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Frontiers in Immunology 2023The induction of an effective immune response is critical for the success of mRNA-based therapeutics. Here, we developed a nanoadjuvant system compromised of Quil-A and...
The induction of an effective immune response is critical for the success of mRNA-based therapeutics. Here, we developed a nanoadjuvant system compromised of Quil-A and DOTAP (dioleoyl 3 trimethylammonium propane), hence named QTAP, for the efficient delivery of mRNA vaccine constructs into cells. Electron microscopy indicated that the complexation of mRNA with QTAP forms nanoparticles with an average size of 75 nm and which have ~90% encapsulation efficiency. The incorporation of pseudouridine-modified mRNA resulted in higher transfection efficiency and protein translation with low cytotoxicity than unmodified mRNA. When QTAP-mRNA or QTAP alone transfected macrophages, pro-inflammatory pathways (e.g., NLRP3, NF-kb, and MyD88) were upregulated, an indication of macrophage activation. In C57Bl/6 mice, QTAP nanovaccines encoding Ag85B and Hsp70 transcripts (QTAP-85B+H70) were able to elicit robust IgG antibody and IFN- ɣ, TNF-α, IL-2, and IL-17 cytokines responses. Following aerosol challenge with a clinical isolate of significant reduction of mycobacterial counts was observed in lungs and spleens of only immunized animals at both 4- and 8-weeks post-challenge. As expected, reduced levels of were associated with diminished histological lesions and robust cell-mediated immunity. Interestingly, polyfunctional T-cells expressing IFN- ɣ, IL-2, and TNF- α were detected at 8 but not 4 weeks post-challenge. Overall, our analysis indicated that QTAP is a highly efficient transfection agent and could improve the immunogenicity of mRNA vaccines against pulmonary , an infection of significant public health importance, especially to the elderly and to those who are immune compromised.
Topics: Animals; Mice; Mycobacterium avium; Mycobacterium tuberculosis; Interleukin-2; RNA; RNA, Messenger
PubMed: 37359562
DOI: 10.3389/fimmu.2023.1188754 -
Critical Reviews in Microbiology 2015This review addresses the survival and persistence of Mycobacterium avium subsp. paratuberculosis (MAP), the causative pathogen of Johne's disease (JD), once it has left... (Review)
Review
This review addresses the survival and persistence of Mycobacterium avium subsp. paratuberculosis (MAP), the causative pathogen of Johne's disease (JD), once it has left its ruminant host. JD has significant economic impact on dairy, beef and sheep industries and is difficult to control due to the long-term sub-clinical nature of the infection, intermittent or persistent MAP shedding during and after this period, inadequate test effectiveness, and the potential for MAP to exist for extended periods outside the host. The role that environmental factors play in the persistence and spread of MAP and consequent disease is assessed. Published risk factor analysis, organism survival across various environmental media and conditions, presence and spread in ruminant and non-ruminant wildlife, and the general potential for survival and multiplication of MAP ex-host both on and off-farm are discussed and knowledge gaps highlighted. An inclusive approach to disease management that takes into account the persistence and transport of the causative organism in on-farm soils and waters, land use and management, dispersal by domestic and non-domestic host species, as well as general animal husbandry is required on those farms where more traditional approaches to disease management have failed to reduce disease prevalence.
Topics: Animals; Cattle; Cattle Diseases; Environment; Mycobacterium avium subsp. paratuberculosis; Paratuberculosis; Prevalence; Risk Factors; Sheep; Sheep Diseases; Soil Microbiology
PubMed: 24670062
DOI: 10.3109/1040841X.2013.867830 -
Blood Apr 2015
Topics: Aged; Anti-Bacterial Agents; Fatal Outcome; Female; Humans; Macrophages; Mycobacterium avium; Pancytopenia; Tuberculosis
PubMed: 26016000
DOI: 10.1182/blood-2015-01-620195 -
Diagnostic Microbiology and Infectious... Jun 2017Mycobacterium avium complex (MAC) was originally composed of 2 species, M. avium and M. intracellulare. However, several new species closely related to M. intracellulare...
Mycobacterium avium complex (MAC) was originally composed of 2 species, M. avium and M. intracellulare. However, several new species closely related to M. intracellulare have recently been identified. In addition, M. avium has been further subdivided into 4 subspecies. The aim of this study was to determine the proportion of different MAC species recovered from respiratory specimens and to elucidate the clinical relevance of these species. Clinical isolates, from 219 patients, that had been initially identified as M. avium or M. intracellulare by non-sequencing methods were reidentified using multilocus sequence typing, and the clinical significance of the identified species was then investigated. Of 91 isolates originally identified as M. intracellulare, 75 (82%) were confirmed to be M. intracellulare, 8 (9%) isolates were identified as M. chimaera, and 4 (4%) isolates each were identified as "M. indicus pranii" and M. yongonense. The 128 isolates originally designated as M. avium were determined to be M. avium subsp. hominissuis. Of the 219 patients, 146 (67%) met the diagnostic criteria for MAC lung disease, and for each MAC species, the proportion of patients meeting these criteria was as follows: M. intracellulare (54/75, 72%), M. chimaera (3/8, 38%), "M. indicus pranii" (3/4, 75%), M. yongonense (2/4, 50%), and M. avium subsp. hominissuis (84/128, 66%). In summary, multilocus sequence typing of respiratory isolates initially identified as MAC revealed that, although most isolates were M. avium subsp. hominissuis or M. intracellulare, approximately 7% were newer MAC members, with clinical evidence supporting their potential pathogenicity for humans.
