IS 5887-7: Methods for Detection of Bacteria Responsible for Food Poisoning, Part 7: General Guidance on Methods for Isolation and Identification of Shigella

12 Pages·1999·1.4 MB·English

by  Bureau of Indian Standards

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“The Right to Information, The Right to Live” “Step Out From the Old to the New”
IS 5887-7 (1999): Methods for Detection of Bacteria
Responsible for Food Poisoning, Part 7: General Guidance on
Methods for Isolation and Identification of Shigella [FAD
15: Food Hygiene, Safety Management and Other Systems]
“!ान $ एक न’ भारत का +नम-ण”
Satyanarayan Gangaram Pitroda
““IInnvveenntt aa NNeeww IInnddiiaa UUssiinngg KKnnoowwlleeddggee””
“!ान एक ऐसा खजाना > जो कभी च0राया नहB जा सकता हहहहै””ै”
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“Knowledge is such a treasure which cannot be stolen”

IS5887( Part7): 1999

Indian Standard
METHODS ROF NOITCETED FO AIRETCAB
ELBISNOPSER ROF FOOD GNINOSIOP
PART 7 GENERAL GUIDANCE ON METHODS FOR ISOLATION AND
IDENTIFICATION OF W/GE/X/I
ICS 07. 100.30

0 BIS 1999
BUREAU OF INDIAN STANDARDS
MANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI 110002

June 1999 Price Group 3
dooF ygoloiborciM lanoitceS ,eettimmoC FAD 46
FOREWORD
sihT naidnI dradnatS was detpoda yb eht uaeruB fo naidnI Standards, retfa eht tfard dezilanif yb eht dooF
ygoloiborciM lanoitceS eettimmoC had neeb devorppa yb eht dooF and erutlucirgA noisiviD .licnuoC
lareveS smsinagro-orcim gnitanimatnoc doof evig esir ot lacinilc ,smotpmys hcihw edulcni lanimodba pain,
nausea, ,gnittimov aeohrraid and semitemos .aixeryp gninosioP hguorht doof is deziretcarahc yb eht evisolpxe
erutan htiw hcihw eht smotpmys rucco ni esiwrehto yhtlaeh .slaudividni hcuS erutan fo doof gninosiop spleh
ni gnitaitnereffid eht snoitidnoc fo skaerbtuo fo enrob-doof suoitcefni sesaesid hcihw yllareneg daerps revo
a doirep fo lareves .syad sihT standard srevoc eht dohtem rof noitalosi and noitacifitnedi fo elbisnopser
ShigeNa
rof doof .gninosiop
sihT naidnI dradnatS was yllanigiro dehsilbup ni 1970 and tsrif desiver ni 1976 nehw ti was split otni parts:
traP III gnirevoc eht sdohtem rof htob and On a weiver yb eht lacinhcet eettimmoc
Salmonella Shigella.
elbisnopser rof gnitalumrof standards ni eht aera fo doof ,ygoloiborcim eht traP III is gnieb desiver ot etarapes
eht .snoisivorp ehT snoisivorp fo Salmonella have neeb dengila htiw 0SI 6579 : 1993 and deniater ni traP ;3
saerehw eht snoisivorp fo evah neeb derevoc ni traP 7 fo eht standard.
