Biology 216 - Lect 28 - Bacillus I. Bacillus - Gm+, sporeforming, large aerobic or facultative rods, rough colonies Many produce antibiotics: Bacitracin from B. licheniformis Viable spores recovered from amber. Examples: B. subtilis - cloning host B. stearothermophilus - autoclave test (resistant and grows best at 80 C) B. thurengiensis - spore associated protein (parasporal protein crystal) Dissolve lepidopteran gut -> gut liquify -> blood -> paralysis Cloned to Ps and to certain plants B. sphaericus (effective vs anopheles mosquito) B. cereus - food intoxication, large number in food, spore assoicated, diarrhea... II. B. anthracis - anthrax A. Bacteriology - boxcar shape, must differentiate from B. cereus Historically - Koch's postulates; Biological warfare 1. Spores - Persist in fields for years (record is 60 yrs) - growth in carcass of victim Remove carcass, turn under soil, drop below ID50 Spores in dead tissue (special media needed to see in vitro) 2. Capsule - protein, single serotype, antiphagocytic 3. Toxin - 3 components - none alone are toxic; injected together simulates disease - plasmid encoded. Lethal factor, edema factor, protective antigen a. PA + LF -> CNS respiratory failure, death b. PA + EF -> Edema (adenylate cyclase) PA probably helps other gain access to cells Death by systemic buildup of toxin; Edema, CNS and respiratory failure, hypovolemic, renal.. Toxins plasmid encoded: Pasteur cured by accident at 42C -> live attenuated vaccine. B. Pathogenesis - zoonosis (man gets from diseased animal) 1. Disease in lower animals - rare in US (occassional outbreak) - economic loss in S. Amer, Afric, Asia Outbreaks by tannery (imported hides, elephant ivory) -> human or drain to fields Herbivore - ingest spores from soil B. anthracis penetrates intestinal mucosa -> bloody diarrhea, septicemia, 80 % fatal Death in hours from first symptoms; capillaries of organs engorged with bacteria Rarely see bacteria in blood with disease - this is exception; Toxin -> rapid death 2. Disease in humans - accidental host Transmission - contact with infected parts (hides, wool, bone); Rare in US due to animal control 3 major disease manifestations based upon route of entry: a. Cutaneous - single site (95 % of cases in US) Spores through cut, germinate, multiply, itch, no pain Eschar - local edema, induration, dark necrotic center ulcerates, malignant pustule (pathognomonic) Spontaneous healing, loosens, falls off, scar b & c. Pulmonary and GI - More severe, less common - similar to animal disease. Considered in biological warfare (US opposes, since cheap and easy for poor country) C. Diagnosis 1. Clinical - Eschar (pathognomonic), history of exposure, occupation 2. Lab - Direct exam (Gm stain, Fl-Ab) Culture and ID (specimen is eschar, sputum) -> BAP, distinguish from B. cereus (biochems) D. Treatment 1. Antibiotics immediately since rapid progression 2. Excission of lesion contraindicated -> spread E. Control 1. Discard infected animal carcasses, turn soil under 2. Decontaminate imported animal materials (ethylene oxide gas) 3. Vaccine for animals - effective, but not universally used. Live attenuated + toxoid of PA or spore vaccine for animals.