Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China

Abstract Background Paratuberculosis (PTB), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is difficult to diagnose in the early stages and poses substantial challenges in prevention, control, treatment, and eradication. A well-defined animal model can help identify disease markers and...

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Main Authors: Meng-Yuan Li, Wei-Kang Meng, Wei Ma, Yu-Lin Ding, Bo Yang, Wei-Hong Zhao, Hasi Bayaer, Alateng Bagen, Rui-Bin Chen, Siqin Tunala, Rong Zhang, Chen-Guang Du, Li Zhao, Yong-Hong Liu
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Language:English
Published: BMC 2025-04-01
Series:BMC Veterinary Research
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Online Access:https://doi.org/10.1186/s12917-025-04765-1
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author Meng-Yuan Li
Wei-Kang Meng
Wei Ma
Yu-Lin Ding
Bo Yang
Wei-Hong Zhao
Hasi Bayaer
Alateng Bagen
Rui-Bin Chen
Siqin Tunala
Rong Zhang
Chen-Guang Du
Li Zhao
Yong-Hong Liu
author_facet Meng-Yuan Li
Wei-Kang Meng
Wei Ma
Yu-Lin Ding
Bo Yang
Wei-Hong Zhao
Hasi Bayaer
Alateng Bagen
Rui-Bin Chen
Siqin Tunala
Rong Zhang
Chen-Guang Du
Li Zhao
Yong-Hong Liu
author_sort Meng-Yuan Li
collection DOAJ
description Abstract Background Paratuberculosis (PTB), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is difficult to diagnose in the early stages and poses substantial challenges in prevention, control, treatment, and eradication. A well-defined animal model can help identify disease markers and serve as a platform for vaccine and drug development. This study used sheep as a ruminant model for experimental MAP infection research. Methods Nine 3-month-old lambs with negative MAP antigen and antibody were divided into three groups (control group A and inoculated groups B and C). The inoculated groups were challenged with sheep-derived type II MAP. After exposure, we recorded clinical signs, assessed fecal shedding, tested blood MAP levels, and performed fecal cultures. We also measured MAP-specific antibodies and monitored IFN-γ and IL-10 responses in vivo. At 255 days after inoculation, we performed autopsy, tissue culture, pathomorphological observation, and bacterial organ burden (BOB) testing. Results All six sheep in groups B and C were infected, regardless of the challenge dose and exhibited emaciation; two had intermittent soft stools. Intermittent MAP shedding in feces was observed from 60 to 255 days after exposure. Typical MAP colonies formed after 4–6 weeks of fecal and tissue culture, and Ziehl–Neelsen staining showed positive results. In the groups challenged with MAP, some blood samples tested positive for MAP and MAP-specific antibodies were detected in some serum samples. IFN-γ response was significantly higher in groups B and C than that in group A from day 60 post-exposure, whereas the IL-10 response was higher than that in group A from day 120 post-exposure. In the infected groups, the ileal lesions were the most severe and were classified as grade 3 PTB granulomatous inflammation (multibacillary lesions). BOB levels varied across different tissues. Conclusions To the best of our knowledge, this is the first experimental MAP challenge study on sheep in China. Polymerase chain reaction detection was more sensitive than MAP culture, whereas enzyme-linked immunosorbent assay was less sensitive for detecting MAP-specific antibodies. IFN-γ and IL-10 responses may serve as targets for monitoring PTB progression. The severity of ileal lesions and acid-fast bacilli grading play crucial roles in the understanding of infection dynamics. Currently, early PTB diagnosis requires a combination of multiple sample types and detection methods.
