Vous êtes sur la page 1sur 13

FOS 4222/5225 Study Guide Exam 3- Revised Spring 2011 Sporulation Chapter 3 1.

Complete table below to characterize different stages of sporulation Spore Marker Morphology Resistance Sigma Other Stage Factor Associated Factors 0. Initiated by Vegetative cell None A quorum sensing 1. Axial filaments None A,H Master transcription factor SpoA-P 2. Differential Asymmetric Amylase, E, F protease, gene septum devides antibotics motility, expressions cell, and other alkaline genes phosphatase markers 3. Septum Mother cell K, G Double hydrolysis membrane membrane surrounds forespore and dehydrates 4. Synthesis of Cortex surrounds Spore acid K Coat proteins cortex, SASP the forespore, soluble produced, (spore acid forms from proteins cortex complete soluble peptidoglycan (SASP) proteins) between provide UV forespore resistance membranes 5. Spore coat Core dehydration Gamma coat complete forms on continues and radiation outside of metabolic and forespore dormancy begins chemical membrane resistance 6. Uptake of Ca, Dehydration Spores are Dipicolinic Mg, complete heat DPA acid is accumulate resistant and very specific to dipicolinicdpa dormant, spore formers, stain and it has two reisistant carboxyl groups on it, 7. Autolysins: Free spore spores released from mother cell

Sample questions Match most likely event association with the sporulation stage. A. Forespore surrounded by 2 membranes 1. __D._Stage II 2. __A._Stage III B. Spore coat formation C. Autolysin synthesis and mother cell lysis 3. __E_Stage IV 4. __B._Stage V D. Septum formation E. Synthesis of SASP, cortex 5. __C._Stage VII Matching (only one correct answer). 1. __D__Spore coat A. Heat resistance 2. B. SASPs (not irradiation resistant)B. DNA 3. E.Cortex C. RNA 4. __C__Germination D. Chemical resistance 5. __A.__Core E. Peptidoglycan spo genes encode sigma factors sigma factors provide promoter specificity to RNA polymerase resistance is due to a variety of factors: core dehydration and cations, SASPs, DAP, coat impermeability

Cronobacter(Enterobacter) sakazakii 1. What group of people is susceptible to this disease and why? a. High suseptability for infants, high fatality after causing meningitis in infants. Symptoms include poor feeding response, irritability, jaundice, grunting while breathing, unstable body temperature. C. sakazakii survive for years in dry formula b. 2. How does food become contaminated with this bug? a. contamination occurring post-production during dry blending Yersinia enterocolitica ask the professor about this one 1. What serovars are important to disease caused by this organism? How does geographic distribution factor into the prevalence of different serogroups? a. Yersinia entercolitica has the biotypes or serogroups that differ in metabolization of organic subtrates, O (LPS), H(flagella) and K (CPS) antigens. these are important because they provide usful tools to subdivide this species in a way that relates to pathologic significance. O:3 isolated from humans, from different parts of the world, there are serotypes that differ in europe 2. What foods are associated with this disease, and why is pork a particular problem? a. In beef or refrigerated foods, psychotrophic (beef, lamb, poultry, icecream, freaquently in milk)only pigs have biovar 4 serovar 0:3 (from tonsils, tongue, and gut, not in muscle) 3. What characteristics are common to acute disease? To autoimmune disease? a. Watery to mucoid diarrhea, fever, abdominal pain, bloody stools, ulcerations

