Immunopathogenesis and Diagnostics to Control Tuberculosis in Cattle and Wildlife

A special issue of Pathogens (ISSN 2076-0817). This special issue belongs to the section "Immunological Responses and Immune Defense Mechanisms".

Deadline for manuscript submissions: closed (1 June 2022) | Viewed by 30022

Special Issue Editors


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Guest Editor
Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, IA, USA
Interests: pathology; mycobacteria; bovine tuberculosis; vaccines; zoonotic diseases

E-Mail Website
Guest Editor
Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, IA, USA
Interests: Intracellular pathogens; veterinary immunology; cell-mediated immune responses; zoonoses; brucellosis; bovine tuberculosis; vaccines; diagnostics

E-Mail Website
Guest Editor
Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, IA, USA
Interests: One Health; zoonoses; mycobacterial diseases; transboundary diseases; wildlife diseases

Special Issue Information

Dear Colleagues,

Mycobacterium bovis has the broadest host range of any member of the Mycobacterium tuberculosis complex. It is the cause of tuberculosis in most mammals, including humans, in whom it can cause disease which is clinically indistinguishable from that caused by M. tuberculosis, the more common cause of human tuberculosis. Due to both human and animal health concerns, many countries initiated bovine tuberculosis eradication programs in the early to mid-20th century. In spite of long-standing and expensive programs, eradication remains elusive, in part due to wildlife reservoirs of M. bovis. Although tuberculosis likely spilled over into wildlife from cattle, in many locations, it is now spilling back into cattle from wildlife. Generally accepted wildlife reservoirs are the European badger (Meles meles) in the UK and Republic of Ireland, the brushtail possum (Trichosurus vulpecula) in New Zealand, white-tailed deer (Odocoileus virginianus) in the US, wild boar (Sus scrofa) in the Iberian Peninsula, and Cape buffalo (Syncerus caffer) in Africa. Numerous other wildlife species are susceptible, including some endangered species such as the Iberian lynx (Lynx pardina) and African wild dog (Lycaon pictus).

Notwithstanding advances in the understanding bovine tuberculosis, the century-old tuberculin skin test, with some modifications, remains the preferred method of diagnosis in many countries. Blood-based assays, such as interferon-gamma release assays, complement skin testing but are not rapid, nor inexpensive, and lack the sensitivity and specificity necessary for a stand-alone test. Much remains unknown about the process of disease development in cattle and wildlife, especially the immune response and how this response can be used to develop improved diagnostic assays.

Key questions remain around the definition of a successful, protective immune response, as well as correlates of protection. Critical interactions between host and pathogen occur at the level of the granuloma, the lesion which exemplifies tuberculosis. Can responses measured in blood tell us what is happening at the host–pathogen interface of the granuloma? How can we examine what is happening at this interface?

This Special Issue of Pathogens is focused on the immunopathogenesis and diagnosis of M. bovis infection in cattle, the eponymous host species, and the many susceptible wildlife species, especially those that may serve as maintenance hosts and reservoirs of disease. We invite you to submit primary research articles and review articles representing recent advances in our knowledge of tuberculosis immunopathogenesis and diagnosis in the numerous affected host species.

Dr. Mitchell V. Palmer
Dr. Paola M. Boggiatto
Dr. Carly Kanipe
Guest Editors

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Published Papers (7 papers)