Topics: Aged; DNA, Bacterial; Female; Humans; Lung Diseases; Male; Middle Aged; Multilocus Sequence Typing; Mycobacterium avium; Mycobacterium avium Complex; Mycobacterium avium-intracellulare Infection
PubMed: 28291631
DOI: 10.1016/j.diagmicrobio.2017.02.017 -
Revue Scientifique Et Technique... Apr 2016The species Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis are the causal agents, respectively, of tuberculosis and paratuberculosis in animals....
The species Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis are the causal agents, respectively, of tuberculosis and paratuberculosis in animals. Both mycobacteria, especially M. bovis, are also important to public health because they can infect humans. In recent years, this and the impact of tuberculosis and paratuberculosis on animal production have led to significant advances in knowledge about both pathogens and their host interactions. This article describes the contribution of genomics and functional genomics to studies of the evolution, virulence, epidemiology and diagnosis of both these pathogenic mycobacteria.
Topics: Animals; Evolution, Molecular; Genomics; High-Throughput Nucleotide Sequencing; Molecular Epidemiology; Mycobacterium avium; Mycobacterium bovis; Tuberculosis; Virulence
PubMed: 27217180
DOI: 10.20506/rst.35.1.2429 -
Journal of the American Veterinary... Dec 2019
Topics: Alligators and Crocodiles; Animals; Citrobacter; Dermatitis; Male; Mycobacterium avium
PubMed: 31793839
DOI: 10.2460/javma.255.12.1349 -
Molecules (Basel, Switzerland) Apr 2023subspecies (MAP) is a chronic, contagious, and typically life-threatening enteric disease of ruminants caused by a bacterium of the genus Mycobacterium, but it can... (Review)
Review
subspecies (MAP) is a chronic, contagious, and typically life-threatening enteric disease of ruminants caused by a bacterium of the genus Mycobacterium, but it can also affect non-ruminant animals. MAP transmission occurs through the fecal-oral pathway in neonates and young animals. After infection, animals generate IL-4, IL-5, and IL-10, resulting in a Th2 response. Early detection of the disease is necessary to avoid its spread. Many detection methods, viz., staining, culture, and molecular methods, are available, and numerous vaccines and anti-tuberculosis drugs are used to control the disease. However, the prolonged use of anti-tuberculosis drugs leads to the development of resistance. Whereas vaccines hamper the differentiation between infected and vaccinated animals in an endemic herd. This leads to the identification of plant-based bioactive compounds to treat the disease. Bioactive compounds of and have been evaluated for their anti-MAP activity. Based on the MIC50 values, Ursolic acid (12 µg/mL) and Solasodine (60 µg/mL) were found to be suitable for anti-MAP activity.
Topics: Animals; Cattle; Mycobacterium avium subsp. paratuberculosis; Paratuberculosis; Ocimum sanctum; Solanum; Ruminants; Cattle Diseases
PubMed: 37110723
DOI: 10.3390/molecules28083490 -
BMC Microbiology Apr 2021Mycobacterium avium subsp. paratuberculosis (Map) causes Johne's disease (JD), a chronic enteritis widespread in ruminants, resulting in substantial economic losses,...
BACKGROUND
Mycobacterium avium subsp. paratuberculosis (Map) causes Johne's disease (JD), a chronic enteritis widespread in ruminants, resulting in substantial economic losses, especially to the dairy industry. Understanding the genetic diversity of Map in Australia will assist epidemiological studies for tracking disease transmission and identify subtype characteristics for use in development of improved diagnostic typing methods. Here we investigated the phylogenetic relationships of 351 Map isolates and compared different subtyping methods to assess their suitability for use in diagnostics and accuracy.
RESULTS
SNP-based phylogenetic analysis of 228 Australian isolates and 123 publicly available international isolates grouped Type S and Type C strains into two distinct lineages. Type C strains were highly monomorphic with only 20 SNP differences separating them. Type S strains, when aligned separately to the Telford strain, fell into two distinct clades: The first clade contained seven international isolates while the second clade contained one international isolate from Scotland and all 59 Australian isolates. The Australian Type B strain clustered with US bison strains. IS1311 PCR and Restriction Enzyme Analysis (REA) intermittently generated incorrect results when compared to Long Sequence Polymorphism (LSP) analysis, whole genome SNP-based phylogenetic analysis, IS1311 sequence alignment and average nucleotide identity (ANI). These alternative methods generated consistent Map typing results. A published SNP based assay for genotyping Map was found to be unsuitable for differentiating between Australian and international strain types of Map.
