title: Optimisation of the Caenorhabditis elegans model for studying the pathogenesis of opportunistic Acinetobacter baumannii.
abstract: This study aimed to increase the sensitivity of Caenorhabditis elegans as an infection model for detection of minor differences in virulence or fitness between different Acinetobacter baumannii strains with known resistance and virulence mechanisms. Selected A. baumannii strains and mutants, comprising wild-type strains (ATCC 17978 and 19606), colistin-resistant strains (ATCC 19606 ΔlpxA and ATCC 19606 ΔlpxC), a clinical encapsulated isolate (AB307-0294), an imipenem-resistant strain (ATCC 17978 Δomp33-36) and an sRNA knock-out strain (ATCC 17978 Δ13573), were employed in developing killing and fertility assays in a C. elegans infection model. Because virulence levels of the strains were known, they could be used to assess assays in the nematode model for their ability to discriminate between degrees of virulence. The model was validated by microscopic analysis and in a murine sepsis infection model. The fertility assay, specifically utilising nematode growth medium, was able to detect virulence differences between the wild-type strains, ATCC 19606 ΔlpxA and isolate AB307-0294. Moreover, modification of an alternative culture medium by incremental changes in osmolarity facilitated detection of subtle virulence differences between isogenic mutants (ATCC 17978 Δomp33-36 and 17978 Δ13573). The success of the proposed fertility model depends on establishing a balance between optimal C. elegans reproduction and environmental stress leading to maximum pathogen-induced damage. This invertebrate model may reduce the need for mammalian in vivo studies of A. baumannii resistance and pathogenicity and may additionally be validated for the study of other low-virulence bacterial pathogens.
authors.affiliation: Servicio de Microbiología, Instituto de Investigación Biomédica (INIBIC), Complejo Hospitalario Universitario A Coruña (CHUAC), As Xubias s/n, 15006 A Coruña, Spain.
authors.forename: J A
authors.initials: JA
authors.lastname: Vallejo
authors.affiliation: Servicio de Microbiología, Instituto de Investigación Biomédica (INIBIC), Complejo Hospitalario Universitario A Coruña (CHUAC), As Xubias s/n, 15006 A Coruña, Spain. Electronic address: alejandro.beceiro.casas@sergas.es.
authors.forename: A
authors.initials: A
authors.lastname: Beceiro
authors.affiliation: Servicio de Microbiología, Instituto de Investigación Biomédica (INIBIC), Complejo Hospitalario Universitario A Coruña (CHUAC), As Xubias s/n, 15006 A Coruña, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Rumbo-Feal
authors.affiliation: Centro Andaluz de Biología del Desarrollo (CABD-CSIC), Departamento de Biología Molecular e Ingeniería Bioquímica, Universidad Pablo de Olavide, Seville, Spain.
authors.forename: M J
authors.initials: MJ
authors.lastname: Rodríguez-Palero
authors.affiliation: Veterans Administration Western Healthcare System, Departments of Medicine and Microbiology & Immunology, The University at Buffalo-State University of New York, Buffalo, NY, USA.
authors.forename: T A
authors.initials: TA
authors.lastname: Russo
authors.affiliation: Servicio de Microbiología, Instituto de Investigación Biomédica (INIBIC), Complejo Hospitalario Universitario A Coruña (CHUAC), As Xubias s/n, 15006 A Coruña, Spain. Electronic address: german.bou.arevalo@sergas.es.
authors.forename: G
authors.initials: G
authors.lastname: Bou
keywords: Acinetobacter baumannii; Caenorhabditis elegans; In vivo models; Pathogenesis; Resistance; Virulence