Shigella
nI gnitroper eht tluser fo a tset ro sisylana edam ni ecnadrocca htiw this standard, fi eht lanif ,eulav devresbo
ro ,detaluclac is ot eb dednuor ,ffo ti shall eb enod ni ecnadrocca htiw SI 2 : 1960 seluR‘ rof gnidnuor ffo
laciremun seulav ( revised )’
SI 5887 (Part 7) : 1999
Indian Standard
METHODS ROF NOITCETED FO AIRETCAB
ELBISNOPSER ROF FOOD GNINOSIOP
PART7 GENERAL GUIDANCE ON METHODS FOR ISOLATION AND
IDENTIFICATION OF SWGELLA
1 EPOCS evitisop ( noitpecxe ni Sh. dysenteriae 1, that is,
esadixo - ,evitagen etartic - ,evitagen
Sh. shiga ),
sihT standard ( traP 7 ) sebircserp eht dohtem rof
shows evitatnemref noitcaer ni s’nosfieL-hguH ,tset
noitalosi and noitacifitnedi fo
Shigella. tnemref ‘sugars’ tuohtiw gas noitcudorp ( snoitpecxe
era emos strains fo ehT suneg
Sh.Jexneri 6). Shigella
2 REFERENCES
has neeb dedividbus no eht basis fo lacimehcoib
ehT gniwollof naidnI Standards niatnoc snoisivorp sretcarahc and lacigolores gnitset otni ruof ,spuorgbus
hcihw hguorht ecnerefer ni this ,txet etutitsnoc puorgbus A gniniatnoc 10 ,sepytores
(Sh. dysenteriae,
noisivorp fo this standard. At eht emit fo ,noitacilbup ,)ol-l puorgbus B ( gniniatnoc 6 -ores
Sh. flexneri,
eht snoitide detacidni erew .dilav llA standards era ,sepyt ,6-1 and owt ,stnairav X and y ,) puorgbus C
tcejbus ot noisiver and seitrap ot stnemeerga desab ( gniniatnoc 15 ,sepytores 51-1 ) and
Sh. boydii,
no this standard era degaruocne ot etagitsevni eht puorgbus D ( gniniatnoc 2 ,smrof esahp I
Sh. sonnei,
ytilibissop fo gniylppa eht tsom tnecer snoitide fo eht and II .) ehT setalosi gnignoleb ot eht suneg
Shigella
standards detacidni :woleb nac eb desufnoc htiw esoht gnignoleb ot
Hafnia,
and eht ,cinegoreana -non
SI .oN Providence, Aeromonas
Title
elitom and esotcal-non gnitnemref
Escherichia coli.
1070 : 1992 tnegaeR edarg retaw (third ehT lufesu lacimehcoib sretcarahc hcihw elbane
noitacifitnedi fo a tcepsus strain as era nevig
revision ) Shigella
ni elbaT 1.
5404 : 1984 edoC fo ecitcarp rof handling fo
samples rof lacigoloiborcim sisylana eallegihS era hcum less evitcaer than and an
E. coli
(Jrst revision ) etalosi that secudorp dica yltpmorp ( that is nihtiw
24 )sruoh morf a ediw yteirav fo ,setardyhobrac
6850 : 1973 Agar, lacigoloiborcim edarg
yldetbuodnu is ton a ,revewoH yna msinagro
Shigella.
:1586 1973 taeM ,tcartxe lacigoloiborcim edarg hcihw is yllacimehcoib a neve fi ti seod ton
Shigella
etanitulgga ni eht elbaliava gnipyt ,ares must eb -er
6852 : 1973 eliB salts, lacigoloiborcim edarg
dedrag as a Shigella and tnes ot a ecnerefer yrotarobal
6853 : 1973 ,enotpeP lacigoloiborcim edarg rof noitamrifnoc and .gnipytores
7004 : 1973 tsaeY ,tcartxe lacigoloiborcim edarg 6 AIDEM
7128 : 1973 esoetorP ,enotpep lacigoloiborcim 6.1 Nutrient Broth
edarg
xiM and evlossid yb gnitaeh 10 g enotpep ( ees
7590 : 1975 ,enitaleG lacigoloiborcim edarg SI 6853 ,) 10,g taem tcartxe SI 6851 ,) and 5 g
(see
muidos edirolhc ni 1000 lm .retaw nehW ,looc adjust
3 GNILFMAS
Hp ot 7.5 ot 7.6. evomeR etatipicerp yb noitartlif
selpmaS shall eb deldnah ni ecnadrocca htiw eht hguorht retlif .repap eziliretS yb gnivalcotua at
erudecorp nevig ni SI 5404. 120°C rof 15 .setunim
4 YTILAUQ STNEGAERFO 6.1.1
Nutrient Agar
Unless deificeps ,esiwrehto erup slacimehc shall eb oT eht stneidergni an ni 6.1, add agar ( ees SI
6850 )
deyolpme ni stset and dellitsid retaw ( ees SI 1070 ) ni hcus a noitartnecnoc as lliw yfidilos and ecudorp
