Mycobacterium avium subsp. paratuberculosis (MAP) in Goat Milk in the Semiarid Region of the Brazilian Northeast - Molecular Characterization

Authors

DOI:

https://doi.org/10.22456/1679-9216.125071

Abstract

Background: Goat farming has been on the rise in Brazil in recent years. Overall, 93% of the national herd is concentrated in the Northeast, with the state of Paraíba being the largest goat milk producer in the country. Considering Mycobacterium avium subsp. paratuberculosis (MAP) as a sanitary issue for the development of animal farming with risks for human health and that is a notifiable disease, this research was structured with the objective of confirming the presence and performing a molecular characterization of MAP in goat milk destined for processing plants in the semiarid region of the Brazilian Northeast.

Materials, Methods & Results: Samples from 179 production units and 5 collective bulk tanks and 4 samples of pasteurized goat milk were analyzed through Real-time polymerase chain reaction (qPCR). Genetic material (DNA) for MAP was found in the goat milk sample from 1 production unit (1/179). From this positive sample, 9 lactating goats were identified in the original property, 7 of which showed MAP DNA in milk samples (77.77%). The characterization of the nucleotide sequence detected in the positive sample has 99% identity with KJ173784.

Discussion: One sample (1/179), from the production units, had MAP genetic material (DNA) detected using the molecular test. Samples from these production units represent the milk from all lactating goats from each producer. Therefore, it was possible to identify from which farm the samples originated, allowing individual animals to then be tested, with milk samples collected from 9 goats and MAP DNA detected in 7 of them (77.77%) via PCR. Control and/or prevention programs need this type of surveillance in reason that it allows the tracking of possible foci from milk samples collected from dairy products or cooling stations. The use of PCR to detect MAP foci via goat milk is thus advantageous because samples are obtained in a non-invasive manner, with faster results when compared to the culture technique. The low detection via PCR in goat milk may be related to factors such as the small amount of MAP eliminated and the intermittent excretion in asymptomatic animals, as also false-positive samples. Samples from the collective bulk tanks was negative. It is possible that the combination of milk from all the properties diluted the amount of MAP. This suggests that the sensitivity of the PCR can be improved if the samples are obtained from the pooled milk from the same property. In some regions of Brazil, for example, showed the frequency of Zona da Mata region of the state of Minas Gerais, Brazil, found 1.94% of positive samples (9/464) and 9.76% (4/41) of properties with at least 1 positive sample for MAP. Different results to what were found in the semiarid region of Paraíba, where climate and production characteristics are different. Goats are susceptible to 3 strains: type “S” (Sheep), “Bison type” and type “C” (Cattle). Previous contact with this species may explain the similarity between the strain found in goat milk and those detected from bovine samples. This must also be taken into consideration during diagnosis and upon implementation of control measures for paratuberculosis in goats. Mycobacterium avium subsp. paratuberculosis was recorded for the first time in goat milk in the semiarid region, which may reveal a potential biological risk to humans and suggests the need for active surveillance of the agent.

Keywords: paratuberculosis, Johne’s disease, caprine, milk, PCR.

Downloads

Download data is not yet available.

Author Biographies

João Paulo de Lacerda Roberto, UFCG

He holds a degree in Veterinary Medicine from the Federal University of Campina Grande (2013), a Master's degree in Animal Science from the UFCG (2015), a PhD in Veterinary Medicine from the UFCG and Substitute Professor of Animal Ecology and Animal Evaluation at the Federal University of Campina Grande. He was a member of the Collegiate of the Postgraduate Program in Animal Science and is currently a member of the Collegiate of the Postgraduate Program in Animal Science and Health. He has experience in the areas of Preventive Veterinary Medicine, Inspection and Technology of Animal Products, Microbiological Analysis of Food of Animal Origin and Water, Sanitary Surveillance, Quality Control, Training of Food Handlers and Public Health.

Clécio Henrique Limeira, UFCG

He holds a degree in Veterinary Medicine from the Federal University of Campina Grande (2009), a specialization in Animal Health Defense from the Federal University of Lavras (2012) and a Master's degree in Veterinary Medicine from the Federal University of Campina Grande - UFCG (2015). PhD in Animal Science and Health - UFCG (2021). He has experience in the field of Veterinary Medicine, with an emphasis on Preventive Veterinary Medicine, Parasitic Diseases, Animal Health Defense, Inspection and Technology of Products of Animal Origin and Animal Production.

