PUBLIC REVIEW DRAFT DRAFT UGANDA STANDARD  
  DUS DEAS 968 
First  Edition  
2019-mm-dd 
 
 Reference number  
DUS DEAS 968: 2019 
© UNBS 2019 
   
Disposable adult diapers — Specification  
 
 PUBLIC REVIEW DRAFTDUS DEAS 968: 2019 
ii                © UNBS  2019 - All rights reserved 
 Compliance with this standard does not, of itself confer immunity from legal obligations  
A Uganda Standard does not purport to include all necessary provisions of a contract. Users are 
responsible for its correct applicati on 
© UNBS 2019 
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilised in any 
form or by any means, electronic or mechanical, including photocopying and microfilm, without prior written 
permission from UNB S. 
Requests for permission to reproduce this document should be addressed to 
The Executive Director 
Uganda National Bureau of Standards 
P.O. Box 6329 
Kampala 
Uganda 
Tel: +256 417 333 250/1/2 
Fax: +256 414 286 123 
E-mail: info@unbs.go.ug  
Web: www.unbs.go.ug  
 PUBLIC REVIEW DRAFTDUS DEAS 968: 2019 
© UNBS  2019 - All rights reserved                iii National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry of Trade, Industry and 
Cooperatives established under Cap 327, of the Laws of Uganda, as amended.  UNBS is mandated to co-
ordinate the elaboration of standards and is  
(a) a member of International Organisation for Standardisation (ISO) and  
(b) a contact point for the WHO/FAO Codex Alimentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TBT Agreement of the World Trade Organisation (WTO). 
The work of preparing Uganda Standards is carried out through Technical Committees. A Technical 
Committee is established to deliberate on standards in a given field or area and consists of representatives of 
consumers, traders, academicians, manufacturers, government and other stakeholders. 
Draft Uganda Standards adopted by the Technical Committee are widely circulated to stakeholders and the 
general public for comments. The committee reviews the comments before recommending the draft standards 
for approval and declaration as Uganda Standards by the National Standards Council.  
This Draft Uganda Standard, DUS DEAS 968: 2019 , Disposable adult diapers — Specification , is identical 
with and has been reproduced from an International Standard, DEAS 968 : 2019, Disposable adult diapers — 
Specification , and is being proposed for adoption as a Uganda Standard. 
The committee responsible for this document is Technical Committee UNBS/TC 7,  Textiles, leather, paper 
and related products , Subcommittee SC 1 , Textiles and related products.  
Wherever the words, “ East African Standard " appear, they should be replaced by "Uganda Standard."  DEAS  968: 201 9 
ICS nn.nnn. nn 
 
