Anaerobiosis

- the
rE.>~i*able svstem
for aerophilic/

MERCK


Complete System for the cultivation

of anaerobic und microaerophilic/
capneic microorganisms

Anaerobic microorganisms have'long been known as constituents of the normal bacterial

flora of human and animal organisms . Both their pathogenic significance in medicine and
their important role in food hygiene have, however, long been underestimated . Attention
has accordingly been focused an "aerobic microbiology" in both of these areas .
During the past few years the importance of anaerobic microorganisms as pathogenic
agents responsible for infectious diseases and the role they play in the microbial spoilage
of food and water have been better appreciated .
Extremely different spedra of anaerobic organisms are of importance for the examination
of food and in the clinical microbiology .
Organisms belonging to the genus Clostridium play a significant part in both areas . Patho-
genic representatives of the genera Baderoides, Fusobaderium, Peptococcus and Pepto-
streptococcus are very important in the clinical (both human and veterinary) field . In some
cases these microorganisms differ greatly as regards their morphologies, Gram-staining
properties and their natural habitats, but they all share one common feature - in addition
to certain demands they make an the culture medium - they require an anaerobic (i . e.
pradically oxygen-free) atmosphere for growth and reproduction during all phases of the
investigation . The increasing attention paid to anaerobes is partly due to the fad that the
possibilities for cultivating these microorganisms have been improved, particularly during
the past twenty years. It should, however, be pointed out that all the methods used so far
display certain weaknesses.

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Infections caused by microaerophilic/capneic pathogens play a special role both in human

and veterinary medicine . Infedions due to these microorganisms are of considerable med-
ical importance, clinical diagnosis should therefore always be confirmed by detection of Q)0)
the pathogenic agent . The culture of these microorganisms is, however, difficult and has in Ö C' N
the past met with serious technical problems .
The spedrum of microaerophilic/capneic microorganisms includes the genera Neisseria, ® C7 ~ c
Campylobader and Haemophilus . Diseases caused by these pathogenic agents are
O „~ ro t
1 . of extreme clinical importance (e . g . Neisseria meningitidis and Haemophilus influenzae :
meningitis), PO ~cN .oc
2. of epidemiological interest (e .g. Campylobader fetus : zoonosis) and O
3. of legal significance (e . g. Neisseria gonorrhoea ; the German Epidemics Law stipulates .
that this disease has to be reported) . 94
Campylobacter spp . are also becoming increasingly important in food hygiene and Legio- PW
I

nella spp. in environmental samples . (D

Microbiolgy is constantly confronted with microaerophilic/capneic microorganisms in the U) 0

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field of medicine, the examination of foodstuffs and in the food industry .
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 The Anaerocult® products (A, A mini, P, IS, C and C mini) are all systems for producing
anaerobic or microaerophilic/capneic conditions and are based an a common basic
principle .
The Anaerocult® reagent mixture consists of components that real with water to
chemically bind all or part of the atmospheric oxygen (0 2) from a given volume of air
(anaerobic jar or special incubation bog) and simultaneously release carbon dioxide
(CO 2). The resulting atmosphere is rich in carbon dioxide and either devoid of oxygen
(anaerobic) or oxygen deficient (microaerophilic/capneic) .

Anaerocult® functions without a catalyst

Simple and Safe to use

Sufficient CO 2 partial pressure
Quick and reliable establishment of the desired 0 2 and CO2 partial pressures
Flexibility to the number of samples to be tested
Economical performance

Anaerotest ®

Anaerotest® is a test strip for checking the anaerobic atmosphere in sealed containers
used for incubation . The blue readion zone of this test strip contains a dye (methylene
blue) that occurs in its oxidised form in an air-filled (i . e. aerobic) environment . An anaero-
bic environment causes a chemical redudion of the dye, which changes colour to white .
The readion is reversible so that when oxygen is introduced the readion zone becomes
blue once more .
The Anaerotest® test strip does not change colour when employed with Anaerocult ® C
and C mini due to the residual oxygen content of the atmosphere . Anaerotest ® is therefo-
re not suitable for the use with Anaerocult ® C and C mini .

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m Anaeroclip ®

Anaeroclip ® is a simple mechanical fastening for the special incubation bags of Anaero-
cult® A mini, P, IS and C mini .
The Anaeroclip® fastener means that the user can seal the bags without a special foil sea-
ling instrument .
Anaeroclip ® sealing bars can be used several times .
 Annerobic jar und plate basket

The anaerobic jar simply consists of two functional parts :

a lid and a base. lt is extremely easy to use and convenient .

