Difco™ Hycheck

13 Pages·2002·0.15 MB·English

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L005259.qxd - HYCHECK.qxd 12/09/2002 10:14 AM Page 1
Difco™ Hycheck™
The hygiene contact slide with the
unique hinge for monitoring microbial
contamination on surfaces or in liquids. L005259
2002/08
INTENDED USE In D/E Neutralizing Agar, sodium thioglycollate
Hycheck™is a hygiene contact slide used to assess neutralizes mercurial compounds. Sodium
the microbiological contamination of surfaces or thiosulfate neutralizes iodine and chlorine.
fluids. Sodium bisulfite neutralizes formaldehyde and
glutaraldehyde. Lecithin neutralizes quaternary
SUMMARYANDEXPLANATION ammonium compounds. Polysorbate 80
neutralizes phenols, hexachlorophene, formalin
Monitoring the microbial flora of environmental
and, with lecithin, ethanol. Bromcresol purple is a
surfaces, walls, ceilings and equipment is an impor-
colorimetric indicator.
tant stage in achieving good manufacturing prac-
tices in factories handling foods, cosmetics or In Violet Red Bile Glucose Agar, bile salts and
pharmaceuticals.1-3To maintain good hygiene crystal violet inhibit gram-positive bacteria.
standards in hotels and restaurant kitchens, micro- Glucose fermenters produce red colonies with
biological contamination must also be monitored.4 red-purple halos in the presence of neutral red, a
Methods to monitor the environmental flora have pH indicator.
been described using either swabbing techniques5 In Rose Bengal Chloramphenicol Agar,
or contact plates.6Contact slides were created to monopotassium phosphate provides buffering
monitor the microbial flora of liquids (e.g. urine, capacity. Magnesium sulfate provides necessary
milk) and equipment surfaces in the clinical and trace elements. Rose bengal is a selective agent
food industries.1Contact slides are statistically that inhibits bacterial growth and restricts the size
comparable to swab and contact plates for surface and height of bacterial colonies of the more
sampling.1The Hycheckcontact slides were devel- rapidly growing molds. The restriction in growth
oped for the testing of fluids and surfaces for of molds aids in the isolation of slow-growing
microbial cleanliness. fungi by preventing overgrowth by more rapidly
Hycheckis a double-sided, hinged plastic paddle growing species. Rose bengal is taken up by yeast
containing two agar surfaces. The agar surface and mold colonies, thereby facilitating their
extends above the paddle allowing for contact with recognition and enumeration. Chloramphenicol
test surfaces. The hinged paddle allows the agar inhibits bacteria.
surface to be easily held against each test area In Plate Count Agar with TTC, triphenyl tetra-
during sampling. The surface area of the paddle is zolium chloride (TTC) is a redox indicator.
clearly divided into seven units of one centimeter
each to allow direct counting of microbial density REAGENTS
per unit area.
Formulae:
The Hycheckrange of hygiene contact slides
D/E Neutralizing Agar
consists of seven media combinations designed to
meet the various needs for monitoring different Approximate Formula* Per Liter Purified Water
types of microbial contamination. Pancreatic Digest of Casein . . . . . .5.0 g
Yeast Extract . . . . . . . . . . . . . . . . . .2.5 g
PRINCIPLES OF THE PROCEDURE Dextrose . . . . . . . . . . . . . . . . . . . . .10.0 g
Pancreatic digest of casein and other peptones pro- Sodium Thioglycollate . . . . . . . . . .1.0 g
vide carbon and nitrogen. Yeast extract provides vita- Sodium Thiosulfate . . . . . . . . . . . . .1.0 g
mins, cofactors, nitrogen and carbon. Papaic digest of Sodium Bisulfite . . . . . . . . . . . . . . .2.5 g
soybean meal provides nitrogen, vitamins and miner- Lecithin . . . . . . . . . . . . . . . . . . . . . .7.0 g
als. Dextrose (glucose) provides fermentable carbohy- Polysorbate 80 . . . . . . . . . . . . . . . . .5.0 g
drate. Sodium chloride maintains osmotic Bromcresol Purple . . . . . . . . . . . . . .0.02 g
equilibrium. Agar is the solidifying agent. Agar . . . . . . . . . . . . . . . . . . . . . . . .15.0 g
L005259.qxd - HYCHECK.qxd 12/09/2002 10:14 AM Page 2
Tryptic Soy Agar Rose Bengal Chloramphenicol Agar (RBCA)
Approximate Formula* Per Liter Purified Water Approximate Formula* Per Liter Purified Water
Pancreatic Digest of Casein . . . . .15.0 g Papaic Digest of Soybean Meal . . .5.0 g
Papaic Digest of Soybean Meal . . .5.0 g Dextrose . . . . . . . . . . . . . . . . . . . . .10.0 g
Sodium Chloride . . . . . . . . . . . . . . .5.0 g Monopotassium Phosphate . . . . . .1.0 g
Agar . . . . . . . . . . . . . . . . . . . . . . . .15.0 g Magnesium Sulfate . . . . . . . . . . . . .0.5 g
Rose Bengal . . . . . . . . . . . . . . . . . . .0.05 g
Violet Red Bile Glucose Agar Agar . . . . . . . . . . . . . . . . . . . . . . . .15.0 g
Approximate Formula* Per Liter Purified Water Chloramphenicol . . . . . . . . . . . . . . .0.05 g
Yeast Extract . . . . . . . . . . . . . . . . . .3.0 g *Adjusted and/or supplemented as required to
Peptone . . . . . . . . . . . . . . . . . . . . . .7.0 g meet performance criteria.
