Micro final

Are Micrococcus and Staphylococcus gram pos cocci that is arranged in pairs, clusters, or tetrads

yes

Are Micrococcus and Staphylococcus cat pos

yes

What media does MICROCOCCACEAE AND STAPHYLOCOCCACEAE grow on

SBA ----------Growth

CNA/PEA-------Growth

What the difference between STAPHYLOCOCCUS and MICROCOCCUS

STAPHYLOCOCCUS

Gluc. fermenter

Facultative  Yes

Microdase  -

 (Mod.oxidase)

Bacitracin   R

Lysostaphin  S

MICROCOCCUS

 Glucose oxidizer

 Facultative  No

 Microdase    +

  (Mod. oxidase)

 Bacitracin   S

 Lysostaphin  R

What are some organisms of staph

AUREUS

EPIDERMIDIS

SAPROPHYTICUS

What are the characteristics of aureus

•Habitat:

 + normal flora,

   skin/resp.tract

•Colonies on SBA

•Cream/white/

•yellow/gold

•B-hemolytic

•Biochemical ID

•Coagulase  +

•Mannitol Salt (MSA)  +

•DNase/Thermo-nuclease +

•Rapid ID

 Latex aggl. + (Staphaurex)

Gram positive cocci, pair & clusters

•PENICILLIN G RESISTANCE

  *B-lactamase + 

resistant to ampicillin and cephalosporins

What are staph aureus infections

•Wounds/Lesions

•scalded skin syndrome/sties

•Impetigo/carbuncles/furuncles

•Urinary Tract Infections (UTI)

•Food poisoning--stool not cultured

•Systemic Infections/Toxic Shock

•Blood/CSF, Osteomyelitis

•Nosocomial/Iatrogenic Pneumonia

What are the are the characteristics of STAPH EPIDERMIDIS

•Habitat:

•normal skin and respiratory flora

•Colonies on SBA:

white/opaque

•Biochemical ID:

•Coagulase -

•Growth on MSA &

  Mannitol -

•Novobiocin S (urine)

Coag Negative Staph Colonies
White or cream, Non-hemolytic

staph latex neg

What are the infection of STAPH EPIDERMIDIS

•Prosthesis-associated infections

•IV catheters 

•CSF shunts 

•total joints (knee/hip/elbow)

•heart valve surgery

Bacteremia/Endocarditis

What are the are the characteristics of STAPH SAPROPHYTICUS

•Habitat:

•normal skin flora

•Colonies on SBA

•Cream/white,

  opaque

non-hemolytic

COAG NEGATIVE

•Biochemical ID

•Coagulase -

•Growth on MSA &

  Mannitol +/-

•Novobiocin R

staph latex neg

What are the are the characteristics of MICROCOCCUS SPECIES

•Habitat:Usually nonpathogenic; may be transient skin flora

•Gram stain: gram pos. Cocci, pairs, clusters and tetrads

•Colonies on SBA: often yellow

•Biochemical ID:

  Glucose oxidizer

    Modified oxidase(Microdase) + 

  Bacitracin (0.04 U)         S

  Lysostaphin (200ug/ml)    R

How is STREPTOCOCCACEAE different from staph

•Gram positive cocci

•arranged in pairs and chains

•catalase negative

What are the infections of S. pyogenes -Grp. A

•strep throat

•wounds: pyoderma/ impetigo/ necrotizing fasciitis

•scarlet fever

•strep toxic shock

•throat and/or wound infection sequellae:

• rheumatic fever

• glomerulonephritis

What are the infections of S. agalactiae-Grp. B in women and newborns

•Newborns

•septicemia

•meningitis

•Women

•septicemia

•puerperal sepsis

•UTI

What are the infections of Grp. D Enterococcus faecalis/faecium

•UTI

•Wounds/below the waist

•Endocarditis: most common cause of nosocomial endocarditis

resistant to Penicillin

vancomycin resistance

What are the infections of Grp. D)  S. bovis

•endocarditis

•wounds

•colon cancer sepsis

•Abiotrophia

•”satelliting strep”

