Free Standard provided by BIS via BSB Edge Private Limited to BELLA -

NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on

(Reaffirmed 2005)
(Reaffirmed!2013)!
(Reaffirmed 2018)
(Reaffirmed 2022)
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) - 1976

Indian Standard
METHODS FOR
DETECTION OF BACTERIA RESPONSIBLE
FOR FOOD POISONING
PART II ISOLATION, IDENTIFICATION AND
ENUMERATION OF STAPHYLOCOCCUS AURfUS
AND FAECAL STREPTOCOCCI

( First Revision)
Food Hygiene, Sampling and Analysis Sectional Committee, AFDC 36
Chairman Representing
DR RANJIT SEN Serologist to the Government of India ( DGHS ),
Calcutta
Members
AGRICULTURAL MAR K E TIN G Directorate of Marketing & Inspection ( Ministry of
ADVISER TO THE GOVERNMENT Agriculture & Irrigation), Faridabad
OF INDIA
SHRI T. V. MATHEW ( Alternate)
SHRI V. N. AMBLE Institute of Agricultural Research Statistics (ICAR).
New Delhi
SHRI K. S. KRISHNAN ( Alternate)
DR G. C. DAS Health Officer, Corporation of Calcutta
DR P. K. DATTA All India Institute of Hygiene and Public Health,
Calcutta
SHRI SUKUMAR DE National Dairy Research Institute ( ICAR ), Kamal
DR C. A. MULAY ( Alternate)
SHRI O. P. DHAMIJA Export Inspection Council of India, New Delhi
DIRECTOR Central Food Laboratory, Calcutta
SHRI C. T. DWARKANATH Central Food Technological Research Institute
( CSIR ), M ysore
DR M. A. KRISHNASWAMY (Alternate)
EXECUTIVE HEALTH OFFICER Municipal Corporation of Greater Bombay
MUNICIPAL ANALYST ( Allernate )
HEALTH OFFICER Corporation of Madras
COL KEWAL KRISHNA Health Department, Municipal Corporation of Delhi
DR A. D. KUMAR ( Allernat, )

( Continued on page 2 )

@ Copyright 1977
INDIAN STANDARDS INSTITUTION
This publication is protected under the Indian Copyright Act (XIV of 1957) and
reproduction in whole or in part by any means except with written permission of the
publisher shall be deemed to be an infringement of c:opyright under the said Act.
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) - 1976

( Continued from page I )

Members ReprlStntinl
DR ( SMT) S. KHOSJ.A Department of Health & Family Planning, Govern-
ment of Punjab, Chandigarh
DR P. K. KYMAL Food & Nutrition Board ( Ministry of Agriculture &
Irrigation), New Delhi
DR O. N. AGARWALA (Alternate)
MAJ V. A. NARAYANAN Defence Food Research Laboratory (Ministry of
Defence ), M ysore
DR G. M. VERMA ( Alternate)
PUBLIC ANALYST (FOOD AND Government of West Bengal, Calcutta
WATER)
PUBLIC ANALYST ( BACTERIO-
LOGY) (Alternate)
DR A. N. RAI CHAWDHURI National Institute of Communicable Diseases, Delhi
MAJ-GEN D. C. SACHDEVA Directorate General of Armed Forces Medical Services
(Ministry of Defence ), New Delhi
SENIOR MEDICAL OFFICER Northern Railway, New Delhi
(HEALTH)
DR S. B. SINGH Public Analyst, Government of Uttar Pradesh,
Lucknow
SlIRI N. SRINIVASAN Public Analyst, Government of Tamil Nadu, Madras
DR M. R. SUBBARAM Directorate of Sugar & Vanaspati (Ministry of
Agriculture & Irrigation)
SHRI 1. A. SIDDIQI ( Alternate )
DR T. A. V. SUBRAMANIAN Vallabhbhai Patel Chest Institute, Delhi
DR M. C. SWAMINATHAN Directorate General of Health Services ( Ministry of
Health & Family Planning ), New Delhi
SHRI D. ·S. CHADHA ( Alternate)
COL R. N. TANEJA Quartermaster General's Branch, Army Headquarters,
New Delhi
LT-COL D. D. VOHRA (Alternate)
SHRI P. C. VIN The Coca-Cola Export Corporation, New Delhi
SHRI J. D. CONTRACTOR ( Alternate)
8HRI T. PURNANANDAM, Director General, lSI ( Ex-officio Memb,r )
Deputy Director ( Agri & Food)
Secretary
SHRI S. K. SUD
Deputy Director ( Agri & Food ), lSI

