Virulent Escherichia coli strains for chicks bind fibronectin and type II collagen.

125I-fibronectin and 125I-collagen (type II) binding was detected in Escherichia coli strains isolated from chickens and poults. High fibronectin binding-strains also bind the 29 kD aminoterminal fragment of fibronectin. Binding properties in strain CK28 were partially characterized. The highest binding of 125I-fibronectin and 125I-collagen for strain CK28 was obtained with bacteria grown at 33 degrees C. Binding of 125I-fibronectin, its 125I-29 kD fragment, and 125I-collagen, was very rapid, reaching a maximum in 5 min. Binding of 125I-fibronectin and 125I-collagen was considerably inhibited by preincubation of bacteria with unlabelled fibronectin and unlabelled type I collagen respectively, but not inhibited with human immunoglobulin G or bovine serum albumin. Inhibition experiments showed that the reversibility of 125I-fibronectin binding was estimated at approximately 50%, while reversibility for 125I-collagen binding was higher than 90%. Receptors for fibronectin, its 29 kD fragment, and collagen were released from the bacterial surface by treatment at different temperatures, and surface material released at 100 degrees C inhibited binding. There was cross-inhibition for both fibronectin and collagen binding when unlabelled fibronectin and unlabelled collagen were used as inhibitors, suggesting that binding receptors for both proteins may be closely located.

Association of virulence markers with animal pathogenicity of Escherichia coli in different models.

Employing chicken and several strains of mice, different routes (intraperitoneal, subcutaneous) of infections and isogenic pairs of strains, association of virulence markers with animal pathogenicity was studied in Escherichia coli. Mouse virulence of avian strains was less significant than the lethality for chicks of human strains. LD50 in various animals did not differ significantly. Strains with antigen K1 were more virulent for mice than their K1- derivatives. Loss of haemolysin (Hly), mannose resistant haemagglutinating capacity or antigen K5 less markedly decreased the virulence. As opposed to other virulence factors, increased virulence of K1+ strains could also be demonstrated in mouse sepsis assay based on bacterial counts in the liver. Loss of Hly alone did not influence the persistence in the liver, however, these strains killed less mice. Aerobactin acts together with other factors, it is not per se a virulence factor. In organotropic experiments 19 strains out of 36 belonging to serotypes O7:K1:H-, O18:K1:H-, O78:H- and spontaneously agglutinable K1+ cultures, caused ophthalmitis with purulent discharge, and 4 out of 22 strains that belonged to serotype O78:H- induced uncoordinated movement of mice. Because of its special organotropic affinity to the brain and as it caused two epidemics of meningitis among newborns in Hungary, serotype O78:H- has a special pathogenic property and differs from other O78 strains that were isolated in other countries.

Comparative infectivity for axenic and specific-pathogen-free chickens of O2 Escherichia coli strains with or without virulence factors.

Adhesion to epithelial respiratory cells, iron acquisition, and production of K1 polysaccharide capsules have been proposed as potential virulence factors of avian Escherichia coli. These factors were studied by inoculating groups of axenic or specific-pathogen-free (SPF) chickens intratracheally with O2 E. coli strains after previous challenge with a wild strain of infectious bronchitis virus (IBV). In all experiments, the association between IBV and an E. coli strain endowed with the three virulence factors previously mentioned resulted in the most severe pathological effects, as measured by mortality, weight gains, lesions, and reisolation of E. coli from internal organs. An E. coli strain devoid of virulence factors was able only to induce mild pathological effects restricted to the respiratory tract when combined with IBV. Both E. coli strains were more invasive in axenic chickens than in SPF chickens. These results confirm the probable involvement of the three factors studied in the pathogenic properties of avian E. coli. This model can be used to assess the role of virulence factors, by comparing pairs of positive and negative isogenic strains.

Presence and expression of aerobactin genes in virulent avian strains of Escherichia coli.

Virulent and nonvirulent isolates of avian Escherichia coli were tested for the presence of aerobactin genes by colony hybridization with a specific gene probe constructed from plasmid pABN1 (A. Bindereif and J. B. Neilands, J. Bacteriol. 153:1111-1113, 1983). Positive hybridization with the gene probe was highly correlated with virulence, as measured by the 50% lethal dose of the strains for chicks. Evidence for the expression of aerobactin genes in the virulent strains was obtained by demonstrating their susceptibility to cloacin DF13, which binds to the same receptor that binds aerobactin, and their ability to produce aerobactin, as revealed by cross-feeding the E. coli mutant WO987 (aroB fepA iuc iut+), which is unable to synthesize but capable of taking up aerobactin. We suggest that the production of aerobactin is involved in the virulence of avian septicemic E. coli.

Adhesive properties and iron uptake ability in Escherichia coli lethal and nonlethal for chicks.

Escherichia coli that are virulent for poultry usually result in a respiratory disease, which is frequently followed by a general infection. Adhesiveness of E. coli to epithelial cells and iron-uptake ability of E. coli could be involved in different steps of the disease. These properties were studied in 59 E. coli strains originating from poultry, with reference to lethality for day-old chicks. Adhesive properties were found in 64% of the lethal strains and in only 23% of the nonlethal strains. The ability to grow in limited iron conditions was strongly correlated with lethality. Fifty-two percent of the lethal E. coli strains, but none of the nonlethal strains, possessed both adhesive and iron-uptake abilities. It is suggested that these two properties play a role in the virulence of E. coli for poultry.

[Qualitative and quantitative analysis of the bacterial flora in the trachea of healthy chickens]. / Analyse qualitative et quantitative de la flore bactérienne dans la trachée du poulet sain.

The bacterial flora of the trachea of 17 to 38 day old chickens was quantitatively and qualitatively studied in two groups of animals from two different sources: an experimental disease-free flock and a commercial flock. In both groups of chickens, the aerobic flora consisted predominantly of Escherichia coli (E. coli), Streptococcaceae, Micrococcaceae and members of the genus Lactobacillus. Some differences between the two groups of animals were observed at the species level. Strictly anaerobic bacteria did not appear until the third week. No bacterial group seemed to predominate during the sampling period. The constant presence of Lactobacilli and E. coli in the trachea suggests that these organisms could play a prominent role in the bacterial ecosystem of the trachea.

Escherichia coli colonization of the trachea in poultry: comparison of virulent and avirulent strains in gnotoxenic chickens.

In chickens, virulent Escherichia coli strains express their pathogenicity in the respiratory tract. A quantitative comparison of tracheal colonization by virulent and avirulent E. coli was carried out in gnotoxenic chickens after intestinal implantation. Two-week-old axenic chicks reared in isolators were inoculated per os with various associations of identified E. coli strains. No clinical sign of disease was observed in any of the chicks, despite the presence of virulent strains in all the intestines and most of the tracheas. The virulent organism reached greater population sizes in the trachea and feces of monocontaminated chicks and of chicks contaminated simultaneously with a virulent and an avirulent strain. In holoxenic chicks, identified virulent and avirulent strains were outnumbered by the E. coli population of the intestinal flora previously established and could not be recovered from the tracheas of most chicks.