Rafael Rodrigues Soares, UFCG

He holds a degree in Veterinary Medicine from the State University of Maranhão (2012), a master's degree in Animal Science from the State University of Maranhão (2015) and a doctoral degree from the Federal University of Campina Grande. He has experience in Veterinary Medicine, with emphasis on Preventive Medicine, working mainly on the following subjects: applied bacteriology, Baixada Maranhão, domestic ruminants, Maranhão, semiarid and Leptospira sp. Member of the Research Group on Communicable Diseases at UFCG, which works in the line of research: Epidemiology and Control of Leptospirosis in the Semi-Arid Region of Northeast Brazil.

João Pessoa Araújo Júnior, Unesp

Graduated in Veterinary Medicine from the Faculty of Agrarian and Veterinary Sciences of Jaboticabal - UNESP (1992), Master's in Biological Sciences (Microbiology) from the University of São Paulo (1994), PhD in Biological Sciences (Microbiology) from the University of São Paulo (1998) ) and Associate Professor in Microbiology at Universidade Estadual Paulista (2006). He is currently an adjunct professor at the Department of Microbiology and Immunology, Institute of Biosciences, Universidade Estadual Paulista Júlio de Mesquita Filho. He is also responsible for the virology and molecular diagnostics laboratory. He has experience in the field of Animal Virology, with an emphasis on diagnostic techniques for Infectious Diseases of Animals.

Camila Dantas Malossi, Unesp

Graduated in Biological Sciences - Bachelor and Licentiate - from the University of São Paulo. She undertook a Scientific Initiation at the Department of Parasitology of the Institute of Biomedical Sciences at the University of São Paulo. Master of Science with emphasis on Biology of the Host-Pathogen Relationship at the Institute of Biomedical Sciences of the University of São Paulo. PhD in Biotechnology from the Botucatu Institute of Biosciences of the São Paulo State University "Júlio de Mesquita Filho". She currently works for FUNDIBIO (Foundation of the Biosciences Institute) as a senior technician at the Molecular Diagnostics Laboratory, based at IBTec, Unesp-Botucatu. She is manager of the Veterinary Molecular Diagnostics Laboratory-LDMVET. She has expertise in parasitology, virology and microbiology with an emphasis on Molecular Biology and Diagnostics.

Severino Silvano dos Santos Higino, UFCG

Graduated (2007), Masters (2010), Doctorate (2012) and Post-Doctorate (2015) in Veterinary Medicine from the Federal University of Campina Grande (UFCG). He is currently an Adjunct Professor (Class C, Level II) at the Center for Rural Health and Technology at the Federal University of Campina Grande, a permanent member of the Postgraduate Program in Animal Health and Science (PPGSCA) and a collaborator in the Postgraduate Program in Animal Science (PPGCA) of the Federal University of Campina Grande in the area of animal health. He has experience in Preventive Veterinary Medicine with emphasis on Infectious Diseases of Animals, working mainly in the epidemiology of animal Leptospirosis. Member of the research group on communicable diseases at the Federal University of Campina Grande working in the line of research: Epidemiology and control of Leptospirosis in the semi-arid region of Northeast Brazil.

Sérgio Santos Azevedo, UFCG

Graduated in Veterinary Medicine from the Federal University of Paraíba, PB, in 2000, Master's in Experimental Epidemiology Applied to Zoonoses from the University of São Paulo, SP, in 2002, and PhD in Experimental Epidemiology Applied to Zoonoses from the University of São Paulo, SP, in 2006. Holds a CNPq Research Productivity Scholarship, level 1A, and is an Associate Professor (level 4) at the Academic Unit of Veterinary Medicine (UAMV) at the Rural Health and Technology Center (CSTR) at the Federal University of Campina Grande (UFCG) . Coordinator of the Preventive Veterinary Medicine area at UAMV/CSTR/UFCG in the period 2018/2019. He is currently Administrative Coordinator at UAMV / CSTR / UFCG. Member of the Board of Directors of the Iberoamerican Society of Veterinary Epidemiology and Preventive Medicine (SIEVMP). Coordinator of the Postgraduate Course in Veterinary Medicine at UFCG, in the 2013-2016 quadrennium. He serves as a consultant for several national and international journals in the areas of Animal Epidemiology and Infectious and Parasitic Diseases. He is an ad hoc consultant at CNPq, CAPES, FAPEAM, FACEPE, ANII (Uruguay) and CSIC (Uruguay). He has experience in Veterinary Medicine, with emphasis on Preventive Veterinary Medicine, working mainly on the following topics: animal epidemiology, epidemiology of infectious and parasitic diseases, animal leptospirosis, zoonoses, epidemiological studies.