Food Microbiology Subcommittee, AFDC 36: 7

Convener
DR RANJIT SEN Serologist to the Government of India ( DGHS ),
Calcutta
Members
AGRICULTURAL MARKETING Directorate of Marketing & Inspection ( Ministry of
ADVISER TO THE GOVERNMENT Agriculture & Irrigation ), Faridabad
OF INDIA
DIRECTOR OJ' LABORATORIES ( Alternate)
M.u G. S. BALI Defence Food Research Laboratory (Ministry of
Defence ), M ysore
811BI K. C. RAPoN ( Alttrnau )
( Continued on page 11 )
2
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) -1976

Indian, Standard
METHODS FOR
DETECTION OF BACTERIA RESPONSIBLE
FOR FOOD POISONING
PART II ISOLATION, IDENTIFICATION AND
ENUMERATION OF STAPHYLOCOCCUS AUREUS
AND FAECAL STERPTOCOCCI

( First Revision)

o. F'ORE WORD
0.1 This Indian Standard ( Part II ) ( First Revision) was adopted by
the Indian Standards Institution on 13 December 1976, after the draft
finalized by the Food Hygiene, Sampling and Analysis Sectional
Committee had been approved by the Agricultural and Food Products
Division Council.
0.2 Several micro-organisms contaminating food give rise to clinical
symptoms. These are abdominal pain, nausea, vomitting, diarrhoea and
sometimes pyrexia. A well-known exception is that of botulism where
the symptoms are those of difficulty in swallowing, diplopia, aphonia and
difficulty in respiration. Poisoning through food is characterized by the
explosive nature with which the symptoms OCcur in otherwise healthy
individuals. Often several persons after having consumed a particular
item of food, develop symptoms that serve as important guide in suspect-
ing food poisoning. Such explosive nature of food poisoning helps in
differentiating conditions from those of out-breaks of food-borne
infectious diseases which generally spread over a period of several days.
The micro-organisms causing food poisoning belong to bacteria, protozoa
and helminths, fungi and viruses. However, this standard covers the
method for detection and estimation of important bacteria responsible
for food poisoning food-borne diseases.
0.3 This standard was first published in 1970. It is being revised in
parts covering methods of detection and estima tion of various bacteria'
separately. This has been done with a view to making each part more
comprehensive including various details of the methods. It is expected
that publication of these methods in parts will facilitate better

3
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) • 1976

implementation and adoption of the standard by concerned orga-

nizations. This will also make review and revision easier. The salient
features of this revision are:
a) Besides detection, estimation procedures for various organisms
where applicable have been incorporated; and
b) Methods of identification have been updated.
0.4 In reporting the result of a test or analysis made in accordance with
this standard, if the final value, observed or calculated, is to be
rounded off, it shall be done in accordance with IS : 2-1960*.

1. SCOPE
1.1 This standard (Part II ) prescribes method for isolation, identi-
fication and enumeration of Staphylococcus atlreus and faecal streptococci
in foods.
2. SAMPLING AND QUALITY OF REAGENTS
2.1 Sampling - For microbiological examination the samples should be
handled carefully. For this purpose, IS : 5404-1969t shall be followed.
2.2 Quality of Reagents - Unless specified otherwise, pure chemicals
shall be employed in tests and distilled water (see IS: 1070-1960:1: ) shall
be used where use of water as a reagent is intended.
NOTE - 'Pure chemicals' shall mean chemicals that do not contain impurities
which affect the results of analysis.