•AKA: NVS/ NDS/ TDS

Cause endocarditis

What are the infections of Strep pneumo

•pneumonia

•septicemia

•meningitis-elderly

children: meningitis & otitis media

What are the infections of Strep viridans

•endocarditis

•dental caries

What media do STREPTOCOCCI  grows on

•SBA/CA------ Growth

•CNA/PEA-----Growth

What are the are the characteristics of S. pyogenes -Grp. A

beta hemolysis (wide)

Bacitracin - S

PYR - + (pink)

Optochin Taxo P - R

tiny, soft or granular colonies

What are the are the characteristics of S. agalactiae-Grp. B

beta hemolysis( narrow)

Bacitracin -R

Camp - +

Hippurate + (purple)

Optochin Taxo P - R

soft, translucent  colonies

What are the are the characteristics of Enterococcus

Alpha, gamma, or beta hemolysis

Bacitracin -R

PYR - + (Pink)

Bile - +

NACL - +

Optochin Taxo P - R

medium,   grayish colonies

What are the are the characteristics of Group D strep (bovis)

gamma or alpha hemolysis

Bacitracin -R

Bile - +

NACI neg

medium,   grayish colonies

What are the are the characteristics of Viridans strep

alpha or gamma hemolysis (wide)

Bacitracin -R

Optochin Taxo P - R

tiny granular or   soft, white/gray colonies

Bile solubility neg

What are the are the characteristics of Strep pneumo-

Wide alpha hemolysis

Bacitracin -R

Optochin Taxo P - S

Bile solubility -+

small or larger,   mucoid; “pennies on a plate”

Are NEISSERIA and MORAXELLA gram neg cocci that are arrange singlely or in  pairs (diplococci) with adjacent sides flattened and oxidase positive and catalase positive

tes

What media do Neisseria grow on

–SBA/MTM------------Variable Growth

–CA---------------------Growth

What special media is use to recover gonococcus & meningococcus

–MTM (Modified Thayer Martin)

What is MTM (Modified Thayer Martin)

•Chocolate based with hemin/NAD

•Vancomycin--inhibits Gram pos cocci

•Colistin--inhibits Gram neg bacilli

•Nystatin--inhibits yeasts & molds

•Trimethoprim--inhibits swarming of  Proteus (highly motile gnb)

What is another media that is similar to MTM

•NYC (New York City) Media

•Martin-Lewis Media

What is JEMBEC

–Rectangular plate with MTM or ML

–Well for CO2 tablet

–Placed in Plastic bag and sealed

What kind of media is use to identify Neisseria

CTA (Cystine Trypticase Agar

What is CTA and its procedure

–Biochemical ID based on sugars fermented: glucose,  maltose, sucrose or lactose (fructose)

–Indicator of sugar used: phenol red

–Stab upper third of tube

–Cap and incubate in air for 24 hrs.

–Yellow = positive   Red = negative

What are the are the characteristics of Neisseria gonorrhoeae

AKA  Gonococcus

–Grows on CA or MTM; requires CO2

–Usually no growth on SBA since lot to  lot cysteine levels vary

–CTA sugars: ferments glucose only

–Test for Penicillin resistance since strains may be B-lactamase +

Grayish, beige, shiny round colonies

What are the infections of GONOCOCCUS

Gonorrhea (STD)

–cervicitis/PID

–urethritis

–pharyngitis

–proctitis

–ophthalmia neonatorum

–Complications: epididymitis, PID, sterility

DGI/DGC

–disseminated infection

–septicemia with petechial rash

–endocarditis

–gonococcal arthritis

What kind of specimen is collected for GONOCOCCUS

Endocervical--females

Urethra-males

Anorectal, throat, conjunctiva

Avoid cotton and calcium alginate swabs

Dacron or rayon swabs best

What the different between a male urethral and a female cervical in a GONOCOCCUS DIRECT SMEARS

