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1.
Vector Borne Zoonotic Dis ; 15(9): 539-44, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26378973

ABSTRACT

Currently, no vaccine has been developed to protect humans from naturally acquired heterologous Orientia tsutsugamushi infections. To enhance the validity of vaccine candidates, we are developing a murine chigger challenge model with the O. tsutsugamushi Lc-1-infected Leptotrombidium chiangraiensis Line-1. To this end, an intraperitoneal (i.p.) murine challenge model using an O. tsutsugamushi Lc-1 isolate was developed for eventual validation of the chigger challenge model. We have determined that the murine lethal dose that kills 50% of the challenged mice (MuLD50) of a liver/spleen homogenate developed from O. tsutsugamushi Lc-1-infected ICR Swiss mice to be 10(-6.9). Employing different inoculum doses of this homogenate, the bacterial load using quantitative real-time PCR (qPCR) was determined to range from 60 to 1.6 × 10(5) genome equivalent copies (GEC)/µL of liver and 33.4 to 2.2 × 10(5) GEC/µL of spleen tissue. The clinical outcomes relative to homogenate dose levels followed a dose-dependent pattern. The successful development and characterization of the O. tsutsugamushi Lc-1 i.p. challenge model will assist in the development and validation of a mouse chigger challenge scrub typhus model.


Subject(s)
Orientia tsutsugamushi/physiology , Scrub Typhus/microbiology , Trombiculidae/microbiology , Animals , Bacterial Load , Disease Models, Animal , Female , Humans , Injections, Intraperitoneal , Lethal Dose 50 , Liver/microbiology , Mice , Mice, Inbred ICR , Orientia tsutsugamushi/genetics , Spleen/microbiology
2.
J Med Entomol ; 51(3): 658-60, 2014 May.
Article in English | MEDLINE | ID: mdl-24897859

ABSTRACT

Orientia tsutsugamushi is a pathogen transmitted by Leptotrombidium that causes scrub typhus. To develop an infection mouse model, a mite-derived isolate of O. tsutsugamushi was established from a laboratory-maintained colony of Leptotrombidium chiangraiensis (O. tsutsugamushi Lc-1). This Lc-1 isolate was initially presented to ICR (CD-1) mice by feeding an infected Lc chigger on the ear of a mouse. Once the Lc-1 was adapted to the ICR mice, quantitative real-time polymerase chain reaction was used to investigate O. tsutsugamushi genomic equivalent copies in tissues and sera. Furthermore, times to onset of the signs of infection are reported in this study. This study provides information useful for future research on this host-pathogen interaction and the associated vaccine efficacy trials.


Subject(s)
Orientia tsutsugamushi/physiology , Scrub Typhus/microbiology , Trombiculidae/microbiology , Animals , Disease Models, Animal , Liver/microbiology , Mice , Mice, Inbred ICR , Real-Time Polymerase Chain Reaction , Spleen/microbiology
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