3. GENERAL CHARACTERISTICS
3.1 Staphylococcus aureus - Aerobic, Gram-positive cocci in clusters,
usually, but not always producing a golden yellow coloured colonies on
nutrient agar ( 4.2 ) and blood agar ( 4.3 ), and shiny black colonies with
or without narrow grey-white margin when grown on Baird-Parker
medium (4.5). Suspect colonies must show coagulase activity.
3.2 Faecal Streptococci - Aerobic, Gram-positive cocci usually in
pairs or short chains, producing small pink colonies on MacConkey agar
( 4.7) and colonies which are dark red or having red or pink centres
when grown on ethyl violet azide dextrose agar (4.6). Growth is
obtained also at 44°C.
4. MEDIA
4.1 Nutrient Broth - Mix and dissolve by heating 10 g peptone
(see IS: 6853-1973§), 10 g meat extract (see IS: 6851-197311), and 5 g
sodium chloride in 1000 ml water. When cool, adjust pH to 7'5 to 7'6.
Remove precipitate by filtration through filter paper. Sterilize by
autoclaving at 120°C for 15 minutes .
• Rules for rounding off numerical values ( revised).
tCode of practice for handling of food samples for microbiological analysis.
tSpecification for water, distilled quality ( reviud).
§Specification for peptone, microbiological grade.
IISpecification for meat extract, microbiological grade.
4
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) - 1976

4.2 Nutrient Agar - To the medium as in 4.1, add agar ( see IS: 6850-
1973* ) in such a concentration as will solidify and produce a sufficiently
firm surface when poured in sterile petri dishes. The concentration of
agar to be added varies from batch to batch and should be adjusted
accordingly. Usual concentrations required vary from 1-5 to 3 percent.
Dissolve the agar in the nutrient broth and sterilize by autoclaving at
120·C for 15 minutes. Plates and slopes are prepared from sterile
nutrient agar.

4.3 Blood Agar - Melt sterile nutrient agar as prepared in 4.2 and hold
between 50 to 55°C in a water-bath. Add sterile blood free from preser-
vatives to give a concentration of 10 percent. Mix well and pour plates.
Horse blood is commonly used but when not available, that of sheep,
human or rabbit may be used.

4.4 Salt MediulD

4.4.1 Cooked Meat Medium - Mince 500 g fresh beef heart and place it
in 500 ml of alkaline boiling water containing 1'5 ml of 1 N sodium
hydroxide solution. Simmer for 20 minutes. Drain off the liquid
through muslin filter while still hot and partially dry the meat in the
cloth or on filter papers. To 500 ml of liquid filtered from the cooked
meat add 2' 5 g, peptone (see IS : 6853-1973t) and 1'25 g sodium chloride.
Steam at 100°C for 20 minutes and add 1 ml concentrated hydrochloric
acid and filter. Bring the r ea ction of the filtrate to pH 8-2 and steam
again at 100°C for 30 minutes; adjust pH to 7'8. Place meat in test-tubes
or in about 30 ml screw-capped bottles to a depth of about 2'5 cm and
cover with 10 ml of the broth obtained. Autoclave at 120°C for
20 minutes. A layer of sterile paraffin may be added to cover the
surface. .

4.4.1.1 For salt medium - Add further 10 percent sodium chloride to

the broth ( 4.4.1), adjust the pH to 7'8 and proceed as in 4.4.1.

4.5 Baird-Parker MediulD - Dissolve by boiling in 950 ml of water,

10 g tryptone (see IS: 7127-1973:):),5 g meat extract (see IS : 6851-1973§),
1 g yeast extract (see IS : 7004-197311), 12g sodium pyruvate, 12 g glycine,
5 g lithium chloride and 15 to 30 g agar (see IS: 6850.1973*). Distribute
in 95 ml amounts into sterilized flasks (or screw-capped bottles) and
sterilize at 120°C for 15 minutes. The final pH should be 6'8 to 7'2.
*Specification for agar, micr~biolo.gical. grade.
tSpecificati?n for peptone, ml~robl(?logl~al grade.
tSpccificatlOn for tryptone, mlcro~)lolo~lcal.grade.
§Specification for meat extract, m~crob~olog~cal grade.
IISpecification for yeast extract, microbiOlogical grade.

5
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on

IS : 5887 ( Part II) -1976

To 95 ml of the molten medium ( 4.5) kept at 45° to 50°C, add 5 mt
of Bacto-Er tellurite enrichment which has been prewarmed to 45° to 50·C.
Mix well and pour on to sterilized petri dishes. The plates should be
made freshly before use and should not be stored longer than 48 hours
before use. The plates, before use, should be well-dried by keeping in an
incubator at 50°C for about 30 minutes, with the lid removed and agar
surface downward.