b. Duration days to 3 weeks 4. Define sequelae, intrussusseption. a. Autoimmune sequelae follows intestinal symptoms, reactive arthritis, mostly O:3 serotype with HLA-B27 (molecular mimicry?), b. Intussussception a chronic disease, which is rare with ulceration progressing to perforation.. Can infect all tissue types: also cases nephritis carditis, thyroiditis, the slipping of one part of the intestine into the other 5. What does Yersinia have in common with Listeria in terms of growth conditions? a. They are both cold tolerant organisms, grow in RTE foods 6. Describe the pathogenesis: what is the targeted cell type for invasion, how does it enter, multiply, a. M cell in small intestine, intracellular and extracellular pathogen. Yersinia uses the Three type secretion system to disrupt and kill macrophages: b. Reduces brush borders on villi and forms abscesses in blood vessels, liver, spleen, c. 7. What cell type is damaged and how does this contribute to disease? a. Macrophage cells. USES TTSS: prevents phagocytosis and cytokine response, disrupts actin and induces apoptosis. b. 8. Why is this both an intracellular and extracellular pathogen? a. Because Yersinia has both TTSS,which is the extracellular pathogenic component enabling it to inject the toxins right into the m cell, and b. They are specialized enterocytes on intestinal surfaces 9. List Chromosomal vs. plasmid virulence factors and the function of each. a. Inv and Yst are on chromosome b. Plasmid encodes 10. How do Ca++ levels play a role in this disease? a. Low calcium signal: bacteria to stop growing, to produce yops (Yersinia outer membrane proteins), lower calcium removes plug 11. What are the specific roles of urease, M cells, crypts, Inv, YadAYstenterotyoxin and macrophagesin this disease? a. Heat stable entertoxin(Yst) Elevates cGMP and increases fluid and electrolyte imbalance in intestine b. Urease acid tolerance based on urease:catabolized urea to NH4 to raise cellular pH c. Crypts separation point between the microvilli d. Inv invasin is Yersinias way to get entry into m cell>> both Ivasin and adhesin recognize the m cells as surface proteins and bind to it. And deposit the Yops (secretion of the yops are also required to partner up with the chaperones, which are small acidic proteins that specifically bind only one partner e. YadA adhesin is Yersinias way to adhere to the Mcell> f. Macrophages 12. What group of proteins are required for construction of the Type Three Secretion System (TTSS) in Yersinia ?

a. TTSS translocon, attaches to the cell, the yopvirulon enters the target site by the Yersinia, the Yop effectors are translocated across the plasma membrane, guided by the translocatorsYopB, YopD, and LcrV. b. The Yopvirulon. When Yersinia are placed at 37C in a rich environment, the Ysc secretion channel is installed. Proteins YscD, -R, -S, -T, -U, and -V are localized in the inner membrane (IM), whereas YscC and YscP are exposed at the bacterial surface. Lipoprotein YscW stabilizes YscC. YscN belongs to the family of ATPases. A stock of Yop proteins is synthesized, and some of them are capped with their specific Syc chaperone. As long as there is no contact with a eukaryotic cell, a stop-valve, possibly made of YopN, TyeA, and LcrG, blocks the Ysc secretion channel. On contact with a eukaryotic target cell, the bacterium attaches tightly by interaction between its YadA and Invadhesins and -integrins, and the secretion channel opens. The Yops are then transported through the Ysc channel, and the Yop effectors are translocated across the plasma membrane, guided by the translocatorsYopB, YopD, and LcrV. c. The Ysc/Yopsattach to the host, which are the TTSS apparatus proteins, and the TTSS effector proteins are just the Yops. 13. What molecules are secreted by TTSS in these species and what do they do to macrophages? 14. How does temperature regulate virulence gene expression? a. When the virulence factor enters the gut lumen and its 37 degrees with a ph change, the bacterium enters a log phase and enterotoxins are produced. b. Enterocytes, lymphocytes with low calcium and the same temp the log phase begins producing ysc and yops Escherichia coli: 1. Describe characteristics that distinguish the different types of E. coli from each other. Which of these(EPEC, ETEC, EIEC, DAEC, EaggEC, EHEC O157:H7) use toxins, which are invasive, what are unusual disease symptoms? How about TTSS and other virulence factors, small vs. large intestine (hint: A table would probably be helpful) Toxins Symptoms Virulence Small vs. factors large intestine EPEC No toxic Watery Attaches Small effect, diarrhea, and more vomiting effacing effectors, and fever (AE) bundle lesions on forming intestinal pili (BFP) epithelial cells ETEC HeatWatery labile diarrhea, toxin (LT) mild and heat-

stable (ST) EIEC Invasive actin tails like shigella but no shigatoxin No toxin Mild Not diarrhea in invasive children Persistent diarrhea in children Symptoms Bloody Large due to diarrhea intestine shiga-like and toxin hemolytic STX uremic syndrome (HUS) Watery diarrhea



