ABSTRACT
Rodentibacter (R.) heylii is frequently detected in laboratory rodents. Repeats in toxin (RTX) toxins are considered important virulence factors of this major murine pathogen. We evaluated the virulence of a R.heylii strain negative for all known RTX toxin genes and Muribacter (M.) muris, a commensal in mice, in experimental infections of C57BL/6 and BALB/c mice. Experimental intranasal infection with 108 CFU of the pnxI-, pnxII- and pnxIII- R. heylii strain resulted in 75% and 100% mortality in C57BL/6 and BALB/c mice, respectively. In early losses, multiple internal organs were infected and purulent bronchopneumonia was the main pathology. Intranasal application of M. muris did not result in mortality or severe weight loss. Immunoproteomics led to the identification of a surface-associated and specific immunogen, which was designated as R. heylii immunogen A (RhiA) and which was exclusively recognised by sera obtained from mice infected with this R. heylii pathotype. RhiA is a 262.6 kDa large protein containing long imperfect tandem repeats and C-terminal RTX consensus sequences. Immunohistochemical analysis confirmed that this R.heylii pathotype expresses RhiA in the lower respiratory tract. In summary, this study describes a specific immunogen in a virulent R. heylii, strain which is an excellent antigen for pathotype-specific serological screenings and which might carry out RTX-related functions.
Subject(s)
Bacterial Proteins/immunology , Pasteurellaceae Infections/veterinary , Pasteurellaceae/immunology , Rodent Diseases/microbiology , Animals , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Toxins/genetics , Bacterial Toxins/immunology , Consensus Sequence , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Pasteurellaceae/chemistry , Pasteurellaceae/genetics , Pasteurellaceae/pathogenicity , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/mortality , Protein Domains , Rodent Diseases/mortality , Tandem Repeat Sequences , Virulence , Virulence Factors/chemistry , Virulence Factors/genetics , Virulence Factors/immunologyABSTRACT
Bacterial pathogens cause severe infections worldwide in livestock and in humans, and antibiotic resistance further increases the importance of prophylactic vaccines. Inactivated bacterial vaccines (bacterins) are usually produced via incubation of the pathogen with chemicals such as formaldehyde, which is time consuming and may cause loss of immunogenicity due to the modification of structural components. We evaluated low-energy electron irradiation (LEEI) as an alternative method to generate a bacterin. Rodentibacter pneumotropicus, an invasive Gram-negative murine pathogen, was inactivated with LEEI and formaldehyde. LEEI resulted in high antigen conservation, and LPS activity was significantly better maintained when compared with formaldehyde treatment. Immunization of mice with LEEI-inactivated R. pneumotropicus elicited a strong immune response with no detectable bacterial burden upon sublethal challenge. The results of this study suggest the inactivation of bacteria with LEEI as an alternative, fast and efficient method to generate bacterial vaccines with increased efficacy.
ABSTRACT
The pathogens Rodentibacter (R.) pneumotropicus and R. heylii as well as the commensal Muribacter (M.) muris are frequently isolated in mice. In this study, a MALDI-TOF MS database was extended with spectra of well characterized strains of these species. Compared to a multiplex PCR, all examined out-of-sample isolates were correctly identified.