Male urethral

–many segs

–extra- and intra- cellular gram negative diplococci

–diagnostic, culture not required

Female cervical

–many segs/other cells/bacteria

–extra- and intra- celular gram negative diplococci

–not diagnostic, must perform a culture or probe

What are the are the characteristics of NEISSERIA MENINGITIDIS

AKA meningococcus

–Grows on SBA, CA or MTM

–Growth enhanced by CO2, but may  not require CO2 for growth

–CTA sugars: ferments glucose and maltose

What kinds of infections dose MENINGOCOCCUS have

nBacterial meningitis

–Encapsulated strains

–Asymptomatic human carriers

–Most common cause  of meningitis in young adults

Septicemia: meningococcemia

–petechial rash/lesions & arthritis

DIC/Waterhouse-Friderichsen Synd.

What is the SPECIMEN PROCESS for MENINGOCOCCUS

CSF

–centrifuge and plate sediment on SBA and CA

–prepare direct smears

–Hold CSF at Room temperature (never  at 4⁰C)

Pharyngeal: plate on CA & MTM

Blood culture: incubate ASAP

What kind of specimen is use for MENINGOCOCCUS

Specimens: CSF / Serum / Urine

CSF most common specimen for direct serological ID

Methodologies: (detect antigen)

–ELISA/EIA

–Latex Agglutination

What does an CSF infected with MENNGOCOCCUS look like in an direct smear

Gram Stain

–Increased SEGS

–Intracellular and extracellular gram negative diplococci, adjacent sides flattened

What are the BIOCHEMICAL ID of N. lactamica

–Glucose +  Maltose +  Lactose +

What are the BIOCHEMICAL ID of nN. sicca

–Glucose+  Maltose + Sucrose +

What are the are the characteristics of M. CATARRHALIS

Grows on SBA and CA; variable growth on MTM

Grows at RT on nutrient agar

No growth on MAC/EMB

Colonies: convex, cream-colored or grayish-beige

Oxidase and Catalase positive

Butyrate esterase positive

CTA sugar fermentation:  none

What are some Common Opportunistic infections for M.Cat

–Otitis media/Sinusitis/ Bronchitis/Conjunctivitis

–Pneumonia/Meningitis/Bacteremia

Are HAEMOPHILUS Tiny, gram negative bacilli and coccobacilli, often pleomorphic

      Require X and/or V factors

yes

What media do HAEMOPHILUS grow on

–CA---------------------Growth; enhanced by    CO₂

What does the x factor contain

–Hemin/Hematin/

    Porphyrins

What doe the v factor contain

–NAD/NADP (yeast

    extract/potato)       

What does the X and V factor contain

Rabbit/Horse blood

What are X and V strips

streak unknown on MH; place X, V , XV strips on streak; incubate and observe for growth around strips to determine X/V requirements

What is an Haem ID Quad Plate

each quad has X, V, or X & V factors. Streak unknown on each quad; incubate & observe for growth and B-hemolysis on blood quad

Which HAEMOPHILUS SPECIES grows on x and v

–Haemophilus   influenzae

–Haemophilus aegyptius

–Haemophilus haemolyticus

Which HAEMOPHILUS SPECIES grow on x

H. ducreyi

Which HAEMOPHILUS SPECIES grows on V

–H. parainfluenzae

–H. parahaemolyticus

What are the are the characteristics of HAEMOPHILUS INFLUENZAE

grayish/beige colonies with mousy odor on CA only. Smear: tiny, pleomorphic gram negative bacilli/coccobacilli

–H. flu requires both X and V factors

–Beta-lactamase + strains are resistant to Penicillin, Ampicillin, and Cephalosporins