There is na need to insert or refill catalysts . The use of

manometers and valves are unnecessary .
P k'

The jar is constructed of high quality materials which

ensure that sealing is leakproof and that the lid always fits
properly.

Volume :
W

with integrated holding clip for the Anaerotest ® strip

Diameter : approximately 90 mm
Height : approximately 200 mm
Capacity : 12 Standard Petri dishes


Anaerocult ® A - for the anaerobic

incubation of up to 12 Petri dishes
in an anaerobic jar

Only water has to be added to start the reaction .

rr --

The atmospheric oxygen is not reacted with hydrogen to

H2O but is chemically bound to the finely divided iron
powder in the reagent mixture .
• No catalyst is required .

e- Use
The special paper ensures rapid, unobstruded diffusion of
water and atmospheric oxygen into the reagent mixture as
well as release of CO 2 into the anaerobic vessel . This
results in a fall of the oxygen concentration and a high
CO2 content . Na further steps are required .

Additional reagents and auxiliaries are not needed.

Anaerocult® C - for the specific

incubation of microaerophilic/
capneic microorganisms

The special formulation of the reagent mixture used in

Anaerocult ® C guarantees that the gas concentrations re-
commended in the literature are produced in the anaero-
bic jar:
CO2 8 -10 vol
02 5 - 6 vol
C
At an incubation temperature of 42°C (the ideal tempera-
ture for Campylobader jejuni, C . coli) the temperature in
the anaerobic jar does not increase because, unlike systems
employing a catalyst, the amount of energy released is
low . Damage of the microorganisms is therefore excluded .

Oxygen-hydrogen mixtures can be dangerous (oxyhydro-

gen gas readion) . The critical 02/H2 ratio is not attained
with either Anaerocult ® C or any of the other Anaerocult ®
produds .
 Anaerocult ® A mini - for the
anaerobic incubation of four
Petri dishes

Anaerocult ® P - for the

anaerobic incubation of single
Petri dishes

Low Sample amount can be incubated immediately . The Anaerocult® sachet is moistened with water and
placed together with the Petri dishes into the special in-
cubation bog and then sealed.


Anaerocult ® C mini - for the

incubation of one or two Petri
dishes in a micronerophilic/
bb„ ._capneic atmosphere

Anaerocult ® IS - for the annerobic

incubation of identification
systems und sensitivity teste

The transparent special incubation bag enables a visual Flexible to the respective number of samples to be tested .
evaluation of the anaerobic or microaerophilic/capneic
growth in the closed anaerobic system at any given time .
The cultivated plates are never exposed to atmospheric
oxygen.
 Ordering Information

Cat. No. Product Package size

Products for 13 829 Anaerocult ® A 10 sachets

the cultivation 1 611 Anaerocult" A mini 25 sets'
13 807 Anaerocult" P 25 Sets'
of anaerobic and 16 819 Anaerocult" IS 25 Sets'
mit nerophilic/ 16 275 Anaerocult ® C 25 sachets
capneic 13 682 Anaerocult ® C mini 25 sets
15112 Anaerotest® 50 test strips
microorganisms 14226 Anaeroclip 25
7 040 Plate basket 1
16387 Anaerobic jar 1

Culure medin 5452 Anaerobic agar acc. to BREWER 500 g

16331 Anaerobic blood agar
acc. to CDC2 (base) 500 g
16 332 Anaerobic fermentation medium
acc. to CDC2 (base) 500 g
10895 Anaerobic TVLS culture medium (base) 500 g
10886 Blood agar (base) 500 g
13 825 Brain heart agar 500 g
10493 Brain heart broth 500 g
2 248 Campylobacter selective agar (base) 500 g
5410 Clostridium agar (RCM) 500 g
5 411 Clostridium culture medium (RCM) 500 g
11 699 Clostridium differential broth 500 g
10455 Columbia agar (base) 500 g
10 928 Cooked meat broth 100 tablets
8191 Fluid thioglycollate medium 500 g
16761 Fluid thioglycollate medium G 500 g, 5 kg
11 709 Haemophilus selective agar (base) 500 g
5 464 Liver broth 500 g
15 045 Meat liver agar 500 g
10 235 SPS agar 500 g
7881 Standard 1 nutrient agar 500 g, 5 kg
10 864 Sulfite iron agar (base) 500 g
10 728 THAYER-MARTIN agar (base) 500 g
8190 Thioglycollate broth 500 g, 5 kg
16330 Thioglycollate medium (enriched)
acc. to CDC 2 (base) 500 g
11 972 TSC agar (base) 500 g
5 264 TSN agar 500 g