© EAC 2019                                                                                                                              First Edition 2019  
 DRAFT EAST AFRICAN STANDARD  
Disposable adult  diaper s — Specification  
EAST AFRICAN COMMUNITY  DEAS 968: 2019 
ii © EAC 2019 – All rights reserved  
 Copyright notice  
This EAC document is copyright -protected by EAC. While the reproduction of this document by 
participants in the EAC standards development process is permitted without prior permission from EAC, 
neither this document nor any extract from it may be reproduced, stored or transmitted in any form for 
any other purpose without prior written permission from EAC.  
Requests for permission to reproduce this do cument for the purpose of selling it should be addressed 
as shown below or to EAC’s member body in the country of the requester:  
© East African Community 2018  — All rights reserved  
East African Community   
P.O. Box 1096 , 
Arusha  
Tanzania  
Tel: + 255 27 2 16210 0 
Fax: + 255 27 2162190  
E-mail: eac@eachq.org  
       Web: www.eac -quality.net  
Reproduction for sales purposes may be subject to royalty payments or a licensing agreement.  
Violators may be prosecuted . DEAS 968: 201 9 
© EAC 2019 – All rights reserved  iii 
 Contents  Page 
Foreword  ................................ ................................ ................................ ................................ ............................  iv 
1 Scope  ................................ ................................ ................................ ................................ ......................  1 
2 Normative references  ................................ ................................ ................................ ............................  1 
3 Terms and definitions  ................................ ................................ ................................ ...........................  1 
4 Requirements  ................................ ................................ ................................ ................................ .........  3 
4.1 Materials  ................................ ................................ ................................ ................................ .................  3 
4.1.1  Absorbent filler  ................................ ................................ ................................ ................................ ...... 3 
4.1.2  Top Sheet (the layer which contacts adult’s skin)  ................................ ................................ .............  3 
4.1.3  Back sheet  ................................ ................................ ................................ ................................ ..............  3 
4.1.4  Fastening device (closure system)  ................................ ................................ ................................ ...... 3 
4.1.5  Breathability  ................................ ................................ ................................ ................................ ...........  3 
4.2 Key performance requirements  ................................ ................................ ................................ ...........  3 
4.3 Microbiological requirements  ................................ ................................ ................................ ..............  4 
5 Packaging and marking  ................................ ................................ ................................ ........................  4 
5.1 Packaging  ................................ ................................ ................................ ................................ ...............  4 
5.2 Marking  ................................ ................................ ................................ ................................ ...................  4 
6 Sampling  ................................ ................................ ................................ ................................ ................  4 
Annex  A (normative)  Rate of absorption and rewet test  ................................ ................................ ...............  6 
A.1 Scope and Purpose:  ................................ ................................ ................................ ..............................  6 
A.2 Principle  ................................ ................................ ................................ ................................ .................  6 
A.3 Apparatus and materials  ................................ ................................ ................................ ......................  6 
A.4 Saline solution  ................................ ................................ ................................ ................................ ....... 6 
A.5 Saline dosage  ................................ ................................ ................................ ................................ ........  6 
A.7 Calculation of results  ................................ ................................ ................................ ............................  8 
Annex  B (normative)  Microbiological Examination  ................................ ................................ .......................  9 
B.1 Apparatus and equipment  ................................ ................................ ................................ ....................  9 
B.2 Media and reagents  ................................ ................................ ................................ ...............................  9 
B.2.1  General  ................................ ................................ ................................ ................................ ...................  9 
B.2.2  Bacteriological peptone  ................................ ................................ ................................ ........................  9 
B.2.3  Plate count agar  ................................ ................................ ................................ ................................ ..... 9 
B.2.4  Neutral red -bile salt peptone glucose medium  ................................ ................................ ..................  9 
B.2.5  Fluid soybean -casein digest medium  ................................ ................................ ...............................  10 
B.2.6  Centri mide agar medium  ................................ ................................ ................................ ....................  10 
B.2.7  Pseudomonas agar  medium for detection of fluorescein  ................................ ...............................  10 
B.2.8  Pseudomonas agar  medium for detection of pyocyanin  ................................ ................................  11 
B.3   Preparation of Test Suspension  ................................ ................................ ................................ ....... 11 
B.4  Procedure  ................................ ................................ ................................ ................................ .............  11 
B.4.1   Total viable b acterial count  ................................ ................................ ................................ ...............  11 
B.4.2  Examination for the presence of Enterobacteriaceae.  ................................ ................................ .... 11 
B.4.3  Examination for the presence of Staphylococcus aureus . ................................ .............................  11 
B.4.4  Examination for the presence of Pseudomonas aeruginosa  ................................ ..........................  11 
 DEAS 968: 2019 
iv © EAC 2019 – All rights reserved  
 Foreword  
Development of the East African Standards has been necessitated by the need for harmonizi ng requirements 
governing quality of products and services in the East African Community. It is envisaged that through 
harmonized standardization, trade barriers that are encountered when goods and services are exchanged 
within the Community will be remove d.  
The Community has established an East African Standards Committee  (EASC)  mandated to develop and 
issue East African Standards  (EAS) . The Committee is composed of representatives of the National 
Standards Bodies in Partner States, together with the repr esentatives from the public and private sector 
organizations  in the community .  
East African Standards are developed through  Technical Committees that are representative of key 
stakeholders including government, academia, consumer groups, private sector an d other interested parties. 
Draft East African Standards are circulated to stakeholders through the National Standards Bodies in the 
Partner States. The comments received are discussed and incorporated before finalization of standards, in 
accordance with the Principles and procedures for development of East African Standards.  
East African Standards are subject to review, to keep pace with technological advances. Users of the East 
African Standards are therefore expected to ensure that they always have the latest versions of the standards 
they are implementing .  
The committee responsible  for this document is Technical Committee EASC /TC 061, Textiles, Textile 
products and Accessories  
Attention is drawn to the possibility that some of the elements of this docu ment may be subject of patent 
rights. EAC shall not be held responsible for identifying any or all such patent rights.  
 DRAFT EAST AFRICAN STANDARD  DEAS 968:2019 
 
