations, were analyzed using Epi-info 7 and WinEpiscope 2.0. Seroprevalence in AGID test was 41.14% (490/1191). In WB, 51.47% (613/1191) of animals were seropositive and nPCR detected 69.44% (827/1191) positive animals. Hence, WB was more sensitive (P < 0.001) than AGID. However, nPCR detected more positive animals than AGID (P < 0.001) and WB (P < 0.001). The analysis of mammary glands revealed that 105 out of 1096 nanny goats presented alterations, of which 101 presented altered consistency, 16 presented elevated temperatures and 60 had enlarged retromammary lymph nodes. There was significant statistic difference (P < 0.05) only when comparing the results of serological tests with MG alterations.

Discussion: In general, AGID technique is most frequently used when screening flocks for the disease due to the practicality and low cost this test presents. However, the results demonstrated that AGID detected the lowest number of positive animals. This low sensitivity that the test presented may be attributed to its antigen-antibody interaction mechanism, considering that agar gel precipitation requires multiple interactions. In addition, WB was more effective than AGID in detecting antibodies. On the other hand, nPCR was important for the detection of infected animals that serological tests failed to detect. The intermittence of immunological response observed in the serological tests may be explained by the variation of antibodies levels that may occur during life. Likewise, viral compartmentalization would justify the intermittent detection of proviral DNA. Hence, the results can be influenced by the viral intermittence, test sensitivity, late seroconversion and statistic values that can be calculated (sensitivity, specificity, positive predictive value, negative predictive level and kappa). Crossing the results of the diagnostic tests with the different mammary gland alterations, it was shown that there was a statistically significant difference (P <0.05) only in the comparison of the results of the serological tests with GM alterations. Everything indicates that the humoral or cellular immune system being on stimulus is more propitious to find these changes. In conclusion, WB was more sensitive than AGID and, considering that nPCR can detect a larger number of animals infected with the goat lentivirus, it must be associated with a sensible serological test, such as Western Blot. In addition, infected animals have alterations in MG, which is more frequent in cases with positive serological results.

Downloads

Download data is not yet available.

References

Alves S.M., Teixeira M.F.S., Pinheiro R.R., Alves F.S.F., Lima A.M.C., Farias D.A., Santos V.W.S., Azevedo D.A.A., Martins G.R. & Aguiar T.D.F. 2018. Seroepidemiological study of maedi-visna in sheep in Ceará, Rio Grande do Norte, Paraíba, and Sergipe States. Semina: Ciências Agrárias. 39(5): 2017-2028.

Andrioli A. 2001. Vírus da Artrite Encefalite Caprina: PCR e isolamento viral de amostras de sêmen, fluido uterino e embriões. 68f. Belo Horizonte, MG. Tese (Doutorado em Ciência Animal) - Programa de Pós-graduação em Ciência Animal, Universidade Federal de Minas Gerais.

Andrioli A., Gouveia A.M.G., Martins A.S., Pinheiro R.R. & Santos D.O. 2006. Fatores de risco na transmissão do lentivírus caprino pelo sêmen. Pesquisa Agropecuária Brasileira. 41: 1313-1319.

Benavides J., Fuertes M., García-Pariente C., Otaola J., Delgado L., Giraldez J., Marín J.F.G., Ferreras M. C. & Pérez V. 2013. Impact of maedi visna in intensively managed dairy sheep. The Veterinary Journal. 197: 607-612.

Bertoni G., Zahno M.L., Zanoni, R., Vogt H.R., Peterhans E., Ruff G., Cheevers W.P., Sonigo P. & Pancino G. 1994. Antibody reactivity to the immunodominant epitopes of the caprine arthritis-encephalitis virus gp38 transmembrane protein associates with development of arthritis. Journal of Virology. 68(11): 7139-7147.

Blacklaws B.A. 2012. Small ruminant lentiviruses: Immunopathogenesis of visna-maedi and caprine arthritis and encephalitis virus. Comparative Immunology, Microbiology & Infectious Diseases. 35(3): 259-269.

Braz G.F. 2013. Lentivirus de pequenos ruminantes: caracterização genética e antigênica de isolados de caprinos e ovinos do Brasil. 79f. Belo Horizonte, MG. Tese (Doutorado em Ciência Animal) – Programa de Pós-graduação em Ciência Animal, Universidade Federal de Minas Gerais.

Brodie S.J., De La Concha-Bermejillo A., Snowder G.D. & Demartini J.C. 1998. Current concepts in the epizootiology, diagnosis and economic importance of ovine progressive pneumonia in North America: A review. Small Ruminant Research. 27(1): 1-17.