shall eb desu erehw esu fo retaw as a tnegaer is a yltneichfus mrif ecafrus nehw deruop ni elirets irtep
.dednetni dishes. ehT noitartnecnoc fo agar ot eb dedda seirav
morf hctab ot hctab and sdeen ot eb detsujda
5 LARENEG SCITSIRETCARAHC
.ylgnidrocca Usual noitartnecnoc deriuqer yrav morf
,ciboreA evitagen-marG ,sdor ,elitom-non -esalatac 1.5 ot 3 .tnecrep evlossiD eht agar ni tneirtun htorb

1
IS 5887 (Part 7 ) : 1999
and ezhirets yb gnivalcotua at 120°C rof 15 .setunim muinomma etartic ( neerg selacs ) ni 100 lm fo
eraperP setalp and sepols as .deriuqer elirets ,retaw htiw .gnitaeh
6.2 Kauffmann-Muller’s Tetrathionate Broth )c Solution B - evlossiD 10 g muidos -yxosed
etalohc ni 100 lm fo elirets .retaw
oT 90 lm fo elirets tneirtun htorb )1.6( add htiw
citpesa snoituacerp and suounitnoc ,noitatiga 5 g )d eziliretS snoitulos A and B at 60°C ni a
muiclac etanobrac hcihw has ylsuoiverp neeb htab-retaw rof eno .ruoh roF gniraperp eht
deredwop and dezilirets ni eht ,evalcotua 10 lm fo a etalohcyxosed etartic agar muidem tlem
50 tnecrep noitulos fo muidos etahplusoiht 100 lm fo eht agar esab ni a htab-retaw and
( ylsuoiverp dezilirets yb gnimaets ,) 2 lm muissatop add 5 lm hcae fo eht snoitulos A and B ni eht
enidoi/edidoi noitulos ( evlossid yb gnidnirg ni a redro ,nevig using etarapes .settepip xiM llew
ratrom 5 g fo muissatop edidoi ni 20 lm retaw and neht retfa hcae .noitidda looC eht ebut at 50 ot
add ylwols htiw tnatsnoc gnidnirg 4 g ;enidoi peek .C’S5 ruoP otni dezilirets irtep dishes ( 12 ot
eht noitulos ni krad dereppots-ssalg elttob ,) 1 lm fo 15 lm ni hcae ) and wolla ot .tes yrD eht ecafrus
0.1 tnecrep suoeuqa noitulos fo tnaillirb neerg and fo eht muidem ni eht rotabucni erofeb .esu
5 lm fo elirets xo elib ro 1 g elibfo salt ( ees SI 6852 .)
etubJ’siD ni 10 lm seititnauq ni elirets tset .sebut ehT 6.5 MacConkey Agar Medium
muidem shall ton eb detcejbus ot noitazilirets retfa Mix 5 g sodium taurocholate ro elib salts ( ees
noitaraperp as eht etanoihtartet yam esopmoced no SI 6852 ,) 20 g fo enotpep ( SI 6853 ,) 5 g muidos
.gnitaeh edirolhc and 15 ot 30 g agar ( ees SI 6850 ,) htiw
1000 lm .retaw maetS litnu eht sdilos era .devlossid
6.3 Selenite F Broth
looC ot tuoba 5O“C, and at this erutarepmet adjust
evlossiD 4 g muidos dica ,etineics 5 g enotpep ( ees noitcaer ot Hp 7.6 ot 7.8. evalcotuA at 120°C rof
SI 6853 ,) 4 g ,esotcal 9.5 g muidosid negordyh 15 nim and retlif elihw toh hguorht a doog edarg fo
etahpsohp and 0.5 g muidos negordyhid etahpsohp retlif ,repap ro a gulp fo nottoc depparw ni ezuag and
ni 1000 lm fo elirets retaw ( retaw devalcotua at 120°C decalp ni eht .lennuf Adjust noitcaer fo eht etartlif
rof 30 setunim ) htiw elirets snoituacerp and etubirtsid ot Hp 7.3 at 50°C ro Hp 7.5 at moor .erutarepmet
eht hsiwolley noitulos ni 10 lm stnuoma ni elirets Add 100 lm fo 10 tnecrep suoeuqa noitulos fo esotcal
deppac-wercs .selttob maetS at 100°C rof 30 .setunim ( ro 10 g esotcal ) and 3.5 lm fo 2 tnecrep noitulos fo
ehT muidem shall ton eb .devalcotua erehT yam eb a lartuen der ni 50 tnecrep .lonahte xiM ,ylhguoroht
slight der etatipicerp ni eht ,muidem tub this seod ton etubirtsid otni dezilirets sksalf and ezilirets ni eht
ereferetni htiw eht noitca fo eht .muidem ehT Hp fo evalcotua at 120°C rof 15 .setunim roF ,esu tlem ni
eht muidem as deraperp shall eb 7.1 and eht ytitnauq eht ,remaets ruop otni elirets irtep dishes and wolla
fo setahpsohp dedda yam eb deirav ylthgils ot eveihca ot .tes
this.
