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Lecture PowerPoint to accompany Foundations in Microbiology Sixth Edition Talaro Chapter 20 The Gram-Negative Bacilli of Medical Importance Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. • Large, diverse group of non-spore-forming bacteria • Wide range of habitats – large intestines (enteric), zoonotic, respiratory, soil, water • Most are not medically important; some are true pathogens, some are opportunists. • All have a lipopolysaccharide outer membrane of cell wall – endotoxin. 2 3 Aerobic Gram-Negative Nonenteric Bacilli • Pseudomonas and Burkholderia – an opportunistic pathogen • Brucella and Francisella – zoonotic pathogens • Bordetella and Legionella – mainly human pathogens • Alcaligenes – opportunistic pathogen 4 Pseudomonas: The Pseudomonads • Small Gram-negative rods with a single polar flagellum • Free living – primarily in soil, sea water, and fresh water; also colonize plants and animals • • • • • Important decomposers and bioremediators Frequent contaminants in homes and clinical settings Use aerobic respiration; do not ferment carbohydrates Produce oxidase and catalase Many produce water soluble pigments. 5 6 Pseudomonas aeruginosa • Common inhabitant of soil and water • Intestinal resident in 10% normal people • Resistant to soaps, dyes, quaternary ammonium disinfectants, drugs, drying • Frequent contaminant of ventilators, IV solutions, anesthesia equipment • Opportunistic pathogen 7 Pseudomonas aeruginosa • Common cause of nosocomial infections in hosts with burns, neoplastic disease, cystic fibrosis • Complications include pneumonia, UTI, abscesses, otitis, and corneal disease • Endocarditis, meningitis, bronchopneumonia • Grapelike odor • Greenish-blue pigment (pyocyanin) • Multidrug resistant • Cephalosporins, aminoglycosides, carbenicillin, polymixin, quinolones, and monobactams 8 9 Other Gram-Negative Aerobic Rods • Genera Burkholderia, Acinetobacter, Stenotrophomonas • Similar to pseudomonads • Wide variety of habitats in soil, water, and related environments • Obligate aerobes; do not ferment sugars • Motile, oxidase positive • Opportunistic 10 Burkholderia • Burkholderia cepacia – active in biodegradation of a variety of substances; opportunistic agent in respiratory tract, urinary tract, and occasionally skin infections; drug resistant • B. pseudomallei – generally acquired through penetrating injury or inhalation from environmental reservoir; wound infections, bronchitis and pneumonia, septicemia 11 Acinetobacter and Stenotrophomonas • Acinetobacter baumanii – nosocomial and community acquired infections; wounds, lungs, urinary tract, burns, blood; extremely resistant – treatment with combination antimicrobials • Stenotrophomonas maltophilia – forms biofilms; contaminant of disinfectants dialysis equipment, respiratory equipment, water dispensers, and catheters; clinical isolate in respiratory soft tissue, blood ,CSF; high resistance to multidrugs 12 Brucella and Brucellosis • Tiny Gram-negative coccobacilli • 2 species: – Brucella abortus (cattle) – Brucella suis (pigs) • Brucellosis, malta fever, undulant fever, and Bang disease – a zoonosis transmitted to humans from infected animals • Fluctuating pattern of fever –weeks to a year • Combination of tetracycline and rifampin or streptomycin • Animal vaccine available • Potential bioweapon 13 Francisella tularensis and Tularemia • Causes tularemia, a zoonotic disease of mammals endemic to the northern hemisphere, particularly rabbits • Transmitted by contact with infected animals, water and dust or bites by vectors • Headache, backache, fever, chills, malaise and weakness • 10% death rate in systemic and pulmonic forms • Intracellular persistence can lead to relapse • gentamicin or tetracycline • Attenuated vaccine • Potential bioterrorism agent 14 Bordetella pertussis • Minute, encapsulated coccobacillus • Causes pertussis or whooping cough, a communicable childhood affliction • Acute respiratory syndrome • Often severe, life-threatening complications in babies • Reservoir – apparently healthy carriers • Transmission by direct contact or inhalation of aerosols 15 Bordetella pertussis • Virulence factors – receptors that recognize and bind to