Acinetobacter baumannii is an important emerging pathogen in health care-acquired infections and is responsible for severe nosocomial and community-acquired pneumonia. Currently available mouse models of A. baumannii pneumonia show poor colonization with little to no extrapulmonary dissemination. Here, we describe a mouse model of A. baumannii pneumonia using a clinical isolate (LAC-4 strain) that reliably reproduces the most relevant features of human pulmonary A. baumannii infection and pathology. Using this model, we have shown that LAC-4 infection induced rapid bacterial replication in the lungs, significant extrapulmonary dissemination, and severe bacteremia by 24 h postintranasal inoculation. Infected mice showed severe bronchopneumonia and dilatation and inflammatory cell infiltration in the perivascular space. More significantly, 100% of C57BL/6 and BALB/c mice succumbed to 10 ⁸ CFU of LAC-4 inoculation within 48 h. When this model was used to assess the efficacy of antimicrobials, all mice treated with imipenem and tigecycline survived a lethal intranasal challenge, with minimal clinical signs and body weight loss. Moreover, intranasal immunization of mice with formalin-fixed LAC-4 protected 40% of mice from a lethal (100× 100% lethal dose) intraperitoneal challenge. Thus, this model offers a reproducible acute course of A. baumannii pneumonia without requiring additional manipulation of host immune status, which will facilitate the development of therapeutic agents and vaccines against A. baumannii pneumonia in humans.

Acinetobacter baumannii is an important human pathogen due to its multi-drug resistance. In this study, the genome of an ST10 outbreak A. baumannii isolate LAC-4 was completely sequenced to better understand its epidemiology, antibiotic resistance genetic determinants and potential virulence factors. Compared with 20 other complete genomes of A. baumannii, LAC-4 genome harbors at least 12 copies of five distinct insertion sequences. It contains 12 and 14 copies of two novel IS elements, ISAba25 and ISAba26, respectively. Additionally, three novel composite transposons were identified: Tn6250, Tn6251 and Tn6252, two of which contain resistance genes. The antibiotic resistance genetic determinants on the LAC-4 genome correlate well with observed antimicrobial susceptibility patterns. Moreover, twelve genomic islands (GI) were identified in LAC-4 genome. Among them, the 33.4-kb GI12 contains a large number of genes which constitute the K (capsule) locus. LAC-4 harbors several unique putative virulence factor loci. Furthermore, LAC-4 and all 19 other outbreak isolates were found to harbor a heme oxygenase gene (hemO)-containing gene cluster. The sequencing of the first complete genome of an ST10 A. baumannii clinical strain should accelerate our understanding of the epidemiology, mechanisms of resistance and virulence of A. baumannii.

Multidrug-resistantAcinetobacter baumanniistrains have increasingly resulted in nosocomial outbreaks worldwide, leaving limited options for treatment. To date, little has been reported on the antimicrobial susceptibilities and genomic profiles ofA. baumanniistrains from hospital outbreaks in the Greater Los Angeles area. In this study, we examined the susceptibilities and genetic profiles of 20 nonduplicate isolates ofA. baumanniifrom nosocomial outbreaks in Los Angeles County (LAC) and determined their mechanisms of fluoroquinolone resistance. Antibiotic susceptibility testing indicated that the majority of these LAC isolates were not susceptible to 14 of the 17 antibiotics tested, with the exception of doxycycline, minocycline, and tigecycline. In particular, all isolates were found to be resistant to ciprofloxacin. Genomic DNA analysis revealed eight epidemiologically distinct groups among these 20A. baumanniiisolates, consistent with antibiotic susceptibility profiles. Sequencing analysis confirmed that concurrent GyrA and ParC amino acid substitutions in the “hot spots” of their respective quinolone resistance-determining regions were primarily responsible for the high-level ciprofloxacin resistance of these isolates. Antibiotic susceptibility testing using two efflux pump inhibitors suggested that the presence of efflux pumps was only a secondary contributor to ciprofloxacin resistance for some of the isolates. In summary, the present study has revealed good correlation between the antibiotic susceptibility profiles and genetic fingerprints of 20 clinical isolates from nosocomial outbreaks in Los Angeles County and has determined their mechanisms of fluoroquinolone resistance, providing an important foundation for continued surveillance and epidemiological analyses of emergingA. baumanniiisolates in Los Angeles County hospitals.