Gram Negative Cocci Lecture Notes PDF
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These lecture notes cover Gram-negative cocci, focusing on Neisseria and Moraxella. The document details their characteristics, virulence factors, and associated diseases. It also includes laboratory diagnosis methods used to identify these bacteria.
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LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores GRAM NEGATIVE COCCI MP–attachtoneutrophils,epithelialcells;resist...
LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores GRAM NEGATIVE COCCI MP–attachtoneutrophils,epithelialcells;resist R NeisseriaandMoraxella effects of host IgG ColonialCharacteristics(growprimarilyonCAP) eisseria N (will not grow on BAP) Gram Reaction and Morphology: Mostly Gram (-) ○ Small Diplococci (coffee-bean or kidney bean) except the ○ Gray to tan ○ Translucent and raised following species which appearBACILLI: ○ Requires CYSTEINE for growth ○ N. bacilliformis ○ Pathogenic strains –smaller and raised ○ N. elongate ○ AHU(AtypicalorArginine,Hypoxanthine, ○ N. weaveri Uracil Auxotypes) Aerobic and Capnophilic More fastidious Pathogenic species are fastidious Smaller Most are Catalase(+); Oxidase(+) Nutritionally variant strains Occur in DGI (disseminated Non-motile; non-sporeformers gonococcal infections) , Habitat:Respiratory Tract; Urogenital Tract Asymptomatic urethritis in males (S) to penicillin They can be found inside or outside neutrophils Neisseria gonorrhoeae TI; causes acutepyogenicinfectionofmucusmembraneof S Neisseria meningitidis endocervix, urethra, anal canal, pharynx, conjunctiva Gonorrhea (“clap”) an be part of transient flora, Carrier state of asymptomatic C ○ Males:Urethritis (oropharynx and nasopharynx) ○ Females:Cervicitis Diseases Complications: ○ Meningitis ○ Pelvic Inflammatory Disease (females) Meningcoccemia – dissemination ○ Fitz Hugh Curtis Syndrome (females) in the blood Perihepatitis-outercoveringorthe ○ S/S: Petechial rash capsule of the liver is inflamed ○ Disseminated Intravascular Coagulopathy ○ Salpingitis (females) ○ Development ofWATERHOUSE ○ Ectopic Pregnancy FRIEDERICHSEN SYNDROME ○ Ophthalmia Neonatorum Hemorrhage of adrenal glands in Pag buntis si mother pwede niya children mahawaan si baby ng OTHER DISEASES: gonorrhoeae sa mata ○ Pneumonia ○ Hematogenous Dissemination ○ Arthritis (disseminated gonococcal infections) ○ Conjunctivitis ○ Septic arthritis ○ Otitis media OTE: the infection is not just limited in genitaltract, it also N irulence factor:CAPSULE V affects theupper respiratory tract,throatand evenanal canal Colonial Characteristics ○ Medium Sized Virulence factor: PILI(forattachmentofliningof ○ Gray ucosa) m ○ Convex ○ Pathogenic strains:T1 and T2 ○ Encapsulated strains are MUCOID ○ Non-pathogenic strains: T3, T4, and T5 ○ Are able to grown in CAPaswellason BAP OTHER VIRULENCE FACTOR: Green tingeunderneath BAP Endotoxin (LOS); for dissemination Protein II (opa – adhere to host epithelial and erotyping of meningitidis is based on the capsule S phagocytic cells; Capsule (A, B, C, W135, X, Y) BASTA 4TH YEAR STUDENTS |1 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores erotype A -seen inunderdeveloped countries S Serotype B, C, Y -US Serotype W135 -Invasive OTHER VIRULENCE FACTOR: Endotoxin - Responsible for the dissemination (rashes) LABORATORY DIAGNOSIS SPECIMENS: Urogenital specimens Anorectal Oropharyngeal (or throat swabs) Conjunctival specimens (in cases of octaminutorum) CSF (meningitis) OTHER SPECIES Blood (disseminated cases, meningococcemia) .cinerea- may be mistaken forN. gonorrheabut N 1 RAM STAINING AND MORPHOLOGY G can grown in BAP, MHA; Colistin (S) Direct Gram Stain of urogenital specimens ○ N. gonorrhoeae= Colistin (R) Gr(-) intracellular diplococci- forgonorrhea N. flavescens-yellow pigmented - MALE: Ifthedischargeofthemalehas N. sicca- dry colonies, bread crumb like; wrinkled Gr(-) intracellular diplococci with N. weaveri**- may be isolated in humans; presentin numerous neutrophil (presumptive of oral cavity of dogs (-) NO3 gonorrhea) Direct Gram Stain of CSF- meningitis 2 CULTURE: - CAP (S BA-can be usedforN.meningitidisbut not for N. gonorrhea); - TraditionalPathogensdonot growat22C;prefer body temperature - prefer growing in enriched media (w/blood;iron) - Can also use Selective media (CAP na may added na antibiotics) for isolation of pathogenic Neisseria; this includes: arely cause disease but may cause disease such as R Thayer Martin (contains vancomycin, colistin, meningitis, endocarditis, bacteremia, pneumonia, empyema, nystatin) ocular infections Modified Thayer Martin (vancomycin, colistin, nystatin, and trimethoprim) enerally less fastidious, easy to grow in ordinary culture G Martin Lewis(vancomycin, colistin, anisomycin) media unlikeN. gonorrhoeaeandN. meningitidis New York City Agar (vancomycin, colistin, trimethoprim, amphotericin B) Moraxella catarrhalis ancomycin- prevent growth ofGr(+)microorganisms V r(-) diplococci; non-motile, non fermenters G Colistin- prevent growth of otherGr(-)microorganisms Formerly known asBranhamella catarrhalis Trimethoprim- preventswarmingofproteus Nystatin, Anisomycin, Amphotericin B - prevent (+)in Tests: Oxidase, DNAse, Butyrate Hydrolysis growth offungi Commensal of upper respiratory tract; opportunistic pathogen Use of Transport Media Causes:upperrespiratorytractinfections;otitismedia ➔ JEMBEC (James E. Martin Biological environmental chamber) (3rd most common cause), sinusitis, endocarditis, ◆ Transport media for gonorrhea meningitis, bacterial tracheitis ➔ Trans-Grow treptococcus pneumoniae(most common) S ➔ Gono-Pak Haemophilus influenzae(2nd most common) Moraxella catarrhalis(3rd most common) olonies: GRAYISH WHITE, opaque withaHOCKEY C PUCKconsistency;WAGON WHEELappearance BASTA 4TH YEAR STUDENTS |2 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores 9 GAMMA GLUTAMYL AMINOPEPTIDASE Fordiscrepant resultsofN. meningitidis . meningitidis may be misidentified as N. gonorrhoeae on N Carbohydrate test which means that carbohydrate test may give an inaccurate test result. It is misdiagnosed and misidentified as gonorrhea sincethereisonlyfermentationin the glucose tube. But if you are sure that the isolate is N. meningitidis. To address discrepancy, you can do the gamma glutamyl 3 OXIDASE TEST aminopeptidase test. Oxidase Reagent:(1% ( +) result:N. meningitidis=YELLOW dimethyl-p-phenylenediamine dihydrochloride) (-) result:N. gonorrhoeae Expected Result:PURPLE NB:Avoid Using Iron or Nichrome Loops 10 HYDROXYPROLYLAMINOPEPTIDASE ○ Cause False (+) result Useful forN. gonorrhea OTE:AllmembersofNeisseriaincludingMoraxella N 11 SEROLOGIC TESTING catarrhalisareOxidase (+) N. meningitidisserogroups * Use applicator stick to fish out colonyorswab;oryou can directly add the reagent to the plate to check out purple coloration 4 CATALASE TEST All are Catalase (+) except: N. elongata and N. bacilliformis Species GLUCOSEMALTOSELACTOSESUCROSEFRUCTOSE 5 SUPEROXOL TEST . N Useful forN. gonorrhea + - - - - gonorrhoeae Reagent:30% H2O2 N. Expected