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spelling doaj-art-cf47e77e5b6144c2a1a8906e65aebfac2025-08-20T01:47:32ZengBMCBMC Veterinary Research1746-61482025-04-0121111510.1186/s12917-025-04765-1Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in ChinaMeng-Yuan Li0Wei-Kang Meng1Wei Ma2Yu-Lin Ding3Bo Yang4Wei-Hong Zhao5Hasi Bayaer6Alateng Bagen7Rui-Bin Chen8Siqin Tunala9Rong Zhang10Chen-Guang Du11Li Zhao12Yong-Hong Liu13College of Veterinary Medicine, Inner Mongolia Agricultural UniversityCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityAnimal Disease Control Center of OrdosCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityOtok Banner Animal Disease Prevention and Control CenterOtok Banner Animal Disease Prevention and Control CenterOtok Banner Animal Disease Prevention and Control CenterOtok Banner Animal Disease Prevention and Control CenterOtok Banner Animal Disease Prevention and Control CenterCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityCollege of Veterinary Medicine, Inner Mongolia Agricultural UniversityAbstract Background Paratuberculosis (PTB), caused by Mycobacterium avium subsp. paratuberculosis (MAP), is difficult to diagnose in the early stages and poses substantial challenges in prevention, control, treatment, and eradication. A well-defined animal model can help identify disease markers and serve as a platform for vaccine and drug development. This study used sheep as a ruminant model for experimental MAP infection research. Methods Nine 3-month-old lambs with negative MAP antigen and antibody were divided into three groups (control group A and inoculated groups B and C). The inoculated groups were challenged with sheep-derived type II MAP. After exposure, we recorded clinical signs, assessed fecal shedding, tested blood MAP levels, and performed fecal cultures. We also measured MAP-specific antibodies and monitored IFN-γ and IL-10 responses in vivo. At 255 days after inoculation, we performed autopsy, tissue culture, pathomorphological observation, and bacterial organ burden (BOB) testing. Results All six sheep in groups B and C were infected, regardless of the challenge dose and exhibited emaciation; two had intermittent soft stools. Intermittent MAP shedding in feces was observed from 60 to 255 days after exposure. Typical MAP colonies formed after 4–6 weeks of fecal and tissue culture, and Ziehl–Neelsen staining showed positive results. In the groups challenged with MAP, some blood samples tested positive for MAP and MAP-specific antibodies were detected in some serum samples. IFN-γ response was significantly higher in groups B and C than that in group A from day 60 post-exposure, whereas the IL-10 response was higher than that in group A from day 120 post-exposure. In the infected groups, the ileal lesions were the most severe and were classified as grade 3 PTB granulomatous inflammation (multibacillary lesions). BOB levels varied across different tissues. Conclusions To the best of our knowledge, this is the first experimental MAP challenge study on sheep in China. Polymerase chain reaction detection was more sensitive than MAP culture, whereas enzyme-linked immunosorbent assay was less sensitive for detecting MAP-specific antibodies. IFN-γ and IL-10 responses may serve as targets for monitoring PTB progression. The severity of ileal lesions and acid-fast bacilli grading play crucial roles in the understanding of infection dynamics. Currently, early PTB diagnosis requires a combination of multiple sample types and detection methods.https://doi.org/10.1186/s12917-025-04765-1ParatuberculosisMycobacterium avium subsp. paratuberculosisSheepInfection model
spellingShingle Meng-Yuan Li
Wei-Kang Meng
Wei Ma
Yu-Lin Ding
Bo Yang
Wei-Hong Zhao
Hasi Bayaer
Alateng Bagen
Rui-Bin Chen
Siqin Tunala
Rong Zhang
Chen-Guang Du
Li Zhao
Yong-Hong Liu
Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China
BMC Veterinary Research
Paratuberculosis
Mycobacterium avium subsp. paratuberculosis
Sheep
Infection model
title Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China
title_full Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China
title_fullStr Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China
title_full_unstemmed Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China
title_short Sheep challenged with sheep-derived type II Mycobacterium avium subsp. paratuberculosis: the first experimental model of paratuberculosis in China
title_sort sheep challenged with sheep derived type ii mycobacterium avium subsp paratuberculosis the first experimental model of paratuberculosis in china
topic Paratuberculosis
Mycobacterium avium subsp. paratuberculosis
Sheep
Infection model
url https://doi.org/10.1186/s12917-025-04765-1
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