© EAC 2019  – All rights reserved  1 
 Disposable adult  diapers  — Specification  
1 Scope   
This draft East African Standard specifies requirements  and test methods  for disposab le adult  absorbent 
products for managing incontinence including adult diapers, adult briefs , adult under pads (inserted in pants) 
and others.  
2 Normative references   
The following documents are referred to in the text in such a way that some or all of thei r content constitutes 
requirements of this document.  For dated references, only the edition cited applies. For undated references, 
the latest edition of the referenced document (including any amendments) applies.  
ISO 139,  Textiles — Standard atmospheres fo r conditioning and testing  
ISO 3071,  Textiles — Determination of pH of aqueous extract   
ISO 4833 -2, Microbiology of the food chain — Horizontal method for the enumeration of microorganisms — 
Part 2: Colony count at 30 C by the surface plating technique   
ISO 7218 , Microbiology of food and animal feeding stuffs — General requirements and guidance for 
microbiological examinations (2nd Edition)   
ISO 11948 -1, Urine -absorbing aids — Part 1: Whole -product testing  
ISO 21149, Cosmetics — Microbiology — Enumeration a nd detection of aerobic mesophilic bacteria  
ISO 22717, Cosmetics — Microbiology — Detection of Pseudomonas aeruginosa  
ISO 22718, Cosmetics — Microbiology — Detection of Staphylococcus aureus  
3 Terms and definitions   
For the purposes of this document, the f ollowing terms and definitions apply.  
ISO and IEC maintain terminological databases for use in standardization at the following 
addresses:  
— ISO Online browsing platform: available at http://www.iso.org/obp  
3.1  
Incon tinence products  
hygienic products specifically developed to help manage bladder or bowel control problems with the capability 
to absorb urine and prevent stool and fluid from leaking out   
2 © EAC 2019 – All rights reserved  
 3.2  
absorbent filler material  
the material at the core of the diape r that absorbs fluids  
3.3  
Top sheet run -off 
The unabsorbed volume of test solution while running across the surface of inclined specimen  
3.4  
Rewet  
The mass of test solution which returns to surface under specific pressure after a certain amount of that being 
absorbed by the specimen  
3.5  
Leakage  
The mass of test solution which penetrates the leak -proof carrier film under specific pressure after a certain 
amount of that being absorbed by specimen  
3.6 
Rate of Absor bency (ROA)  
Time required to fully absorb  test fluid into the test product.  A specified quantity of simulated urine is 
discharged at a prescribed rate under specified  conditions onto a test specimen of an absorbent article with a 
nonwoven cover. The time  taken for the entire liquid dose to pen etrate the nonwoven is measured  
3.7 
Retention Capacity  
A measure of a product’s capacity to hold fluid without leaking and rewetting the skin  
3.8 
disposable adult diaper  
disposable hygienic pad  for adults having capability to absorb  urine  and prevent stool an d fluid from leaking  
out  
© EAC 2019 – All rights reserved  3 
  
