Ex vivo bioluminescence imaging of late gestation ewes following intrauterine inoculation with lux-modified Escherichia coli

Hormonal Profiles of Late Gestation Ewes Following Intra-uterine Inoculation With and Without Lux-modified Escherichia coli

“…Since we used photonic bacteria in both trials (Trials 1 and 2) of this experiment, we report both studies here despite the confounding influences of mixed species contamination in Trial 1 since we are able to screen for our E. coli pAK1-lux bacteria and isolate its presence. Thus while we cannot confirm definitively that preterm delivery in treated and Control groups in Trial 1 was due solely to our E. coli pAK1-lux inoculation and not the other contaminating organisms, we were able to identify the presence of E. coli pAK1-lux by photonic imaging [16] in these inoculated ewes and report these results for this reason. Furthermore, we have removed the ewes with identified mixed species bacterial concentrations from the analysis of the treated groups in Table 2, and from the treated and Control ewes in the analysis of hormonal data reported in Figs.…”

“…Furthermore, we have removed the ewes with identified mixed species bacterial concentrations from the analysis of the treated groups in Table 2, and from the treated and Control ewes in the analysis of hormonal data reported in Figs. 2 and 3 (See also Table for "n" in relation to treatment groups as influenced by experimental procedures and ewe outcomes).As reported in a companion study [16] for Trial 2, following further refinement of the inoculation procedures used in Trial 1, none (0%; n=5) of the Controls preterm delivered. However 60% (n=3 of 5) of the 1.2 × 10 6 CFU/ml-treated ewes and 60% (n=3 of 5) of the 5.6 × 10 6 CFU/ml-treated ewes preterm delivered.…”

“…As reported in a companion study [16] for Trial 2, following further refinement of the inoculation procedures used in Trial 1, none (0%; n=5) of the Controls preterm delivered. However 60% (n=3 of 5) of the 1.2 × 10 6 CFU/ml-treated ewes and 60% (n=3 of 5) of the 5.6 × 10 6 CFU/ml-treated ewes preterm delivered.…”

“…Hormone evaluation post intra-uterine inoculation with photonemitting Escherichia coli (MM294 transformed with plasmid pAK1-lux) in late gestation ewes: Escherichia coli MM294 (Migula) Castellani and Chalmer (ATCC # 33625) was chosen for Experiment 2 because of its higher transformation host strain capabilities than the E. coli used in Experiment 1 and is described in a companion study which details the in vivo photonic imaging of this E. coli strain within the uterus of the ewe [16]. Delivery preterm in ewes occurred between 48 and 120 h post-inoculation in Trials 1 and 2, while ewes maintaining pregnancy without indications of fetal distress experienced uninterrupted maintenance of pregnancy to study termination (i.e., 144 h post-inoculation).…”

“…Ewes were assigned to the following treatment groups: Trial 1: Control, 1 ml sterile LB (CT1; n=5); 4 × 10 6 CFU/ml E. coli-pAK1-lux (n=5); or 20.0 × 10 6 CFU/ml E. coli-pAK1-lux (n=5); Trial 2: Control, 1 ml sterile LB (CT2; n=5); 1.2 × 10 6 CFU/ml E. coli-pAK1-lux (n=5); or 5.6 × 10 6 CFU/ml E. coli-pAK1-lux (n=5). The inoculation dose resulted from pre-inoculation photon emissions calculated for utilization in the companion study which details the in vivo photonic imaging of this E. coli strain within the uterus of the ewe [16]. It was identified that 72.7% (8/11 for Trials 1 and 2 combined) of ewes were inoculated intra-cotyledon, while 27.3% (3/11 for Trials 1 and 2 combined) were inoculated intra-amniotic.…”