Clebert José Alves, UFCG

He holds a degree in Veterinary Medicine from the Federal University of Paraíba (1985), a master's degree in Experimental Epidemiology Applied to Zoonoses from the University of São Paulo (1991) and a PhD in Experimental Epidemiology Applied to Zoonoses from the University of São Paulo (1995). He is currently a Full Professor at the Federal University of Campina Grande and leads the Communicable Diseases Research Group and integrates a research group with researchers from the University of São Paulo / Faculty of Veterinary Medicine and Animal Science. He has experience in Veterinary Medicine, with emphasis on Animal Epidemiology, working mainly on the following subjects: leptospira, isolation, sheep, agglutinins and goats.

References

Alburquerque P.P.F., Santos A.S., Souza Neto O.L., Kim P.C.P., Cavalcanti E.F.T.S.F., Oliveira J.M.B., Mota R.A. & Pinheiro Jr. J.W. 2017. Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil. Brazilian Journal of Microbiology. 48: 113-117. DOI: 10.1016/j.bjm.2016.10.010. DOI: https://doi.org/10.1016/j.bjm.2016.10.010

Bauman C.A., Jones-Bitton A., Jansen J., Kelton D. & Menzies P. 2019. Evaluation of bulk tank milk PCR and bulk tank milk modified ELISA tests for the detection of paratuberculosis at the herd level in goat and sheep dairies in Ontario, Canada. Journal of Dairy Science. 102: 511-520. DOI: 10.3168/jds.2018-15020. DOI: https://doi.org/10.3168/jds.2018-15020

Dimareli-Malli Z., Mazaraki K., Stevenson K., Tsakos P., Zdragas A., Giantzi V., Petridou E., Heron I. & Vafeas G. 2013. Culture phenotypes and molecular characterization of Mycobacterium avium subsp. paratuberculosis isolates from small ruminants. Research in Veterinary Science. 95: 49-53. DOI: https://doi.org/10.1016/j.rvsc.2013.03.010

Favila-Humara L.C., Chávez-Gris G.G., Carrillo-Casas E.M. & Hernández-Castro R. 2010. Mycobacterium avium subsp. paratuberculosis detection in individual and bulk tank milk samples from bovine herds and caprine flocks. Foodborne Pathogens and Disease. 7: 351-355. DOI: 10.1089/fpd.2009.0374. DOI: https://doi.org/10.1089/fpd.2009.0374

Freitas T.D., Azevedo S.S., Silva M.L.C.R., Garino Jr. F., Santos C.S.A.B., Clementino I.J., Amaral F.R. & Alves C.J. 2015. Epidemiological characterization and risk factors associated with Mycobacterium avium subsp. paratuberculosis infection in dairy goats in the Brasilian semiarid region. Semina: Ciências Agrárias. 36: 267-276. DOI: 10.5433/1679-0359.2015v36n1p267. DOI: https://doi.org/10.5433/1679-0359.2015v36n1p267

Galiero A., Turchi B., Pedonese F., Nuvoloni R., Cantile C., Colombani G., Forzan M., Cerri D., Bandecchi P. & Fratini F. 2017. Serological, culture and molecular survey of Mycobacterium avium subsp. paratuberculosis in a goat flock in Tuscany. Folia Microbiologica. 62: 471-477. DOI: 10.1007/s12223-017-0518-7. DOI: https://doi.org/10.1007/s12223-017-0518-7

Gouy M., Guindon S. & Gascuel O. 2010. SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building, Molecular Biology and Evolution. 27(2): 221-224. DOI: 10.1093/molbev/msp259. DOI: https://doi.org/10.1093/molbev/msp259

Governo do Estado da Paraíba. 2015. Secretaria de Desenvolvimento Humano. Agricultores familiares com pertinência ao programa leite da paraíba em setembro de 2015, 25p. Disponível em: https://paraiba.pb.gov.br/diretas/secretaria-de-desenvolvimento-humano/fornecedores-paa-leite/>.

