Mycobacterium bovis Strain Ravenel Is Attenuated in Cattle
Abstract
:1. Introduction
2. Methods
2.1. Mycobacterium Tuberculosis Variant Bovis Challenge Strains
2.2. Cattle Studies: Treatment Groups and Aerosol MBO Challenge Procedures
2.3. IFN-γ Whole Blood Assays
2.4. Delayed Type Hypersensitivity (DTH) Responses (Skin Test Procedures)
2.5. Whole Genome Sequencing of MBO Ravenel
2.6. Genome Comparisons
2.6.1. Identification of Single Nucleotide Polymorphisms (SNPs) in MBO Ravenel
2.6.2. Region of Difference (RD) and Gene Presence/Absence Analysis
3. Results
3.1. Experimental Infection of Cattle
3.2. Whole Genome Sequencing
3.3. Genome Comparisons
SNPs from MBO Ravenel versus Virulent MBO Strains
4. Discussion
4.1. Ravenel Elicits Robust Cell-Mediated Immune Response but Is Attenuated
4.2. Ravenel Carries SNPs in Genes Encoding Cell Wall Integrity
4.3. Ravenel Carries SNPs in Genes Associated with Respiration or Lipid Metabolism
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Treatment Group | Mean Disease Score a | ||
---|---|---|---|
Tracheobronchial Lymph Nodes | Mediastinal Lymph Nodes | Lung | |
Non-infected | 0 ± 0 (0/5) | 0 ± 0 (0/5) | 0 ± 0 (0/5) |
MBO strain Ravenel infected | 0 ± 0 (0/5) | 0 ± 0 (0/5) | 0.04 ± 0.04 (1/5) |
MBO strain 95-1315 infected | 1.4 ± 0.2 * (5/5) | 1.4 ± 0.2 * (5/5) | 1.2 ± 0.3 * (5/5) |
Treatment Group | Quantitative Culture (CFU/g) a | Qualitative Culture b | ||
---|---|---|---|---|
Tracheobronchial Lymph Node | Mediastinal Lymph Node | Lung | ||
Non-infected | 0 ± 0 | 0/5 | 0/5 | 0/5 |
MBO Ravenel infected | 427 ± 1246 | 4/5 | 1/5 | 0/5 |
MBO 95-1315 infected | 28,141 ± 9063 * | 5/5 | 4/5 | 3/5 |
Position | Gene Name | Gene Identifier | Mycobrowser Classification | Functional Annotation | DNA Change (Protein Change) |
---|---|---|---|---|---|
4305161 | mycP1 | MB3913C | Intermediary metabolism and respiration | membrane-anchored mycosin mycp1 (serine protease) (subtilisin-like protease) (subtilase-like) (mycosin-1) | C374T (T125I) |
4283808 | espH | MB3897 | Cell wall and cell processes | esx-1 secretion-associated protein | G307A (A103T) |
4229553 | mmpL8 | MB3853C | Cell wall and cell processes | conserved integral membrane transport protein | G2662A (V888I) |
4208072 | aftB | MB3835C | Cell wall and cell processes | possible arabinofuranosyltransferase | C1450T (H484Y) |
2015643 | eccC5 | MB1812 | Cell wall and cell processes | esx conserved component eccc5. esx-5 type vii secretion system protein | C1520T (T507M) |
1129357 | rpfB | MB1036 | Cell wall and cell processes | Probable resuscitation-promoting factor | A788G (E263G) |
1715469 | pks5 | MB1554C | Lipid metabolism | Probable polyketide synthase | G1363A (G455S) |
3262864 | fadD29 | MB2974C | Lipid metabolism | fatty-acid-amp ligase fadd29 (fatty-acid-amp synthetase) (fatty-acid-amp synthase) | A692G (N231S) |
3929690 | fadE29 | MB3573C | Lipid metabolism | Probable Acyl-CoA dehydrogenase | T1079G (V360G) |
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Hadi, S.A.; Brenner, E.P.; Palmer, M.V.; Waters, W.R.; Thacker, T.C.; Vilchèze, C.; Larsen, M.H.; Jacobs, W.R., Jr.; Sreevatsan, S. Mycobacterium bovis Strain Ravenel Is Attenuated in Cattle. Pathogens 2022, 11, 1330. https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens11111330
Hadi SA, Brenner EP, Palmer MV, Waters WR, Thacker TC, Vilchèze C, Larsen MH, Jacobs WR Jr., Sreevatsan S. Mycobacterium bovis Strain Ravenel Is Attenuated in Cattle. Pathogens. 2022; 11(11):1330. https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens11111330
Chicago/Turabian StyleHadi, Syeda A., Evan P. Brenner, Mitchell V. Palmer, W. Ray Waters, Tyler C. Thacker, Catherine Vilchèze, Michelle H. Larsen, William R. Jacobs, Jr., and Srinand Sreevatsan. 2022. "Mycobacterium bovis Strain Ravenel Is Attenuated in Cattle" Pathogens 11, no. 11: 1330. https://0-doi-org.brum.beds.ac.uk/10.3390/pathogens11111330