UGANDA STANDARD
 
    US ISO 21527-2
First Edition
2009-mm-dd
 
Reference numbe r
     US ISO 21527-2:2008
 © UNBS 2009
  
Microbiology of food and animal feeding stuffs — Horizontal 
method for the enumeration of yeasts and moulds — Part 2: 
Colony count technique in products with water activity less 
than or equal to 0.95 
 US ISO 21527-2: 2008 
 
© UNBS  2009 – All rights reserved ii 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 application  
© UNBS 2009 
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, in cluding photocopying and microfilm, without prior written 
permission from UNBS. 
Requests for permission to reproduce this document should be addressed to 
The Executive Director 
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Tel: 256 41 505 995 
Fax: 256 41 286 123 
E-mail: unbs@infocom.co.ug
Web: www.unbs.go.ug  
      US ISO 21527-2:2008  
© UNBS  2009 - All rights rese rved                iii National foreword 
Uganda National Bureau of Standards (UNBS) is a parastatal under the Ministry  of Tourism, Trade and 
Industry established under Cap 327, of the Laws of U ganda. 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 A limentarius Commission on Food Standards, and  
(c) the National Enquiry Point on TBT/SPS Agr eements 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 gi ven field or area and consists of representatives of 
consumers, traders, academician s, manufacturers, governme nt and other stakeholders. 
Draft Uganda Standards adopted by the Technical Committ ee 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 Uganda Standard, US ISO 21527-2, Microbiology of food and animal feeding stuffs — Horizontal
method for the enumeration of yeasts and moulds — Part 2: Colony count technique in products with water
activity less than or equal to 0.95,  is identical with and has been reproduced from an International Standard,
ISO 21527-2:2008, Microbiology of food and animal feeding st uffs — Horizontal method for the 
enumeration of yeasts and moulds — Part 2: Colony count technique in products with water activity less than 
or equ al to 0.95, and adopted as a Uganda Standard.  
This standard was developed under the subcommittee on fruits and vegetables by the Food and agriculture Standards Technical Committee (UNBS/TC 2). 
Wherever the words, "International Standard" a ppear, they should be replaced by "Uganda Standard." 
  
 
  
 Reference number
ISO 21527-2:2008(E)
© ISO 2008
  
 INTERNATIONAL 
STANDARD  ISO
21527-2
First edition
2008-07-01
Microbiology of food and animal feeding 
stuffs — Horizontal method for the enumeration of yeasts and moulds — 
Part 2: 
Colony count technique in products with 
water activity less than or equal to 0,95 
Microbiologie des aliments — Méthode horizontale pour le 
dénombrement des levures et moisissures — 
Partie 2: Technique par comptage des colonies dans les produits à 
activité d'eau inférieure ou égale à 0,95 
 ISO 21527-2:2008(E) 
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ii © ISO 2008 – All rights reserved
  ISO 21527-2:2008(E) 
© ISO 2008 – All rights reserved  iiiForeword 
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies 
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO 
technical committees. Each member body interested in a subject for which a technical committee has been 
established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. 
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards 
adopted by the technical committees are circulated to the member bodies for voting. Publication as an 
International Standard requires approval by at least 75 % of the member bodies casting a vote. 
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent 
rights. ISO shall not be held responsible for identifying any or all such patent rights. 
ISO 21527-2 was prepared by Technical Committee ISO/TC 34, Food products , Subcommittee SC 9, 
Microbiology . 
ISO 21527 consists of the following parts, under the general title Microbiology of food and animal feedings 
stuffs — Horizontal method for the enumeration of yeasts and moulds : 
⎯ Part 1: Colony count technique in products with water activity greater than 0,95 
⎯ Part 2: Colony count technique in products with water activity less than or equal to 0,95 
This part of ISO 21527, together with ISO 21527-1, cancel and replace ISO 7698:1990, ISO 7954:1987 and 
ISO 13681:1995.  ISO 21527-2:2008(E) 
iv © ISO 2008 – All rights reserved
 Introduction 
Because of the large variety of food and feed products, the applications of the horizontal method specified in 
ISO 21527 (all parts) may not be appropriate for certain products. In this case, different methods, which are 
specific to these products, may be used if absolutely necessary for justified technical reasons. Nevertheless, 
every attempt shall be made to apply the horizontal method as specified in ISO 21527 (all parts) as far as possible. 