6.6 Nutrient Agar Medium for Motility Test
6.4 Desoxycholate Citrate Agar Medium
ehT muidem stsisnoc fo 3 g taem tcartxe ( ees
)a Agar esaB - evlossiD yb gnitaeh 20 g taem
SI 6851 ,) 10 g enotpep ( ees SI 6853 ,) 5 g muidos
tcartxe ( ees SI 685 1 ) ni 200 lm ,retaw and
,edirolhc and 4 ot 5 g agar ( ees SI 6850 ,) devlossid ni
ekam eht noitulos just enilakla ot
1000 lm .retaw Adjust Hpeht ot 7.5 ot 7.6. tuP eht
nielahthplonehp htiw 50 tnecrep muidos
devlossid muidem otni tset sebut ot llif a trap fo eht
edixordyh .noitulos lioB and retlif hguorht
ebut and ecalp otni this a glass ebut nepo at htob
retlif .repap Adjust ot Hp 7.3, ekam up ot
.sdne enO dne fo eht glass gnibut shall tcejorp morf
200 lm and add 20 g esoetorp enotpep ( ees
evoba eht ecafrus fo eht agar rof ton less than 1.5
SI 7128 .) nI rehtona ,lessev evlossid 90 g agar .mc eziliretS eht sebut htiw eht muidem at 120°C rof
( ees SI 6850 ) ni 3 700 lm retaw yb gnimaets
15 ,setunim and neht .looc ehT ycnetsisnoc fo eht
rof eno .ruoh retliF eht agar ,noitulos add ot
agar dluohs eb tfos tub ton .diuqil sihT is deveihca
ti eht taem enotpep-tcartxe noitulos and .xim
yb gniretla eht tnuoma fo agar ,desu fi .yrassecen
Add 5 lm fo 2 tnerep lartuen der noitulos ni
50 tnecrep lonahte and 40 g .esotcal ekaM 6.7 Medium for Hugh-Leifson’s Test
up ot 4 sertil htiw .retaw ,xiM elttob yletarucca
evlossiD yb gnitaeh ni 1 000 lm ;retaw 2 g enotpep
ni stol fo 100 lm and ezilirets ni eht evalcotua
ees( SI 6853 ;) 5 g muidos ;edirolhc 0.3 g muissatopid
yb eerf gnimaets rof eno ruoh and neht at 120°C
negordyh ;etahpsohp and 3 g agar ( ees SI 6850 .)
rof 10 .setunim
Adjust Hp ot 7.1, retlif and add lomyhtomorb ,eulb
)b Solution A - evlossiD 17 g muidos ,etartic 15 lm fo 0.2 tnecrep cilohocla .noitulos eziliretS at
17 g muidos etahplusoiht and 2 g cirref 115°C rof 20 .setunim Add elirets noitulos fo esoculg
2
IS 5887 traP( )7 : 1999
ot evig a lanif noitartnecnoc fo 1 .O .tnecrep xiM and etubirtsiD otni elirets tset sebut gniniatnoc detrevni
etubirtsid ni ,sebut 14 mm x 100 ,mm adding 3 ot 4 lm Durham’s noitatnemref ,sebut and maets rof 30 .nim
rep .ebut s’edardnA rotacidni noitulos is deraperp yb adding
1 N muidos edixordyh noitulos ot 0.5 tnecrep suoeuqa
6.8 TSI Medium for H2S Test noitulos fo dica nishcuf litnu eht ruoloc fo eht rotacidni
taeH ot evlossid ni 1000 lm ,retaw 3 g taem tcartxe noitulos is just .wolley
( ees SI 685 1 ), 3 g tsaey tcartxe ( ees SI 7004 ,) 20 g 6.13 Medium for Dihydrolase and Decarboxylase
enotpep ( ees SI 6853 ,) 1 g ,esoculg 10 g ,esotcal
Activity
10 g ,esorcus 0.2 g suorref etahplus ( ,40SeF 0zH7 ,)
5 g muidos ,edirolhc 0.3 g muidos etahplusoiht evlossiD htiw ,taeh 5 g enotpep ( ees SI 6853 ,) 5 g
( 0zH5,302S2aN ) and 15 ot 30 g agar ees( SI 6850 ;) taem tcartxe ees( SI ,)1586 5 gm ,laxodiryp and 0.5 g
add 12 lm fo 0.2 tnecrep lonehp der ,noitulos xim and esoculg ni 1 000 lm fo .retaw HptsujdA ot 6.0 and .
