Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Genomic characteristics of sequenced M. microti strains

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques:

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: IS 6110 insertion element profiles and type III-A CRISPR-Cas locus in M. microti strains. (a) IS 6110 elements (black bars) identified along the genome of each M. microti strain. The number of IS 6110 elements is indicated at the top of each coloured bar. The genome size is in bp. (b) Type III-A CRISPR-Cas locus in M. microti strains as compared to M. tuberculosis H37Rv. Repeats are coloured in grey; spacers that are identical between M. tuberculosis H37Rv and M. microti strains are coloured in black, while those that are specific either to M. tuberculosis H37Rv or to M. microti strains are coloured in yellow and orange, respectively.

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques: CRISPR

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Phospholipase C locus and ESX-5 type VII protein secretion in M. microti strains. (a) Phospholipase C locus in M. microti strains. (b) Immunoblot secretion analysis of PE_PGRS proteins, PPE41 and EsxN, in M. microti strains. OV254-C corresponds to the OV254 strain complemented with the ppe38-71 locus from M. tuberculosis . SigA was used as a loading and lysis control. M, Molecular mass.

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques: Western Blot, Lysis, Control

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Polymorphism signals detected within M. microti strains. (a) Intersections of called variants (blue bars) within M. microti strains when compared to the genome of OV254. Black circles in the bottom matrix layout indicate the strains that are part of each intersection. The number of called variants within each intersection set is indicated at the top of each black bar. (b, c) Functional categories of genes affected by variants specific to strain ATCC 35782 (b), and by variants common to strains 94-2272, Maus III and Maus IV (c). The number and percentage of genes within each category are indicated.

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques: Functional Assay

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Regions of difference (RDs and MiDs) in M. microti strains

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques:

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Polymorphism signals detected between M. microti strains and M. tuberculosis H37Rv. (a) Functional categories of genes affected by variants specific to strain OV254 and/or strain ATCC 35782 as compared to M. tuberculosis H37Rv. The number and percentage of genes within each category are indicated. (b) Maximum-likelihood phylogenetic tree of M. microti strains. Branch lengths are proportional to nucleotide substitutions detected relative to M. tuberculosis H37Rv, used as outgroup strain for phylogenetic reconstruction. Support values obtained from 1000 bootstrap replicates are indicated in brown as percentages. Scale bar represents the number of substitutions per site.

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques: Functional Assay

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Virulence evaluation of M. microti strains relative to the control strains BCG Pasteur and M. tuberculosis H37Rv in SCID mice. A total of 13 SCID mice per group were infected intravenously via the lateral tail vein with 1×10 6 c.f.u. in 200 µl of PBS (1x). (a) Bacterial load in the spleen of infected mice after 1 day of infection. N =3 mice per group. The results are shown as the mean± sd . (b) Percentage of weight change of infected SCID mice over time. N =10 mice per group. Coloured lines and surrounding areas represent the mean± sd . (c) Percentage of survival of infected SCID mice over time. N =10 mice per group. Mice were killed when reaching the humane endpoint, defined as a weight loss of more than 20 %, in accordance with ethics committee guidelines.

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques: Control, Infection

Journal: Microbial Genomics

Article Title: Pathogenomic analyses of Mycobacterium microti, an ESX-1-deleted member of the Mycobacterium tuberculosis complex causing disease in various hosts

doi: 10.1099/mgen.0.000505

Figure Lengend Snippet: Protective efficacy of the vaccine candidates in the guinea pig low-dose infection model. Eight guinea pigs per group were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 µl of PBS (1x) and challenged 12 weeks after immunization by the aerosol route with 10–20 c.f.u. M. tuberculosis /lungs. Bacterial load (a), histological analysis of consolidation (b), and representative photomicrograph images of group mean lesion pathology from sections (c) of lungs and spleen of guinea pigs vaccinated with M. microti strains ATCC 35782, ATCC 35782::ESX-1 or BCG Danish or unvaccinated (saline control) after 4 weeks of infection with M. tuberculosis . (a) N =8 guinea pigs per group; mean± sem . One-way ANOVA test followed by Tukey HSD post-hoc test: ns, not significant; ***, P <0.001. (b) N =8 guinea pigs per group; mean± sem . Kruskal–Wallis test followed by Dunn post-hoc test: ns, not significant; *, P <0.05; ***, P <0.001. (c) Haematoxylin and eosin stained tissue sections of formalin-fixed and paraffin-embedded lung (×15 objective) and spleen (×25 objective).

Article Snippet: Guinea pigs (eight per group) were vaccinated once subcutaneously on the nape with 5×10 4 c.f.u. in 250 μl of PBS (1x) M. microti strain ATCC 35782, ATCC 35782::ESX-1 or BCG Danish, or inoculated with saline as a control.

Techniques: Infection, Aerosol, Saline, Control, Staining