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Research

Jump to: Review

15 pages, 2496 KiB  
Article
Mycobacterium bovis Strain Ravenel Is Attenuated in Cattle
by Syeda A. Hadi, Evan P. Brenner, Mitchell V. Palmer, W. Ray Waters, Tyler C. Thacker, Catherine Vilchèze, Michelle H. Larsen, William R. Jacobs, Jr. and Srinand Sreevatsan
Pathogens 2022, 11(11), 1330; https://doi.org/10.3390/pathogens11111330 - 11 Nov 2022
Cited by 1 | Viewed by 2115
Abstract
Mycobacterium tuberculosis variant bovis (MBO) has one of the widest known mammalian host ranges, including humans. Despite the characterization of this pathogen in the 1800s and whole genome sequencing of a UK strain (AF2122) nearly two decades ago, the basis of its host [...] Read more.
Mycobacterium tuberculosis variant bovis (MBO) has one of the widest known mammalian host ranges, including humans. Despite the characterization of this pathogen in the 1800s and whole genome sequencing of a UK strain (AF2122) nearly two decades ago, the basis of its host specificity and pathogenicity remains poorly understood. Recent experimental calf infection studies show that MBO strain Ravenel (MBO Ravenel) is attenuated in the cattle host compared to other pathogenic strains of MBO. In the present study, experimental infections were performed to define attenuation. Whole genome sequencing was completed to identify regions of differences (RD) and single nucleotide polymorphisms (SNPs) to explain the observed attenuation. Comparative genomic analysis of MBO Ravenel against three pathogenic strains of MBO (strains AF2122-97, 10-7428, and 95-1315) was performed. Experimental infection studies on five calves each, with either MBO Ravenel or 95-1315, revealed no visible lesions in all five animals in the Ravenel group despite robust IFN-γ responses. Out of 486 polymorphisms in the present analysis, 173 were unique to MBO Ravenel among the strains compared. A high-confidence subset of nine unique SNPs were missense mutations in genes with annotated functions impacting two major MBO survival and virulence pathways: (1) Cell wall synthesis & transport [espH (A103T), mmpL8 (V888I), aftB (H484Y), eccC5 (T507M), rpfB (E263G)], and (2) Lipid metabolism & respiration [mycP1(T125I), pks5 (G455S), fadD29 (N231S), fadE29 (V360G)]. These substitutions likely contribute to the observed attenuation. Results from experimental calf infections and the functional attributions of polymorphic loci on the genome of MBO Ravenel provide new insights into the strain’s genotype-disease phenotype associations. Full article
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16 pages, 2979 KiB  
Article
Protein Levels of Pro-Inflammatory Cytokines and Chemokines as Biomarkers of Mycobacterium bovis Infection and BCG Vaccination in Cattle
by Hamza Khalid, Anouk van Hooij, Timothy K. Connelley, Annemieke Geluk and Jayne C. Hope
Pathogens 2022, 11(7), 738; https://doi.org/10.3390/pathogens11070738 - 29 Jun 2022
Cited by 2 | Viewed by 2832
Abstract
Bovine tuberculosis (bTB), caused by Mycobacterium bovis, is a globally prevalent infectious disease with significant animal welfare and economic impact. Difficulties in implementing test-and-slaughter measures in low- and middle-income countries (LMICs) and the underperformance of the current diagnostics establish a clear need [...] Read more.
Bovine tuberculosis (bTB), caused by Mycobacterium bovis, is a globally prevalent infectious disease with significant animal welfare and economic impact. Difficulties in implementing test-and-slaughter measures in low- and middle-income countries (LMICs) and the underperformance of the current diagnostics establish a clear need to develop improved diagnostics. Adaptive immunity biomarkers other than IFNγ could be useful as suggested by various gene expression studies; however, a comprehensive assessment at the protein level is lacking. Here, we screened a range of chemokines and cytokines for their potential as biomarkers in samples from M. bovis experimentally challenged or naive animals. Although serum concentrations for most proteins were low, the pro-inflammatory markers, IL-2, CXCL-9, IP-10 and CCL4, in addition to IFNγ, were found to be significantly elevated in bovine tuberculin (PPDb)-stimulated whole blood supernatants. Further assessment of these molecules in BCG-vaccinated with or without subsequent M. bovis challenge or naive animals revealed that PPDb-specific IL-2 and IP-10, in addition to IFNγ, could discriminate naive and BCG-vaccinated from M. bovis challenged animals. Moreover, these proteins, along with CCL4, showed DIVA potential, i.e., enabling differentiation of M. bovis-infected animals from BCG-vaccinated animals. Combined analysis of cytokines and chemokines could also accurately identify M. bovis infection with strong correlations observed between PPDb-specific IFNγ, IL-2 and IP-10 levels. This provides proof of concept for utilizing multiple biomarker signatures for discrimination of animals with respect to M. bovis infection or BCG vaccination status. Full article
