In Vivo Tracking of Bacterial Colonization in Different Murine Models Using Bioluminescence: The Example of Salmonella

Abstract

Applications of bioluminescence for the in vivo study of pathogenic microorganisms are numerous, ranging from the quantification of virulence gene expression to measuring the effect of antimicrobial molecules on the colonization of tissues and organs by the pathogen. Most studies are performed in mice, but recent works demonstrate that this technique is applicable to larger animals like fish, guinea pigs, ferrets, and chickens. Here, we describe the construction and the utilization of a constitutively luminescent strain of Salmonella Typhimurium to monitor in vivo and ex vivo the colonization of mice in the gastroenteritis, typhoid fever, and asymptomatic carriage models of Salmonella infection.

From intestine to beyond: Salmonella entry factors display distinct transcription pattern upon infection in murine models

“…enterica ser. Typhimurium wild-type strainAmerican Type Culture Collection att Tn7-NoP:: luxCDABE promoterless transcriptional lux fusion inserted in the att Tn7 site of strain 14028 (Cm r )[51] att Tn7-sig70c35:: luxCDABE transcriptional fusion sig70c35:: lux inserted in the att Tn7 site of strain 14028 (Cm r )[51] att Tn7-P pefI-srgC :: luxCDABE transcriptional fusion P pefI-srgC :: lux inserted in the att Tn7 site of strain 14028 (Cm r )this work att Tn7-P pagN :: luxCDABE transcriptional fusion P pagN :: lux inserted in the att Tn7 site of strain 14028 (Cm r )this work att Tn7-P invF ::: luxCDABE transcriptional fusion P invF :: lux inserted in the att Tn7 site of strain 14028 (Cm r )this work MC1061 E. coli hsdR mcrB araD 139 Δ ( araABC-leu )7679 ΔlacX 74 galU galK rpsL thi [52] CC118 E. coli Δlac X74 galE galK phoA 20 thi , r psE rpoB argE (Am) recA 1( λpir )[53]plasmids pCS26-sig70c35:: luxCDABE plasmid-borne construction allowing constitutive expression of the luxCDABE luciferase system (Cm r )[33] pUC18R6K-mini-Tn7T-PacIdelivery vector with a λpir -dependent replication (Cb r )[33] pHSG415- tnsABCD helper plasmid with temperature-sensitive replication (Cb r )[33] pACYC177cloning vector (Cb r , Kan r )[54] pACSdiApACYC177 containing S . Typhimurium 14028 sdiA ORF and its RBS (Cb r )[26]…”

“…Results are expressed in photons per second per square centimetre per steradian (p s −1 cm −2 sr −1 ), that is, radiance. Then, bacterial loads of the different organs were determined as previously described [ 51 ]. Two different schedules were tested in the typhoid fever model in order to assess all the kinetics of S. Typhimurium infection.…”

“…All animal experimentations were performed in the Infectiology of Farm, Model and Wildlife Animals Facility (PFIE, Centre INRAE Val De Loire: https://doi.org/10.15454/1.5572352821559333E12; member of the National Infrastructure EMERG'IN). Inocula were prepared as previously described [51]. The typhoid-like and the gastroenteritis models were used as described [36,56].…”

Respuesta histomorfométrica de la mucosa del intestino delgado en cuyes (Cavia porcellus) de engorde desafiados con Salmonella enterica var. Typhimurium

“…El agente causal más frecuentemente aislado en infecciones por Salmonella en cuyes es la especie Salmonella enterica serovar Typhimurium (SeVT) [7,11,25,28,30], patógeno Gram negativo de la familia Enterobacteriaceae, que se transmite principalmente por la vía fecal-oral a través del alimento contaminado con heces de cuy infectados, de roedores o pájaros silvestres [21,25,45], desarrollando una colonización a nivel intestinal seguida de una infección sistémica [4,24]. Uno de los signos clínicos más evidente es la presentación de diarreas en animales y postración, seguido de aborto (en hembras), aunque también puede presentarse una infección subclínica convirtiendo al cuy en un portador asintomático de Salmonella, transmitiendo la enfermedad a otros animales y al hombre [15].…”

Direct monitoring of meropenem therapeutic efficacy against Klebsiella pneumoniae respiratory infection by bioluminescence imaging

“…Bioluminescence imaging (BLI) has been used to detect light-producing live cells within small animals [27][28][29][30]. The BLI system involves engineering of a luminescent micro-organism through the expression of a lux operon either from a transformed plasmid or, more stably, through the insertion of the luxCDABE genes into bacterial genome [31].…”