Result:Vigorous bubbling + + - - - meningitidis 6 ARBOHYDRATE UTILIZATION C N. lactamica + + +(ONPG+) - V In the diagnosis of Neisseria, traditionally if you wantto confirm or determine the species, the presumptive of M.catarrhalis - - - - - organisms, you have to do carbohydrate utilization. GRAM NEGATIVE BACILLI se ofCystineTrypticaseSoyAgarBasewith U 1% Concentrationfor Carbohydrates; Family Enterobacteriaceae (Order Enterobacterales) ○ To check fermentation of that particular sugar NOTE:“Enterics” shortcut ni sir sa Enterobacteriaceae/Enterobacterales pH Indicatorphenol red; Used for Species ID; GENERALITIES No CO2 incubation(24-72 hours incubation) ○ Lead to False (+) result ramReactionandMorphology:Gram(-)coccobacilliin G (+) result:YELLOWcolor in tube pairs, singles ○ Coccobacilli – short, plump bacilli 7 BUTYRATE HYDROLYSIS For ID ofMoraxella OnBAP:coloniesappearlarge,gray,smooth,andmostare (+) result:BLUE non/gamma hemolytic Facultative Anaerobes 8 NITRATE REDUCTION Glucose fermenters; (+)forMoraxella ○ This is the reason why on Triple Sugar Iron (TSI), the buttisalwaysyellow.Themembers BASTA 4TH YEAR STUDENTS |3 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores of Enterics would always have yellow/acid ○ lesiomonasistheonlyoxidase(+)memberof P utts. b the enterics. Xylose Fermentersexcept: Shigella Most are Motile except: Klebsiella, Shigella & Yersinia Catalase (+)except: Shigella dysenteriae (-) which are nonmotile at 25 and 37 deg C. Oxidase (-)except: Plesiomonas (+) Most are nitrate reducers except: Photorhabdus and ○ To distinguish Enterics from other gm (-) Xenorhabdus; some biotypes of Pantoea agglomerans; bacteria like Vibrio, Pseudomonas, etc., some species of Serratia and Yersinia Enterics are oxidase (-). Rapid Lactose Fermenters Late Lactose Fermenters Non Lactose Fermenters (18-24 hours) (>24 hours) ONPG (+) nzymes present: E nzymes present: E Beta-galactosidase Beta-galactosidase Lactose permease scherichia E itrobacter C almonellaexceptS. arizonae S Klebsiella Shigella sonnei ShigellaexceptS. sonnei Enterobacter Salmonella arizonae Providencia Hafnia ** Edwarsiella Serratia ** Morganella Yersinia enterocolitica** Proteus Yersinia ** may be NLF Note:Generallyspeaking,thenonlactosefermentersareusuallythepathogenic/opportunisticmembersofentericssincenandiyan sina Salmonella, Shigella, Yersinia. Whilenonpathogenicanglactose fermenters. Hydrogen Sulfide (+) Rapid Urease (+) Late Urease (+) Deaminase (+) Voges Proskauer (+) almonella S roteus P itrobacter C roteus P antoea P Proteus Providencia Klebsiella Providence Serratia Citrobacter freundii Morganella Enterobacter Morganella Hafnia Edwarsiella Yersinia Enterobacter Serratia Klebsiella ( +) Blackening on TSI bletoproduceUrease D A elay in production of R /A(redslant/acidbutt) A ble to produce acetoin Mnemonic:SPaCEd enzyme within 4-8 hrs Urease enzyme after reaction on LIA Mnemonic:PSHEK Mnemonic:PPM 18-24 hrs Mnemonic:PPM Mnemonic:CKEYS Rememberthe relationshipbetweenVP (+) and Methyl Red (+) organisms: If VP (+), MR (-); If VP (-), MR (+) LABORATORY DIAGNOSIS ○ pH Indicator:Phenol Red (NEG = Red) . GRAM STAINING 1 unction: Detection of fermentation, gas production(in F 2. USE OF SELECTIVE AND DIFFERENTIAL MEDIA the form of cracks or spaces on agar), and H2S EMB, MacConkey, HEA, SSA, XLD, BSA production 3. USE OF TRIPLE SUGAR IRON (TSI) AGAR Inoculation is done by stab-streak method Composition: Note: If there is blackeningonthebutt,youcansafely ○ 10 parts lactose; 10 parts sucrose; 1 part glucose