ciliated respiratory epithelial cells – toxins that destroy and dislodge ciliated cells • Loss of ciliary mechanism leads to buildup of mucus and blockage of the airways. • Vaccine – DTaP- acellular vaccine contains toxoid and other Ags 16 17 Alcaligenes • Live primarily in soil and water • May become normal flora • A. faecalis – most common clinical species – isolated from feces, sputum, and urine – occasionally associated with opportunistic infections – pneumonia, septicemia, and meningitis 18 Legionella pneumophila and Legionellosis • Widely distributed in water • Live in close association with amebas • 1976 epidemic of pneumonia afflicted 200 American Legion members attending a convention in Philadelphia and killed 29 • Legionnaires disease and Pontiac fever • Prevalent in males over 50 • Nosocomial disease in elderly patients • Fever, cough, diarrhea, abdominal pain, pneumonia fatality rate of 3-30% • Azithromycin 19 20 Enterobacteriaceae Family • Enterics • Large family of small, non-spore-forming Gram-negative rods • Many members inhabit soil, water, decaying matter, and are common occupants of large bowel of animals including humans. • Most frequent cause of diarrhea through enterotoxins • Enterics, along with Pseudomonas sp., account for almost 50% of nosocomial infections. 21 22 • Facultative anaerobes, grow best in air • All ferment glucose, reduce nitrates to nitrites, oxidase negative, and catalase positive. • Divided into coliforms (lactose fermenters) and non-coliforms (non-lactose fermenters) • Enrichment, selective and differential media utilized for screening samples for pathogens 23 24 25 Antigenic Structures and Virulence Factors Complex surface antigens contribute to pathogenicity and trigger immune response: • H – flagellar Ag • K – capsule and/or fimbrial Ag • O – somatic or cell wall Ag – all have • Endotoxin • Exotoxins 26 27 Coliform Organisms and Diseases 28 Escherichia coli: The Most Prevalent Enteric Bacillus • Most common aerobic and non-fastidious bacterium in gut • 150 strains • Some have developed virulence through plasmid transfer, others are opportunists. 29 Pathogenic Strains of E. coli • Enterotoxigenic E. coli causes severe diarrhea due to heat-labile toxin and heat-stable toxin – stimulate secretion and fluid loss; also has fimbriae • Enteroinvasive E. coli causes inflammatory disease of the large intestine. • Enteropathogenic E. coli linked to wasting form infantile diarrhea • Enterohemorrhagic E. coli, O157:H7 strain, causes hemorrhagic syndrome and kidney damage; ID 100 cells 30 Escherichia coli • Pathogenic strains frequent agents of infantile diarrhea – greatest cause of mortality among babies • Causes ~70% of traveler’s diarrhea • Causes 50-80% UTI • Coliform count - indicator of fecal contamination in water 31 Other Coliforms Clinically important mainly as opportunists • Klebsiella pneumoniae– normal inhabitant of respiratory tract, has large capsule, cause of nosocomial pneumonia, meningitis, bacteremia, wound infections and UTIs • Enterobacter sp. – UTIs, surgical wounds • Serratia marcescens – produces a red pigment; causes pneumonia, burn and wound infections, septicemia and meningitis • Citrobacter sp. – opportunistic UTIs and bacteremia 32 Noncoliform Lactose-Negative Enterics • Proteus, Morganella, Providencia • Salmonella and Shigella 33 Opportunists: Proteus and Its Relatives Proteus, Morganella, Providencia – ordinarily harmless saprobes in soil, manure, sewage, polluted water, commensals of humans and animals – Proteus sp. - swarm on surface of moist agar in a concentric pattern – involved in UTI, wound infections, pneumonia, septicemia, and infant diarrhea – Morganella morganii and Providencia sp. involved in similar infections • All demonstrate resistance to several antimicrobials. 