Cavalcante R.F.R.A., Andrioli A., Pinheiro R.R., Sousa K.C., Veras A.K.A., Lopes T.A. & Sousa S.D. 2013. Detecção do vírus da Artrite Encefalite Caprina por nested PCR e nested RT-PCR em ovócitos e fluido uterino. Arquivos do Instituto Biológico. 80: 381–386.

Celer Junior V., Celer V., Némcová H., Zanoni R.G. & Peterhans E. 1998. Serologic diagnosis of ovine lentiviruses by whole virus ELISA and AGID test. Journal of Veterinary Medicine Series B. 45(3): 183-188.

Dantas T.V.M., Araújo S.A.C., Pinheiro R.R., Aragão M.A.C., Silva J.B.A., Ricarte A.R.F., Ribeiro A.L. & Teixeira M.F.S. 2008. Desenvolvimento e padronização de um ELISA indireto para diagnóstico de Maedi Visna em ovinos. Ciência Animal Brasileira. 9(1): 181-187.

Gouveia A.M. 1994. Padronização de microtécnica de imunodifusão em gel de agarose para diagnóstico de lentivírus da pneumonia progressiva ovina (OPP), maedi-visna (MVV) e artrite encefalite caprina (CAEV). Sobral (Mimeografado). 4p.

Grimberg J., Nawoschik S., Belluscio L., Mckee R., Turck A. & Eisenberg A. 1989. A simple and eficiente non-organic procedure for the isolation of genomic DNA from blood. Nucleic Acids Research. 17: 8390.

Hasegawa Y.M., Lara M.C.C.S.H., Gaeta N.C., Marques J.A., Ribeiro B.L.M., Rossi R.S. & Gregory L. 2017. Transmissibilidade de Lentivírus de Pequenos Ruminantes para cabritos e cabras adultas por meio de sêmen infectado experimentalmente. Pesquisa Veterinária Brasileira. 37: 805-812.

IBGE - Instituto Brasileiro de Geografia e Estatística. Censo Agropecuário 2014. Disponível em: . [Accessed online in September 2018].

Lara M.C.C.S.H., Birgel Junior E.H., Gregory L. & Birgel E.H. 2005. Aspectos clínicos da artrite-encefalite dos caprinos. Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 57(6): 736-740.

Martínez-Navalón B., Peris C., Gómes E.A., Peris B., Roche M.L., Caballero C., Govena E. & Berriatua E. 2013. Quantitative estimation of the impact of caprine arthritis encephalitis virus infection on milk production by dairy goats. Veterinary Journal. 197(2): 311-317.

Michiels R., Mael E.V., Quinet C., Adjadj N.R., Cay A.B. & De Regge N. 2018. Comparative Analysis of Different Serological and Molecular Tests for the Detection of Small Ruminant Lentiviruses (SRLVs) in Belgian Sheep and Goats. Viruses. 10: 696.

Minguijón E., Reina R., Pérez M., Polledo L., Villoria M., Ramírez H., Leginagoikoa I., Badiola J.J., García-Marín J.F., Andrés D., Luján L., Amorena B. & Juste R.A. 2015. Small ruminant lentivirus infections and diseases. Veterinary Microbiology. 181(1-2): 75-89.

Office International des Epizooties - OIE. 2004. Manual of standards diagnostic tests and vaccines. World Organization for Animal Health. 5th edn. Paris: OIE, 1178p.

Panneum S. & Rukkwamsuk T. 2017. Diagnosis of Caprine Arthritis Encephalitis Virus infection in dairy goats by ELISA, PCR and Viral Culture. Polish Journal of Veterinary Sciences. 20(2): 347-353.

Peixoto R.M., Andrioli A., Pinheiro R.R., Alves F.S.F., Santos V.W.S., Sousa M.M., Azevedo D.A.A., Damasceno E.M. & Teixeira M.F.S. 2018. Mycoplasma agalactiae em rebanhos leiteiros no estado do Ceará em associação com o vírus da artrite encefalite caprina. Acta Scientiae Veterinariae. 46: 1533.

Peterson K., Brinkhof J., Houwers D.J., Colenbrander B. & Gadella B.M. 2008. Presence of pro-lentiviral DNA in male sexual organs and ejaculates of small ruminants. Theriogenology. 69: 433-442.

Pinheiro R.R., Olortegui C.D.C., Gouveia A.M.G., Araújo S.C. & Andrioli A. 2006. Desenvolvimento de dot-blot para detecção de anticorpos para o vírus da Artrite Encefalite Carina em Caprinos. Revista Portuguesa de Ciências Veterinárias. 101: 557-558.