4.6 Ethyl Violet Azide Dextrose Broth - Dissolve in 1 000 ml

water 20 g tryptose, 5 g dextrose, 2'7 g dipotassium phosphate, 2'7 g
monopotassium phosphate, 5 g sodium chloride, 0'4 g sodium azide
and 0'000 83 g ethyl violet. Distribute into tubes. Sterilize at 120°C
for 15 minute,. The final pH should be 7'0.

4.6.1 Ethyl Violet Azide Dextrose Agar - To medium as in 4.6, add 15 to

30 g agar (see IS: 6850-1973*), and dissolve by heating. Sterilize at
120°C for 15 minutes, and pour on to sterilized petri dishes.

4.7 MacConkey Agar Medium - Mix 5 g sodium taurocholate or

bile salts (see IS: 6852-1973t), 20 g peptone (see IS: 6853-1973! ),
5 g sodium chloride and 15 to 30 g agar (see IS: 6850-1973*), with
I 000 ml water. Steam until the solids are dissolved. Cool to about
50°C, and at this temperature adjust reaction to pH 7'6 to 7'8. Auto-
clave at 120°C for 15 minutes and filter while hot through a good grade
of filter paper, or a plug of cotton wrapped in gauze and placed in the
funnel. Adjust reaction of the filtrate to pH 7'3 at 50°C or pH 7'5 at
room temperature. Add 100 ml of 10 percent aqueous solution of
lactose (or 10 g lactose) and 3'5 ml of 2 percent solution of neutral
red in 50 percent ethanol. Mix thoroughly, distribute into flasks and
sterilize in the autoclave at 120°C for 15 minutes. For use, melt in the
steamer, pour into sterile petri dishes and allow to set.

5. PROCEDURE FOR ISOLATION

5.1 Staphylococcus aureus- Inoculate sample on blood agar (4.3) and in
salt medium ( 4.4 ) and on Baird-Parker medium (4.5), if available.
5.1.1 Incubate the blood agar and salt medium at 37°C overnight and
the inoculum in Baird-Parker medium at 37°C for at least 30 hours.
From the salt medium, make subcultures on the one or both solid media
mentioned for overnight incubation at 37°C in case of blood agar, and for
30 hours in case of Baird-Parker medium .
• Specification for agar, microbiological grade.
tSpecification for bile salts, microbiological grade.
:t:Specification for peptone, microbiological grade.

6
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) • 1976

5.1.2 Examine not less than 5 of the suspect colonies of Staphylococcus

on the solid media and mark out as many suspect colonies as possibJe
to investigate. A portion of each colony picked with a straight nichrome
wire may be used for preliminary coagulase test using the slide technique.
The remainder of the colony is streaked out on blood agar medium for
incubation at 37°C to check purity and to proceed with identification.
If preliminary slide testing is avoided, the suspect colony is checked for
purity as just described. Pure colonies are stained by Gram's method
and tested for coagulase by the slide and tube methods.
5.2 Faecal Streptococci - To estimate the number of faecal strepto-
cocci in the sample, the procedures given in 8.2 shall be followed.

6. TESTS FOR IDENTIFICATION

6.1 Staphylococcus aureus

6.1.1 Gram's Stain - The stain consists of: (a) 0'5 percent methyl
violet or crystal violet in water; (b) iodine solution (1 percent iodine
and 2 percent potassium iodide in water); and (c) counterstain
( 0'1 g neutral red, 0'2 ml of 1 percent acetic acid and 100 ml water ).
On a clean grease-free slide, very light and thin smear covering a
small area is made directly from liquid culture and in clean tap water if
from solid media. The smear is fixed by passing to and fro over a flame
and cooled. Cover the smear with the stain (a) for 30 seconds, pour off
the stain and wash with (b) and then cover with (b) and allow to
remain for 30 seconds. Wash off with ethanol until the dye ceases to
stream out. Wash in running tap water and apply (c) for about one
minute. Wash in tap water and dry for examination.
6.1.2 Colonial Character - By growth on nutrient agar (4.2), blood
agar (4.3) and/or on Baird-Parker medium ( 4.5 ), as described in 3.1.
6.1.3 Coagulase Test - This test may be carried out by the following
methods. The tube method shall be preferred:
a) Slide method - Emulsify a portion of the suspect colony in normal
saline or water. Mix this with a straight wire dipped in human
or rabbit plasma. Coagulase-positive staphylococci produce
visible clumping immediately.
b ) Tube method - Emulsify a single suspect colony from a 24 hour
growth of blood agar medium (4.3) in I ml citra ted
rabbit plasma diluted 1 in 5 in 0'85 percent saline. The test is
usually carried out in narrow tubes. Place in an incubator or

