Integrons and antibiotic resistance in phylogenetic group B2 Escherichia coli.

It has been reported that Escherichia coli B2 phylogenetic group strains are more susceptible to antibiotics, especially to quinolones, and tend to carry less integrons than other phylogenetic groups in commensal environments. To gain a better understanding of the relationships between antibiotic resistance, integrons, and phylogenetic groups in an environment with high antibiotic selective pressure, we compared these characteristics in three selected groups of urinary tract infection E. coli isolated in a university hospital (G1, G2, and G3). The isolates were fully susceptible to antibiotics, resistant to amoxicillin and cotrimoxazol, or resistant to amoxicillin, cotrimoxazol, and nalidixic acid in the G1, G2, and G3 group, respectively. The prevalence of B2 isolates was significantly lower in the most resistant G3 group (22.6%) than in susceptible G1 (57.8%, p < 0.001) and G2 groups (50%, p < 0.01). In contrast, the prevalence of B2 isolates was not significantly different between G1 and G2 groups. The prevalence of integrons was nil in G1 isolates but very high in G2 (94.3%) and G3 (87.5%) isolates, and integrons were equally distributed among the phylogenetic groups. We propose a step-by-step mechanism for the emergence of antibiotic resistance in E. coli. Under very low selective pressure, resistance emerges without integrons. When the antibiotic pressure increases, quinolone and integron-mediated resistance occurs outside phylogenetic group B2. With strong antibiotic selective pressure, integrons are highly prevalent and widespread regardless of the phylogenetic group.

Characteristics of human intestinal Escherichia coli with changing environments.

To investigate if the characteristics of human intestinal Escherichia coli are changing with the environment of the host, we studied intestinal E. coli from subjects having recently migrated from a temperate to a tropical area. We determined the phylogenetic group, the prevalence of the antibiotic resistance, the presence of integrons and the strain diversity in faecal isolates from 25 subjects originally from metropolitan France and expatriated to French Guyana. These characteristics were compared with those of 25 previously studied Wayampi Amerindian natives of French Guyana and from 25 metropolitan French residents. The three groups of subjects were matched for age and sex, had not taken antibiotics for at least 1 month, nor had been hospitalized within the past year. In all, the characteristics of intestinal E. coli from Expatriates were intermediate between those found in residents from metropolitan France and those found in natives of French Guyana. Prevalence of carriage of resistant Gram-negative bacteria in Expatriates was intermediate between French residents and Wayampi as were the prevalence of integrons in E. coli (12.3% versus 16.3% and 7.8% respectively), and the intra-host diversity of E. coli (2.3 strains/subject versus 1.9 and 3.1, respectively); lastly, in Expatriates, the prevalence of carriage of phylogenetic group B2 strains was lower than in French residents (16% versus 56%, P = 0.005), while carriage of phylogenetic group A strains was lower than in Wayampi (56% versus 88%, P = 0.03). Our results suggest that the composition of the commensal intestinal flora of humans is not static but changes dynamically in response to new environmental conditions.

Polyvalent 23 epitope polysaccharide pneumonia vaccine induced effective protection through strain-adapted effector mechanisms as demonstrated by the different cytokine responses in mice challenged with two different strains of Streptococcus pneumoniae.

We used a Balb/c mouse model of pneumococcal pneumonia to investigate the protection mechanisms induced by immunization with a polyvalent 23 epitope polysaccharide pneumonia vaccine. Groups of mice were injected x 4 times s.c. within one month, with this vaccine preparation. Mice were subsequently challenged at day 45, with a lethal, intratracheal inoculum of two strains of Streptococcus pneumoniae - either a highly virulent and strongly immunogenic serotype 3 strain (P4241), or a less virulent and weakly immunogenic serotype 19F strain (P15986). The intratracheal S. pneumoniae challenge-induced lethality, antibody response, bacterial clearance, and cytokine secretions were monitored to analyze the strain-adapted effector mechanisms. Pulmonary levels of TNFalpha, IL-6, IL-1 beta, MIP-1 alpha, KC, MCP-1/JE and MIP-2 cytokines were determined up to 48 hours post-infection. Survival rates were 82% and 100% among vaccinated animals challenged at day 45 with P4241, and P1598 mice respectively, and 0% in non-vaccinated mice (p<0.001). Survival was associated with a rapid bacterial clearance from blood and lungs, which similar for the two strains. Immunization induced a serotype-specific antibody response. Kinetics of the cytokine profile in the lung following intratracheal inoculation with the 4241 strain was different in animals vaccinated 45 days previously, compared to naïve, control mice. Generally speaking the bacterial-induced inflammatory cytokine response induced with the 4241 strain was much weaker in vaccinated animals than in control mice. The only cytokines showing a greater increase in vaccinated mice compare to control animals were IL-1 beta, KC and MCP-1. Production of TNFalpha and IL-6 was lower in vaccinated animals than in controls. At variance with the previous bacteria strain-induced cytokine profile, infection with the P15986 strain induced a strong inflammatory response, with a substantial increase in all the cytokine tested, which was similar in vaccinated and in naïve, control animals, except for MIP-1 alpha, which was the only mediator significantly more produced by vaccinated animals than by naïve, control mice following P15986 infection. The distinct cytokine profiles, which were observed in this study depending upon the two strains of S. pneumoniae used for challenge, demonstrated that protection against each strain was obtained through a different defence strategy.

