Response of the chick embryo to live and heat-killed Campylobacter jejuni injected into the yolk sac.

Graded doses of live and heat-killed cells of Campylobacter jejuni were injected into the yolk-sac of 5-day-old chick embryos, and the 50% lethal dose (LD50) was determined 7 days later. A strain dependent virulence was seen. In the diluted series of cultures the LD50 values for live campylobacter ranged from 10(6) c.f.u. beyond the last dilution showing growth, that is to less than one organism per embryo. When the 22 strains were tested as heat-killed cells, the chick embryo LD50 values retained the same relative order of toxicity obtained with viable cells, but the LD50 values were increased by +1 to +4 log units. Heat-killed cells from strains known to be invasive, but non-toxigenic, were still lethal for the embryos, suggesting that viability was not solely necessary for virulence. Semi-pure lipopolysaccharide from a non-virulent strain of C. jejuni was not toxic to the embryos, but semi-pure and ultracentrifuge-purified lipopolysaccharide from the most lethal campylobacter strains gave LD50 values in the order of 3.0 micrograms lipopolysaccharide per ml (0.6 microgram per embryo) in the yolk-sac assay. No relationship between serotype and lethality was seen. Injection into the yolk-sac appears to be an easy, rapid and reproducible in vivo assay of the virulence of C. jejuni.

Effect of egg storage upon the survival of Campylobacter jejuni in laboratory-infected fertile poultry eggs.

Fertile eggs were infected by Campylobacter jejuni in the laboratory by a temperature differential method of inoculation, which resulted in up to 10% of the hatched birds carrying C. jejuni in the intestine. When infected eggs were stored for 5 1/2 days before incubation, the infection rate of the eggs had decreased to 20% or less when set, and no infected chicks were hatched. Inoculation of eggs after 8 days in storage also failed to yield infected chicks. In all cases, the hatch ratio was no different from that of uninfected control eggs.

Laboratory infection of chicken eggs with Campylobacter jejuni by using temperature or pressure differentials.

Fertile chicken eggs were infected in our laboratory with Campylobacter jejuni suspensions by using temperature or pressure differential methods of inoculation. After 2 days of incubation, over 90% of the eggs carried C. jejuni when iron was present in the inoculum. This percentage declined rapidly until by day 8, less than 10% of the eggs were detectably infected. However, up to 11% of hatched, healthy chicks carried C. jejuni in their intestinal tracts. The isolated organisms were of the same serotype as the initial inoculum. C. jejuni was recovered without difficulty when the intestinal tracts of chicks were enriched, but recovery from early dead-in-shell or infertile eggs was poor. This poor recovery and the rapid decline of C. jejuni after 2 days of egg incubation suggest that the vibrio is sensitive to some part of the incubating egg or to the temperature of prolonged incubation. It was impossible to predict which eggs would yield infected chicks on the basis of the number of organisms taken up by each egg, and no correlation existed between the number of organisms taken up and the efficiency of the hatch, i.e., the hatch ratio. If iron was omitted from the inoculum broth, the egg infection rate at day 2 was lower.

Escherichia coli variants for gas and indole production at elevated incubation temperatures.

Two strains of Escherichia coli were subjected to heat and cold-storage treatments to determine the stability of the fecal E. coli characteristics of gas production from lactose and indole production at elevated incubation temperatures. No variants were detected with repeated sublethal heat treatment. A high incidence of variants was observed with extended cold storage of the organisms in liquid and semisolid media, especially with poor nutrient composition, and in the absence of cryoprotective agents. The indole characteristic at elevated temperature was more stable than the production of gas from lactose. The critical temperature at which both gas production from lactose and the indole characteristic were lost was 44.5 degrees C. It appeared that the variants resulted from increased temperature sensitivity of the formic hydrogen lyase and tryptophanase enzymes, respectively.