When ISO 21527 (all parts) is next reviewed, account will be taken of all information then available regarding 
the extent to which the horizontal method has been followed and the reasons for deviations from this method in the case of particular products. 
The harmonization of test methods cannot be immediate, and for certain groups of products International 
Standards and/or national standards may already exist that do not comply with the horizontal method as specified in ISO 21527 (all parts). It is hoped that when such standards are reviewed they will be changed to comply with ISO 21527 (all parts) so that eventually the only remaining departures from this horizontal method will be those necessary for well-established technical reasons. 
 INTERNATIONAL STANDARD ISO 21527-2:2008(E)
 
© ISO 2008 – All rights reserved  1Microbiology of food and animal feeding stuffs — Horizontal 
method for the enumeration of yeasts and moulds — 
Part 2: 
Colony count technique in products with water activity less 
than or equal to 0,95 
WARNING — It is essential that enumeration of moulds is carried out with the greatest care to protect 
the operator and to prevent contamination of the atmosphere with mould spores. 
1 Scope 
This part of ISO 21527 specifies a horizontal method for the enumeration of viable osmophilic yeasts and 
xerophilic moulds in products intended for human consumption or feeding of animals that have a water activity 
less than or equal to 0,95 (dry fruits, cakes, jams, dried meat, salted fish, grains, cereals and cereal products, 
flours, nuts, spices and condiments, etc. [Annex A]), by means of the colony count technique at 25 °C ± 1 °C 
(Reference [3]). 
This part of ISO 21527 does not apply to dehydrated products with water activity less than or equal to 0,60 
(dehydrated cereals, oleaginous products, spices, leguminous plants, seeds, powders for instant drinks, dry products for domestic animals, etc.) and does not allow the enumeration of mould spores (Reference [3]). 
Neither the identification of fungal flora nor the examination of foods for mycotoxins lie within the scope of this 
part of ISO 21527. The method specified in this part of ISO 21527 is not suitable for enumeration of halophilic 
xerophilic fungi (i.e. Polypaecilum pisce, Basipetospora halophila) such  as may be found in dried fish. 
2 Normative references 
The following referenced documents are indispensable for the application 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 6887 (all parts), Microbiology of food and animal feeding stuffs — Preparation of test samples, initial 
suspension and decimal dilutions for microbiological examination  
ISO 7218, Microbiology of food and animal feeding stuffs — General requirements and guidance for 
microbiological examinations  
ISO 8261, Milk and milk products — General guidance for the preparation of test samples, initial suspensions 
and decimal dilutions for microbiological examination  
ISO/TS 11133 (all parts), Microbiology of food and animal feeding stuffs — Guidelines on preparation and 
production of culture media 
ISO 21527-1, Microbiology of food and animal feedings stuffs — Horizontal method for the enumeration of 
yeasts and moulds — Part 1: Colony count technique in products with water activity greater than 0,95  ISO 21527-2:2008(E) 
2 © ISO 2008 – All rights reserved
 3 Terms and definitions 
For the purposes of this document the terms and definitions given in ISO 21527-1 and the following apply. 
3.1 
osmophilic yeast 
xerophilic mould 
fungus which is capable of growth at a water activity less than or equal to 0,95 
4 Principle 
4.1 Surface-inoculated plates are prepared using a specified selective culture medium. Depending on the 
expected number of colonies, a specified quantity of the sample (if the product is liquid), or of an initial 
suspension (in the case of other products), or decimal dilutions of the sample/suspension are used. 
Additional plates can be prepared under the same conditions, using decimal dilutions of the test sample or of 
the initial suspension. 
4.2 The plates are then aerobically incubated at 25 °C ± 1 °C for 5 d to 7 d. If necessary, the agar plates 
are left to stand in diffuse daylight for 1 d to 2 d. 
4.3 Colonies/propagules are then counted and, if required (to distinguish yeast colonies from bacterial 
colonies), the identity of any doubtful colonies is confirmed by examination with a binocular magnifier or 
microscope. 