assumethatthebuttisalsoyellowbecauseEntericsare UsefulfordetectionofY.enterocolitica able to ferment glucose. because they can fermentsucrose ○ H2S Indicator: Sodium Thiosulfate and TSI REACTIONS Ferrous Sulfate BASTA 4TH YEAR STUDENTS |4 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores K / K LDC/Decarboxylase (+) Klebsiella R / A Deaminase (+) PPM K / K + LDC (+) and H2S production Salmonella K / A LDC (-) and deaminase (-) Shigella K / A + LDC (-) and deaminase (-) with H2S Citrobacter OTE: The yellow butt in R/A is due to the production of N (Control - K/K - K/A - A/AG - A/AG+ - K/AG+) glucose. Only deamination is present, so there is no A / A 2 or more sugars fermented scherichia, Klebsiella, E production of cadaverine (product of decarboxylase; kaya Enterobacter naging purple butt sa K/K). The acidity of the butt will be retained, kaya yellow pa rin siya. A / A G or more sugars fermented 2 . coli, Klebsiella, E with gas Enterobacter, Serratia If LDC (+) = may cadaverine A / A G + 2 or more sugars fermented Citrobacter freundii with gas and H2S production 5. IMVIC TESTS a. Indole Test K / A Glucose only fermented higella, some Hafnia, S ○ Detection of tryptophanase enzyme Yersinia pestis ○ Medium:Tryptone Broth, SIM ○ Indicator: Kovac’s or Ehrlich’s Reagent K / A G lucose only fermented with G organella, M gas Providencia, Hafnia, (p-dimethylaminobenzaldehyde) Citrobacter diversus ○ (+) result:Red color (indication of indole prod.) ○ Useful to differentiate P. vulgaris (+) from P. K / A G + Glucose only fermented with Salmonella mirabilis (-), and E.coli (+) from Klebsiella (-) gas and H2S production K / A + lucose only fermented and G Proteus, Salmonella H2S production K / K o sugars fermented N seudomonas, P (Non-fermenter; Non-Enteric) Acinetobacter b. M ethyl Red and Voges Proskauer Test (Glucose B: KIA (Kligler’s Iron Agar) - 2 sugars only (Glucose and N Metabolism test) Lactose only) ○ Medium: MRVP Broth (contains glucose) ○ Determineswhatpathwaytheorganismutilized 4. USE OF LYSINE IRON AGAR (LIA) for metabolism Detection of Lysine Deamination and Decarboxylation Mixed Acids pathway = MR (+) Composition: Glucose; Lysine Butylene Glycol pathway = VP (+) ○ pH Indicator:Bromcresol Purple ○ H2S Indicator:Ferric Ammonium Citrate Methyl Red- detects mixed acids; pH of 4.4 or less not reliable - not all H2S producers Indicator Reagent:methyl red may produce blackening Expected Result:red color Inoculated using stab-streak method Incubated for 18-24 hours Voges Proskauer - detects acetoin (by-product of butylene glycol pathway); pH of 5.5 or more Reagents: alpha naphthol and KOH LIA REACTIONS (Barritts);alphanaphtholin40%KOHand creatine (Coblentz) Expected Result:red color Slant = deamination (aerobic rxn); Butt = decarboxylation (anaerobic rxn) BASTA 4TH YEAR STUDENTS |5 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores These are all alkaline byproducts edium: Decarboxylase Medium (Moeller's) with M bromcresol purple, glucose and amino acid; ○ Control tube - contains medium and glucose but no amino acid ○ Medium Overlaid with mineral oil ○ It is overlaid w/ mineral oil because the decarboxylase test is an anaerobic reaction (occuring in an anaerobic environment) c. Citrate Utilization Result: Purple ○ Determinesiftheorganismusescitrateassole Useful to differentiate Klebsiella from Enterobacter source of carbon ○ Medium:Simmon Citrate Agar Slant ○ pH Indicator:Bromthymol blue ○ (+) result: color change from green to blue; growth 9. PHENYLALANINE DEAMINASE Enzyme that removes amino group from