What specimens are use for HAEMOPHILUS INFLUENZAE

CSF, Serum,  or Urine

What are the infections of H. FLU

Encapsulated H. influenzae

–Children 6 mos. to 6yrs.:meningitis,    septicemia, epiglottitis/tracheitis

– Adults: pneumonia & arthritis

–Also ear, eye, nose infections

nnon-encapsulated strains:

–otitis media

–sinusitis

–conjunctivitis

–pneumonia (elderly, or underlying lung problem)

bronchitis

What are the are the characteristics of HAEMOPHILUS DUCREYI

Requires X factor only: grows slowly

gram negative coccobacilli in chains    “school of fish” arrangement

What cause HAEMOPHILUS DUCREYI

chancroid, genital ulcers

Are Enterobacteriaceae an Diverse family of gram negative bacilli (gnb) and Aerobic; facultative anaerobes, so they grow well on most  anaerobic primary isolation media

yes

What is the common Characteristics of Enterics

All ferment glucose

Most are oxidase negative (1 exception)

All are catalase positive

Most reduce nitrates  to nitrites

All except Klebsiella, Shigella and Yersinia are motile at 25/35⁰C;  Yersinia is motile at 25⁰C, but nonmotile at 35⁰C.  Flagellar arrangement is generally peritrichous

What media is use to test oxidase

SBA or CA

Somatic O antigen-

heat stable cell wall lipopolysacharide

Flagellar H antigen

heat labile protein

Capsular K  ( Vi)

antigen--heat labile polysaccharide

What kind of infection do E. coli and Kleb.  pneumoniae cause

UTI, wound and respiratory infections, meningitis,  septicemia, etc.

What are the are the characteristics of Edwardsiella

•rarely isolated in US

•found mainly in tropics or subtropics

•causes infections associated with aquatic environments

•warm and cold blooded reservoir hosts

What are the infections of Edwardsiella

•gastroenteritis

•wound infections : trauma or accidents in fresh water/aquaria

•meningitis

What media dose Enterics grow on

SBA and CA: excellent growth: large, gray colonies

MAC/EMB: excellent growth

THIO: Growth throughout  (facultatives)

What are the infections for Y. enterocolitica

•Enterocolitis (most  = infants/children)

•Ileitis (mesenteric adenitis)

•Septicemia, and transfusion  sepsis

How is Y. enterocolitica transmitted

•Contaminted food,  H₂O

•fecal-oral???

What are the infections for Yersinia pestis

•Causes the Plague:

–bubonic

–septicemic

–pneumonic

What are the transmissions of Yersinia pestis

–infected flea bite

–direct contact: rodents

What are some stool pathogens

•Salmonella

•Shigella

•E. coli O157:H7

What are some less common stool pathogens

•Edwardsiella tarda

•Yersinia enterocolitica

•Other  strains of E. coli

What media is Selective for enterics and Differential  based on Lactose utilization

MAC or EMB

What is MAC

–Indicator of lactose fermentation:  neutral red

–Lactose positive = bright pink/red 

–Lactose negative = clear/colorless

–Inhibitor of gram positives:  bile salts/crystal violet

What is EMB

•Eosin Methylene Blue  (EMB)

–Indicators of Lactose fermentation:  eosin and methylene blue

–Lactose positive = dark purple

–Lactose negative = clear/colorless

–Inhibitors of gram positives:  eosin and methylene blue

•Note : Levine’s EMB also has sucrose in it.  Some gnr use sucrose and produce a green sheen on  Levine’s formulation.