4.4 The number of yeasts and moulds per gram or per millilitre of sample is calculated from the number of 
colonies/propagules/germs obtained on plates chosen at dilution levels producing countable colonies. Moulds and yeasts are counted separately, if necessary. 
5 Diluent and culture medium 
For current laboratory practice, see ISO/TS 11133 (all parts). 
5.1 Diluent 
5.1.1 General 
See ISO 6887 (all parts), ISO 8261 and the specific International Standard dealing with the product 
concerned. 
The use of a diluent containing a sufficient amount of solute [e.g. a 20 % to 35 % (mass concentration) 
solution of glycerol or D-glucose] is recommended to minimize osmotic shock to xerophilic mould and 
osmophilic yeast cells when serial dilutions are made prior to plating (References [1], [3]). 
NOTE It is possible to add surface-active agents such as sodium poly(oxyethylene)sorbitanmonooleate  1) [0,05 % 
(mass concentration)] to diluents to reduce clumping of mould spores and conidia (Reference [3]). 
Except for specific preparation of the test sample, the use of 0,1 % (mass concentration) peptone water broth 
as diluent is recommended . 
                                                      
1) Tween 80 is an example of a suitable product available commercially. This information is given for the convenience of 
users of this International Standard and does not co nstitute an endorsement by ISO of this product. ISO 21527-2:2008(E) 
© ISO 2008 – All rights reserved  35.1.2 Composition of 0,1 % (mass concentration) peptone water broth 
Enzymatic digest of animal or vegetal tissues 1,0 g 
Water 1 000 ml 
5.1.3 Preparation of 0,1 % (mass concentration) peptone water broth 
Dissolve the components in the water, by heating if necessary. 
If necessary, adjust the pH so that, after sterilization, it is 7,0 ± 0,2 at 25 °C. 
5.2 Culture medium 
5.2.1 Dichloran 18 % (mass concentration) glycerol agar (DG18) (References [4], [5], [6])  
5.2.1.1 Composition 
Casein enzymatic digest 5,0 g 
D-Glucose (C6H12O6) 10,0 g 
Potassium dihydrogenphosphate (KH2PO4) 1,0 g 
Magnesium sulfate (MgSO4 · H2O) 0,5 g 
Dichloran (2,6-dichloro-4-nitroaniline) 0,002 g 
Glycerol anhydrous 220 g 
Agar 12 g to 15 g a 
Chloramphenicol 0,1 g 
Water, distilled or deionized 1000 ml 
a Depending on the gel strength of the agar. 
5.2.1.2 Preparation 
5.2.1.2.1 General 
Suspend all the ingredients except chloramphenicol in the water and bring to the boil to dissolve completely. If 
necessary, adjust the pH (6.4) so that after sterilization it is 5,6 ± 0,2 at 25 °C. 
Add 10 ml of a 1 % (mass concentration) solution of chloramphenicol in ethanol and mix. Dispense the 
medium in quantities into suitable containers (6.5) of suitable capacity. Sterilize by autoclaving at 121 °C for 15 min. 
Immediately cool the medium in a water bath (6.3) maintained at a temperature of 44 °C to 47 °C. Cool to 
below 50 °C and dispense 15 ml amounts into sterile Petri dishes (6.6). 
Allow the medium to solidify, and dry, if necessary, the surface of the plates as described in ISO 7218 and 
ISO/TS 11133 (all parts). 
Use immediately, or store in the dark, according to ISO/TS 11133 (all parts) until required. 
CAUTION — Avoid exposure of the medium to light, since cytotoxic breakdown products can result in 
underestimation of mycoflora in samples. ISO 21527-2:2008(E) 
4 © ISO 2008 – All rights reserved
 5.2.1.2.2 Optional addition of chlortetracycline hydrochloride 
Where bacterial overgrowth may be a problem, chloramphenicol (50 mg/l) and chlortetracycline (50 mg/l) are 
recommended. In this case, prepare the basic medium as described above, with only chloramphenicol 50 mg, dispense it in quantities of 100 ml and sterilize. Since it is relatively unstable, freshly prepare also a 0,1 % (mass concentration) solution of chlortetracycline hydrochloride in water and sterilize by filtration. Just prior to use, add 5 ml of this solution aseptically to 100 ml of the basic medium, and pour plates. Gentamicin is not 
recommended, as it has been reported to cause inhibition of some yeast species (Reference [3]). 