7
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS : 5887 ( Part II ) • 1976
preferably in water-bath at 37°0. Observe every hour to note
clotting of plasma. Reading should be carried out for as long as
possible, preferably avoiding overnight incubation. Positive
control with a known coagulase-positive strain of Staphylococcus
and a control of the diluted plasma without inoculum should be
included in the test. .
NOTE I - If the slide method gives a negative result, the tube method
shall be carried out.
NOTE 2 - False positive results may occur on the slide t~st. A small
number of strains give a positive slide test with a negative tube test due to
the production of • bound' coagulase alone.

6.2 Faecal Streptococci

6.2.1 Gram's Stain - See 6.1.1.
6.2.2 Colonial Character- By growth on MacConkey agar (4.7) at 44°C
and ethyl violet azide dextrose agar (4.6.1) at 37°C, as described
in 3.2.
7. PHAGE TYPING
7.1 A single colony of coagulase-positive strain of Staphylococcus tested by
the tube method may be maintained on nutrient agar (see 4.2) slopes,
and sent for phage typing.
NOTE - Presently phage typing facilities are available at the Staphylococcus Phage
Typing Centre, Department of Microbiology, MauJana Azad Medical College,
Bahadur Shah Zafar Marg, New Delhi 110002.

8. ENUMERATION
8.1 Staphylococcus aureus - Since the presence of small numbers of
coagulase-positive staphylococci does not necessarily indicate association
of the isolate with food poisoning, estimation of approximate number of
organisms per gram of suspect food form more valid determination if the
suspect strains are to indicate food poisoning. While a rough estimate
may be made on direct smear of the material stained by Gram's method
( 6.1.1 ), quantitative estimation shalI always be made by the procedure .
given in 8.1.1.
8.1.1 Twentyfive to fifty grams of the sample is taken in a sterile
blender jar and to this is added diluting fluid to have dilution of 10-1,
The diluting fluid shall be peptone (see IS: 6853-1973* ) 0'1 percent in
water sterilized at 120·C for 20 minutes, final pH 6'8 :±: 0-1 or 3'4 percent
potassium dihydrogen phosphate (KH 2 P0 4 ) in water, pH adjusted to
7'2 and sterilized at 120°C for 20 minutes. Blend at 8000 to 10 000
rev fmin for 2 minutes. Alternatively, macerate the sample with diluting

.Specification for peptone, microbiological grade.