Dextrose . . . . . . . . . . . . . . . . . . . . .10.0 g
Sodium Chloride . . . . . . . . . . . . . . .5.0 g Precautions:For Laboratory Use
Bile Salts No. 3 . . . . . . . . . . . . . . . . .1.5 g
STORAGE
Neutral Red . . . . . . . . . . . . . . . . . . .0.03 g
Crystal Violet . . . . . . . . . . . . . . . . . .0.002 g Store Hycheckat 2 – 8ºC with the arrow on the
Agar . . . . . . . . . . . . . . . . . . . . . . . .15.0 g carton label pointing up in a vertical position.
Plate Count Agar (PCA) Expiration Date: The expiration date applies to
Approximate Formula* Per Liter Purified Water the product in its intact container when stored as
Pancreatic Digest of Casein . . . . . .5.0 g directed. Do not use a product if it fails to meet
Yeast Extract . . . . . . . . . . . . . . . . . . 2.5 g specifications for identity and performance.
Dextrose . . . . . . . . . . . . . . . . . . . . . .1.0 g
Agar . . . . . . . . . . . . . . . . . . . . . . . .15.0 g PROCEDURE
Plate Count Agar (PCA) with TTC contains 0.1 g/L Materials Provided:One type is provided per
in addition to the above ingredients. package of the following:HycheckD/E
Neutralizing Agar, HycheckDisinfection Control,
HycheckEnterobacteriaceae, HycheckPlate Count
Agar with TTC, Hycheck Total Count, Hycheck
Yeasts and Molds and HycheckYeasts and Molds
with TTC.
HYCHECK:THECOMPLETERANGE
The Hycheckrange of hygiene contact slides consists of seven products designed to meet the various
needs for monitoring different types of microbial contamination. All the slides contain two agar surfaces.
Most products have one agar to give a non-selective total count. The other agar surface is designed to
identify or select a specified group of microorganisms. The exceptions to this are Plate Count Agar with
TTC and D/E Neutralizing products which have the same medium on both sides, and the Total Count
product which has two non-selective media.
METHOD OF USE: GENERAL
1 2
REMOVESLIDE SAMPLING
Loosen cap on the tube and remove a. Surfaces
the HycheckSlide from the container
Hold the terminal spike against the surface to
taking care not to touch the agar
be tested. Press down on the spike to bend
surface. Examine for any
the paddle around the hinge line. Gently
dehydration or contamination.
lower the slide and press the agar into
contact with the test surface, still holding the
slide by the cap.
2
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3
Apply a firm and even pressure on the test LABEL–Enter the site, location, date and time.
surface for a few seconds taking care not to Also indicate whether the sample has been
smear the agar over the test area. taken before or after cleaning.
4
Repeat the procedure using the second agar INCUBATION–Incubate in an upright
surface on an area adjacent to the initial test position at the temperature indicated in the
site. Replace slide in the container and instructions for the particular product.
close tightly. However, alternative times and temperatures
b. Liquids for incubation may be used according to indi-
vidual requirements.
Immerse the HycheckSlide into the test fluid so 5
that the agar surface becomes totally covered. READINGRESULTS–Remove slide from
If insufficient liquid is available, pour over the the tube and count the number of colonies
surface of the slide. Allow to drain. Replace the on each agar surface. Reflected light facili-
slide in the container and close tightly. tates reading.