EHEC O157:H7

A. EPEC bfp, TTSS, secretes the translocatedintimin receptor (TIR) into host cell B. EHEC - shiga-toxin = Stx C. ETEC because it makes the LT and ST toxins D. Eagg/Ec

E. EIEC enteroinvasive E. Coil F. DAEChides from cytokine response

2. Check out Brett Findlays video http://www.hhmi.org/biointeractive/media/ecolilg.mov a. Studies for vaccine have to show they are very protective against diseases. 3. What are A/E lesions and are they only found with EPEC? a. Attaching and effacing lesions on epithelial wall b. EPEC and 4. Describe the different stages of A/E lesion formation. a. Injects effector proteins, EspB and Tir, into the host cells by type III sectretion machinery and they mondify cellular function and lead the signaling events by its direct biding with cytoskeletal proteins. TIR interacts with intimin on bacterial surface to produce intimate attachment. b. Pedestal forms due to polymerization of host actin c. Diarrhea due to loss of microvilli and tight juctions, host inflammatory response (cytokines) ion secreation, genes encoded on chromosome and plasmid 5. What are the roles of BFP pili, TTSS, Esps, TIR, Intimin, and actin in lesion formation? a. Bundle-forming Pili (BFP) attach to host cells b. TTSS secretes treanslocatedintimin receptor (TIR) into host cell c. TIR binds intimin protein on bacteria surface, secretes ESPS proteins to form pedestals by activating actin, becomes the receptor for intimin d. EPS effectors damage host cells loss of microvilli, decreased adsorption, ion secretion-diarrhea e. Intimin protein on bacteria provides intimate adherence to host by pedestal formation following TIR/Intimin binding. Actin polymerization does this formation. 6. Why is the production of shiga toxin more important than serovar in defining EHEC? a. Shiga toxin is the cause of HUS by entering the bloodstream and damaging vascular endothelial cells (e.g. those in the kidney), because the production of the shiga toxin is what identifies it as EHEC. EHEC itself has many different serovars that can cause the same disease. 7. What are Stx1/Stx2 and how do they work? a. Shiga toxin = stx b. vasculotoxin targeting vascular endothelial cells i. blood vessels and kidneys are targets because they have stx receptor Gb3. stx damages ribosomes to inhibit protein synthesis and cause cell death. renal tube cells express TNF to up-regulate expression of toxin receptor Gb3. LPS contributes to increase ii. Stx work by biding receptor to blood vessel and macrophage, produces lesion binds renal tube cells, induces cytokines, cytokines induce more receptors, LPS may enhance response. (RTEC = renal tube endothelial cells)