ebut gnizilirets yb gnivalcotua at 115°C rof 20 .setunim add 5 lm fo 0.2 tnecrep noitulos fo losercomorb elprup
ruoP otni elirets tset ebut and looc ot mrof a epols and 2.5 lm fo 0.2 tnecrep noitulos fo loserc .der eziliretS
htiw peed butts. ni an evalcotua at 115’C rof 20 .setunim ediviD this
esab muidem otni ruof lauqe ,semulov and ot hcae fo
6.9 Medium for Urease Test eht eerht add eht evitcepser onima sdica ( ,eninigra
enisyl and enihtinro ) as a lanif noitartnecnoc fo
evlossiD ni 1 000 lm ,retaw 1.5 g enotpep ( ees
1 tnecrep fi eht mrof-I fo eht onima dica is desu ro as
SI 6853 ,) 5 g muidos ,edirolhc 15 ot 30 g agar ( ees
2 tnecrep fi eht mrof& fo eht onima dica is .desu ehT
SI 6850 ,) 2 g muissatopid negordyh etahpsohp and
htruof touqila fo eht esab muidem seod ton niatnoc
add 6 lm fo 1 : 500 suoeuqa noitulos fo lonehp .der
onima dica and sevres as .lortnoc etubirtsiD eht ruof
Adjust Hp ot 6.8 ot 6.9 and ezilirets yb gnivalcotua at
aidem ni 1 ot 1.5 lm stnuoma otni small dezilirets ,sebut
1 20°C rof 15 .setunim nehW eht noitulos has delooc
67 mm x 10 ,mm and reyal htiw elirets diuqil niffarap
ot tuoba SO“C, add a elirets noitulos fo esoculg ot evig
ot a thgieh fo tuoba 5 .mm HpehT fo eht lanif muidem
a lanif noitartnecnoc fo 0.1 tnecrep and add 100 lm fo
dluohs eb ,detsujdaer fi ,yrassecen ot 6.0. eziliretS ni
20 tnecrep noitulos fo aeru ylsuoiverp dezilirets yb
an evalcotua at 115°C rof 10 .setunim
ztieS .noitartlif etubirtsiD ni elirets tset sebut ot mrof
peed .sepols
6.13.1 An evitanretla muidem erehw eht esab muidem
sniatnoc 5 g enotpep ( ees SI 6853 ,) 3 g tsaey tcartxe
6.10 Simmon’s Citrate Agar
( ees SI 7004 ,) 1 g esoculg and 10 lm fo 0.2 tnecrep
evlossiD ni 1000 lm ,retaw 5 g muidos ,edirolhc 0.2 g losercomorb elprup ni 1 000 lm retaw yam eb .desu
muisengam etahplus ( 0,H7,40SgM ,) 1 g muinomma ehT Hp is detsujda ot 6.7 and eht gniniamer erudecorp
negordyhid ,etahpsohp 1 g muissatopid negordyh rof noitaraperp fo eht muidem is as nevig ni 6.13.
,etahpsohp 2 g muidos ,etartic and 15 ot 30 g agar
ees( SI 6850 ;) add 40 lm fo 1 ni 500 lomyhtomorb 6.14 Medium for Malonate Test

eulb rotacidni .noitulos eziliretS at 120°C rof 15 ,nim evlossiD yb ,gnitaeh fi ,yrassecen ni 1 000 lm ,retaw
ebut and slant os that ereht is lauqe ttub and .epols 1 g tsaey tcartxe ( ees SI 7004 ,) 2 g muinomma
etahplus ( ,40S2)4HN 0.6 g fo muissatopid negordyh
6.11 Medium for Indole Production
,etahpsohp 0.4 g muissatop negordyh@ ,etahpsohp
ehT muidem stsisnoc fo 20 g enotpep ( ees SI 6853 ) 2 g muidos ,edirolhc 3 g muidos ,etanolam 0.25 g
and 5 g muidos edirolhc ni 1000 lm ;retaw Hptsujda esoculg and 0.025 g lomyhtomorb .eulb esnepsiD ni
ot 7.4. ecalP ni sebut ni 5 lm noitrop and ezilirets at tuoba 3 lm noitrop otni dezilirets tset sebut and
120°C rof 15 .setunim ehT muidem dluohs eb detset evalcotua at 120°C rof 15 .setunim ehT lanif Hp
htiw a strain fo muiretcab nwonk ot ecudorp .elodni dluohs eb 6.7 f 0.1.
