bioRxiv Science⌕ Search

bioRxiv · 10.1101/2022.06.06.494966

High rifampicin peak plasma concentrations accelerate the slow phase of bacterial decline in tuberculosis patients: evidence for heteroresistance

Abstract

BackgroundAntibiotic treatments are often associated with a late slowdown in bacterial killing. This separates the killing of bacteria into at least two distinct phases: a quick phase followed by a slower phase, the latter of which is linked to treatment success. Current mechanistic explanations for the in vitro slowdown are either antibiotic persistence or heteroresistance. Persistence is defined as the switching back and forth between susceptible and non-susceptible states, while heteroresistance is defined as the coexistence of bacteria with heterogeneous susceptibilities. Both are also thought to cause a slowdown in the decline of bacterial populations in patients and therefore complicate and prolong antibiotic treatments. Reduced bacterial death rates over time are also observed within tuberculosis patients, yet the mechanistic reasons for this are unknown and therefore the strategies to mitigate them are also unknown. Methods and FindingsWe analyse a dose ranging trial for rifampicin in tuberculosis patients and show that there is a slowdown in the decline of bacteria. We show that the late phase of bacterial killing depends more on the peak drug concentrations than the total drug exposure. We compare these to pharmacokinetic-pharmacodynamic models of rifampicin heteroresistance and persistence. We find that the observation on the slow phases dependence on pharmacokinetic measures, specifically peak concentrations are only compatible with models of heteroresistance and incompatible with models of persistence. The quantitative agreement between heteroresistance models and observations is very good [Formula]. To corroborate the importance of the slowdown, we validate our results by estimating the time to sputum culture conversion and compare the results to a different dose ranging trial. ConclusionsOur findings indicate that higher doses, specifically higher peak concentrations may be used to optimize rifampicin treatments by accelerating bacterial killing in the slow phase. It adds to the growing body of literature supporting higher rifampicin doses for shortening tuberculosis treatments.

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Martinecz, A., Boeree, M. J., Diacon, A. H., Dawson, R., Hemez, C., Aarnoutse, R. E., Abel zur Wiesch, P.. 2022-06-07. High rifampicin peak plasma concentrations accelerate the slow phase of bacterial decline in tuberculosis patients: evidence for heteroresistance. https://doi.org/10.1101/2022.06.06.494966

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Characterization of Lactobacillus rhamnosus-Derived Extracellular Vesicles and Their Cellular Cytoprotective Effects Against Trimethylamine-Induced Oxidative Stress in Caco-2 Cells

Dietary trimethylamine (TMA) underwent gut microbiota-mediated catabolism and subsequent hepatic oxidation to generate trimethylamine N-oxide (TMAO), a key metabolic factor linked to vascular and colorectal diseases. While the systemic effects of circulating TMAO were heavily investigated, the direct localized cellular interactions of its precursor, TMA, at the intestinal epithelial level required further elucidation. This study characterized extracellular vesicles isolated from Lactobacillus rhamnosus (Lr-EVs) and evaluated their cytoprotective potential in an in vitro human intestinal Caco-2 cell model under TMA-induced stress. Exposure to a high concentration of TMA (20 mM) significantly reduced Caco-2 viability to 78.8%, triggered intracellular reactive oxygen species (ROS) accumulation, and compromised tight junction integrity as observed via the disruption of Zonula Occludens-1 (ZO-1) alignment. Co-treatment with Lr-EVs (100-300 g/mL) counteracted this cellular toxicity, enhancing cell survival up to 172%. Quantitative PCR analysis revealed that Lr-EVs suppressed the transcriptional upregulation of the pro-inflammatory gene IL-1{beta}, restored the expression of the metabolic regulator SIRT1, and stabilized transcript levels of key mitochondrial-encoded electron transport chain genes (ND6, CYTB, CO1, and ATP6). Additionally, Lr-EVs exhibited intrinsic radical scavenging activities in cell-free ABTS and DPPH assays. These preliminary in vitro findings demonstrated that Lr-EVs mitigated TMA-induced cellular injury, oxidative stress, and junctional disruption at the transcriptomic level, supporting further investigation of probiotic-derived EVs as prospective cell-free functional ingredients.

microbiology↗

Metagenomic Insights into the Fecal Microbiome, Resistome, and Mobilome of Beef Cattle in Cow-Calf Operations in Florida, United States

Food-producing animals are a focus of One Health approaches to antimicrobial resistance (AMR) as reservoirs of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs). We hypothesized that microbial composition, ARGs profiles, and mobile genetic element (MGE) profiles would differ by operation size and animal age group. Fecal samples from 162 cows and calves were combined into 54 composite samples, each comprising three animals of the same age group. Across operation sizes, Bacillota (51.4-64.9%), Bacteroidota (12.0-17.3%), Pseudomonadota (10.3-16.5%), and Actinomycetota (3.3-10.9%) were predominant phyla. Alpha diversity did not differ among operation sizes; however, cows exhibited greater observed richness, Shannon diversity, and Simpson diversity than calves (all P < 0.001). Microbial composition did not differ significantly among operation sizes (PERMANOVA, R{superscript 2} = 0.065, P = 0.609) but differed significantly between cows and calves (R{superscript 2} = 0.200, P < 0.001). Seventy-five ARGs representing 14 resistance classes were identified. Normalized ARG hit rates were higher in small operations (0.444) than in medium (0.278) and large operations (0.222; P = 0.088), whereas calves exhibited a higher estimated marginal mean normalized ARG hit rate than cows (0.383 vs. 0.247 hits/million reads; P < 0.001). Of 1,054 curated ARG hits, 23.9% occurred on predicted plasmid contigs, 0.9% on predicted viral contigs, and 7.1% on contigs containing insertion sequences. Overall findings demonstrate differences in fecal microbiome diversity and ARG abundance between cows and calves and indicate that predicted plasmids are associated with a notable proportion of ARG hits in Florida beef cow-calf operations.

microbiology↗

A global post-transcriptional regulator directs Acinetobacter baumannii carbon preference and gut colonization

Acinetobacter baumannii is a multidrug-resistant pathogen that colonizes the gut, increasing the risk of subsequent infection and transmission. Recent work showed that A. baumannii catabolizes ornithine, a non-preferred carbon source, to colonize the gut. However, the mechanisms of carbon preference were unknown in A. baumannii. Here, experimental evolution identified the conserved RNA-binding protein CsrA as a regulator of carbon preference in A. baumannii, a previously unreported CsrA function. RNA immunoprecipitation sequencing (RIP-Seq) revealed hundreds of CsrA-associated transcripts. A. baumannii CsrA directly bound and repressed expression of the ast arginine/ornithine catabolic operons. The Pseudomonas aeruginosa CsrA homolog RsmA repressed arginine/ornithine catabolic operon expression, suggesting this CsrA function extends to other bacterial taxa. Finally, CsrA was required for competitive fitness during A. baumannii gut colonization. These findings uncover a previously unrecognized role for a global regulator in A. baumannii carbon preference and persistence in the gut, a critical reservoir for multidrug-resistant pathogens.

microbiology↗