1. enterobacteriaceae (upload)

7/23/2019 1. Enterobacteriaceae (Upload)

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Gram Negative

Bacteria:

INTRODUCTIONINTRODUCTION

TOTO

ENTEROBACTERIACEAE


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Study Outline /Review

uic! A""e""ment #For Residual Knowledge GNB # Types & Short overview, Position of

Enterobacteriaceae among them, ut picture!

Enter$%acteriaceae # "istory, #e$nition,

%haracteristics, %lassi$cation, mportance,#iseases, 'ntigenic structures & (irulencefactors!

GI di"ea"e" # Terminologies, )rief

mechanisms of pathogenesis! &a% D' # 'pproach for the & other disorders

*with em()a"i"on Enterobacteriaceae+!


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Categories of GNB

*+ Related t$ Enteric tract:

Enterobacteriaceae - Esch coli, Salmonella (ibrionaceae - Vibrio cholerae Pseudomonadaceae - Pseudomonas aeroginosa )acteroidaceae - Bacteroides fragillis Helicobactor. Campylobacter

,+ Related t$ Re"(irat$ry tract: Haemophilus, Legionella, Bordetella,

Acinetobacter Enterobacteriaceae - Klebsiella

-+ Related t$ Ur$genital tract:

ardnerella, Haemophilus ducreyi,Acinetobacter Enterobacteriaceae - Proteus, /PE%, C!granulomatis

.+ Related t$ Nerv$u" Sy"tem: Haemophilus, Pseudomonas Enterobacteriaceae - 01E%, Klebsiella, Serratia


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Categories of GNB (contd.)

+ Related t$ 0u"cul$"!eletal "y"tem: Pseudomonas,Acinetobacter Enterobacteriaceae - E! coli, Klebsiella, Serratia

1+ Related t$ C2S: 3ACE4 gr$u( 2Haemophilus,Actinobacillus

actinomycetemcomitans, Cardiobacteriumhominis, Ei"enella corrodensand Kingellaspp!3

N$n53ACE4 gr$u(- Pseudomonas,

Enterobacteriaceae 6Enterobacter, Citrobacter,

Pro#identia, Serratia, Proteus, Salmonella, E!coli7

8+ Related t$ 9$$n$"i" 2'nimal sources3: Brucella, $rancisella, pasteurella,

Enterobacteriaceae - %ersinia


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Enter$%acteriaceae

Scienticcla""icati$nD$main: Bacteria

;)ylum: ;r$te$%acteria

Cla"": Gamma(r$te$%acteria

Order: Enter$%acteriale"


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n the 4899s, the $rst member Serratiamarcescenswas discovered by )i:io in 48;6 ona dish of talian barley 2Polenta3!

Klebsiella and Proteus were discovered in4889s!

n 4856, Theobald Smith distinguished


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n 4567, =tto Rahn $rst proposed the

name Enterobacteriaceae family with asingle genus, Enterobacter and 44;species!

n 45>>, )orman, Stuart & ?heeler

de$ned the family characteristics!

n 457;, there were 44 genera and ;@species!

n 458A, ;; genera, @5 species! n ;99>, >9 genera, and ;99 species!

't present, >.8 genera and ?,@@

"(ecie"+


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Enter$%acteriaceae is a compleB andheterogeneous bacterial family

consisting of large number of relatedgenera that are found in the small &mostly large intestines 2colons3 ofhuman, animals & insects, and also in

soil, plants, water, foods & decayingmatters having the following

di"tingui")edc)aracteri"tic":5

DE


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4! 0)C ram 0egative )acilli.Rods ;! Facultative anaerobes 5555 Cf+Pseudomonas,

Helicobacter pylori

6! Either 1otile2peritrichous Dagella3ORnon

motile 5555555 Cf+ Vibrio, Pseudomonas,H! pylori>! Ferment #glucose *#eBtrose+ 2with production

of


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1+ Reduce 0itrate *NO-5+ to 0itrite 2NO,532(ery few eBceptions, e!g! Photorhabdusspp!3

555 &ycobacteriaalso reduce Nitrateto 0itrite+

7! Some have capsule 2Klebsiella3 or slimelayer!

8! Some erment &act$"e along with orwithout other substances!

5! 0on sporeforming!

49! "ave pili or $mbriae!

44! Sharing and de$nitive antigenic

characteristics!