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16 pages, 488 KiB  
Article
Culture-Independent PCR Detection and Differentiation of Mycobacteria spp. in Antemortem Respiratory Samples from African Elephants (Loxodonta Africana) and Rhinoceros (Ceratotherium Simum, Diceros Bicornis) in South Africa
by Wynand J. Goosen, Charlene Clarke, Léanie Kleynhans, Tanya J. Kerr, Peter Buss and Michele A. Miller
Pathogens 2022, 11(6), 709; https://doi.org/10.3390/pathogens11060709 - 20 Jun 2022
Cited by 5 | Viewed by 2684
Abstract
Since certain Mycobacterium tuberculosis complex (MTBC) members, such as M. bovis, are endemic in specific South African wildlife reserves and zoos, cases of clinically important nontuberculous mycobacteria (NTM) in wildlife may be neglected. Additionally, due to the inability of tests to differentiate [...] Read more.
Since certain Mycobacterium tuberculosis complex (MTBC) members, such as M. bovis, are endemic in specific South African wildlife reserves and zoos, cases of clinically important nontuberculous mycobacteria (NTM) in wildlife may be neglected. Additionally, due to the inability of tests to differentiate between the host responses to MTBC and NTM, the diagnosis of MTBC may be confounded by the presence of NTMs. This may hinder control efforts. These constraints highlight the need for enhanced rapid detection and differentiation methods for MTBC and NTM, especially in high MTBC burden areas. We evaluated the use of the GeneXpert MTB/RIF Ultra, the Hain CMdirect V1.0 line probe assay, and novel amplicon sequencing PCRs targeting the mycobacterial rpoB and ku gene targets, directly on antemortem African elephant (n = 26) bronchoalveolar lavage fluid (BALF) (n = 22) and trunk washes (n = 21) and rhinoceros (n = 23) BALF (n = 23), with known MTBC culture-positive and NTM culture-positive results. Our findings suggest that the Ultra is the most sensitive diagnostic test for MTBC DNA detection directly in raw antemortem respiratory specimens and that the rpoB PCR is ideal for Mycobacterium genus DNA detection and species identification through amplicon sequencing. Full article
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9 pages, 438 KiB  
Communication
The Risk of False-Positive Serological Results for Paratuberculosis in Mycobacterium bovis-Infected Cattle
by Anna Didkowska, Monika Krajewska-Wędzina, Daniel Klich, Kinga Prolejko, Blanka Orłowska and Krzysztof Anusz
Pathogens 2021, 10(8), 1054; https://doi.org/10.3390/pathogens10081054 - 19 Aug 2021
Cited by 6 | Viewed by 3481
Abstract
Both bovine tuberculosis (BTB) and paratuberculosis (paraTB) continue to cause significant economic losses in cattle breeding; in addition, their etiological agents have zoonotic potential. Although the diagnostics of both diseases are still being improved, problems still remain, such as the potential for cross-reactivity [...] Read more.
Both bovine tuberculosis (BTB) and paratuberculosis (paraTB) continue to cause significant economic losses in cattle breeding; in addition, their etiological agents have zoonotic potential. Although the diagnostics of both diseases are still being improved, problems still remain, such as the potential for cross-reactivity to the antigens used in tests. The aim of the present study was to confirm whether animals known to harbor Mycobacterium bovis antibodies are at increased risk of yielding positive results in paraTB serotesting and, additionally, to verify the accuracy of three commonly used methods for confirming M. bovis infection: ELISA, the tuberculin skin test (TST), and the presence of gross lesions. Material was collected from 98 dairy cattle suspected of BTB due to TST-positive results. During postmortem examination, gross lesions were assessed visually. Blood, lymph nodes, and TB-suspected organs were collected. Serum was obtained from the collected blood and tested serologically for TB and paraTB. The tissues underwent standard microbiological testing for M. tuberculosis complex. Among the 98 TST-positive individuals, tuberculous gross lesions were detected in 57 (58.1%), MTBC were isolated in 83 (84.7%), and the ELISA test was positive for 21 (21.4%). None of the lesions characteristic for paraTB were detected. The chance of obtaining a positive TB result by ELISA was seven times higher using the ELISA-paraTB method; hence, there is a significant risk of obtaining false-positive serological results for paraTB in M. bovis-infected cattle. However, the hypothesis that infection of M. bovis or prior TST performance may have boosted the host immune response and therefore increased the sensitivity of the paraTB-ELISA cannot be excluded. Full article
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Review