iii. 8. What are ATR proteins? How are they protective? a. Acid tolerant response(ATR) EHEC is uniquely tolerant to acidic environments because they grow at pH of 4.0-4.5 b. Promotes expression of acid-shock proteins for survival at low pH c. Provides cross-protection for tolerance of heat, radiation and antibiotics 9. What are the reservoirs for EHEC? Why were the outbreaks so serious? a. Undercooked ground beef, Humans, cider, sprouts, berries, also green onions and spinach. Not horses, deer. b. The outbreaks were so serious because the undercooked hamburger meat, transmission between people, 10. How did the Walkerton outbreak differ from most outbreaks in the U.S.? a. Contamination happened in the ground water, which came back positive in water purification plant. Testing occurred because of all the cases and tests came back positive but officials did not stop the water purification process. 11. Why are children more at risk for HUS? a. outbreaks in apple cider, in zoo animals and children like to put their hands in their mouth, leading cause of kidney failure in children and the elderly, still building natural flora b. Children have more receptors for the shiga toxin 12. Describe differences and similarities between EPEC and EHEC? a. EHEC pathogens do not have BFP because they colonize in the large intestine. EHEC produce shiga-like toxins similar to toxins produced by Shigella. Shiga toxins include stx1 and stx2 and produce symptoms of bloody diarrhea and hemolytic uremic syndrome (HUS) b. Both have TIR phosphorylation, and have a co infection 13. How is E. coli TTSS different from that in Yersinia? a. In EC TTSS there are no yops as found in Yersinia b. E coli utilizes actin filaments and Yersinia doesnt c. Yersinia targets M cells while e. coli targets any epithelial cell Shigellaspp.: 1. Which Shigellaspecies are in Group A-D y Group A- S. dysenteriae (makes Stx+ and causes epidemics) y Group B- S. flexneri (most common) y Group C- S. boydii y Group D- S. sonnei 2. Why does S. dysenteriaecause more serious disease? y Because it is the only one that produces Stx+ 3. What are the hallmarks of shigellosis symptoms? y Bloody diarrhea y Low infectious dose 4. Why is Shigella difficult to detect in food?

y y y y y

Shigella is generally present in low numbers The source of contamination is rarely identified Sensitive to physical attributes: pH, salt, fat content Natural micro flora typically overgrow shigella Physiological stress can hamper recovery

5. What causes the bloody diarrhea? y Because shigella forms micro abscesses, which will form large abscesses and ulcerations in the colon. When the dead cells begin to slough off, it produces blood and mucous in the stools. 6. What is HUS and Reiters syndrome? y HUS- Hemolytic Uremic Syndrome, is a disease characterized by hemolytic anemia, acute renal failure (uremia) and a low platelet count. Some patients will develop chronic kidney disease. y Reiters Syndrome- is reactive arthritis, is classified as an autoimmunecondition that develops in response to an infection in another part of the body. 7. What genes are on the virulence plasmid and what proteins are encode? y Virulence Plasmid- is very large, contains the genes necessary for the invasion (ics A, mxi, ipa, spa genes) 8. How does Shigella use TTSS differ? y 9. How does actin tail formation in Shigella differ from Listeria? y Shigella uses Icsprotein which catalyzes the polymerization of actin. (Listeria uses ActA) 10. What are mxi, spa,icsA, actin tails, aerobactin, virR/BF, Stx, ShET1/2 y Mxi- makes up the TTSS apparatus with Spa y Spa- makes up the TTSS apparatus with Mxi y iscA- protein that catalyzes the polymerization of actin, and forms the actin tail. y Actin Tails- expressed asymmetrically on one pole of the bacterial surface. Provides force for motility y Aerobactin-a bacterialsiderophore , it is a virulence factor enables the sequestering of iron in iron-poor environments. y virR- represses the virulence genes at 37C, its target is vir B y virB- encodes the transcriptional activator of ipa, mxi/spa y virF- binds the DNA as an antagonist to virR *All virs are part of virulence gene regulation y Stx- Shigatoxin, only found in S. dsysenteriae y ShET1/2- ?

