Biology 216 - Lect 21 Salmonella I. Salmonella - range from gastroenteritis to enteric fevers (typhoid) A. Bact - Tribe Salmonellae (Most H2S+) - Salmonella (man and animals): Arizona (now Salmonella - reptiles and man); Citrobacter (animals and soil) 1. Classification schemes - Kauffmann/White - O-Ag (2500 species) Ewing and CDC - clinical basis (3 species) - S. enteritidis, S. cholerae-suis, S. typhi (man) Kauffmann O-Ag as serovar: Ex. S. enteritidis serovar pullorum DNA homology - One species, 6 subgroups (no correlation with clinical) Zinsser uses one species: S. enterica subspecies xxxx 2. Biochemical - Lac-, H2S+, very bile resistant (media and gall bladder) 3. Other - Vi capsule (S. typhi - intracellular survival and antiphagocytic) Rough vs smooth associated with virulence (resist complement mediated lysis) Enterotoxin - CT related by a few strains. S. enteritidis genetics - Ames test. Range of hosts: S. typhi (man); others have animal reservoirs. B. Pathogenesis virulence: S. enteritidis < S. cholerae-suis < S. typhi Genetics of cellular invasion well characterized. Chromosomal (invasion) and plasmid (plasmid enhance virulence) 1. Gastroenteritis - "stomach flu" - food poisoning (infection) Estimated 2 million/yr in US a. Reservoir - animal GI (normal); fowl, cattle, turtles (300K in 1970), dogs/cats (1 %) Subspecies often reflects reservoir: Ex. S. enteritidis serovar pullorum (chicken) Subspecies vary in virulence: Ex. serovar Dublin Foods contaminated during processing: chicken, milk, eggs. Classic cutting board. b. Ingestion:ID50 approx 10(6) - 10(9), colonization (sm and lg);invasion of mucosal cells c. Symptoms - 8 - 24 hr incubation, fever, headache, myalgia, diarrhea (mild) d. Blood, mucus, pus rare; Self limiting; may secrete for months or years (1 % of cases). 2. Enteric fevers (typhoid fever is more severe) - invasion not restricted to GI. Animal models: mouse and tissue culture. a. Reservoir - man only (S. typhi) - controllable by sewage treatment and sanitation. 1900 - 300K cases in US (35K deaths); Today 400 cases/yr - not endemic. b. Transmission - Contaminated food (handlers as carriers; 3 % lifetime carriers) Typhoid Mary (Mary Mallon) - 1900; cook in NY city; offered gall bladder operation, refused, so imprisoned 3 years - 200 cases traced to her. c. Colonization - ID50 approx 10(6); Multiply -> M-cells -> Macs -> messenteric nodes (Peyer's) Note: Macs attached to M-cells stimulate T and B -> IgA -> stimulated B cells at other mucosal surfaces. Induced phagocytosis - ruffling cell surface by actin rearrangement. d. Invasion - Phagocytosed by Macs (survive ie CMI important) -> liver,spleen, skin,bone e. Disease - Gradual onset (2 - 4 weeks); High fever (104 F); delerium, Exhaustion, splenomegaly, rose spots (thick, raised, upper abdomen): all LPS mediated. f. Liver to gall bladder - reseeding small intesting -> diarrhea, intestinal perforation. Colonize gall bladder (survive bile), scarred tissue, stones, 3 % lifetime carrier. g. Complications - spleen rupture, abscess in organs, peritonitis, shock (LPS) GI perforation. h. Duration - 3 - 4 weeks, CMI, 10 % fatality w/o treatment, 10 % relapse, 3 % carriers. C. Diagnosis - Non-typhoidal vs typhoidal 1. Non-typhoidal - High, moderate and enrichment. Feces in acute, but few later, ID. 2. Typhoid - History (travel) a. Culture - Fecal - early and 3rd week (after reseeding); Blood, urine, rose spots during acute infection (weeks 1 - 4). b. ID - biochemical; Serology (Widal agglutination) D. Treatment - Non-typhoidal - self limit, rehydrate, pain relief, antibiotics (little effect, may prolong). Typhoid - Fever control, fluids (sweating), antiinflammation (LPS), antibiotics. E. Prevention and control 1. Sanitation control - water, sewage, Typhoid Carrier Registry, Bladder removal. 2. Prevent recontamination of foods, refrigerate. 3. Vaccine - Typhoid Fever Field trials for new vaccine in endemic countries - Nepal Three vaccines: killed, Vi, ty21 1. Old killed bacteria (phased out)- fever, flu-like 2. Vi capsule - some protection; Protein capsule conjugate - better. 3. Live attenuated (ty21a) in capsule; Auxotroph, penetrate, replicate, die, CMI ty21a as delivery for cloned vaccines to other enteropathies: rotavirus, LT-B, ST.... In trials this has worked to protect from Shigella, not cholera (cloned O-Ag). Genes for sIgA against human sperm (works in mice) for birth control