an amino acid Medium: Phenylalanine Agar with 10% Ferric Chloride Result: Green 10. GELATIN LIQUEFACTION Detection of enzyme gelatinase 6. UREASE TEST Medium: 12% Gelatin Medium: Christensen's Urea Agar; Stuart's Urea Broth ○ This testusuallyhaslongerincubationtime(5 pH Indicator: Phenol Red days) Expected Result: Pink/ Fuschia Result:GelatinLiquefies(putinreforicebathfor15-30 mins before interpret) ○ Before interpreting the results, you put the inoculated gelatin first in the ref before interpreting to avoid any false positive results. ○ Kasi if galing sa incubator yan, mainit so malambotpaunggelatinkayaaakalainmona 7. ONPG TEST (Ortho Nitrophenyl Beta D-Galactopyranoside) positive shea. Determines lactose fermentation Ex. Serratia (Gelatinase (+)) ○ Detection of enzyme: Beta galactosidase Substrate: ONPG 11. OF MEDIUM - Oxidation and Fermentation (o-nitrophenyl-beta-D-galactopyranoside) To determine whether the organism can oxidize or Expected Result: Yellow ferment the sugars Beta galactosidase are produced by our lactose pH Indicator: bromothymol blue fermenters Inoculate 2 tubes (1 open and 1 closed tube) Application of ONPG tests are usually for late lactose ○ If both closed and open tubes are yellow : fermenters, instead of waiting for the fermentation to fermenter happen. Ex. enterics ○ If there is no color change in closeandopen 8. DECARBOXYLASE tubes experienced color change (close is Enzymethatremovesthecarboxylgroupfromanamino green, open is yellow) : oxidizer acid (ornithine, lysine, arginine) ○ No color change occurred (both close and ○ Lysine Decarboxylase >> Cadaverine prodn open are green) : asaccharolytic - not a ○ Ornithine Decarboxylase >> Putrescine fermenter nor and oxidizer ○ ArginineDihydrolase>>Citrulline>>Ornithine > Putrescine ○ Arginine would initially produce Citrulline, which could be broken down into Ornithine, and then Putrescine BASTA 4TH YEAR STUDENTS |6 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores 13. OXIDASE TEST Eenterics are negative except Plesiomonas 12. MUG TEST 4-methylumbelliferyl-Beta-d-glucoronide; Detection of enzyme beta-d-glucoronidase; ○ Useful as a presumptive identification of E.coli ○ Most types of E.coliarepositiveforMUGtest except EHEC Positive Result:Electric blue fluorescence Medium Description Levine's Eosin Methylene Blue Purple colony in EMB E.coli: purple colonies w/green metallic sheen indicates that it is a lactose fermenter Klebsiella: purple mucoid Klebsiella and enterobacter Enterobacter: purple mucoid are mucoid because they are capsulated NLF: colorless colonies MacConkey Agar It is selective and differential LF: pink colonies NLF: colorless Hektoen Enteric Agar More selective than Mac and E. coli: orange w/o black center EMB Has H2s indicator which has Proteus: blue green w/ black center black center for H2s positive Bromthymol blue is the pH Salmonella: blue green w/ black center indicator Shigella: blue green w/o black center Salmonella Shigella Agar Selective and differential for Salmonella: colorless with black center salmonella and shigella Both are NLF, so they will Shigella: colorless w/o black center appear colorless Neutral red as the pH indicator BASTA 4TH YEAR STUDENTS |7 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores o differentiate the two is T throughtheblackcenter(H2S (+)) Xylose Lysine Deoxycholate Agar Also has H2S indicator Salmonella Red w/ black center Salmonellaandshigellawould shigella Red w/o black center appear red colonies on XLD Also differs in H2S Bismuth Sulfite Agar Aka Wilson