6.12 Peptone Water Medium for Carbohydrate 6.15 Gelatin Liquification Test Medium
Fermentation Tests
evlossiD ni 1 000 lm ,retaw 3 g taem tcartxe ( ees
evlossiD 10 g enotpep ( ees SI 6853 ) and 5 g muidos SI 685 1 ,) 5 g enotpep ( ees SI 6853 ,) 120 g nitaleg
edirolhc ni 900 lm retaw and adjust Hpeht at 7.1 ot ( ees SI 7590 ) and 30 g muidos .edirolhc etubirtsiD
7.3 os that retfa noitidda fo 10 lm fo s’edardnA otni dezilirets tset sebut and ezilirets at 120°C rof 12
rotacidni eht Hp dluohs eb 7.5. eziliretS at 115°C rof .setunim ehT Hplanif dluohs eb 7.0.
20 .setunim evlossiD 10 g fo eht etisiuqer sugar ni
7 PROCEDURE FOR ISOLATION
90 lm retaw and maets rof 30 setunim ro ezilirets yb
.noitartlif With elirets ,snoituacerp add 90 lm fo this 7.1 nI ,lareneg all sdoof hcihw evah deviecer yrd taeh
sugar noitulos and 10 lm fo s’edardnA rotacidni ’ ,tnemtaert ,yleman ,noitaccised noitardyhed ro -dwop
noitulos ot 900 lm fo eht elirets enotpep retaw .noitulos ,gnire shall eb dehcirne-erp ni a yrotibihni-non
3
IS 5887 ( Part 7 ) : 1999
muidem hcus as tneirtun htorb ( 6.1). Raw sdoof and neht revoc htiw )b( and wolla ot niamer rof 30 .sdnoces
dehsinif stcudorp hcihw era detcepsus ot eb ylssorg Wash ffo htiw lonahte litnu eht eyd sesaec ot maerts
detanimatnoc retfa gnissecorp dluohs ton eriuqer .tuo Wash ni gninnur tap retaw and ylppa )c( rof tuoba
tnemhcirne-erp and eht erudecorp as nevig ni 7.1.1 eno .etunim Wash ni tap retaw and yrd rof .noitanimaxe
shall eb .dewollof
8.2.2 Test for Motility
7.1.1 erehW tnemhcime-erp is ton ,deriuqer eht elpmas
etaluconI yb stabbing htiw a straight eriw otni eht
is dednelb ni a elirets rednelb raj rof 2 setunim ro
pot fo eht muidem as ni 6.6, eht strain ot eb ,detset
detarecam htiw elirets sand ni a elirets ratrom using
edisni eht glass gnibut ot a htped fo tuoba 5 .mm ekaT
yletamixorppa 200 lm etineles F htorb )3.6( rep
erac that noitaluconi is ton edam no ot eht ecafrus fo
yletamixorppa 25 g fo eno noitrop fo eht ,elpmas and
eht muidem edistuo eht glass .gnibut etabucnI at 37°C
200 lm etanoihtartet htorb )2.6( rep yletamixorppa
rof 18 ot 24 .sruoh elitoM strains shall eb dnuof ot
25 g fo rehtona noitrop fo eht .elpmas etabucnI at
wohs htworg no eht ecafrus fo eht muidem edistuo
37°C rof 24 .sruoh Raw taem samples dluohs eb
eht renni‘ glass ’ebut gnivah dellevart hguorht eht
detabucni at 43°C rehtar than at 37°C.
eritne muidem edisni this renni .ebut fI evitagen no
7.2 noitalosI eht tsrif ,yad peek eht detaluconi ebut at moor
erutarepmet rof a rehtruf 4 ot 6 syad ot ees fi ecnedive
7.2.1 morF eht detabucni elpmas as ni (1.1.7 detabucni
fo ytilitom is .tneserp
at 37°C ) etalp tuo slufpool no ot etalohcyxosed etartic
agar muidem )4.6( and etabucni rof 24 sruoh at 37°C. 8.2.3 Test for Catalase
tI is osla elbasivda ot etalp tuo no ot eht dilos muidem
worG eht strain rof 24 sruoh no tneirtun agar epols
)4.6( slufpool fo hcae fo eht diuqil muidem retfa
)1.1.6( and ruop 1 lm negordyh edixorep revo eht
,gnidnelb tub erofeb gnitabucni eht etalp at 37°C
htworg htiw eht ebut ni a slanting .noitisop esaeleR