5.2.1.2.3 Optional addition of trace elements 
In order for moulds to exhibit their full morphology, particularly any pigments they normally produce, they need 
trace elements that may not be present in DG18. To identify moulds on this medium, add the following trace element solution at 1 ml per litre of the medium, prior to autoclaving: ZnSO
4 · 7H2O 1g; CuSO4 · 5H2O 0,5 g; 
water, distilled or deionized 100 ml (Reference [2]). 
5.2.1.3 Performance testing for the quality assurance of the culture medium 
5.2.1.3.1 General 
DG18 medium is a solid medium. Productivity and selectivity shall be tested according to ISO/TS 11133 (all 
parts) according to the following specifications: 
5.2.1.3.2 Productivity 
Incubation: 5 d at 25 °C ± 1 °C 
Strains: Saccharomyces cerevisiae  ATCC 9763 
Wallemia sebi  ATCC 42694 
Aspergillus restrictus ATCC 42693 
Eurotium rubrum  ATCC 42690 
or strains recorded as equivalent in other fungal collections 
Reference media: media batch SDA (Sabouraud D-glucose agar) already validated 
Control method: quantitative Criteria: productivity ratio, P
R W 0,5 
Characteristic reaction: characteristic colony/propagules/germs according to each species 
5.2.1.3.3 Selectivity 
Incubation: 5 d at 25 °C ± 1 °C 
Strains: Escherichia coli  ATCC 25922 
Bacillus subtilis  ATCC 6633 
or strains recorded as equivalent in other bacterial collections 
Control method: qualitative 
Criteria: total inhibition ISO 21527-2:2008(E) 
© ISO 2008 – All rights reserved  56 Apparatus and glassware 
Disposable apparatus is an acceptable alternative to reusable glassware, provided that it has suitable 
specifications. 
Usual microbiological laboratory equipment (see ISO 7218) and, in particular, the following. 
6.1 Incubator , capable of operating at 25 °C ± 1 °C. 
6.2 Total delivery pipettes , sterile, of nominal capacity 1 ml, and graduated in divisions of 0,1 ml. 
6.3 Water bath , or similar apparatus, capable of operating at 44 °C to 47 °C. 
6.4 pH meter , accurate to 0,1 pH units at 25 °C. 
6.5 Bottles, flasks and tubes , for boiling and storage of culture media, and for making of dilutions. 
6.6 Petri dishes , sterile, in glass or plastic, with a diameter 90 mm to 100 mm. 
6.7 Microscope , for distinguishing yeast from bacterial cells (bright field, of magnification 250 to 
1 000 times). 
6.8 Spreaders , made of glass or plastic (of diameter less than 2 mm and length 80 mm). Diameter should 
not exceed 2 mm in order to minimize the amount of sample adhering to the spreader at the end of the 
procedure. 
6.9 Binocular magnifier , for discriminating and differentiating colonies/cells of yeasts and moulds 
(magnification 6,5 to 50 times). 
7 Sampling 
A representative sample should have been sent to the laboratory. It should not have been damaged or 
changed during transport or storage. The laboratory sample shall not be frozen. 
Sampling is not part of the method specified in this part of ISO 21527. Sampling should be carried out in 
accordance with the specific International Standard appropriate to the product concerned. If there is no specific International Standard, it is recommended that the parties concerned come to an agreement on this 
subject. 
8 Preparation of the test sample 
Prepare the test sample in accordance with ISO 6887 (all parts), ISO 7218, ISO 8261 and the specific 
International Standard dealing with the product concerned. If there is no specific International Standard, it is 
recommended that the parties concerned come to an agreement on this subject. 
9 Procedure 
9.1 Test portion, initial suspension and dilutions 
Prepare the test portion, initial suspension (primary dilution) and further dilutions in accordance with ISO 6887 (all parts), ISO 7218, ISO 8261 and the specific International Standard appropriate to the product concerned. 
Except for specific preparation of the test sample, it is recommended to use 0,1 % (mass concentration) 
peptone water broth (5.1.3) as diluent. Use a peristaltic homogeniser in preference to a blender or shaker. 
