Avian combination-vaccine against e.coli and salmonella
A technology for Escherichia coli and poultry, applied in the field of combined poultry vaccines
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Embodiment 1
[0029] Construction of Escherichia coli Mutant with AROA Gene Deletion
[0030] I) Receptor
[0031] The parental organism was an avian E. coli isolate isolated from a clinical case of avian colibacillosis submitted to the Veterinary Laboratories Agency (VLA), Addlestone, Surrey, UK in 1995 and serotyped at the VLA. The parental strain was selected for its characteristics of persistence, invasiveness, persistence and pathogenicity in day-old SPF chickens, and antibiotic susceptibility in vitro. Recipient strains were generated by combining transformed donor (E. coli K12 S17λpir carrying PNG101, deletion of aroA carrying 100 bp) and wild-type parental strain (wild-type E. coli isolate EC34195).
[0032] II) Missing features
[0033] The aroA gene, which encodes 3-phosphoenolpyruvylshikimate-5-phosphate synthase, an enzyme of the general aromatic biosynthetic pathway, is located at the end of the promoter of serC adjacent to the serC-aroA operon. Deletion of the aroA gene in ...
Embodiment 2
[0047] Preparation of master species
[0048] Escherichia coli aroA-strain (constructed in Example 1) was cultured once on a typical soybean agar plate and then three times in a typical soybean broth. Cultures were distributed into glass vials, sealed and lyophilized.
Embodiment 3
[0050] Evaluation of the efficacy of combinations of E. coli gene-deleted mutant microorganisms and Salmonella typhimurium gene-deleted mutant microorganisms against E. coli and Salmonella infection in chickens
[0051] In the evaluation, the Escherichia coli gene-deleted mutant microorganism used was the Escherichia coli aroA-strain described in Examples 1 and 2, and the used Salmonella typhimurium gene-deleted mutant microorganism was prepared as described in US 6,231,871 B1 Salmonella typhimurium STM-1.
[0052] For evaluation, 103 SPF white Leghorn chickens, regardless of sex, were divided into 5 groups, 2 groups of 25 birds each, 2 groups of 24 birds each and 1 control group (no vaccination, no change) of 5 birds. Chickens with their wings removed were forced to be placed in designed isolation chambers. Each experimental group was housed in two isolation rooms with 12 or 13 chickens in each isolation room.
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