4 Requirements  
4.1 Materials  
All materials used for making  absorbent products  shall not harm the skin in contact. None of the components 
in the products, including additives, should be listed in any Regulatory Agency as being “un safe’’  
4.1.1 Absorbent filler  
a) it shall allow for air circulation and be comfortable for the baby user. 
b) it shall be clean, free from harmful foreign materials, lumps, splits, holes and protruding  points when 
visually examined; and  
c) it shall be arranged in a m anner that will draw in and trap all moisture rapidly and efficiently and keeps  
it away from the adult’s skin.  
4.1.2 Top Sheet (the layer which contacts adult’s skin)  
a) shall be of material that helps absorption, and shall have no harmful effect ; and  
b) shall c over the absorbent filler completely and prevent the filler from reaching the adult’s skin or 
clothes under normal handling . 
4.1.3 Back sheet  
There shall be an outer cover to prevent direct contact of the absorbent filler with the adult’s clothing and to 
prevent li quid leakage out of the absorbent product.  
4.1.4 Fastening device (closure system ) 
There shall be a suitable device for fastening the absorbent products for secure use. The closure system 
regardless of how its functionality is achieved should allo w for multiple fastening and unfastening. This 
promotes better fit and allows for easy check of wetness without having to dispose off and replace unsoiled 
product  
4.1.5 Breathability  
There should be a  minimum air flow sufficient to release trapped body hea t or gaseous body per spiration in 
the pelvic region  
4.2 Key performance requirements  
Adult absorbent products  shall comply with  the performance requirements  given in Table 1 when tested in 
accordance with the methods specified therein.  
Table  1 — Characteri stics of disposable adult absorbent products  
Characteristics  All size categories  Test methods  
Rate of absorption , s, (max)  85 Annex A  
Rewet under load , g, (max)  5 Annex A   
4 © EAC 2019 – All rights reserved  
 Absorptive capacity (min)  1200  ISO 11948 -1 
pH  6 – 8.5 ISO 3071  
 
4.3 Microbiolog ical requirements  
a) the total viable bacterial count, when determined in accordance with B.4.1 shall not exceed 100 CFU / 
g; and  
b) when tested in accordance with ISO 21149 , disposable diapers shall be free from E. coli , 
Staphylococcus aureus, and Pseudomonas a eruginosa and C. albicans in 1 g of product   
5 Packaging and marking  
5.1 Packaging  
Disposable adult absorbent products shall be packed in a suitable waterproof package that shall protect them 
from any form of contamination and damage.  Packaging for shipmen t shall be in accordance with the 
manufacturer's standard practice and in a manner readily accepted by  the market.  Within the shipping carton, 
units shall be packed in manner designed to minimize damage during shipment due to rough or improper 
handling.  
5.2 Marking  
On both the primary and secondary packages, the disposable adult absorbent product packs shall be marked 
with legible and indelible pre -printed marking or a securely affixed and durable label bearing the following 
information:  
a) name of products;  
b) name and address of the manufacturer and/or importer/distributor (if applicable);  
c) number of pieces in the pack;  
d) instruction for use, storage and disposal ; 
e) date of manufacture and expiry;  
f) batch/ lot number; and  
g) country of origin.  
6 Sampling  
6.1 Lot 
In any c onsignment , all packages of the adult absorbent product  belonging to one batch of manufacture or 
supply shall constitute a lot.  
6.2 Scale of sampling  
6.2.1 Samples shall be tested from each lot ascertaining its conformity to the requirements of this 
specif ication.   
© EAC 2019 – All rights reserved  5 
 6.2.2 The number of packages to be selected from a lot sha ll be in accordance with Table 2. 
Table 2 –scale of sampling  
Number of packages in a lot  Number of packages to be selected  
Up to 250  6 
251-500 8 
501-1000  11 
1001 -2500  15 
2501 -5000  20 
5001 and above  30 
 
6.2.3 The bulk packages and packag es shall be selected at random  
6.3 Number of tests  
6.3.1 Each package selected as per table 2 shall be inspected for packaging and marking requirements  
6.3.2 Disposable adult diapers  selected as per tab le 2 shall be examined for requirements stipulated in 
clause 4  
  
6 © EAC 2019 – All rights reserved  
 Annex  A 
(normative)  
 