C)aracteri"tic"

6c$ntd+7


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0em%er" $Enter$%acteriaceae

T)ere are >.8genera and ?,@@"(ecie"

Num%er" are increa"ing r$m time t$ time

1. Escherichia Esch. coli2. Salmonella S. typhi S. paratyphi.

3. Shigella S. dysentery S! sonni,

4. Klebsiella K. pneumonae

5. ProteusP. mirabilis'!%ersinia %! pestis,%! enterocolitica

(!Enterobacter E! cloacae

)! Citrobacter C! freundi


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Ot)er mem%er":

*! Serratia S! marcescens+! Hafnia H! al#ei

! &organella &! morganii

-! Calymmatobacterium C!granulomatis

.! Er/inia

'! Pectinobacterium

(! Ed/ard siella

)! Plesiomonas etc!

*@species cause F@ of the humandiseases, whereas ,-5, species consist

FFof all Enterobacteriaceae isolatesfrom human


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Shigellawith no Flagella (Atrichous)


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Escherichia coli with PeritrichousFlagella


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Vibrio choleraewith Polar MonotrichousFlagella


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Campylobacter jejuni

with Amphitrichous flagella


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Helicobacter curved rod with Lophotrichous Flagella


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Traditi$nal Cla""icati$n: )ased on disease production, )ased on site of the disease

6Clinical cla""icati$n7 )ased on


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On The Basis of Disease

ProductionNormal floraOROpportunistic (potential)

pathogen :

Most of them, e.g.E.coli,Enterobacter,

Citrobacter,Proteus

Primary pathogen:

Salmonella,Shigella,Yersinia,


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Clinical Classification

Primarily Intestinal Pathogen

Shigella,Yersinia enterocolitica

Intestinal & Extra-intestinal PathogenSalmonella,Esch. coli,

Extra-intestinal Pathogen-

Klebsiella,Proteus, Yersinia pestis


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On The Basis of

Lactose Fermentation

Lactose fermenters LF!:Esch coli, Klebsiella(except K.

rhinoscleromatis),Enterobacter,

Citrobacter,

Late Lactose ferments LLF! :Shigella sonni,Providencia, Serratia,

Lactose non"fermenters NLF!:Salmonella, Shigella (except S. sonni),

Proteus,Klebsiella rhinoscleromatis,Yersinia


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On The Basis of #otility

$anging drop, %oft agar!

#otile:

Salmonella, Esch. coli, Proteus

Non"motile:

Shigella, Klebsiellaetc.


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#otility in %oft %emi"solid! &gar #edia

Left 'Right: #otile( Center:Non"motile


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Modern Classification

TRIBES

E'am(le" $Enter$%acteriaceaeN$ NA0E

IE"c)eric)ie

ae Escherichia,Shigella

II Edward"ieleae

Ed/ardsiella

IIISalm$nellac

eae Salmonella

I2Citr$%acter

eae Citrobacter

24le%"iellace

aeKlebsiella, Enterobacter,

Hafnia, Serratia

2I ;r$teeae

Proteus, &organella,

Pro#idencia


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The antigenic structures are used todiGerentiate organisms within a genus orspecies!

Three maHor classes of antigens are foundI O 2=hne "auch3 # Somatic or cell wall

antigens, 3 2"auch3 #Flagellar antigens and 4 2Kapsel3 # %apsular 6or 2irulence7

antigens!

Plays important role in epidemiology,classi$cation as well as serotyping!

Antigenic

Structure"


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"eat stable =uter component 2polysaccharide part or

side chain3 of the


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"eat labile!

%onstructed from proteins!

'ntigenic variation is due to the diGerence in'mino 'cid seJuences!

'ntibody against " proteins - IgG+

The Dagellar $lament is composed of a singleprotein, Flagellin!

1olecular weights of Flagellin range from 66,999to @9,999 among diGerent bacterial species!

n some bacterial genera, such as Salmonella,many varieties of Dagellar antigen have beenidenti$ed!


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3eat la%ile$rrelatively heat stable 67!

Polysaccharides in nature 2antiphagocytic3!

Found on the surface of many bacterialspeciesI

Klebseilla %ertain strain of Esch! Coli

Salmonella typhi2Klie 'g3,

certain Citrobacter2Klie 'g3 etc

=ften constitutes the outermost layer of thecell!

)oiling for 4A minutes to 4 hour would destroythe K antigen and unmas = antigens!

Ca("ular 4 $r2i Antigen"


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4 $r2iAntigen" 6c$ntd+7


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Determinant" $;at)$genicity

'bility to coloni:e, some coloni:ation factors areI

'dhesins, Pili or Fimbria En:ymesI g' protease, /rease etc!

nvasiveness

ToBinsI EndotoBin 2


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)irulence Factors contd(!