METHOD OF USE FOR SPECIFIC HYCHECK PRODUCTS
Enterobacteriaceae READINGRESULTS
Side 1 -Coated with Violet Red Bile Glucose Agar, Tryptic Soy Agar:Count the total number of
a clear red medium used for the enumeration of colonies on the slide after 48 h incubation to
Enterobacteriaceae in foods. The presence of obtain the total bacterial count of the test
Enterobacteriaceae,coliforms, Salmonellae, surface.
Klebsiellaor Citrobacterspp. in raw foodstuffs is an
Violet Red Bile Glucose Agar:Enterobacteriaceae
indicator of faecal contamination. Their presence
will ferment the glucose in the medium to
after processing may indicate a failure in the manu-
produce acid. The resulting drop in pH of the
facturing procedure.
medium causes bile salts to precipitate around the
Side 2 - Coated with Tryptic Soy Agar, this clear red colonies as a red opaque halo. Non-glucose
medium will give a total aerobic bacterial count of fermenters remain colorless on the medium and
the test surface. show no bile halo. Most gram-positive organisms are
INCUBATION inhibited on the medium. Count the red colonies
Incubate in an upright position at 35 – 37ºC for surrounded by a precipitate of bile to obtain the
24 h. Examine the slide again after a further 24 h Enterobacteriaceae count.
incubation. Other times and temperature of incu-
bation may be used dependent upon the require-
ments of the environment tested.
TYPICALCULTURALRESPONSE
Organism Tryptic Soy Agar V.R.B.G.A. (24 hours)
(24 hours)
*Enterobacter aerogenes Good growth Good growth
ATCC™ 13048
*Escherichia coli Good growth of cream Good growth of red colonies
ATCC 25922 moist colonies surrounded by an opaque red halo
Proteus mirabilis Good growth Good growth
NCTC™ 11938
*Shigella sonnei Good growth Good growth
ATCC 25931
*Salmonella typhimurium Good growth of cream Good growth of red colonies
ATCC 14028 moist colonies surrounded by an opaque red halo
*Staphylococcus aureus Good growth of cream/ Growth inhibited
ATCC 25923 yellow colonies
3
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Organism Tryptic Soy Agar V.R.B.G.A. (24 hours)
(24 hours)
*Enterococcus faecalis Good growth of small Growth inhibited
ATCC 19433 white colonies
*Cultures listed above may be available as Microtrol™ (UK) or Bactrol™ Plus (USA) Disks.
Hycheck - Yeasts and Molds
incubation and then on the Rose Bengal
Side 1 -Coated with Rose Bengal Chloramphenicol
Chloramphenicol Agar after 120 h incubation.
Agar, a pink agar recommended for the selective
Other times and temperature of incubation may
isolation of yeasts and molds from environmental
be used dependent upon the requirements of
materials and foodstuffs. The pH of the medium
the environment tested.
is near neutral for improved growth of
acid-sensitive strains. READINGRESULTS
Side 2 -Coated with Tryptic Soy Agar, a clear Tryptic Soy Agar:Record the total number of
colorless medium recommended in the European colonies on the Tryptic Soy Agar to obtain the
PharmacopoeiaandThe United States Pharmaco- total aerobic bacterial count.
poeiafor microbial limits testing. The medium will Rose Bengal Chloramphenicol Agar:Count the
give a total aerobic count. total number of colonies on the agar to obtain
an assessment of the fungal contamination
INCUBATION
(colonies of yeasts appear pink due to uptake of
Incubate in an upright position at 30ºC. Record Rose Bengal). Bacterial growth is inhibited on
the count on the Tryptic Soy Agar after 48 h this medium.
TYPICAL CULTURALRESPONSE
Organism Tryptic Soy Agar Rose Bengal Chloram-
(48 hours) phenicol Agar (5 days)
Candida albicans Good growth of white Good growth of pink smooth
ATCC™2091 raised colonies colonies
Aspergillus niger Good growth appearing Good growth appearing white
NCPF™ 2275 white and filamentous and filamentous and becoming
and becoming black black after 5 days incubation
after 5 days incubation
Saccharomyces cerevisiae Good growth Good growth
NCYC™ 1211
*Serratia marcescens Good growth Inhibited
ATCC 8100
*Esherichia coli Good growth of cream Growth restricted
ATCC 25922 moist colonies
*Staphylococcus aureus Good growth of cream/ Growth restricted
ATCC 25923 yellow colonies
*Streptococcus pyogenes Good growth Inhibited
ATCC 19615
*Cultures listed above may be available as Microtrol™ (UK) or BactrolPlus (USA) Disks.