Rate of absorption and rewet test  
A.1 Scope and Purpose:  
a) To measure the ability of an absorbent products to accept and retain synthetic urine (saline solution) 
under simu lated in -use conditions of load and pressure.  
b) To determine amount of time required for an absorbent article to absorb a fixed quantity of a test 
solution  
A.2 Principle  
The method measures the ability of the absorbent products to accept and retain 0.9% sali ne solution under 
simulated in -use conditions of load and pressure  
A.3 Apparatus and materials  
a) a rigid cover plate with a weight as shown below. Dimensions of the plate (200 mm ± 2 mm) x (70 mm 
± 2 mm). Inner diameter of cylinder 40 mm ± 2 mm, total Weigh t: 6300 g ± 10 g representing a 
pressure of 4.41 kPa ± 0.05 kPa (0.64 psi ± 0.007 psi) for all sizes  
b) tray; 
c) filter paper; having a diameter of 110  mm and conditioned together with the test samples  
d) graduated cylinder (1 ml graduation);  
e) stopwatch;  
f) ruler (at l east 2 cm longer as absorbent core of the sample, 1mm graduation); and  
g) a pen.  
A.4 Saline solution  
Add 9 g sodium chloride, and 1 g of blue dye into 200 mL distilled water at ambient temperature after which 
the solution is made up to 1 Liter. The pH shall b e between 6.2 – 6.7. When not within this range, adjust by 
using 0.1 mol/L sodium hydroxide solution or acetic acid as relevant. The temperature of the solution shall be 
37 ºC ± 2 ºC, and shall be maintained in a water bath during testing.  
A.5 Saline dosag e 
Measure out the volume of saline solution for each of the products being tested as follows:  
a) adult disposable diaper (brief) of size: Medium to XX -Large, Single dose of 200 ml   
© EAC 2019 – All rights reserved  7 
 b) adult disposable diaper (brief) of size: Small, Single dose of 100 ml  
c) all sizes  of adult disposable under -pads/ underpants: Single dose of 100 ml  
A.6 Sample preparation and Set -up 
a) conditioning: Bring samples to moisture equilibrium in the standard atmosphere for testing nonwovens 
in accordance with ISO 139. Equilibrium is considered to have been reached when the increase in 
mass of the specimen in successive weighings made at intervals of not less than 2 hours does not 
exceed 0.25 % of the mass of the specimen;  
b) take 5 test specimens  randomly from the test samples;  
c) weigh a stack of dry  filter paper and record as weight W1. The stack should have a dry weight of 
about 10.0 grams;  
d) mark the loading point in the middle of the absorbent core. Note that the absorbent core does not 
cover the full length of the diaper but it is more concentrated  in the front of the diaper. Therefore, the 
middle of the absorbent core will not be the same as the middle of the diaper: Measure the length and 
width of the absorbent core. Mark the midpoint, which will be the loading point;  
e) place core with top sheet fac ing upward on the tray;  
f) place rigid cover plate, ensure the plate is centered towards the width of the diaper core and the 
cylinder opening is placed over marked loading point. Diaper should be stretched showing minimum 
amount of wrinkles;  
g) gently place wei ghts on the plate (ideally use weight in a ring form to allow for equally applied 
pressure;  
h) fill the measuring cylinder with respective amount of saline solution;  
i) gently pour the saline onto the diaper starting the stop watch at the same time with the pour ing; 
j) stop the stopwatch as soon as the saline solution has been absorbed by the diaper (all the wet sheen 
has disappeared) and record the result in seconds as the rate of absorption;  
Rewet test  
a) leave the diaper undisturbed for 10 min;  
b) lift the weight and c over plate and p lace the filter paper stack on the absorption point of the diaper;  
c) replace the cover plate and the weight for 2 min on top of the filter paper stack;  
d) after 2 min, remove the weight and cover plate and immediately determine and record the we ight of 
the wet filter paper stack as W2; and  
e) repeat the procedure on the remaining test specimens  recording each result separately;   
8 © EAC 2019 – All rights reserved  
 
 
Fig A.3: apparatus and set up  
A.7 Calculation of results  
A.7.1  Calculate as the rate of absorption in s, the mean time f or absorption recorded in (j) above.  
A.7.2  REWET (g) = W2 (Wet filter paper weight) - W1 (Dry filter paper weight). Report the average of the 5 
products tested  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  
© EAC 2019 – All rights reserved  9 
 Annex  B 
(normative)  
 