8
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS: 5887 ( Part II ) ·1976

fluid in a sterile mortar with sterile sand. Make serial ten-fold dilutions
with the diluting fluid in duplicate series up to 10-6 • Streak 0·1 ml from
each tube evenly on to blood agar (4.3) and/or Baird-Parker medium
(4.5). Incubate the blood agar plates at 37°0 overnight and the
Baird-Parker plates at 37°0 for at least 30 hours. Enumerate the colonies
which are as described in 3.1. These colonies are to be confirmed as
being S. aureus by the coagulase test described in 6.1.3. The number of
viable colonies per gram of sample is determined by multiplying the
dilution factor( s ) and dividing by the mass of the sample.
8.2 Faecal Streptococci
8.2.1 Plate Count - Take 25 to 50 g of the sample in a blender
jar and add diluting fluid (8.1.1) to have dilution of 10- 1 • Blend
at 8000 to 10000 rev/ min for 2 minutes. Alternatively, macerate with
diluting fluid (8.1.1) in a sterile mortar with sterile sand. Make
serial ten-fold dilutions with the diluting fluid in duplicate series up to
10- 6 • Streak 0·1 ml from each tube evenly on to the ethyl violet azide
dextrose agare 4.6.1) and incubate at 370 0 for 48 hours. Enumerate
the colonies which are as described in 3.2 and confirm these by Gram's
stain (6.2.1) and by growth at 44°0 in MacOonkey agar (4.7) for
typical small pink colonies. The number of viable colonies per gram of
sample is determined by multiplying hy the dilution factor( s) and
dividing by the mass of the sample.
8.2.2 Enterococci Index - Obtain serial dilutions of the sample as
in 8.2.1. Transfer, with a fresh sterile pipette, a measured volume of 1 ml
of the homogenized mixture and of the five following serial dilutions of
both dilution series in triplicate to the tubes of 10 ml of ethyl violet
azide dextrose broth (4.6). Start with the highest dilution and
proceed to the' lowest, filling and emptying the pipette three times
before transferring the 1 mI portions to the tubes of medium ( see 4.6 ).
When the number of enterococci is assumed to be very small, start by
transferring, using a sterile 10 ml pipette, 10 mI of the homogenized
mixture in triplicate to 10 ml of double strength broth which contains
twice the amounts of the ingredient as in 4.6 in 1000 ml of water.
Incubate at 37"C for 48 hours. Record as positive the tubes which have
developed turbidity (growth) and the growth having been confirmed as
being faecal streptococci as described in 6.2. Using Table I, obtain
the most probable number ( MPN) of faecal streptococci per gram of the
sample. Use for the calculation the results from three dilutions, select-
ing the highest dilution showing three positive tubes below which no sets
with a smaller number of positive tubes occur, and the two following
higher dilutions. The number obtained from Table I has to be
multiplied by the lowest dilution factor, that is, that of the first set of
tubes, to obtain the most probable number of faecal streptococci
per gram of the sample. For example, when dilution 100 ( = 10 ml of

9
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS I 5887 ( Part D ) .. 1976
macerate), 10- 1 and 10-1 are found to give the following numbers of
positive tubes: 2, 2, 1, the MPN is 2'8 bacteria per gram, and when the
dilutions 100, 10- 1, 10- 2 , 10- 3 , 10-' and 10-& are found to give the
following numbers of positive tubes: g, 3, 3, 2, 0, 0, the MPN is 9'3
( 3, 2, 0), multiplied by the dilution factor 101 , that is, 9 '3 X 101
bacteria. per gram, The MPN is reported as the average of the results
obtained from each of the duplicate dilution series.

TABLE 1 MOST PROBABLE NUMBER ( MPN) OF FAECAL

STREPTOCOCCI
( Clause 8,2,2 )

NUMBER OF MPN NUMBER OF MPN NUMBER OF MPN

POSI~'IVE TUBES POSITIVE TUBES POSITIVE TUBES
PER DILUTION PER DILUTION PER DILUTION
r------A.----,
s
,------'-------, r----.A--~
100 10-
1 10- 100 10- 1 10- 1 10· 10- 1 lO- a

(1) (2) (3) (4) (1) (2) (3) (4) (I) (2) (3) (4)

0 0 0 0'30 1 1'1 2 2 3 4'2

0 0 1 0'30 1 2 I'S 2 3 0 2'9
0 0 2 0'60 I 3 1'9 2 3 I 3'6
0 0 3 0'90 2 0 1'1 2 3 2 4'4
0 1 0 0'30 1 2 1 I'S 2 3 3 5'3
0 1 1 0'61 1 2 2 2'0 3 0 0 2'3
0 1 2 0'92 2 3 2'4 3 0 1 3'9
0 1 3 1'2 3 0 1'6 3 0 2 6'4
0 2 0 0'62 3 1 2'0 3 0 3 9'5
0 2 1 0'93 1 3 2 2'4 3 1 0 4'3
0 2 '2 1'2 1 3 3 2'9 3 1 1 7'S
0 2 3 1'6 2 0 0 0'91 3 1 2 12'0
0 3 0 0'94 2 0 1 1'4 3 1 3 16'0
0 3 1 1'3 2 0 2 2'0 3 2 0 9'3
0 3 2 1'6 2 0 3 2'6 3 2 IS'O
0 3 3 1'9 2 0 I'S 3 2 2 21'0
0 0 0'36 2 1 2'0 3 2 3 29'0
0 0'72 2 2 2'7 3 3 0 24'0
0 2 1'1 2 1 3 3'4 3 3 1 46'0
0 3 I'S 2 2 0 2'1 3 3 2 110'0
0 0'73 2 2 1 2'8 3 3 3 110'0
2 2 2 3'S