Hycheck - Yeasts and Molds with TTC produced. This aids counting of normally colorless
or translucent colonies. The product should be
This product is the same as the HycheckYeast
used as described for the standard Yeasts and
and Molds product except that 0.01% triphenyl
Molds Hycheckproduct. Some organisms, e.g.
tetrazolium chloride (TTC) is incorporated into
staphylococci, may grow less profusely on
the Tryptic Soy Agar. In an oxidized state TTC is
the medium containing TTC and some yeasts, e.g.
colorless but on reduction by microorganism
Saccharomyces cerevisiae, are inhibited
growth an insoluble red formazan dye is
completely.
4
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TYPICALCULTURALRESPONSE
Organism Tryptic Soy Agar RoseBengal
PlusTTC(48 hours) Chloramphenicol
Agar(5 days)
Aspergillus niger Good growth Good growth
NCPF™ 2275
*Candida albicans Good growth, red dye reduction Good growth of pink
ATCC™ 2091 smooth colonies
Saccharomyces Growth inhibited Good growth of pink
cerevisiae colonies
NCYC™ 1211
*Escherichia coli Good growth, red dye reduction Growth restricted
ATCC 25922
*Serratia marcescens Good growth Inhibited
ATCC 8100
*Staphylococcus aureus Trace growth, red dye Growth restricted
ATCC 25923 reduction
*Streptococcus pyogenes Good growth Inhibited
ATCC 19615
*Cultures listed above may be available as Microtrol™ (UK) orBactrol Plus (USA) Disks.
Hycheck - Disinfection Control give a total aerobic count. The surface under test
Side 1 -Coated with D/E Neutralizing Agar, a should be sampled with Tryptic Soy Agar before
purple agar gel used for determining the cleaning.
efficiency of sanitization of containers, equipment
and environment surfaces after cleaning. INCUBATION
D/E Neutralizing Agar will neutralize a broad Incubate in an upright position at 35 – 37ºC,
spectrum of antiseptic and disinfectant chemicals examining after 24 and 48 h incubation. Other
including quarternary ammonium compounds, times and temperature of incubation may be used
phenolics, iodine and chlorine preparations, dependent upon the requirements of the environ-
mercurials, formaldehyde and gluteraldehyde. ment tested.
Neutralization allows growth of bacteria which
may be held in bacteriostasis after disinfection READINGRESULTS
but are, nevertheless, still viable. Tryptic Soy Agar: Count the number of colonies
Side 2 -Coated with Tryptic Soy Agar, a clear on the Tryptic Soy Agar to obtain an assessment
colorless medium recommended in the European of the microbial load before cleaning.
Pharmacopoeia andThe United States Pharmaco- D/E Neutralizing Agar:Count the number of
poeiafor microbial limits testing. This medium will colonies on D/E Neutralizing Agar to detect the
number of organisms surviving cleaning and
sanitization.
TYPICAL CULTURAL RESPONSE
Organism Tryptic Soy Agar D/E Neutralizing Agar
(24 hours) (24 hours)
Aspergillus niger Good growth Good growth
NCPF™ 2275
Bacillus subtilis Good growth of Good growth with medium
ATCC™6633 large rough colonies turning yellow around the
colonies
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Organism Tryptic Soy Agar D/E Neutralizing Agar
(24 hours) (24 hours)
Candida albicans Good growth Good growth
ATCC 2091
*Escherichia coli Good growth of Good growth with medium
ATCC 25922 cream moist colonies turning yellow around the
colonies
*Pseudomonas aeruginosa Good growth of Good growth with medium
ATCC 27853 colorless colonies turning yellow around the
colonies
*Staphylococcus aureus Good growth of cream/ Good growth of cream/yellow
ATCC 25923 yellow colonies colonies
*Staphylococcus epidermidis Good growth Good growth
ATCC 12228
*Cultures listed above may be available as Microtrol™ (UK) or BactrolPlus (USA) Disks.
Hycheck - D/E Neutralizing Agar INCUBATION
Sides 1 & 2 -Coated with D/E Neutralizing Agar, Incubate in an upright position at 35 – 37ºC,
a purple agar gel for determining the efficiency examining after 24 and 48 h incubation. Other
of sanitization of containers, equipment and times and temperature of incubation may be
environment surfaces after cleaning. D/E used dependent upon the requirements of the
Neutralizing Agar will neutralize a broad environment tested.
spectrum of antiseptic and disinfectant chemicals
including quarternary ammonium compounds, READINGRESULTS
phenolics, iodine and chlorine preparations, Count the number of colonies on both sides of
mercurials, formaldehyde and gluteraldehyde. the paddle. They should be approximately equal.
