Arid_Recovery_APDP_soil_scats_map

QIIME mapping fil

Sample-specific MIDs for scat samples.

Sample-specific MIDs for scat samples

otu_table_mc2_w_tax

Soil & scat fungal OTU table generated by open-reference OTU picking with the UNITE database

barcodes_AridRecITS_soil

Sample-specific MIDs for soil samples

PGM_AridRecITS_soil.fastq

FASTQ file of PGM reads for Arid Recovery soil samples

Arid recovery soil fungi

IonTorrent PGM run for fungi diversity stud

Rev_barcodes_AridRecITS_soil_scats

6 bp MID at the 5’ end of the ITS5 prime

Type Six Secretion System of <i>Bordetella bronchiseptica</i> and Adaptive Immune Components Limit Intracellular Survival During Infection

<div><p>The Type Six Secretion System (T6SS) is required for <i>Bordetella bronchiseptica</i> cytotoxicity, cytokine modulation, infection, and persistence. However, one-third of recently sequenced <i>Bordetella bronchiseptica</i> strains of the predominantly human-associated Complex IV have lost their T6SS through gene deletion or degradation. Since most human <i>B</i>. <i>bronchiseptica</i> infections occur in immunocompromised patients, we determine here whether loss of Type Six Secretion is beneficial to <i>B</i>. <i>bronchiseptica</i> during infection of immunocompromised mice. Infection of mice lacking adaptive immunity (Rag1^-/- mice) with a T6SS-deficient mutant results in a hypervirulent phenotype that is characterized by high numbers of intracellular bacteria in systemic organs. In contrast, wild-type <i>B</i>. <i>bronchiseptica</i> kill their eukaryotic cellular hosts via a T6SS-dependent mechanism that prevents survival in systemic organs. High numbers of intracellular bacteria recovered from immunodeficient mice but only low numbers from wild-type mice demonstrates that <i>B</i>. <i>bronchiseptica</i> survival in an intracellular niche is limited by B and T cell responses. Understanding the nature of intracellular survival during infection, and its effects on the generation and function of the host immune response, are important to contain and control the spread of <i>Bordetella</i>-caused disease.</p></div

The T6SS modulates virulence and bacterial dissemination.

<p>(A) Groups of Rag1^-/- mice (n = 8) were inoculated with 5x10⁵ CFU of RB50 (blue) or RB50Δ<i>clpV</i> (red) and were monitored for survival. (B) Groups of Rag1^-/- mice (n = 4 per group) were inoculated with 5x10⁵ CFU of RB50 (blue) versus RB50Δ<i>clpV</i> (red) and dissected on day 21 p.i. for bacterial enumeration in respiratory tract and systemic organs. The experiment was performed three times with similar results and a representative data set is shown. (C) Rag^-/- mice were inoculated with 5x10⁵ CFU RB50 (blue) versus RB50Δ<i>clpV</i> (red) and bacteria were enumerated from the spleen, liver, and kidney on days 3, 7, 21, and 35. With the exception of RB50Δ<i>clpV</i> on day 21 (n = 1), three mice were sacrificed per group per timepoint. * denotes p value <0.05. Grey dotted line indicates the limit of detection.</p

Deletion of <i>clpV</i> increases intracellular survival <i>in vitro</i>.

<p>(A) RAW264.7 macrophages were infected with RB50 (blue, n = 4) or RB50Δ<i>clpV</i> (red, n = 4) at an MOI of 100 and bacterial invasion and intracellular survival was determined at 1, 24, and 48 hour after addition of gentamicin. The experiment was conducted five times with similar results and a representative dataset is shown. (B) The cytotoxicity of RAW264.7 macrophages infected with RB50 (blue) or RB50Δ<i>clpV</i> (red) at an MOI of 100 was determined 1 hour and 24 hours after gentamicin application. The average percent cytotoxicity of four wells in three different experiments was measured by (LDH release from a well / LDH release from positive control well) x 100 ±SE is shown. * denotes p value <0.05. Grey line indicates limit of detection.</p