title: Pathophysiology of in-vitro induced filaments, spheroplasts and rod-shaped bacteria in neutropenic mice.
abstract: This study compared the in-vitro properties and in-vivo effects of Escherichia coli filaments, spheroplasts and normal cells in a murine thigh infection model. E. coli was exposed to ceftazidime, meropenem or saline to obtain filaments, spheroplasts or normal bacilli, which were then injected into neutropenic mice. After 24 h, morphology, CFUs, local and circulating endotoxin levels, cytokine levels and mortality were recorded, and correlations between bacterial and host parameters of infection were investigated. Filaments and spheroplasts contained more endotoxin/CFU than controls. Histological studies showed that morphologically altered bacteria changed into rod-shaped cells in the absence of antibiotics. Bacterial spread to the liver was significantly higher in mice challenged with rod-shaped cells, compared with antibiotic-exposed bacteria (p 0.007). Muscle endotoxin levels correlated significantly with circulating interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha, and both pro-inflammatory cytokines were correlated significantly (p 0.011). Despite a tendency toward higher local and systemic concentrations of endotoxin in the filament group, inflammatory responses and survival did not differ between groups. It was concluded that morphologically altered bacteria contain more endotoxin and can regain a rod shape after withdrawal of antibiotics, while non-antibiotic-exposed bacteria show greater spread to the liver. There was a clear intra-individual relationship between local endotoxin, systemic endotoxin, TNF-alpha and IL-6 production, but these parameters did not differ among groups.
authors.affiliation: Department of Medical Microbiology and Infectious Diseases, Canisius-Wilhelmina Hospital, Nijmegen, The Netherlands. jacqbuijs@hotmail.com
authors.forename: J
authors.initials: J
authors.lastname: Buijs
authors.forename: A S M
authors.initials: AS
authors.lastname: Dofferhoff
authors.forename: J W
authors.initials: JW
authors.lastname: Mouton
authors.forename: J W M
authors.initials: JW
authors.lastname: van der Meer
mesh_terms: D000818:Animals; D021461:Animals, Outbred Strains; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002442:Ceftazidime / Q000494:pharmacology; D003520:Cyclophosphamide / Q000009:adverse effects; D003599:Cytoskeleton / Q000378:metabolism; D004195:Disease Models, Animal; D004731:Endotoxins / Q000009:adverse effects / Q000032:analysis / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D004927:Escherichia coli Infections / Q000097:blood / Q000150:complications / Q000378:metabolism / Q000503:physiopathology*; D005260:Female; D015850:Interleukin-6 / Q000097:blood; D008099:Liver / Q000382:microbiology; D000077731:Meropenem; D051379:Mice; D009132:Muscles / Q000378:metabolism; D009135:Muscular Diseases / Q000378:metabolism / Q000503:physiopathology; D009503:Neutropenia / Q000139:chemically induced / Q000150:complications / Q000503:physiopathology*; D013104:Spheroplasts / Q000378:metabolism; D013845:Thienamycins / Q000494:pharmacology; D013848:Thigh / Q000382:microbiology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
chemical_list: D000900:Anti-Bacterial Agents; D004731:Endotoxins; D015850:Interleukin-6; D013845:Thienamycins; D014409:Tumor Necrosis Factor-alpha; D003520:Cyclophosphamide; D002442:Ceftazidime; D000077731:Meropenem