HE,XLD and SS media is use for what

used  for the primary isolation of enteric (gut) pathogens, Salmonella or Shigella, from stool specimens

What do HE, XLD and SS all contain

–All contain bile salts to inhibit normal flora gut enterics

–All contain an H₂S indicator

–All contain an indicator to detect lactose (or other carbohydrate)  fermentation

–Shigella may be inhibited by SS and  XLD

What are the color reactions for HE

–Lactose/Sucrose  fermenter:  yellow or salmon-orange

–Non lactose fermenters (NLF): green or colorless

–Salmonella:  Blue-green with black center

–Shigella:  Green/colorless

•SS (Salmonella-Shigella agar)

–Salmonella:  Colorless with black center

Shigella:  Colorless (except  S. sonnei  which at       48 hrs. is pale pink--why???) late fermenter

What are the color reactions for XLD

–Lactose/Xylose fermenter:  Yellow colonies

–Lactose/Xylose nonfermenter:  Colorless colonies

–Lysine positive:  Red colonies

–Salmonella:  Red with black centers

–Shigella:  Colorless

What is SMAC

•MacConkey with Sorbitol)

–Used to recover E. coli, O157:H7

–Contains sorbitol instead of lactose

–Sorbitol positive:  pink/red colonies

–Sorbitol  negative: Clear/colorless colonies

–E. coli, O157:H7 is sorbitol negative--clear coloniest

What is CIN and what enteric is it use for

–Used to recover Yersinia enterocolitica

–Contains antibiotics to inhibit normal gut enterics

–Contains mannitol, not lactose

–Incubated at 25⁰C

–Yersinia enterocolitica grows best at 25-30⁰C,  and ferments mannitol

–Yersinia enterocolitica :  “Bull’s eye” colonies-thin pink halo  rim with large, bright pink/red center

–Note: mannitol negative enterics: Clear  colonies 

What are three broths that are use to recovery Salmonella and Shigella from stools by inhibiting coliforms

•Gram Negative (GN) Broth   

•Tetrathionate Broth

•Selenite Broth

What is the glucose, lactose and sucrose ratio for tsi

Glucose 1

Lactose 10

Sucrose 10

What is the procedure for TSI

Using an inoculating needle, stab the butt all the way to the bottom

Streak the slant as the needle is pulled out

Loosely cap the tube

What are the interpretation results for TSI

Red/Red (Alk/Alk) : nonfermenter

Red/Yellow (K/A) (Alk/Acid): Ferments Glucose only

5Yellow/Yellow (A/A) (Acid/Acid) = Ferments Lactose and/or Sucrose

Black in Deep (Butt) = H₂S producer

Bubbles/cracks in Butt = Gas produced

What is Kovak and its interpretation

•para-dimethylamino-benzaldehyde

•Interpretation

–Indole + = pink/red

–Indole - = colorless

what is DMAC and its interpretation

•para-dimethyl-      cinnamaldehyde

•Interpretation

–Indole + = blue-green

–Indole - = pink-red

What is IMViC

•I    =   Indole

•M =   Methyl Red

•Vi =   VP  (Voges Proskauer)

•C  =   Citrate

What are the characteristics of Salmonella 

HE = Blue green with black center 

SS = Colorless with black center

XLD = Red with black centers 

TSI = K/A

H2S= pos

I= -

M= - 

Vi= - 

C= + 

Urea = neg

What are the characteristics of Shigella 

HE =Green or colorless

SS =Colorless

XLD = Colorless

TSI = K/A

Indole = variable

motility = nonmotile

Urea = neg

SBA = delicate colonies

What are the characteristics of Edwardsiella Tarda 

TSI = K/A

Indole = pos

Motility = motile

I= + 

M= + 

Vi= - 

C= -

Urea = neg

What are the characteristics of Y. enterocolitia 

• Gram Neg coccobacillus 

• A/A (sick- yellow/ orange) or K/A 

• SUC + 

• LAC – 

• IND +/- 

• Urea + 

• Motile at 25C (not motile at 35) 

• CIN at RT 

• MAC: tiny clear colonies  

• Bulls eye colonies: bright pink center inside a colorless halo 

• Mannitol + 

What are the characteristics of Yersinia pestis 

• Gram negative coccobacilli 

• K/A 

• SUC – 

• IND – 

• Nonmotile 

What are the infections for Shigella  

Shigellosis ( bacillary dysentery) 

How is Shigella  transmitted

fecal oral, contaminated food/ water, only human hosts