CONCLUSION
This is the first phylogenetic analysis of Australian Map isolates. The Type C lineage was highly monomorphic, and the Type S lineage clustered all Australian isolates into one clade with a single Scottish sheep strain. The Australian isolate classified as Type B by IS1311 PCR and REA is likely to be descended from bison and most closely related to US bison strains. Limitations of the current typing methods were identified in this study.
Topics: Animals; Australia; Genetic Variation; Genome, Bacterial; Genotype; Mycobacterium avium subsp. paratuberculosis; Paratuberculosis; Phylogeny; Polymerase Chain Reaction; Polymorphism, Single Nucleotide
PubMed: 33789575
DOI: 10.1186/s12866-021-02140-2 -
Meat Science Dec 2014Nontuberculous mycobacteria (NTM) are ubiquitous, potentially pathogenic organisms that have been isolated from a variety of environmental sources. NTM have been... (Review)
Review
Nontuberculous mycobacteria (NTM) are ubiquitous, potentially pathogenic organisms that have been isolated from a variety of environmental sources. NTM have been isolated from various kinds of food and many studies support the hypothesis that food, especially raw or partially cooked products, plays a role as a source of NTM for humans. Animals with disseminated infection have been diagnosed with NTM not only in the gastro-intestinal tract and intestinal lymph nodes, but also in tissues like muscle and parenchymatous organs. Infected animals may harbor NTM in their tissues even without clinical symptoms and especially minced meat with the possible addition of lymph nodes are considered as potential source of NTM. The purpose of this paper was to review articles concerning the detection of mycobacteria in the muscle tissue and lymph nodes of domestic animals, farmed and free-living game and to summarize methods and techniques for their detection.
Topics: Animals; Animals, Domestic; Animals, Wild; Lymph Nodes; Meat Products; Muscle, Skeletal; Mycobacterium avium; Nontuberculous Mycobacteria
PubMed: 25170820
DOI: 10.1016/j.meatsci.2014.08.001 -
Experimental Parasitology Apr 2019Association of the water- and foodborne pathogen Campylobacter jejuni with free-living Acanthamoeba spp. trophozoites enhances C. jejuni survival and resistance to...
Association of the water- and foodborne pathogen Campylobacter jejuni with free-living Acanthamoeba spp. trophozoites enhances C. jejuni survival and resistance to biocides and starvation. When facing less than optimal environmental conditions, however, the Acanthamoeba spp. host can temporarily transform from trophozoite to cyst and back to trophozoite, calling the survival of the internalized symbiont and resulting public health risk into question. Studies investigating internalized C. jejuni survival after A. castellanii trophozoite transformation have neither been able to detect its presence inside the Acanthamoeba cyst after encystation nor to confirm its presence upon excystation of trophozoites through culture-based techniques. The purpose of this study was to detect C. jejuni and Mycobacterium avium recovered from A. polyphaga trophozoites after co-culture and induction of trophozoite encystation using three different encystation methods (Neff's medium, McMillen's medium and refrigeration), as well as after cyst excystation. Internalized M. avium was used as a positive control, since studies have consistently detected the organism after co-culture and after host excystation. Concentrations of C. jejuni in A. polyphaga trophozoites were 4.5 × 10 CFU/ml, but it was not detected by PCR or culture post-encystation. This supports the hypothesis that C. jejuni may be digested during encystation of the amoebae. M. avium was recovered at a mean concentration of 1.9 × 10 from co-cultured trophozoites and 4.4 × 10 CFU/ml after excystation. The results also suggest that M. avium recovery post-excystation was statistically significantly different based on which encystation method was used, ranging from 1.3 × 10 for Neff's medium to 5.4 × 10 CFU/ml for refrigeration. No M. avium was recovered from A. polyphaga cysts when trophozoites were encysted by McMillen's medium. Since C. jejuni internalized in cysts would be more likely to survive harsh environmental conditions and disinfection, a better understanding of potential symbioses between free-living amoebae and campylobacters in drinking water distribution systems and food processing environments is needed to protect public health. Future co-culture experiments examining survival of internalized C. jejuni should carefully consider the encystation media used, and include molecular detection tools to falsify the hypothesis that C. jejuni may be present in a viable but not culturable state.
Topics: Acanthamoeba; Bacterial Load; Campylobacter jejuni; Coculture Techniques; Culture Media; DNA, Protozoan; Mycobacterium avium; Nucleic Acid Amplification Techniques; Refrigeration; Symbiosis; Trophozoites
PubMed: 30902623
DOI: 10.1016/j.exppara.2019.03.011