11. What is the function of the Ipa proteins? y They are secreted by the TTSS into the cell, they induce uptake by M cell into vaculole, eventually causes cell death 12. Compare Shigella, Campy, and E.coliis terms of disease incidence, causes, infectious dose, symptoms, and pathogenesis. (HINT: you might want to make a table) Incidence Cause Infect. Symptoms Pathogenisis dose Shigella Common in Direct fecal Low Watery Bacteria multiply in the small children oral contact <1000 Diarrhea, intestine, invade M cells via <6yrs. from an Bloody TTSS in large intestine, escapes Endemic to infected stools, and enters macrophages and prisons, individual. Abdominal enterocytes, colonizes and kills mental Can be in Cramps the epithelial cells in the colon, hospitals, water or on Possible spreads through enterocytes via and nursing fresh foods HUS and actin tails, forms abcesses in homes. from the Reiters intestine, dead cells slough off field. Syndrome and produce bloody mucous stool. Campylobacter Mostly Raw Milk is Low Diarrheal Invades intestinal epithelial cell found in the cause of <1000 type illness lines using flagella, may or may young most out associated not transverse the M cells, causes children and breaks. with the secretion of invasion-related young Frequently lower effectors. (Phosphorylates host adults. found in intestinal proteins, Activates Kinases, Incidence chicken. tract. causes Carlease, and alters decreases Veggies and Abdominal microtubules). with age. Shell fish. cramping, Can cause Zoonotic occ. travelers Inflammator diarrhea. y colitis with fever. E.coli Can cause Fecal oral <10 with Diarrhea, Different depending on type. Will illness in contact. Can pathoge vomiting, use TTSS, Stx, BFP, etc. any be found in nic fever, Reference the E. coli section of individual water, meats strains hemorrhagic study guide. that (especially diarrhea, consumes a ground) , HUS, pathogenic apple cider, Reactive strain and arthritis, contaminate Kidney veggies failure

Campylobacter spp.: (campylobacter) 1. How have the sources of Campylobacter disease changed over time?









a. Campylobacter infections are considered to be mainly food borne diseases in which foods of animal origin play an important role. Besides the food-related cases, close contact with pet animals and activities related to recreational waters contribute to the large number of human illnesses each year. What is it the most common cause of sporadic disease? Outbreak disease? a. Vegetables and shellfish are sources of sporadic disease. Raw milk is the most common cause of outbreaks. Unpasteurized raw milk is still a source of outbreaks of campylobacter infection in the U.S. Is the disease more frequent in summer/winter? Why? a. The disease is more frequent in the summer because of improperly handled or cooked foods (mostly poultry) left out in the heat or not refrigerated properly, which is a perfect condition for campylobacter growth leading to outbreaks. What is meant by microaerophilic, zoonotic a. Does not grow in normal atmosphere, sensitive to extrinsic factors b. Zoonotic: many animal reservoirs What is GuillianBarre syndrome and how does it relate to infections caused by Campylobacter. a. Progressive weakness in more than two limbs b. Ascending paralysis leading to respiratory distress, progression for no more than four weeks c. Other signs include mild sensory defects, protein in the cerebrospinal fluid (CSF) How dos C. jejunidiffer from C. coli? a. Clinically indistinquishable, associated with lower G.I. tract b. C.Jejuni acute inflammatory diarrhea in developed countries; typing systems not standardized and include: biotyping, serotyping, bacteriocin sensitivity What is the HS antgen? a. The HS antigen = capsule, whichis a way for serotyping the campylobacter. This enables the bacteria to be classified according to its different components, and this explains some of what the symptoms are or include. Campylobacter HS:19 serotype is common to guillianbarre syndrome How are flagella, toxins, and microtubules important to disease? a. Flagella how it gets to the cell and invades the cell (gives it motility, invasion-translocation) goes through M-cells, adherence to the M Cell, b. Toxins cholera-like enterotoxins, contain three subunitsCytolethal distending toxin, type IV secretion What is difference between type III and type IV secretion? a. Type III secretion uses structural and effector proteins, chaperones to inject the operons, or toxins while the type IV secretion system deliver their effector toxin molecules to the intestinal epithelial cell lines using their flagella. The bacteria using type IV sectretion phosphorylate host proteins, activate kinases and cause Ca release, also, altering microtubules.