thginrevo as tcepsus Shigella seinoloc yam eb nees
fo ,negyxo nwohs as ,selbbub morf negordyh edixorep
eno yad .reilrae
setacidni eneserp fo .esalatac
7.2.2 fI ereht is on tcepsus htworg no eht dilos
muidem retfa 24 ,sruoh etabucnier rof a rehtruf 8.2.4 Test for Oxidase
24 .sruoh Shigella seinoloc no etalohcyxosed etartic oT tneirtun agar )1.1.6( slant fo eht ,erutluc ylhserf
agar shall eb ,euqapo htiw a ssalg-dnuorg ecnaraeppa ,nworg is dedda a wef spord fo ylhserf dexim tset
and htiw neve margins. ,stnegaer 1 tnecrep noitulos fo lohthpan-ahpla ni
95 tnecrep lonahte and lauqe tnuoma fo 1 tnecrep
8 STSET FORIDENTIFICATION
noitulos fo enilinalyhtemid-onima-arap -ordyh
8.1 kciP tuo as ynam tcepsus seinoloc as era elbissop edirolhc ni ,retaw A evitisop noitcaer is detacidni yb
( ton less than 5 ) ot etagitsevni and etalp tuo otni eht ecnaraeppa fo a eulb ruoloc nihtiw owt .setunim

yeknoCcaM agar setalp )5.6( an etabucni thginrevo 8.2.5 Hugh-Leifson k Test
at 37’C ot ekam niatrec fo eht ytirup fo eht .seinoloc
eruP seinoloc era derrefsnart no ot tneirtun agar sepols ehT strain morf hserf tneirtun agar htworg ( ees )1.1.6
)1.1.6( ot niatbo a yvaeh htworg rof rehtruf ,gnitset is debbats otni owt sebut fo muidem ,)7.6( eno fo hcihw
as osla rof gnivreserp eht setalosi rof gnipytores and/ is neht dereyal revo htiw a small tnuoma fo elirets diuqil
ro egahp .gnipyt .niffarap etabucnI htob sebut at 37°C and evresbo
up ot 4 .syad dicA ,noitamrof wolley ,ruoloc ni eht
8.2 Shigella era tcepsus nehw gnimrofnoc ot eht ebut tuohtiw nilfarap setacidni evitadixo noitazilitu
sretcarahc nevig ni elbaT 1.
fo .esoculg dicA ni htob sebut setacidni evitatnemref
8.2.1 Gram k Stain .noitcaer kcaL fo dica ni rehtie ebut setacidni eht strain
as ton gnieb elba ot ezilitu esoculg ylevitadixo ro
ehT stain stsisnoc fo )a( 0.5 tnecrep lyhtem teloiv ro .ylevitatnemref
latsyrc teloiv ni ,retaw )b( enidoi noitulos ( 1 tnecrep
enidoi and 2 tnecrep muissatop edidoi ni retaw ,) and 8.2.6 Testfor H2S Production
)c( niatsretnuoc ( 0.1 g lartuen ,der 0.2 lm fo 1 tnecrep
etaluconI IST muidem )8.6( yb stabbing eht strain otni
citeca dica and 100 lm retaw .)
eht ttub and gnikaerts eht .epols etabucnI at 37°C
On a naelc eerf-esaerg ,edils yrev thgil and niht ,raems and evresbo yliad rof up ot 7 .syad ehT ecneserp ro
gnirevoc a small ,aera is edam yltcerid morf diuqil ecnesba fo gninkcalb ni eht ttub fo eht muidem is ot
erutluc and ni naelc tap retaw morf dilos .aidem ehT eb .dedrocer
raems is dexif yb passing ot and orf revo a emalf and
8.2.7 Testfor &ease
.delooc revoC eht raems htiw eht stain )a( rof
30 ,sdnoces ruop ffo eht stain and wash htiw )b( and etaluconI eht smsinagro morf ruoh-42 detabucni

4
IS 5887 ( Part 7) : 1999
tneirtun htorb )1.6( erutluc ylivaeh revo eht eritne epols Table 1 Useful Characters of Shigellu Organisms
ecafrus fo muidem as ni 6.9 and etabucni at 37°C rof (Clauses5.1,8.2,8.2.11and9.1)
18 ot 24 .sruoh A evitisop esaeru is nwohs yb eht
gnimocebmuidem knip ro der no .noitabucni fI negative, Sl Tests Sub Gr. Sub Gr. Sub Gr.
