Dataset Information

Development, Characterization, and Standardization of a Nose-Only Inhalation Exposure System for Exposure of Rabbits to Small-Particle Aerosols Containing Francisella tularensis.

ABSTRACT: Inhalation of Francisella tularensis causes pneumonic tularemia in humans, a severe disease with a 30 to 60% mortality rate. The reproducible delivery of aerosolized virulent bacteria in relevant animal models is essential for evaluating medical countermeasures. Here we developed optimized protocols for infecting New Zealand White (NZW) rabbits with aerosols containing F. tularensis We evaluated the relative humidity, aerosol exposure technique, and bacterial culture conditions to optimize the spray factor (SF), a central metric of aerosolization. This optimization reduced both inter- and intraday variability and was applicable to multiple isolates of F. tularensis Further improvements in the accuracy and precision of the inhaled pathogen dose were achieved through enhanced correlation of the bacterial culture optical density and the number of CFU. Plethysmograph data collected during exposures found that respiratory function varied considerably between rabbits, was not a function of weight, and did not improve with acclimation to the system. Live vaccine strain (LVS)-vaccinated rabbits were challenged via aerosol with human-virulent F. tularensis SCHU S4 that had been cultivated in either Mueller-Hinton broth (MHB) or brain heart infusion (BHI) broth. LVS-vaccinated animals challenged with SCHU S4 that had been cultivated in MHB experienced short febrile periods (median, 3.2?days), limited weight loss (<5%), and longer median survival times (?18?days) that were significantly different from those for unvaccinated controls. In contrast, LVS-vaccinated rabbits challenged with SCHU S4 that had been cultivated in BHI experienced longer febrile periods (median, 5.5?days) and greater weight loss (>10%) than the unvaccinated controls and median survival times that were not significantly different from those for the unvaccinated controls. These studies highlight the importance of careful characterization and optimization of protocols for aerosol challenge with pathogenic agents.

SUBMITTER: O'Malley KJ

PROVIDER: S-EPMC6652755 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Development, Characterization, and Standardization of a Nose-Only Inhalation Exposure System for Exposure of Rabbits to Small-Particle Aerosols Containing Francisella tularensis.

O'Malley Katherine J KJ Bowling Jennifer D JD Barry Eileen M EM Hazlett Karsten R O KRO Reed Douglas S DS

Infection and immunity 20190723 8

Inhalation of <i>Francisella tularensis</i> causes pneumonic tularemia in humans, a severe disease with a 30 to 60% mortality rate. The reproducible delivery of aerosolized virulent bacteria in relevant animal models is essential for evaluating medical countermeasures. Here we developed optimized protocols for infecting New Zealand White (NZW) rabbits with aerosols containing <i>F. tularensis</i> We evaluated the relative humidity, aerosol exposure technique, and bacterial culture conditions to ...[more]

PMID: 31085702

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Project description:Francisella tularensis is composed of a number of subspecies with varied geographic distribution, host ranges, and virulence. In view of these marked differences, comparative functional genomics may elucidate some of the molecular mechanism(s) behind these differences. In this study a shared probe microarray was designed that could be used to compare the transcriptomes of Francisella tularensis subsp. tularensis Schu S4 (Ftt), Francisella tularensis subsp. holarctica OR960246 (Fth), Francisella tularensis subsp. holarctica LVS (LVS), and Francisella novicida U112 (Fn). To gain insight into expression differences that may be related to the differences in virulence of these subspecies, transcriptomes were measured from each strain grown in vitro under identical conditions, utilizing a shared probe microarray. The human avirulent Fn strain exhibited high levels of transcription of genes involved in general metabolism, which are pseudogenes in the human virulent Ftt and Fth strains, consistent with the process of genome decay in the virulent strains. Genes encoding an efflux system (emrA2 cluster of genes), siderophore (fsl operon), acid phosphatase, LPS synthesis, polyamine synthesis, and citrulline ureidase were all highly expressed in Ftt when compared to Fn, suggesting that some of these may contribute to the relative high virulence of Ftt. Genes expressed at a higher level in Ftt when compared to the relatively less virulent Fth included genes encoding isochorismatases, cholylglycine hydrolase, polyamine synthesis, citrulline ureidase, Type IV pilus subunit, and the Francisella Pathogenicity Island protein PdpD. Fth and LVS had very few expression differences, consistent with the derivation of LVS from Fth. This study demonstrated that a shared probe microarray designed to detect transcripts in multiple species/subspecies of Francisella enabled comparative transcriptional analyses that may highlight critical differences that underlie the relative pathogenesis of these strains for humans. This strategy could be extended to other closely-related bacterial species for inter-strain and inter-species analyses.

Project description:Particle inhalation rate (PIR) is an air pollution exposure metric that relies on age-, sex-, and physical activity-specific estimates of minute respiratory volume (MRV; L/min-kg) to account for personal inhalation patterns. United States Environmental Protection Agency (USEPA)-generated MRV estimates derive primarily from relatively homogenous populations without substantial cardiorespiratory challenges. To determine if these MRV estimates are relevant to populations in generally poor cardiorespiratory health (e.g., the Boston Puerto Rican Health Study (BPRHS) population) or whether population-specific estimates are needed, we 1) estimated population-specific MRVs and compared them to USEPA MRV estimates, and 2) compared exposure distributions and health effect estimates using PIR with population-specific MRVs, PIR with USEPA MRVs, and ambient particle number concentration (PNC). We recruited 40 adults (80% Puerto Rican, mean age = 60.2 years) in Boston with health characteristics similar to the BPRHS population. We measured pulse, oxygen saturation, respiration rate, and inspiratory volume while participants walked, stood, sat, and lay down. Pulse, respiration rate, inspiratory volume, and MRV were greater when participants were walking/standing compared to sitting or lying down. We then calculated MRVs adjusted for age, sex, measured body weight, and physical activity using data from 19 Puerto Rican participants who wore a nose clip or held their nostrils closed. We applied the population-specific and USEPA MRVs to estimate ultrafine particle exposure for participants in the BPRHS (n = 781). We compared exposure distributions and health effect estimates using the PIR with population-specific MRV estimates, PIR with USEPA MRV estimates, and ambient concentrations. We found that while population-specific MRVs differed from USEPA MRVs, particularly for unhealthy participants, PIR exposure distributions and health effect estimates were similar using each exposure metric. Confidence intervals were narrower using the PIR metrics than ambient PNC, suggesting increased statistical efficiency. Even in our understudied population, using USEPA MRVs did not meaningfully change PIR estimates.

Dataset Information

Development, Characterization, and Standardization of a Nose-Only Inhalation Exposure System for Exposure of Rabbits to Small-Particle Aerosols Containing Francisella tularensis.

Publications

Development, Characterization, and Standardization of a Nose-Only Inhalation Exposure System for Exposure of Rabbits to Small-Particle Aerosols Containing Francisella tularensis.

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