Salmonellaenterica 1. What are the O, K, H, Vi antigens?

a. O antigens: lipopolysaccharides (LPS) on outer membrane, specifically the polysaccharides b. H antigens: associated with the flagella c. Vi antigen: capsular occurs only in S.typhi, S. Paratyphi C and S. Dublin d. K antigen is the capsular in other bacterium 2. What are the reservoirs and common food sources for this disease? a. Ubiquitous reservoirs, a variety of animal products, produce contaminated by irrigation water, wildlife an humans. Common in rivers and ponds, high incidence in hrses, influenced by antibiotic practices. b. Poultry and eggs are the primary vehicles of transmission, eggs infected from the ovaries of infected hens. 3. How do you discriminate virulent vs. less virulent Salmonella a. Serotypes that are associated with food. S. typhii produce typhoid fever which are a higher death rate. They become systemic and are diseases are much harder to get rid of. 4. What is the difference between typhoid fever vs.nontypoidgasteroenteritis, a. Typhoid fever = systemic febrile disease (only S.typhi or S. paratyphi strains), Asymptomatic chronic carrier state commonly follow the acute phase of enteric fever = typhoid Mary, fatality 10%, colonize via Fimbrae to host glycoproteins, enters enterocytes and M cells in peyers patches. Causes ulceration (no diarrhea), Type III secretion; membratne ruffling, Ca2+ influx, intracellular growth in vacuoles requires at low pH, LPS modulates cytokines to cause fever, chills, delirium, can have diarrhea/vomiting, goes to liver, then gall bladder, shed in bile b. Non-typhoid = gastroenteritis(Multiple S. entericaserovars) abrupt onset symptoms: nausea, fever, headache, vomiting, diarrhea cramps. Fatality = 1%, severe cases can have bloody diarrhea, aseptic arthritis, reiters syndrome, ankylosing spondylitis. (genetic predisposition), LPS associated autoimmunity: inserts into the hose cell membranes, which are then attached by immune system 5. What proteins comprise its Type III apparatus and what do they secrete? 6. How does salmonella suppress host response? 7. What are the function of PhoQ/PhoP 8. What genes are on the Pathogenicity island? On the plasmid? 9. What vaccines and treatment are available? 10. Why is the cow a better animal model than mice? 11. Compare and contrast TTSS in EPEC, EHEC, Yersinia, and Salmonella in terms of targeted cells, effects on actin and other functions of the targeted cells.


Type of cell targeted for TTSS Enterocyte,

Molecules secreted into cell by TTSS Translocatedintimin receptor secreted

Effects on the cell by TTSS Pedestal formation

Symptoms created by the TTSS Diarrhea due to loss of microvilli and



Campylobacter jejuni

into host cell, binds following tight junctions, host intimin protein on TIR/Intimin inflammatory bacteria surface, binding due to response,(cytokines), secretes Espsproteis actin Ion Secretion to form pedestal polymerization, and bundle forming pili (BFP) Vascular Stx = shiga toxin, Binds Shiga toxins act by endothelial glycolipid inhibiting protein cells receptors on synthesis, causing (vasculotoxin), vascular cells, bloody diarrhea, Stx high number of HUS (hemolytic GB3 receptors Uretic Syndrome) on kidney this can cause cells, induces cortical necrosis. cytokine Stx can also activate response, macrophage and polymorphonuclear neutrophils M cells Secrete Ipa Ipas (A,B,C,D) Epithelial cells in proteins, TTSS app. are effector colon forms Composed of Mxi proteins microabscesses, + Spa protein rearrange host ulcerations, dead actin to cells produces blood promote and mucus in stools. uptake, lyses vacuole, induces apoptosis Microtubules Cytolethal Alters Associated with in enterocytes, distending toxin: microtubules, septic abortions, m cells? cell death, CDT septic arthritis, taken up abcesses, meningitis

Species: Cronobacter




Virulence Factors


Yersinia EPEC EHEC Shigella Campylobacter Salmonella