10
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
IS I 5887 ( Part II ) • 1976
( Continued from page 2 )
Members Representing
DR A. N. BOSE The Bengal Immunity Co Ltd, Calcutta
DR SUBRATA CHAKRAVORTY Bengal Chemical and Pharmaceutical Works Ltd,
Calcutta
DIRECTOR King Institute, Madras
DR A. K. GHOSH Cholera Research Centre ( Indian Council of Medical
Research ). Calcutta
HEAD, DIVISION OF BIOLOGICAL Indian Veterinary Research Institute (ICAR).
PRODUCTS Izatnagar
DR A. P. JOSHI Vallabhbhai Patel Chest Institute, Delhi
DR ( SMT ) V. BAJAJ ( Alternate)
DR M. A. KRISHNASWAMY Central Food Technological Research Institute
( CSIR), Mysore
SHRI C. T. DWARKANATH ( Alternate)
SHRI K. R. NARASIMHAN The Metal Box Company of India Ltd, Calcutta
DR S. C. CHAKRAVORTY ( Alternate)
DR A. N. RAI CHOWDHURY Central Research Institute, Kasauli
DR B. RANGANATHAN National Dairy Research Institute ( ICAR ). Karnal
DR M. V. SANT Haffkine Institute, Bombay
DR SHRINIWAS All India Institute of Medical Sciences, New Delhi
DR N. S. SUBBA RAo Indian Agricultural Research Institute (ICAR),
New Delhi
COL R. N. TANEJA Food Inspection Organization, Quartermaster
General's Branch, Army Headquarters
LT-COL D. D. VORRA ( Alternate)

11
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on
INDIAN STANDARDS
ON
FOOD MICROBIOLOGY

IS:
5401-1969 Methods for detection and estimation of coliform bacteria in foodstuffs
5402-1969 Method for standard plate count of bacteria in foodstuffs
5403-1969 Method for yeast and mould count of foodstuffs
5404-1969 Code of practice for handling of samples for microbiological analysis
5887 ( Part I )-1976 Methods for detection of bacteria responsible for food poisoning:
Part I Isolation, identification and enumeration of Escherichia coli (first
revision)
5887 ( Part II )-1976 Met~ods. for .dete~tion of bacteria :esponsible for food poisoning:
Part II Isolation, IdentificatIOn and enumeration of Staphylococcus aureus and
Faecal streptococci (first revision)
5887 ( Part III )-1976 Methods for detection of bacteria responsible for food poisoning:
Part III Isolation and identification of Salmonella and Shigella (first revision)
5887 ( Part IV )-1976 Methods for detection of bacteria responsible for food poisoning:
Part IV Isolation and identification of Clostridium welchii, Clostridium botulinum
and bacilIus cereus and enumeration of Clostridium welchii and Bacillus
cereus (first revision )
5887 ( Part V )-1976 Methods for detection of bacteria responsible for food poisoning:
Part V Isolation, identification and enumeration of Vibrio cholerae and Vibrio
parahaemolyticus (first revision)
6850-1973 Agar, microbiological grade
6851-1973 Meat extract, microbiological grade
6852-1973 Bile saits, microbiological grade
6853-1973 Peptone, microbiological grade
6854-1973 Methods of sampling and test for ingredients used in media for microbio-
logical work
7004-1973 Yeast extract, microbiological grade
7127-1973 Ttyptone, microbiological grade
7128-1973 Proteose peptone, microbiological grade
7203-1973 Casein hydrolysate ( acid digested ), microbiological grade
7535-1975 Liver extract, microbiological grade
7536-1975 Soluble starch, microbiological grade
7590-1975 Gelatin, microbiological grade
7591-1975 Malt extract, microbiological grade
7801-1975 Trypsine, microbiological grade
 Free Standard provided by BIS via BSB Edge Private Limited to BELLA -
NNALAPATTI([email protected]) 182.74.88.150 [for non-commercial use on