eunitnoc noitabucni rof at tsael 4 .syad suetorP seiceps No. A B&C D
evig a evitisop ,tluser and nac eb desu as .’lortnoc‘ )1( )2( (3) (4) (5)
8.2.8 Phenyl Pyruvic Acid Production i) Gram-negative rods + + +
ii) Motility
Obtain an thginrevo yvaeh htworg fo eht smsinagro
no tneirtun agar .)1.1.6( dnepsuS eht htworg ni iii) Catalase + (a) + +
0.5 lm fo lamron enilas and refsnart ot a erob-ediw tset iv) Oxidase
ebut ( retemaid at tsael 1.5 mc .) Add 0.5 lm fo v) Hugh-Leifson’s test F F F
0.2 tnecrep eninalalynehp-ld ni lamron .enilas xiM
vi) TSI for H,S
and peek gniyl yllatnoziroh rof at tsael eerht sruoh at
vii) Urease
moor .erutarepmet Add a wef spord fo detarutas-flah
viii) Phenyl pyruvic acid
cirref edirolhc .noitulos A evitisop noitcaer is an
etaidemmi peed neerg ruoloc hcihw sedaf no .gnipeek ix) Citrate
Proteus strains evig a evitisop noitcaer and sevres x) Indole varies varies varies
as a .lortnoc (b) (b)
xi) Glucose + + +
8.2.9 Test for Citrate Utilization
xii) Lactose (1 percent) (+)
etaluconI eht strain no ot muidem as ni 6.10 htiw a xiii) Sucrose I+(
gnuoy tneirtun agar )1.1.6( slant erutluc using a xiv) Salicin
straight .eriw etabucnI at 37” rof up ot 4 syad rof htworg
xv) Dulcitol - (c) - (c)
fo eht .smsinagro xvi) Mannitol + +
8.2.10 Test for Indole xvii) Lysine
xviii) A&nine - or + - or + - or +
etaluconI muidem )11.6( htiw a lufpool fo 24 sruoh
htworg ni tneirtun htorb )1.6( and etabuni at 37°C rof xix) Ornithine - (d) +
48 sruoh Add 0.5 lm s’cavoK ,tnegaer deraperp yb xx) Malonate
gnivlossid 10 g edyhedlaznebonimalyhtemidarap ni xxi) Gelatin
150 lm lyma ro iso-amyl lohocla and ot hcihw is dedda
NOTE - Some important exceptions to the above
ylwols 50 lm fo detartnecnoc cirolhcordyh .dica
characters are as follows:
eraperP eht tnegaer ni small seititnauq and erots ni
(a) Sh. dysenteriae 1 (S/I. shiga) is catalase-positive;
.rotaregirfer retfA adding s’cavoK ,tnegaer ekahs eht

(b) Sh. dysenturiae 1 (Sh. shigu), 5%.j lexneri 6 are
ebut ,yltneg eht ecnaraeppa fo a der ruoloc setacidni
always indole-negative, and Sh. dysentertae 2 is always
eht ecneserp fo .elodni
indole-positive; (c) Some strains of Shigella subgroup
A, Sh. jlexneri 6 and subgroup C are dulcitol-positive;
8.2.11 Tests for Fermentation Carbohydrates (d) Sh. boydti 13 is ornithine-decarboxylase-positive.
etaluconI hcae fo muidem as ni 6.12, and eht + = Present or Positive
setardyhobrac detacidni ni elbaT 1 using a 1 tnecrep (+) = Late Positive
noitartnecnoc rof eht ,esotcal and etabucni at 37“C - = Negative or no reaction
rof 18 ot 48 .sruoh droceR eht ecneserp fo dica morf F = Fermentative reaction
knip ruoloc and that fo gas ni eht Durham’s .ebut
8.2.12 Test for Dihydrolase and Decarboxylase
nehW gnimrofrep eht ,tset rof hcae onima dica syawla
Activities
esu )s(niarts fo muiretcab )a( nwonk ot wohs evitisop
,etaluconI using a straight ,eriw hguorht eht diuqil tluser and )s(niarts fo muiretcab )a( nwonk ot wohs
n?ffarap hcae fo eht ruof sebut fo eht muidem as evitagen .tluser esehT evres as .’slortnoc‘
debircsed ni 6.13 morf a ylhserf nworg erutluc ni
tneirtun agar .)1.1.6( etabucnI at 37°C and enimaxe
8.2.13 Test for Utilization of Malonate
yliad rof up ot 4 syad .ylno ehT muidem tsrif semoceb
wolley eud ot dica noitcudorp morf eht ;esoculg ,retal etaluconI eht muidem as ni 6.14 and etabucni rof
fi noitalordyhid ro noitalyxobraced fo eht evitcepser 24 sruoh at 37°C. A evitisop etanolam noitcaer is
onima dica ,srucco eht muidem semoceb teloiv ni detacidni yb a peed eulb ruoloc and a evitagen noitcaer

.ruoloc yb eht degnahcnu hsineerg ruoloc fo eht .muidem With