Blair Agar S. typhi: black with black metallic sheen A selective medium toisolate Salmonella typhi from stool samples 1. Escherichia coli: “Colon Bacillus” ormal flora of colon N IGHLY INVASIVE; Watery H Important human pathogen (UTI’s, CNS infections, to Blood Diarrhea and Colitis; sepsis, endotoxin induced shock, and diarrheal Virulence Factor: Verotoxin; diseases) Shiga like Toxin Uropathogenic UTI: E. coli most common cause of Most Important Strain: Community acquired UTI O157: H7 - Implicated in an outbreak of hemorrhagic diarrhea, colitis DiarrheagenicE.coli Features and hemolytic urenic syndrome (HUS) TEC (Enterotoxigenic E ausesTraveler’s Diarrhea. C -Implicated food: Escherichia coli) Montezuma’s Revenge, hamburgers; deli meats; Turista Production of Heat cheese, lettuce; Labile and Heat Stable Toxin unpasteurized milk, apple Watery diarrhea w/o blood cider - this is quite DEADLY otes:Have a history of N -Selective & Differential travel outside the U.S. (3rd Medium Employed:sorbitol world country where MacConkey sanitation is not that good.) -Expected Colonies: colorless colonies PEC (Enteropathogenic E Infantile Diarrhea; (non-sorbitol fermenter) Escherichia coli) Non-invasive; characteristic -Other Media: lesions that attach and efface Cefixime-Sorbitol Mac; cells ChromAgar O157 otes:usually targets infant N AEC (Enteroadherent E nteroaggregative E. coli - E patients, pediatrics Escherichia coli) “stacked brick pattern”(on cell cultures: Hep2: IEC (Enteroinvasive E higella-like infection; S Diffusely adherent E.coli - Escherichia coli) Dysentery-like or BLOODY “Diffused Pattern” diarrhea symptoms; invasion Implicated in diarrhea & UTIs Sereny Test – Virulence Test 2. Klebsiella otes:usually non-motile; N ormalHabitat:GITractofhumansandanimals;Found N usually late-lactose or non-lactose fermenters in the environment UnlikeE.colithatcanonlybefoundontheGITtract& HEC (Enterohemorrhagic E ka STEC (Shiga-toxin A cannot be normally found in the environment. Escherichia coli) producing Ecoli/ VTEC Causes Opportunistic & Nosocomial infections>>UTI, (Verocytotoxic E.coli) pneumonia & wound infection. BASTA 4TH YEAR STUDENTS |8 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores K. pneumoniae 3. Enterobacter ommon Name: Friedlander’s Bacillus C Cause nosocomial & opportunistic infections Causes Lobar pneumonia; currant jelly like sputum; ○ UTI, respiratory tract infections, wound UTIs infections K. oxytoca Aerogenic Similar infections caused by K. pneumoniae ○ E. aerogenes - Urease (-); (reclassified as K. Unique biochemical test:INDOLE (+) aerogenes) Other Species ○ E. cloacae; E. gergoviae - from respiratory K. granulomatis (STD: granuloma inguinale; Donovan specimens(UREASE POS) Bodies) ○ Cronobacter sakazakii (E. sakazakii) - K. ozaenae - ozena (atrophic rhinitis) production of yellow pigmented colonies @ 25C; milk products, infant milkformula;assoc w/ bacteremia, sepsis, necrotizing colitis 4. Pantoea agglomerans ormerly known as Enterobacter agglomerans F Septicemia; IV fluids contamination Triple Decarboxylase Negative ○ LDC, ODC, & ADH (-) . rhinoscleromatis - rhinoscleroma (granulomatous K diseases of nasal passage) I M V C LDC ODC ADH MUG Test E. coli + + - - + + - + K. Pneumoniae - - + + + - - - K. oxytoca + - + + + - - - E. aerogenes - - + + + + - - E. cloacae - - + + - + + - 5. Serratia pportunistic pathogens associated w/ outbreaks in O ○ Mostly H2S (+) except: P. penneri (-) health care settings (bacteremia, urinary, respiratory tract infections) Indole MR VP Cit LDC ODC