Streptococcus pneumoniae strain-dependent lung inflammatory responses in a murine model of pneumococcal pneumonia.

OBJECTIVE: The inherent properties of an invading bacterium may influence the cytokine profile that is ultimately produced. We determined the alterations in proinflammatory (TNF-alpha, IL-1, and IL-6) and anti-inflammatory cytokine (IL-10) expressions in lung tissues within the first 48 h after infection in mice with pneumonia induced by direct intratracheal inoculation of five different pneumococcal strains. DESIGN: Experimental murine model of Streptococcus pneumoniae pneumonia. SUBJECTS: Female BALB/cby mice aged 8-10 weeks. INTERVENTIONS: Five S. pneumoniae clinical isolates were used in this study. The strains included two serotype 3 strains (P4241 and P30606), two serotype 6 strains (P26772 and P23477), and one serotype 19 strain (P15986). The trachea of anesthetized animals was cannulated via the mouth with a blunt needle, and 50 micro l bacterial suspension of two different inocula (their respective 100% lethal inoculum and the same 10(5) CFU/mouse inoculum of S. pneumoniae strains) were instillated. At predetermined times after pneumococcal infection, i.e., time 0 (preinfection) and 2, 4, 6, 12, 24, and 48 h postinfection in experimental groups, lung tissues were sampled from groups of three mice to quantify lung pro- and anti-inflammatory mediators. The experiments were repeated at least three times. RESULTS: Pneumonia induced by five different pneumococcal isolates resulted in pronounced differences in the local pro- and anti-inflammatory profiles. For example, with a 100% lethal inoculum of S. pneumoniae, the extent and timing of TNF-alpha expression varied greatly among strains, ranging from 2,643 to 10,022 pg/g and from 4 to 48 h, respectively. Moreover, TNF-alpha productions within 48 h postinfection measured by the 48 h area under the curve were differed significantly, ranging from 59,700 to 275,825. These different profiles were not serotype dependent. Comparable results were obtained when IL-1, IL-6, and IL-10 expressions in lung tissues were studied. CONCLUSIONS: These findings confirm that the production of the pro- and anti-inflammatory mediators are critically dependent not only upon the different species of bacteria used to establish the experimental infection but also upon the different strains of a specific bacterial species used, i.e., S. pneumoniae in this study. These substantially different host responses were not serotype dependent. Moreover, the profile of lung pro-and anti-inflammatory cytokines within 48 h postinfection, at least in this pneumonia model, was not related to outcome of animals.

Early bacterial and inflammatory responses to antibiotic therapy in a model of polymicrobial peritonitis in rats.

OBJECTIVE: To assess the consequences of different more or less selective treatments on the microbiological and inflammatory responses within the peritoneum. METHODS: The early effects of various antibiotic regimens were evaluated in a model of polymicrobial peritonitis with specifically prepared organisms. Six regimens (amoxycillin plus gentamicin, pefloxacin, ornidazole, pefloxacin plus ornidazole, imipenem and imipenem plus gentamicin) were evaluated at 24 h and 3 days in a non-fatal model of peritonitis in rats achieved by implantation of a capsule containing Escherichia coli, Bacteroides fragilis and Enterococcus faecalis. RESULTS: Therapies that disregarded several organisms were associated with persistence of the strains and an increased peritoneal inflammatory response within the peritoneum. In contrast, therapies active against Enterobacteriaceae and anaerobes were associated with decreases of all the inoculated organisms and a smaller inflammatory response. CONCLUSION: Therapies that disregarded the microorganisms implicated in peritoneal infection are associated with delayed bacterial eradication. The persistence of these organisms within the peritoneal fluid might be involved in prolonged peritoneal inflammation. Although it disregards enterococci, the standard therapy, represented by therapy against Enterobacteriaceae and anaerobes, demonstrates satisfactory effects towards all the inoculated organisms. This apparent contradiction could be related to mechanisms of bacterial synergy.