Haenlein G.F.W. 2004. Goat milk in human nutrition. Small Ruminant Research. 51: 155-163. DOI: https://doi.org/10.1016/j.smallrumres.2003.08.010

Instituto Brasileiro de Geografia e Estatística (IBGE). 2019. Censo agropecuário: resultados definitivos 2017. Disponível em: <https://censoagro2017.ibge.gov.br/templates/censo_agro/resultadosagro/pecuaria.html>.

Kumar S., Singh S.V., Sevilla I., Singh A.V., Whittington R.J., Juste R.A., Sharma G., Singh P.K. & Sohal J.S. 2008. Lacto-prevalence, genotyping of Mycobacterium avium subspecies paratuberculosis and evaluation of three diagnostic tests in milk of naturally infected goat herds. Small Ruminant Research. 74: 37-44. DOI: 10.1016/j.smallrumres.2007.03.005. DOI: https://doi.org/10.1016/j.smallrumres.2007.03.005

Lombard J.E. 2011. Epidemiology and economics of paratuberculosis. Veterinary Clinics of North America: Food Animal Practice. 27: 525-535. DOI: https://doi.org/10.1016/j.cvfa.2011.07.012

Medeiros J.M.A., Garino Jr. F., Almeida A.P., Lucena E.A. & Riet-Correa F. 2012. Paratuberculose em caprinos e ovinos no Estado da Paraíba. Pesquisa Veterinária Brasileira. 32: 111-115. DOI: 10.1590/S0100-736X2012000200003. DOI: https://doi.org/10.1590/S0100-736X2012000200003

Nebbia P., Robino P., Zoppi S. & Meneghi D. 2006. Detection and excretion pattern of Mycobacterium avium subspecies paratuberculosis in milk of asymptomatic sheep and goats by Nested-PCR. Small Ruminant Research. 66: 116-120. DOI: 10.1016/j.smallrumres.2005.07.049. DOI: https://doi.org/10.1016/j.smallrumres.2005.07.049

Patel A. & Shah N. 2011. Mycobacterium avium subsp. paratuberculosis - incidences in milk and milk products, their isolation, enumeration, characterization, and role in human health. Journal of Microbiology, Immunology and Infection. 44: 473-479. DOI: https://doi.org/10.1016/j.jmii.2011.04.009

Platt A.R., Woodhall R.W. & George Jr. A.L. 2007. Improved DNA sequencing quality and efficiency using an optimized fast cycle sequencing protocol. Biotechniques. 43(1): 58-62. DOI: 10.2144/000112499. DOI: https://doi.org/10.2144/000112499

Rathnaiah G., Zinniel D.K., Bannantine J.P., Stabel J.P., Gröhn Y.T., Collins M. & Barletta R.G. 2017. Pathogenesis, molecular genetics, and genomics of Mycobacterium avium subsp. paratuberculosis, the etiologic agent of Johne’s disease. Frontiers in Veterinary Science. 4: 1-13. DOI: 10.3389/fvets.2017.00187. DOI: https://doi.org/10.3389/fvets.2017.00187

Roberto J.P.L., Limeira C.L., Barnabé N.N.C., Soares R.R., Silva M.L.C.R., Gomes A.A.B., Higino S.S.S., Azevedo S.S. & Alves C.J. 2021. Antibody detection and molecular analysis for Mycobacterium avium subspecies paratuberculosis (MAP) in goat milk: Systematic review and meta-analysis. Research in Veterinary Science. 135: 72-77. DOI: 10.1016/j.rvsc.2021.01.004. DOI: https://doi.org/10.1016/j.rvsc.2021.01.004