Due to the rapid sedimentation of spores in the pipette, maintain the pipette (6.2) in a horizontal (not vertical) 
position when filled with the appropriate volume of initial suspension and dilutions. 
Shake the initial suspension and dilutions in order to avoid sedimentation of microorganism-containing 
particles. ISO 21527-2:2008(E) 
6 © ISO 2008 – All rights reserved
 9.2 Inoculation and incubation 
9.2.1 On to one DG18 agar plate (5.2.1), using a sterile pipette (6.2), transfer 0,1 ml of the test sample if  
liquid, or 0,1 ml of the initial suspension in the case of other products (Clause 8). 
On to a second DG18 agar plate, using a fresh sterile pipette, transfer 0,1 ml of the first decimal (10−1) dilution 
(liquid product), or 0,1 ml of the 10−2 dilution (other products). 
To facilitate enumeration of low populations of yeasts and moulds, volumes of up to 0,3 ml of a 10−1 dilution of 
sample, or of the test sample if liquid, can be spread on three plates. 
Repeat these operations with subsequent dilutions, using a new sterile pipette for each decimal dilution. 
For particulate or solid foods, e.g. nuts or grains, direct plating is recommended. Samples of these types of 
products are surface disinfected in a 0,35 % (1 000 µg/g) sodium hypochlorite solution with a contact time of 2 min, followed by rinsing with sterile distilled water, dried on a sterile paper and then placed on solidified media (References [1, 2]). 
9.2.2  Spread the liquid over the surface of the agar plate with a sterile spreader (6.8) until the liquid is 
completely absorbed into the medium. 
Inoculation of plates by the pour-plate method may also be used, but in this case the equivalence of the 
results shall be validated compared to inoculation on the surface, and the discrimination and differentiation of moulds and yeast are not admissible. The method of spreading out on the surface can give higher enumerations. The spread-plate technique facilitates maximum exposure of cells to atmospheric oxygen and 
avoids any risk of thermal inactivation of fungal propagules. The results can depend on the type of fungi. 
9.2.3 Incubate the prepared plates (9.2.2) aerobically, lids uppermost, in an upright position in the incubator 
(6.1) at 25 °C ± 1 °C for 5 d to 7 d. If necessary, leave the agar plates to stand in diffuse daylight for 1 d to 2 d. 
If the presence of Xeromyces bisporus is suspected, incubate the plates for 10 d. 
It is recommended that the plates be incubated in an open plastic bag in order not to contaminate the 
incubator in the event of dissemination of the moulds out of the dishes. 
9.3 Counting and selection of colonies for confirmation 
After the specified period of incubation, select the dishes (9.2.3) containing < 150 colonies/propagules/germs 
and count these colonies/propagules/germs. If fast-growing moulds are a problem, count 
colonies/propagules/germs after 2 d and again after 5 d to 7 d of incubation. 
NOTE 1 Enumeration methods for yeasts and especially moulds are imprecise because they consist of a mixture of 
mycelium and asexual and sexual spores. Numbers of col ony-forming units depend on the degree of fragmentation of 
mycelium and the proportion of spores able to grow on the plating medium. 
NOTE 2 Non-linearity of counts from dilution plating often occurs , i.e. 10-fold dilutions of samples often do not result in 
10-fold reductions in numbers of colonies recovered on plating media. This has been attributed to fragmentation of mycelia 
and breaking of spore clumps during dilution in addition to competitive inhibition when large numbers of colonies are 
present on plates. 
CAUTION — The spores of moulds disperse in the air with a great facility, handle the Petri dishes with 
care to avoid development of satellite colonies whic h would give an overestimation of population in 
the sample. 
If necessary, carry out an examination with a binocular magnifier (6.9) or with a microscope (6.7) in order to 
distinguish between cells of yeasts or moulds and bacteria from colonies. 
Count the colonies of yeasts and the colonies/propagules of moulds separately, if necessary. For identification of yeast and moulds, select areas of fungal growth and remove for high microscopic 
examination or inoculate on suitable isolation or identification media. ISO 21527-2:2008(E) 
© ISO 2008 – All rights reserved  710 Expression of results and confidence limits 
See ISO 7218. 