H2S DNase (+) Gelatinase (+) P. vulgaris + + - + - - + LDC (+) Citrate (+) P. mirabilis - + - + - + + S. marcesens & S. rubidaea - pigment producers (prodigiosin)-redpigmentw/cisusuallyenhanced@ Providencia room temperature. auses UTIs, diarrhea among travelers, nosocomial C ○ S. marcesens: Malonate (-) : green infections ○ S. rubidaea: Malonate (+) : blue P. stuartii - health care UTIs; wound infectionsinburn units 6. Proteus P. rettgeri- UTI, diarrhea, rapid Urease (+) nown for swarming on blood agar plates K Morganella Morganii UTIs, wound, ear infections, nosocomial infections, Implicated in UTI’s, neonatal sepsis kidney stone formationwhich is usually P. mirabilis Key Characteristics: ○ Swarming Colonies ○ Odor: burnt chocolate cake odor BASTA 4TH YEAR STUDENTS |9 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores Indole MR VP Cit H2S yphoid T Fever: (Incubationperiod:1week Providencia rettgeri + + - + - to 1 month) 1st week: fever, dull Morganella morganii + + - - - frontal headache, malaise, body pain, constipated (“scybalous” stool) / goat 7. Edwardsiella tarda droppings pportunistic pathogen; isolated from the environment O Best sample to be Causes bacteremia, GI infections, wound infections submitted:Blood (aquatic incidents) Other Sx: Bone Marrow Similar IMVC withE.coli but it is H2S: (+) nd to 3rd week: 2 LDC: (+) intestinaltract,“rosespots: TSI: K/AG (+) appear diarrheic stools Indole MR VP Cit H2S Sample to besubmitted: Stool Other Sx: Urine E.coli + + - - + hronic Carrier State - C E.tarda + + - - + may shed in stool and E. tarda produces large amounts of H2S urine for up to 1 year 8. Citrobacter Salmonella Paratyphi Causes typhoid fever, paratyphoid nown for being Citrate (+) K fever, enteric fever Causes nosocomial infections, UTIs, pneumonia, S.paratyphi A= H2S (-) ; LDC (-) meningitis KCN (+) Salmonella Food poisoning (dairy products); Citrate Utilizer Typhimuirum invasive infections Mostly H2S except: Citrobacter koseri/C/diversus Maybe mistaken for Salmonella, Citrobacter is: Salmonella Bacteremia; sepsis; osteomyelitis holerasuis C ○ ONPG (+) ○ LDC (-) Salmonella Pullorum Non motile ○ KCN (+) and Salmonella Gallinarum TRUE PATHOGENS Salmonella Indole MR VP Cit LDC ODC ADH H2S Causes gastroenteritis, enteric fever (Typhoid fever), bacteremia (sepsis) MOT: Ingestion of contaminated water, milk, dairy almonella S - + - + + + + + products, shellfish, poultry products, meats, peanut enteriditis butter, vegetables; animal contact for certain species almonella S - + - - + - - + (reptiles) typhi Carrier state- Gallbladder (asymptomatic patients) Th ere are 2 recognized species for Salmonella almonella S - + - + + + + + ○ Salmonella retorica- with 6 subspecies; more arizonae clinically significant (Subgroup 1) ○ Salmonella bongori- seen in usually isolated almonella typbi - produces curved wedged on TSI or S in cold blooded animals; environment mustache-ike Genus and serotype - are commonly used to ID the bacteria For Confirmatory Testing KauffmanWhiteClassification-serologic(Salmonella Salmonella Serotype Associated disease based on O and H antigen) - use of polyvalent and monovalent O antisera, Vi antigen testing for S.typhi Salmonella Typhi nly harbored by humans O Widal Test- to determine febrile agglutinins or Most important cause of antibodies in the diagnosis of typhoid fever typhoid fever / enteric fever BASTA 4TH YEAR STUDENTS |10 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores Shigella ○ Bubonic, Septicemic and Pneumonic forms elated toEscherichia;biochemicallyinertmembersof