Schwarz D.G.G., Lima M.C., Barros M., Valente F.L., Scatamburlo T.M., Rosado N. & Moreira M.A.S. 2017. Short communication: Passive shedding of Mycobacterium avium ssp. paratuberculosis in commercial dairy goats in Brazil. Journal of Dairy Science. 100: 8426-8429. DOI: 10.3168/jds.2017-12918. DOI: https://doi.org/10.3168/jds.2017-12918

Singh M., Gupta S., Chaubey K., Singh S. & Sohal J. 2019. Profiling of Mycobacterium avium subspecies paratuberculosis in the milk of lactating goats using antigen-antibody based assays. Comparative Immunology, Microbiology and Infectious Diseases. 64: 53-60. DOI: 10.1016/j.cimid.2019.02.006. DOI: https://doi.org/10.1016/j.cimid.2019.02.006

Souza M.C.C., Lima M.C., Braga I.F.E., Schwarz D.G.G., Rodrigues A.N.S., Sales E.B., Fonseca Jr. A.A. & Moreira M.A.S. 2016. Molecular typing of Mycobacterium avium subsp. paratuberculosis (MAP) isolated from dairy goats in Brazil. Small Ruminant Research. 140: 18-21. DOI: 10.1016/j.smallrumres.2016.05.009. DOI: https://doi.org/10.1016/j.smallrumres.2016.05.009

Stevenson K. 2015. Genetic diversity of Mycobacterium avium subspecies paratuberculosis and the influence of strain type on infection and pathogenesis: a review. Veterinary Research. 46: 1-13. Doi: 10.1186 /s13567-015-0203-2. DOI: https://doi.org/10.1186/s13567-015-0203-2

Sukumar B., Gunaseelan L., Porteen K. & Prabu K. 2014. Goat milk as a non-invasive sample for confirmation of Mycobacterium avium subspecies paratuberculosis by IS900 PCR. Journal of Advanced Veterinary and Animal Research. 1: 136-139. DOI: 10.5455/javar.2014.a25. DOI: https://doi.org/10.5455/javar.2014.a25

Thrusfield M. & Brown H. 2018. Surveys. In: Thrusfield M. (Ed). Veterinary Epidemiology. 4th edn. Oxford: Blackwell Science, pp.270-295.

Windsor P.A. 2015. Paratuberculosis in sheep and goats. Veterinary Microbiology. 181: 161-169. DOI: 10.1016/j.vetmic.2015.07.019. DOI: https://doi.org/10.1016/j.vetmic.2015.07.019

World Organization for Animal Health (OIE). 2018. Leptospirosis: Manual of diagnostic tests and vaccines for terrestrial animals. Paris: World Organization for Animal Health.

Additional Files

Published

2022-10-24

How to Cite

Roberto, J. P. de L., Limeira, C. H., Rodrigues Soares, R., Araújo Júnior, J. P., Malossi, C. D., Ullmann, L. S., … Alves, C. J. (2022). Mycobacterium avium subsp. paratuberculosis (MAP) in Goat Milk in the Semiarid Region of the Brazilian Northeast - Molecular Characterization. Acta Scientiae Veterinariae, 50. https://doi.org/10.22456/1679-9216.125071

Issue

Vol. 50 (2022): ARTICLES

Section

Articles

License

Copyright (c) 2022 João Paulo de Lacerda Roberto, Clécio Henrique Limeira, Rafael Rodrigues Soares, João Pessoa Araújo Júnior, Camila Dantas Malossi, Leila Sabrina Ullmann, Maria Luana Cristiny Rodrigues Silva, Severino Silvano dos Santos Higino, Sérgio Santos Azevedo, Clebert José Alves

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

This journal provides open access to all of its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. Such access is associated with increased readership and increased citation of an author's work. For more information on this approach, see the Public Knowledge Project and Directory of Open Access Journals.

We define open access journals as journals that use a funding model that does not charge readers or their institutions for access. From the BOAI definition of "open access" we take the right of users to "read, download, copy, distribute, print, search, or link to the full texts of these articles" as mandatory for a journal to be included in the directory.

La Red y Portal Iberoamericano de Revistas Científicas de Veterinaria de Libre Acceso reúne a las principales publicaciones científicas editadas en España, Portugal, Latino América y otros países del ámbito latino