11 Test report 
The test report shall include at least the following information: 
a) all information necessary for the complete identification of the sample; 
b) the sampling method used, if known; 
c) the test method used, with reference to this part of ISO 21527; d) all operating details not specified in this part of ISO 21527, or regarded as optional, together with details 
of any incidents which may have influenced the test results; 
e) the test results obtained. ISO 21527-2:2008(E) 
8 © ISO 2008 – All rights reserved
 Annex A  
(informative) 
 
Examples of water activity according to matrices 
Table A.1 gives examples of water activity according to the products to be analyzed. 
Table A.1 — Water activity aw according to matrices 
Water 
activity 
aw Food examples/matrices  
W 0,95 Highly perishable foods (fresh and canned fruits, vegetables, meat, fish), milk, cooked sausages, breads, 
foods containing up to 4 % of sucrose or 7 % of NaCl (salt)  
W 0,91 Hard cheeses such as Cheddar, cured meat, fruit juice concentrates containing 55 % sucrose or 12 % NaCl  
W 0,87 Fermented sausage, sponge cakes, dry cheese, margarine, foods containing 65 % sucrose or 15 % NaCl 
(salt) 
W 0,80 Most fruit juice concentrates, condensed milk, syrup, flour, high-sugar cakes, pulses containing 15 % to 17 % 
moisture  
W 0,75 Jam, marmalade, glace fruits, marzipan, marshmallows, cake  
W 0,65 Rolled oats containing 10 % moisture, jelly, molasses, nuts  
W 0,60 Dried fruits containing 15 % to 20 % moisture, caramel, toffee, honey, bars cereals, dog feed, granulated 
food, grains, cereals and cereal products  
W 0,50 Noodles containing 12 % moisture, spices containing 10 % moisture  
W 0,40 Whole egg powder containing 5 % moisture, nougat  
W 0,30 Cookies, crackers, bread crusts containing 3 % to 5 % moisture, base of dehydrated sauce  
W 0,03 Whole milk powder containing 2 % to 3 % moisture, dehydrated soups, instant coffee  
 ISO 21527-2:2008(E) 
© ISO 2008 – All rights reserved  9Bibliography 
[1] A NDREWS , S.,  PITT, J.I. Selective medium for isolation of Fusarium  species and dematiaceous 
hyphomycetes from cereals. Appl. Environ. Microbiol . 1986, 51, pp. 1235-1238 
[2] B ELL, C.,  NEAVES , P.,  WILLIAMS , A.P. Food microbiology and laboratory practice . Blackwell, Oxford, 
2005. 324 p. 
[3] B EUCHAT , L.R. Media for detecting and enumerating yeasts and moulds. In: C ORRY , J.E.L.,  
CURTIS , G.D.W.,  BAIRD, R.M., editors. Handbook of culture media for food microbiology , pp. 369-386. 
Elsevier, Amsterdam, 2003. ( Progress in industrial microbiology , Vol 37) 
[4] B EUCHAT , L.R.,  FRÄNDBERG , E.,  DEAK, T.,  ALZAMORA , S.M.,  CHEN, J., GUERRERO , A.S.,  LÓPEZ -MALO, A.,  
OHLSSON , I., OLSEN , M.,  PEINADO , J.M.,  SCHNURER , J., DE SILONIZ , M.I.,  TORNAI -LEHOCZKI , J. (2001) 
Performance of mycological media in enumerating desiccated food spoilage yeasts: An interlaboratory 
study. Int. J. Food Microbiol.  2001, 70, pp. 89-96 
[5] D EAK, T.,  CHEN, J., GOLDEN , D.A.,  TAPIA, M.S.,  TORNAI -LEHOCZKI , J., VILJOEN , B.C.,  WYDER , M.T.,  
BEUCHAT , L.R. Comparison of dichloran 18 % glycerol (DG18) agar with general purpose mycological 
media for enumerating food spoilage yeasts. Int. J. Food Microbiol . 2001, 67, pp. 49-53 
[6] H OCKING , A.D.,  PITT, J.I. Dichloran-glycerol medium for enumeration of xerophilic fungi from low 
moisture foods. Appl. Environ. Microbiol . 1980, 39, pp. 488-492 
 ISO 21527-2:2008(E) 
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© ISO 2008 – All rights reserved