Supplements 2 249 Campylobacter selective supplement 16 vials3

4032 Fluorocult® TSC supplement 1 vial°
10 729 THAYER-MARTIN supplement 1
(mixture of inhibitors) 16 vials
10730 THAYER-MARTIN Supplement II
(mixture of growth factors) 16 vials
1 1 set consists of 1 special incubation bog and 1 Anaerocult ® suchet (powder mixture)
2 Centers for Disease Control, Atlanta/USA
3 Sufficient for 3 200 ml culture medium
4 Sufficient for 500 ml TSC agar
 References
Altmeyer, M. ; P. Krabisch ;
P . Dorn
Christopher, F . M ; G . C . Smith ;
C . Vanderzant
Costin, J . D . ; W. Fischer ;
M. Kappner ; W. Schmidt ;
H . Schuchmann
Costin, J . D . ; M . Kappner;
W. Schmidt
Fukushima, H . ; R. Nakamura ;
S . litsuka ; Y. Ito ; K. Saito
Haba, J. H . ; M . Mateo ;
M. P . Santamarina ;
E . H . Gimenez
Heizmann, W . R. ; H . Werner
Kellog, D . S . ; K. K. Holmes ;
G . A. Hill
Loesche, W. J .
Newell, D . G. (Ed .)
Pöhn, H.-Ph .
Robinson, D. A. ; D . M . Jones
Stern, N .J . ; A .W . Kotula
Watt, B.
Werner, H .
Zum Vorkommen und zur Verbreitung von Campylo-
bader jejuni/coli in der Jungmastgeflügel-Produktion .
1 . Mitteilung Dtsch . tierärztl . Wschr ., 92 ;
456-459 (1985)
Examination of poultry giblets, raw milk and meat for
Campylobader fetus subsp. jejuni .
Archiv Lebensmittelhyg ., 33 ; 137-176 (1983)
Kultivierung von anaeroben Microorganismen ;
Methode zur Erzeugung eines anaeroben . Milieus .
Forum Mikrobiologie, 5 ; 246-248 (1982)
Anaerocult ® C a new system for cultivation of micro-
aerophilic and capneic microorganisms . Poster, Ist
European Congress of Clinical Microbiology, Bologna,
Oct. 1983
Presence of zoonotic pathogens (Yersinia spp ., Campy-
lobader jejuni, Salmonella spp ., and Leptospira spp .)
simultaneously in dogs and cats.
Zbl . Bakt . Hyg ., 1 . Abt., Orig . B,181 ;
430-440 (1985)
Isolation of Campylobacter jejuni from market chickens :
Effed of temperature and time of incubation .
Zbl . Mikrobiol ., 140 ; 649-652 (1985)
GasPak Plus versus Anaerocult A - Two Carbon Di-
oxide/Hydrogen Systems for Cultivation of Anaerobes .
Zbl . Bakt . Hyg . A 270 ; 511-516 (1989)
Laboratory diagnosis of Gonorrhea . Cumitech . No . 4,
ASM, Washington D . C ., USA (1976)
Oxygen sensitivity of various anaerobic baderia .
Appl . Microbiol ., 18 ; 723-727 (1969)
Campylobader, epidemiology, pathogenesis and bio-
chemistry. Proceedings of an International Workshop
an Campylobader Infedions. March 24-26,1981,
MTP Press Ltd ., Lancaster, England
Infektionsepidemiologische Situation in der Bundes-
republik Deutschland 1984 .
Münch . med . Wschr. 127, Nr. 31 ; 750-752 (1985)
Milkborne Campylobader infedion .
Brit. Med . J ., 282 ;1374-1377 (1981)
Survival of Campylobader jejuni inoculated into
Ground Beef .
Appl . Environm . Microbiol ., 44 ; 1150-1153 (1982)
The influence of carbon dioxide an the growth of
obligate and facultative anaerobes an solid media .
J . Med . Microbiol ., 6 ; 307-314 (1973) .
Anaerobier Infektionen . Stuttgart : Thieme Verlag (1981)
Merck's Anaerocult ® program offers the microbiologist a comprehensive System
for the cultivation of anaerobic and microaerophilic/capneic microorganisms . The
absence of palladium catalysts in the gas generating System improves safety
and reliability and also reduces costs .

The extensive range of culture media and the reagent program for the bio-
chemical differentiation of microorganisms enable the user to work both ration-
ally and economically .

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E . Merck, P.O . Box 4119, Frankfurter Strasse 250, D-6100 Darmstadt 1 231092