R Bubonic - most common form of Family Enterobacteriaceae plague Causes shigellosis/bacillary dysentery (enterocolitis) Pneumonic - most deadly; Manifestations: complication of septicemic form ○ Fever, chills, cramps, diarrhea, pain and ○ S/S: high fever, headaches and buboes, tenesmus 24 - 48 hours after ingestion; hemorrhages (blackish discoloration), rapidly bloody, with pus and mucoid stools developing pneumonia, DIC, shock MOT: ○ Fecal-oral, contaminated food/water, Low Inclusion bodies present :Bipolar bodies infective dose Colonies:Hammered Copper colonies S.flexneri- severe On Broth:Flocculent or Stalactite appearance S.sonnei- most commonly isolated spp Non-motile: @ 25 and 37 C erologic Testing for Salmonella and Shigella - based S Yersinia pseudotuberculosis on O antigen nimal pathogen (pseudotubercles) A acute mesentric lymphadenitis Species I MR VP CIT LDC ODC septicemia A S.dysenteriae V + - - - - Yersinia enterolitica ost common type M B S.flexneri V + - - - - Causes of Yersinosis ( Strong’s) ○ MOT: Ingestion of contaminated food (pork meat, milk) C S.boydii V + - - - - ○ S/S: mimics appendicitis, enteritis, arthritis, ( Newcastle erythema nodosum, septicemia (px with Manchester hemochromatosis) D . sonnei S - + - - - + Sucrose Fermenter (Duval’s) Culture Medium: CIN (Cefsulodin Irgasan Novobiocin) Agar ○ Characteristic Appearance of Colonies: Bull’s Species ADH CATALASE ONPG MANNITOL eye colonies A S.dysenteriae - - - - Species I M V C UREASE B S.flexneri - + - + ( Strong’s) Y. pestis V + - - - C S.boydii - + - + Y. enterocolitica - + - - + ( Newcastle Manchester Y. pseudotuberculosis - + - - + D . sonnei S - + + + Species LDC ODC ADH SUCROSE SORBITOL (Duval’s) Y. pestis - - - - - ersinia Y Zoonotic; Non-motile @ 37 C Y. enterocolitica - + - + + Yersinia pestis Causative agent of plague (Justinian Plaque, Black Y. pseudotuberculosis - - - - - Death, Modern Pandemic) ○ Transmittedbythebiteofratfleas(Xenopsylla cheopsis) ○ Other MOT: Direct contact w/ an infectedperson; handling infected animal tissues caracasses BASTA 4TH YEAR STUDENTS |11 LECTURE 2_BACTERIOLOGY 081524 | Reference: ARFlores OXIDASE (+) ENTERIC lesiomonas shigelloides P Oxidase (+) Enteric enerally susceptible to Vibrio static agent G Formerly under Vibrio group but it is now Enteric Able to be positive to gelatinase (gelatinase +) Associated with diarrhea Extraintestinal infections (neonatal meningitis and septicemia) ○ Inositol fermenter ○ Glucose fermenter ○ Triple Decarboxylase (+) ○ String Test (-) ○ Non-Halophilic GRAM NEGATIVE NON-ENTERIC ORGANISMS THAT ARE AGENTS OF DIARRHEAL DISEASES AND OTHER DISEASES VIBRIO ram (-) straight or curved bacilli but can be G Vibrio cholerae pleomorphic ○ It Looks like a comma shaped bacilli. ommon Name: Komma Bacillus C MOT: ingestion of contaminated water Motile with a single polar flagellum (sheathed) Causative agent of CHOLERA (Asiatic cholera or ○ Monotrichous epidemic cholera) Facultative Anaerobes or Aerobes ○ V. cholerae 01 Catalase(+),Oxidase(+),andNitrateReducersexcept ○ V. cholerae 0139 - Bengal Strain V. metschnikovii Pathogenic stains because they are able to produce toxins Glucose Fermenters; Most are able to ferment sugars Severe Gastroenteritis; Vomiting Rice Water Stools; 10 - 30 bowel movements per such as sucrose, and lactose. day; with flecks of mucus; ○ Sucrose fermenter:V. cholerae Sunken Eyes; Washerwoman’s Hand Skin ○ Lactose Fermenter:V. vulnificus Electrolyte Loss; Dehydration eventually leading to Most species thrive at high con