Neutralization allows growth of bacteria which
may be held in bacteriostasis after disinfection but
are nevertheless still viable.
TYPICAL CULTURAL RESPONSE
Organism D/E Neutralizing Agar
(24 hours, 35 – 37ºC)Sides 1 & 2
Aspergillus niger NCPF™ 2275 Good growth
Bacillus subtilisATCC™6633 Good growth of large rough colonies
Candida albicans ATCC 2091 Good growth
*Escherichia coli ATCC 25922 Good growth of cream moist colonies
*Pseudomonas aeruginosaATCC 27853 Good growth, colorless colonies
*Staphylococcus aureusATCC 25923 Good growth, cream/yellow colonies
*Staphylococcus epidermidisATCC 12228 Good growth
*Cultures listed above may be available as Microtrol™ (UK) or BactrolPlus (USA) Disks.
Hycheck- Total Count be used dependent upon the requirements of the
Both paddle sides have Plate Count Agar surfaces environment tested.
but one agar also incorporates 0.01% triphenyl
READINGRESULTS
tetrazolium chloride (TTC). This aids the
recognition of microorganism colonies due to the Count the number of colonies on both sides of the
formation of the insoluble red formazan dye paddle. They should be approximately equal. A
when the TTC is reduced. few microorganisms may grow less profusely in the
presence of TTC. If the colony count is not equiva-
INCUBATION
lent on the two surfaces then take the larger
Incubate in an upright position at count as representing the microbial load from the
35 – 37ºC, examining after 24 and 48 h incubation. sample under test.
Other times and temperature of incubation may
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TYPICAL CULTURAL RESPONSE
Organism Plate Count Agar Plate Count Agar
(24 hours) with TTC (24 hours)
*Enterococcus faecalis Good growth Good growth
ATCC™ 19433
*Escherichia coli Good growth of cream Good growth, red dye reduction
ATCC 25922 moist colonies
*Proteus vulgaris Good growth Good growth
ATCC 13315
*Salmonella typhimurium Good growth Good growth
ATCC 14028
*Staphylococcus aureus Good growth of cream/ Moderate growth, red dye
ATCC 25923 yellow colonies reduction
*Cultures listed above may be available as Microtrol™ (UK) or BactrolPlus (USA) Disks.
LIMITATIONS
Do not use the HycheckSlide if it is contaminated or the agar medium is significantly dehydrated. The
“Typical Cultural Response” tables indicate the expected reactions with various control strains of
organisms.
AVAILABILITY
We are happy to customize HycheckSlides to individual requirements for a minimum number of 600 slides.
For more information please contact Technical Services in the United States, toll free [phone].
Cat. No. Description Pack Size
290001 Hycheck D/E Neutralizing 10 units
(D/E Neutralizing Agar both sides)
290002 Hycheck Disinfection Control 10 units
(Tryptic Soy Agar and D/E Neutralizing Agar)
290003 Hycheck Enterobacteriaceae 10 units
(Tryptic Soy Agar and Violet Red Bile Glucose Agar)
290004 Hycheck Plate Count Agar with TTC 10 units
(Plate Count Agar with TTC both sides)
290005 Hycheck Total Count 10 units
(Plate Count Agar and Plate Count Agar with TTC)
Primarily for control of Cutting Oils and Process Water.
290006 Hycheck Yeasts and Molds 10 units
(Tryptic Soy Agar and Rose Bengal Chloramphenicol Agar)
290007 Hycheck Yeasts and Molds with TTC 10 units
(Tryptic Soy Agar with 0.01% TTC and Rose Bengal Chloramphenicol Agar)
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Hygienic assessment of
SURFACES

This chart is intended as a guide only; users should always validate their own application.
Microorganism counts with Hycheckare a semi-quantitative guide to the degree of contamination in
the environment being tested.
Monitoring with Hycheckover a period of time gives a good indication of change in the microbial
load at a given site.
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Hygienic assessment of
LIQUIDS
Median Regression Slope (non-selective media)
L
m
er
p
nt
u
o
c
al
eri
act
B
Bacterial Colonies per HycheckContact Slide
This chart is intended as a guide only; users should always validate their own application.
Microorganism counts with Hycheckare a semi-quantitative guide to the degree of contamination in
the environment being tested.
Monitoring withHycheckover a period of time gives a good indication of change in the microbial load at a
given site.
9