Pinheiro R.R., Andrioli A., Martinez P.M., Gouveia A.M.G., Aragão M.A.C. & Martinez P.M. 2010. Avaliação de antígenos para o diagnóstico de lentivírus em rebanho caprino sob programa de controle. Arquivos do Instituto Biológico. 77(1): 133-137.

Pinheiro R.R., Andrioli A., Sider L.H., Santiago L.B., Oliveira E.L., Sousa A.L.M., Alves F.S.F. & Cruz J.C.M. 2012. Lentiviroses em Pequenos Ruminantes: Principais Métodos de Diagnóstico (Documentos 107). Sobral: Embrapa Caprinos e Ovinos, 42p.

Plummer P.J. & Still K.M. 2012. Diseases of the Respiratory System. In: Pugh D.N. & Baird A.N. (Eds). Sheep and Goat Medicine. 2nd edn. St. Louis: Saunders, 640p.

Preziuso S., Sanna E., Sanna M.P., Loddo C., Cerri D., Taccini E., Mariotti F., Braca G., Rossi G. & Renzoni G. 2003. Association of maedi visna virus with Brucella ovis infection in rams. European Journal Histochemistry. 47: 151-158.

Ramírez H., Reina R., Bertolotti L., Cenoz A., Hernández M.M., San Román B. & De Andrés D. 2012. Study of compartmentalization in the visna clinical form of small ruminant lentivirus infection in sheep. BMC Veterinary Research. 8: 1-12.

Ravazzolo A.P., Nenci C., Vogt H.R., Waldvogel A., Obexer-Ruff G., Peterhan E. & Bertoni G. 2006. Viral load, organ distribution, histopathological lesions, and cytokine mRNA expression in goats infected with a molecular clone of the caprine arthritis encephalitis virus. Virology. 350: 116-127.

Reina R., Berriatua E., Lujhn L., Juste R., Shnchez A., de Andre´s D. & Amorena B. 2009. Prevention strategies against small ruminant lentiviruses: an update. The Veterinary Journal. 182: 31-37.

Rodrigues A.S., Brito R.L.L., Pinheiro R.R., Dias R.P., Alves S.M., Souza T.S., Souza K.C., Azevedo D.A.A., Andrioli A., Magalhães D.C.T. & Teixeira M.F.S. 2014. Padronização do Elisa indireto e Western Blot para diagnóstico da artrite-encefalite caprina. Arquivo Brasileiro de Medicina Veterinária e Zootecnia. 66(2) 417-424.

Saltarelli M., Querat G., Konings D.A., Vigne R. & Clements J.E. 1990. Nucleotide sequence and transcriptional analysis of molecular clones of CAEV which generate infectious virus. Virology. 179(1): 347-364.

Souza K.C. 2010. Artrite-encefalite caprina: Infecção experimental via inseminação artificial e acompanhamento clínico e sorológico. 100f. Sobral, CE. Dissertação (Mestrado em Zootecnia) - Programa de Pós-graduação em Zootecnia, Universidade Estadual Vale do Acaraú.

Tariba B., Kostelić A., Šalamon D., Roić B., Benić M., Babić N.P. & Salajpal K. 2017. Subclinical mastitis and clinical arthritis in French Alpine goats serologically positive for caprine artritis-encephalitis virus. Veterinarski Arhiv. 87: 121-128.

Tigre D.M., Campos G.S. & Sardi S.I. 2006. Isolamento e identificação do Vírus da Artrite Encefalite Caprina, a partir do co-cultivo de células mononucleares do sangue com células de membrana sinovial de cabra. Revista de Ciências Médicas e Biológicas. 5: 124-131.

Tyler J.W. & Cullor J.S. 1989. Titers, tests, and truisms: rational interpretation of diagnostic serologic testing. Journal of the American Veterinary Medical Association. 194(11): 1550-1558.

Valas S., Benoit C., Baudry C., Perrin G. & Mamoun R.Z. 2000. Variability and immunogenicity of caprine arthritis-encephalitis virus surface glycoprotein. Journal of Virology. 74(13): 6178-6185.

Published

2019-01-01

How to Cite

Aragão de Azevedo, D. A., Pinheiro, R. R., Souza de Santos, V. W., Damasceno, E. M., Madeira de Sousa, A. L., Araújo, J. F., Andrioli, A., Sider, L. H., Peixoto, R. M., & da Silva Teixeira, M. F. (2019). Comparison of Serological and Molecular Tests for Diagnosis of Caprine Arthritis Encephalitis and Clinical Evaluation of Mammary Glands of Infected Dairy Goats. Acta Scientiae Veterinariae, 47(1). https://doi.org/10.22456/1679-9216.92281

Issue

Section

Articles

Most read articles by the same author(s)