Microbiological Examination  
B.1 Apparatus and equipment  
Use apparatus and equipment complying with the relevant requirements of ISO 7218.  
B.2 Media and reagents  
B.2.1 General  
Ensure compliance with the general requirements for the ingredients and for the preparation of media and 
reagents given in ISO 7218   
B.2.2 Bacteriological peptone  
Peptone               10 g  
Disodium phosphate dodecahydrate         1 g  
Sodium chloride             5 g  
Mono -potassium phosphate          1.5 g  
Dissolve the ingredients in distilled water and make up to 1 L. Adjust the pH value  to be 7.0 ± 0.1 after 
sterilization. Dispense 300 mL volumes into flasks of capacity 500 mL and sterilize by autoclaving at 121 °C ± 
2 °C for 20 min.  
B.2.3 Plate count agar  
Agar               15 g  
Glucose               1 g  
Tryptone               5 g  
Yeas t extract              2.5 g  
Dissolve the ingredients in distilled water, made up to 1litre, and adjust the pH value to 7.2 ± 0.2. Dispense 15 
mL volumes into bottles and sterilize by autoclaving at 121 °C ± 2 °C for 20 min.  
B.2.4 Neutral red -bile salt pep tone glucose medium  
Peptone               20 g  
Glucose               10 g  
Bile salts No. 3             1.5 g   
10 © EAC 2019 – All rights reserved  
 Sodium chloride           5 g  
Neutral red            0.03 g  
Crystal violet            0.002 g  
Dissolve the ingredients in 400 mL of distilled wate r and make up to 500 mL boiling to aid solution. Adjust the 
pH value to 7.4 and filter to a clear solution. Dispense 10 mL volumes into bottles each containing a Durham 
tube and sterilize by autoclaving at 121 °C ± 2 °C for 20 min  
B.2.5 Fluid soybean -casei n digest medium  
Pancreatic digest of casein         17 g 
Papaic digest of soybean meal        3 g  
Sodium chloride           5 g  
Dibasic potassium phosphate        2.5 g  
Dextrose             2.5 g  
Dissolve the ingredients in distilled water and make up to 1 l itre, warming slightly to aid solution. Cool the 
solution to room temperature and adjust the pH value to be 7.3 ± 0.2 after sterilization. Filter to clarify (if 
necessary), dispense into suitable containers, and sterilize by autocl aving at 121 ± 2 °C for 2 0 min.  
B.2.6 Centrimide agar medium  
Pancreatic digest of gelatine        20 g  
Magnesium chloride          1.4 g  
Potassium sulphate          10 g  
Agar             13.6 g  
Cetyltrimethylammonium bromide (Cetrimide)    0.3 g  
Glycerine            10 mL  
Dissol ve all the solid ingredients in distilled water, make up to 1 L, and then add the glycerine. Heat, agitating 
frequently, and boil for 1 min. Adjust the pH value to be 7.2 ± 0.2 after sterilization. Dispense into suitable 
containers and sterilize by autocla ving at 121 °C ± 2 °C for 20 min.  
B.2.7 Pseudomonas agar  medium for detection of fluorescein  
Pancreatic digest of casein         10 g  
Peptic digest of animal tissue        10 g  
Anhydrous dibasic potassium phosphate      1.5 g  
Magnesium sulphate (MgSO4.7H2 O)      1.5 g  
Glycerine            10 mL  
Agar             15 g   
© EAC 2019 – All rights reserved  11 
 Dissolve all the solid ingredients in distilled water, make up to 1 L, and then add the glycerine. Heat, agitating 
frequently, and boil for 1 min. Adjust the pH value to be 7.2 ± 0.2 after ste rilization. Dispense into suitable 
containers and sterilize by autoclaving at 121 °C ± 2 °C for 20 min.  
B.2.8 Pseudomonas agar  medium for detection of pyocyanin  
Pancreatic digest of casein           20 g  
Anhydrous magnesium chloride          1.4 g  
Anhydrou s potassium sulphate          10 g  
Agar               15 g  
Glycerine              10 Ml  
B.3   Preparation of  Test Suspension  
Transfer 300 ml of the sterile soluti on of bacteriological peptone (B .2.2) to a sterile wide -mouthed jar of 
capacity not less than 1  litre and not more than 2  litres. The jar shall have a mouth of diameter not less than 
150 mm and not more than 250 mm, and is fitted with a hermetically closing glass or metal -and-glass lid.  