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25 pages, 455 KiB  
Review
Review on Bovine Tuberculosis: An Emerging Disease Associated with Multidrug-Resistant Mycobacterium Species
by Mohamed Borham, Atef Oreiby, Attia El-Gedawy, Yamen Hegazy, Hazim O. Khalifa, Magdy Al-Gaabary and Tetsuya Matsumoto
Pathogens 2022, 11(7), 715; https://doi.org/10.3390/pathogens11070715 - 21 Jun 2022
Cited by 35 | Viewed by 6822
Abstract
Bovine tuberculosis is a serious infectious disease affecting a wide range of domesticated and wild animals, representing a worldwide economic and public health burden. The disease is caused by Mycobacteriumbovis and infrequently by other pathogenic mycobacteria. The problem of bovine tuberculosis is [...] Read more.
Bovine tuberculosis is a serious infectious disease affecting a wide range of domesticated and wild animals, representing a worldwide economic and public health burden. The disease is caused by Mycobacteriumbovis and infrequently by other pathogenic mycobacteria. The problem of bovine tuberculosis is complicated when the infection is associated with multidrug and extensively drug resistant M. bovis. Many techniques are used for early diagnosis of bovine tuberculosis, either being antemortem or postmortem, each with its diagnostic merits as well as limitations. Antemortem techniques depend either on cellular or on humoral immune responses, while postmortem diagnosis depends on adequate visual inspection, palpation, and subsequent diagnostic procedures such as bacterial isolation, characteristic histopathology, and PCR to reach the final diagnosis. Recently, sequencing and bioinformatics tools have gained increasing importance for the diagnosis of bovine tuberculosis, including, but not limited to typing, detection of mutations, phylogenetic analysis, molecular epidemiology, and interactions occurring within the causative mycobacteria. Consequently, the current review includes consideration of bovine tuberculosis as a disease, conventional and recent diagnostic methods, and the emergence of MDR-Mycobacterium species. Full article
17 pages, 3609 KiB  
Review
The Bovine Tuberculoid Granuloma
by Mitchell V. Palmer, Carly Kanipe and Paola M. Boggiatto
Pathogens 2022, 11(1), 61; https://doi.org/10.3390/pathogens11010061 - 4 Jan 2022
Cited by 13 | Viewed by 7399
Abstract
The bovine tuberculoid granuloma is the hallmark lesion of bovine tuberculosis (bTB) due to Mycobacterium bovis infection. The pathogenesis of bTB, and thereby the process of bovine tuberculoid granuloma development, involves the recruitment, activation, and maintenance of cells under the influence of antigen, [...] Read more.
The bovine tuberculoid granuloma is the hallmark lesion of bovine tuberculosis (bTB) due to Mycobacterium bovis infection. The pathogenesis of bTB, and thereby the process of bovine tuberculoid granuloma development, involves the recruitment, activation, and maintenance of cells under the influence of antigen, cytokines and chemokines in affected lungs and regional lymph nodes. The granuloma is key to successful control of bTB by preventing pathogen dissemination through containment by cellular and fibrotic layers. Paradoxically, however, it may also provide a niche for bacterial replication. The morphologic and cellular characteristics of granulomas have been used to gauge disease severity in bTB pathogenesis and vaccine efficacy studies. As such, it is critical to understand the complex mechanisms behind granuloma initiation, development, and maintenance. Full article
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15 pages, 289 KiB  
Review
Review of Methods Used for Diagnosing Tuberculosis in Captive and Free-Ranging Non-Bovid Species (2012–2020)
by Rebecca Thomas and Mark Chambers
Pathogens 2021, 10(5), 584; https://doi.org/10.3390/pathogens10050584 - 11 May 2021
Cited by 7 | Viewed by 3585
Abstract
The Mycobacterium tuberculosis complex (MTBC) is a group of bacteria that cause tuberculosis (TB) in diverse hosts, including captive and free-ranging wildlife species. There is significant research interest in developing immunodiagnostic tests for TB that are both rapid and reliable, to underpin disease [...] Read more.
The Mycobacterium tuberculosis complex (MTBC) is a group of bacteria that cause tuberculosis (TB) in diverse hosts, including captive and free-ranging wildlife species. There is significant research interest in developing immunodiagnostic tests for TB that are both rapid and reliable, to underpin disease surveillance and control. The aim of this study was to carry out an updated review of diagnostics for TB in non-bovid species with a focus predominantly on those based on measurement of immunity. A search was carried out to identify relevant papers meeting a pre-defined set of inclusion criteria. Forty-one papers were identified from this search, from which only twenty papers contained data to measure and compare diagnostic performance using diagnostic odds ratio. The diagnostic tests from each study were ranked based on sensitivity, specificity, and diagnostic odds ratio to define high performing tests. High sensitivity and specificity values across a range of species were reported for a new antigenic target, P22 complex, demonstrating it to be a reliable and accurate antigenic target. Since the last review of this kind was undertaken, the immunodiagnosis of TB in meerkats and African wild dogs was reported for the first time. Suid species showed the most consistent immunological responses and highlight a potential dichotomy between humoral and cellular immune responses. Full article
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