Respiratory Dysbiosis in Canine Bacterial Pneumonia: Standard Culture vs. Microbiome Sequencing.

Aida I Vientós-Plotts,Aaron C Ericsson,Carol R Reinero,Hansjorg Rindt

doi:10.3389/fvets.2019.00354

Aida I Vientós-Plotts, Aaron C Ericsson + Show 2 more

Open Access

https://doi.org/10.3389/fvets.2019.00354

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Abstract

It is unknown how the respiratory microbiome influences and is influenced by bacterial pneumonia in dogs, as culture of lung samples and not microbial sequencing guides clinical practice. While accurate identification of pathogens are essential for treatment, not all bacteria are cultivable and the impact of respiratory dysbiosis on development of pneumonia is unclear. The study purposes were to (1) characterize the lung microbiome in canine bacterial pneumonia and compare deviations in dominant microbial populations with historical healthy controls, (2) compare bacteria identified by culture vs. 16S rDNA sequencing from bronchoalveolar lavage fluid (BALF) culture-, and (3) evaluate similarities in lung and oropharyngeal (OP) microbial communities in community-acquired and secondary bacterial pneumonia. Twenty BALF samples from 15 client-owned dogs diagnosed with bacterial pneumonia were enrolled. From a subset of dogs, OP swabs were collected. Extracted DNA underwent PCR of the 16S rRNA gene. Relative abundance of operational taxonomic units (OTUs) were determined. The relative abundance of bacterial community members found in health was decreased in dogs with pneumonia. Taxa identified via culture were not always the dominant phylotype identified with sequencing. Dogs with community-acquired pneumonia were more likely to have overgrowth of a single organism suggesting loss of dominant species associated with health. Dogs with secondary bacterial pneumonia had a greater regional continuity between the upper and lower airways. Collectively, these data suggest that dysbiosis occurs in canine bacterial pneumonia, and culture-independent techniques may provide greater depth of understanding of the changes in bacterial community composition that occur in disease.

Highlights

Canine bacterial pneumonia is a common respiratory disorder, occurring as primary disease process, or secondary to aspiration, viral infections [1, 2], immunodeficiency, or a nosocomial event [3]
Twenty bronchoalveolar lavage fluid (BALF) samples were collected from 15 different dogs that met the enrollment criteria, i.e., a diagnosis of bacterial pneumonia based on clinical signs and associated with BALF septic suppurative inflammation or a positive culture result
In contrast to the number of taxa identified based by culture, the DNA detected in patient BALF samples represented between 22 and 185 distinct operational taxonomic units (OTUs)

Summary

Introduction

Canine bacterial pneumonia is a common respiratory disorder, occurring as primary disease process, or secondary to aspiration, viral infections [1, 2], immunodeficiency, or a nosocomial event [3]. The recent development of culture-independent molecular techniques has revealed that in humans [4], cats [5], dogs [6], sheep [7], and likely other host species, the healthy lungs harbor low biomass microbial populations seeded via direct extension from upper airway communities, repeated microaspiration, and inhalation of bacteria in air [8]. These culture-independent methods have reinforced that a lack of cultivable organisms does not necessarily indicate a sterile environment [9]. The current study compared the results of traditional culture-based methods and a targeted sequencing approach applied to 20 BALF samples collected from 15 dogs affected with bacterial pneumonia in a referral veterinary hospital setting

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