'ntigenic phase variationI

'bility to alternately eBpress or not eBpresseither capsule or Dagella

Thus avoid host immune response!

Susceptibility of the host 6NOT a virulence

factor7:astric acid suppression

'nomalies, circumcision etc!

=ther factorsI

En:ymes 2e!g! /rease,0lactamases3Resistance plasmid 2#rug resistance3

)ile solubility 2)ile salt tolerance3

'bility to resist illing inside the cells etc!


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1echanisms inovlved

0('S=0 Salmonella, Shigella, Campylobacter,

Escherichia coli, Entamoeba histolytica!

0=00('S(E 1E%"'0S1ToBin production

Vibrio cholerae, ETE%

#ecreased absorption

Rotavirus, norwal agent, giardia lamblia

E0L1ES

Proteus


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Di"ea"e" ;r$duced4! TI Salmonella, Shigella, Esch! %oli,

Lersinia

;! /TI Esch! %oli, Proteus, Klebsiella,%itrobacter, Providentia!

6! ?ound infectionI Esch! %oli, Proteus,Klebsiella, Enterobacter!

>! PneumoniaI Esch! %oli, Klebsiella, Proteus!

A! )acteraemiaI Salmonella, Klebsiella,

Esch! %oli, Proteus!@! 1eningitisI Esch! %oli, Klebsiella,

Salmonella, Proteus 2rare3


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S(ectrum $ GIT


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S(ectrum $ GITdi"ea"e"

#iarrhea FreJuent &.or loose.Duid stool 2smallintestine3!

#ysentery Stool with blood.pus 2M fever3accompanied by pain, abdominal cramps

2inDammatory disorder of large intestine3! Oranybloody diarrhea!

astroenteritis Syndrome with T symptomsnausea.vomiting, diarrhea, abdominal discomfort!

astritis Syndrome with stomach symptoms i!e!nausea, vomiting, upper abdominal discomfort!


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S(ectrum $ GIT di"ea"e"

Enteritis Syndrome associated withinDammation of small intestine!

Enterocolitis Syndrome associatedwith inDammation of both intestines!

%olitis Syndrome associated withinDammation of large intestine!

Food poisoning - Syndrome of infectionassociated with consumption ofcontaminated food Food associatedinfection!


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DIARR3EA&DISEASES

Em()a"iJing $nENTEROBACTERIACEA

E

&a%+

Diagn$"i"


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;RINCI;&E

t is based on demonstration of thecausative agent by 1.E and isolation

& identi$cation by standardmicrobiological techniJues and alsoby immunological tests and somespecial tests! 1olecular techniJuesare also employed!


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STE;S5*:STE;S5*:

*+ S(ecimen: Acc$rding t$ t)e "ite $

t)e le"i$n:Freshly passed stool or rectal swab, Urine,Sputum, )lood, Pus, %SF & other body Duids

etc!

,+ 0icr$"c$(ic E'aminati$n 60/E7:CRUCIA&

Saline I$dine and 95N (re(arati$n:

For E! histolytica, iardia, Cryptosporidium! iardia & Cryptosporidium both are acidfast in modi$ed 0 staining!

;u" Cell" 0acr$()age RBC:

"elps to detect type of diarrhea


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STE;SSTE;S 6c$ntd+7 # ,:

-+ I"$lati$n K Identicati$n 6Culture7:

Specimen should either be immediatelycultured OR should be placed in transportmedia 2Stuart, 'mies, %ary)lair etc!3 incase of stool!

ORin case of urine, should be stored at .@C,if not inoculated in the media within -@minutes!

ncubation at -8@C aerobically for usualpathogens & at .,@C microaerophilically forCampylobacter!

Selection of culture media depends on

provisional diagnosis&.or information provided bythe referrin h sician! St$$l culture i" $


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STE;SSTE;S 6c$ntd+7 # -:

Culture 0edia:

1ac %onKeyNs 'gar 60A7ShigellaSalmonella 6SS7 Agar

Triple Sugar ron 6TSI7 etc+

L

Thiosulfate %itrate )ilesalts Sucrose 2TCBS3,Cam(y5BAetc! Additi$nally


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STE;SSTE;S 6c$ntd+7 # .:

I"$lated c$l$nie" are identied %y:

1orphology & rowth on the Type of 1edia Staining & 1.E, )iochemical tests 6


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STE;SSTE;S 6c$ntd+7 # :

Immun$l$gical te"t:Serogrouping by antisera,#etection of rota viral 'g, C! di2cile toBin fromstool by


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1. enterobacteriaceae (upload)

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