34 35 Salmonella and Shigella • Well-developed virulence factors, primary pathogens, not normal human flora • Salmonelloses and Shigelloses – some gastrointestinal involvement and diarrhea but often affect other systems 36 Typhoid Fever and Other Salmonelloses • Salmonella typhi – most serious pathogen of the genus; cause of typhoid fever; human host • S. cholerae-suis – zoonosis of swine • S. enteritidis – includes 1,700 different serotypes based on variation on O, H, and capsular antigen • Flagellated; ferments glucose • Resistant to chemicals –bile and dyes 37 Typhoid Fever • Bacillus enters with ingestion of fecally contaminated food or water; occasionally spread by close personal contact; ID 1,000-10,000 cells • Asymptomatic carriers; some chronic carriers shed bacilli from gallbladder • Bacilli adhere to small intestine, cause invasive diarrhea that leads to septicemia • Treat chronic infections with chloramphenicol or sulfa-trimethoprim • 2 vaccines for temporary protection 38 39 Animal Salmonelloses • Salmonelloses other than typhoid fever are called enteric fevers, Salmonella food poisoning, and gastroenteritis. • Usually less severe than typhoid fever but more prevalent • Caused by one of many serotypes of Salmonella enteritidis; all zoonotic in origin but humans can become carriers – cattle, poultry, rodents, reptiles, animal and dairy products – fomites contaminated with animal intestinal flora 40 Shigella and Bacillary Dysentery • • • • Shigellosis – incapacitating dysentery S. dysenteriae, S. sonnei, S. flexneri and S. boydii Human parasites Invades villus of large intestine, can perforate intestine or invade blood • Enters Peyer’s patches instigate inflammatory response; endotoxin and exotoxins • Treatment – fluid replacement and ciprofloxacin and sulfa-trimethoprim 41 42 The Enteric Yersinia Pathogens • Yersinia enterocolitica – domestic and wild animals, fish, fruits, vegetables, and water – bacteria enter small intestinal mucosa, some enter lymphatic and survive in phagocytes; inflammation of ileum can mimic appendicitis • Y. pseudotuberculosis – infection similar to Y. enterocolitica, more lymph node inflammation 43 Nonenteric Yersinia pestis and Plague • Nonenteric • Tiny, Gram-negative rod, unusual bipolar staining and capsules • Virulence factors – capsular and envelope proteins protect against phagocytosis and foster intracellular growth – coagulase, endotoxin, murine toxin 44 45 Yersinia pestis • Humans develop plague through contact with wild animals (sylvatic plague) or domestic or semidomestic animals (urban plague) or infected humans. • Found in 200 species of mammals – rodents, without causing disease • Flea vectors – bacteria replicates in gut, coagulase causes blood clotting that blocks the esophagus; flea becomes ravenous 46 47 Pathology of Plague • ID 3-50 bacilli • Bubonic – bacillus multiplies in flea bite, enters lymph, causes necrosis and swelling called a bubo in groin or axilla • Septicemic – progression to massive bacterial growth; virulence factors cause intravascular coagulation subcutaneous hemorrhage and purpura – black plague • Pneumonic – infection localized to lungs, highly contagious; fatal without treatment 48 • Diagnosis depends on history, symptoms, and lab findings from aspiration of buboes. • Treatment: streptomycin, tetracycline or chloramphenicol • Killed or attenuated vaccine available • Prevention by quarantine and control of rodent population in human habitats 49 Oxidase-Positive Nonenteric Pathogens • • • • • • Pasteurella multocida Haemophilus influenzae H. aegyptius H. ducreyi H. parainfluenzae H. aphrophilus 50 Pasteruella multocida • Zoonotic genus; normal flora in animals • Opportunistic infections • Animal bites or scratches cause local abscess that can spread to joints, bones, and lymph nodes. • Immunocompromised are at risk for septicemia and complications. • Treatment: penicillin and tetracycline 51 Haemophilus • Tiny Gram-negative pleomorphic rods • Fastidious, sensitive to drying, temperature extremes, and disinfectants • None can grow on blood agar without special techniques – chocolate agar. • Require hemin, NAD or NADP • Some species are normal colonists of upper respiratory tract or vagina (H. aegyptius, H. parainfluenzae, H ducreyi). • Others are virulent species responsible for conjunctivitis, childhood meningitis, and chancroid. 52 Haemophilus • H. influenzae – acute bacterial meningitis, epiglottitis, otitis media, sinusitis, pneumonia, and bronchitis – subunit vaccine Hib • H. aegyptius –conjunctivitis, pink eye • H. ducreyi – chancroid STD • H. parainfluenzae and H. aphrophilus – normal oral and nasopharyngeal flora; infective endocarditis 53 54