Aseptically place the towel under test in the solution in the jar, fit the lid, agitate the contents of the jar for 2 
min and then allo w the jar to stand for 10 min. Repeat this agitating and standing procedure twice more. 
Aseptically remove about 100 ml of the test suspensi on for testing as described in B .4 below.  
B.4  Procedure  
B.4.1   Total viable bacterial count  
Into each of three sterile petri dishes aseptically pipette a 1 mL portion of the test suspension. To each dish 
add 15 mL of freshly  melted plate count agar  that has been cooled to 45 °C, and mix well. I ncubate, count and 
calculate the total count as described in ISO 4833  Part 2. From the total viable bacterial count and the mass of 
the sanitary towel, calculate the total viable bacterial count per gram of sanitary towel.  
B.4.2  Examination for the presen ce of Enterobacteriaceae.  
Aseptically add 10 mL of the test suspension to a bottle that contains neutral red -bile salt peptone glucose 
medium (B .2.4). Incubate the bottle for 24 h to 36 h at 37 ± 0.5°C and examine for the presence of 
Enterobacteriaceae as evidenced by the formation of acid and gas.  
B.4.3  Examination for the presence of Staphylococcus aureus .  
Use the media, reagents and procedure described in ISO 22718  to ex amine th e test suspension (see B .3). As 
a control, pipette 0.1 mL of a 1:1000 dilu tion of an 18 h to 24 h culture of Staphylococcus aureus  SATCC Sta 
10 into Staphylococcus  medium and proceed as with the test suspension.  
B.4.4  Examination for the presence of Pseudomonas aeruginosa  
a) Use the media, reagents and procedure described in ISO 22717. Aseptically pipette 10 mL of the test 
suspension into 90 mL of fluid  soybean -casein digests medium (B .2.5) and mix well. Incubate for 24 h 
at 30 °C to 35 °C. By means of an inoculating loop transfer a portion from the 24 h incubated sample 
tube of fluid soybean -casein digest medium to the dry surface of petri dishes each containing 
approximately 2 0 mL of Cetrimide agar medium (B .2.6). Incubate at 30 °C to 35 °C and examine after 
24h, and again after 48 h incubation, for suspect colonies, bearing in mind that in general greenish  
12 © EAC 2019 – All rights reserved  
 fluorescent colonies are typical of Pseudomonas aureginosa  and that in its presence a gram stain 
examined microscopically will reveal gram -negative slender rod -shaped cells.  
b) As a control, add 0.1 ml of a 1:1 000 dilution of a n 18 h to 24 h culture of Pseudomonas aeruginosa  
SATCC Pse 11 mL to 100 mL of fluid  soybean -casein digest medium (B .2.5), and proceed as with the 
test suspension.  
c) If none of the colonies obtained from the test suspension conforms to the description given in i) above 
and the control culture has been satisfactorily recovered, deem the test sample to be free from 
Pseudomonas aeruginosa.  
d) If colonies conforming to the description given in i) above are found, streak representative suspect 
colonies from the Cetr imide agar onto the surfaces of Pseudomonas agar  medium for  the detection of 
flourescein (B .2.7) and Pseudomonas agar medium f or the detection of pyocyanin (B .2.8) to obtain 
isolated colonies.  Cover and invert the petri dishes and incubate at 30 – 35 °C f or at least 3 days.  
Examine the streaked surfaces under ultraviolet light for suspect c olonies, as described in Table B .1.  
Table B .1 — Description of colonies  
Medium  Description of colonies  
Pseudomonas agar for the detection of fluorescein  Generally co lourless to yellowish  
Yellowish fluorescence in ultra violet light  
Pseudomonas agar for the detection of pyocyanin  Generally greenish. Blue fluorescence in ultraviolet light  
 
If any further doubt exists as to the identity of the colonies, obtain final co nfirmation by inoculating the suspect 
colonies to the wells on commercially available diagnostic kits in accordance with the manufacturer’s 
instructions.  
 DEAS 1783 : 2019   
 
© EAC 2019 – All rights reserved