GB2129013A - Supplement for culture media for the growth of campylobacter jejuni - Google Patents

Supplement for culture media for the growth of campylobacter jejuni Download PDF

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Publication number
GB2129013A
GB2129013A GB08325721A GB8325721A GB2129013A GB 2129013 A GB2129013 A GB 2129013A GB 08325721 A GB08325721 A GB 08325721A GB 8325721 A GB8325721 A GB 8325721A GB 2129013 A GB2129013 A GB 2129013A
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United Kingdom
Prior art keywords
supplement
cefoperazon
rifampicin
culture medium
campylobacter jejuni
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GB08325721A
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GB8325721D0 (en
GB2129013B (en
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Jean Paul Butzler
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INST VIRION AG
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INST VIRION AG
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C12Q1/045—Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/18—Testing for antimicrobial activity of a material

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Toxicology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

A supplement for bacteriological culture media for the selective growth of campylobacter jejuni contains cefoperazon as an antimicrobially active substance, preferably together with rifampicin and preferably also amphotericin B and colistin.

Description

SPECIFICATION Supplement for culture media for the growth of campylobacter jejuni The present invention relates to supplements for culture media, for the selective growth of campylo bacterjejuni, which supplements contain at least one antimicrobially active substance.
For a few years it has been known that the bacteria genus campylobacter plays a decisive role in producing gastroenteritis. In particular, the species campylobacter faetus subsp. jejuni, hereinafter referred to briefly as campylobacter jejuni, was found to be particularly dangerous for humans and many animals. There is therefore a need for selectively growing campylobacter jejuni for the purpose of diagnosis and/or the testing of drugs and resistance.
In order to as far as possible prevent other micro-organisms substantially multiplying in addition to campylobacter jejuni on a bacteriological culture medium, certain antimicrobially active substances against which campylobacter jejuni has a higher resistance than other micro-organisms have already been mixed with the culture medium used.
Known supplements for mixing with the culture media contain for example one or more of the following antimicrobially active substances: polymycin, rifampicin, trimethoprim, actidione, colistin, bacitracin, novobiocin, cyloheximide, cephazolin, amphotericin.
Accordingly the present invention provides a supplement for use in culture media, for the selective growth of campylobacter jejuni, said supplement including cefoperazon as an antimicrobially active substance.
Cefoperazon has never previously been used as an antimicrobially active substance in supplements for culture media for the selective growth of campylo bacterjejuni. It has been found that cefoperazon is able to exert a particularly significant selective antimicrobial action in the required sense.
The supplement according to the invention peferably also contains rifampicin as an antimicrobially active substance in addition to cefoperazon, and preferably also additionally amphotericin B and colistin.
The invention further provides a bacteriological culture medium forthe selective growth of campylo bacterjejuni, containing a supplement as defined above, in which substantially 15 9 of cefoperazon are used per ml of total medium.
Preferably 10 > 9 of rifampicin, and possibly additionally about 2 Ag of amphotericin B and about 10 I.E. of colistin are also used per ml of the medium.
Such a composition allows very good growth of campylobacter jejuni, whereas the growth of undesirable accompanying flora is significantly inhibited.
Some examples of the invention will now be described for the purposes of illustration only.
Example 1 A supplement containing 3 parts by weight of cefoperazon and 2 parts by weight of rifampicin was mixed with a normal bacteriological culture medium consisting of 1 litre of Columbia agar and 50 ml of sheep's blood. The concentration was chosen such that each ml of total mixture contained about 15 Fg of cefoperazon and about 10 Fg of rifampicin. This culture medium mixture is indicated hereinafter as "i-A".
For comparison with the state of the art, a second culture medium mixture, indicated hereinafter as "l-B" was prepared, having the following composition: 1 litre of Columbia agar, 50 ml of sheep's blood and a supplement which had previously been judged to be particularly selective, and which for each ml of total mixture comprised 25 I.E. of bacitracin, 50 Fg of cycloheximide, 10 I.E. of colistin,5pg of cephazolin and 5 pLg of novobiocin.
759 sterile petri dishes were covered with the culture medium mixture I-A, and the same number of sterile petri dishes were covered with the second culture medium mixture I-B. All dishes were then incubated with samples of the same human faeces in the normal manner four bacteriological tests, after which all dishes were allowed to stand at 37"C in an atmosphere of 85% nitrogen, 10% carbon dioxide and 5% oxygen.
After 20 hours, the presence of campylobacter jejuni could be determined in 55 dishes (ie 7.24%) containing the culture medium mixture I-A and in 56 dishes (ie 7.37%) containing the culture medium I-B.
Thus, in the case of the supplement as contained in the culture medium mixture I-A, the growth of campylobacter jejuni was practically as good as in the case of the known supplement contained in the second culture medium mixture I-B. Differences were apparent however in the growth of the accompanying flora. Only in 65% of the dishes could an approximately equally strong growth of accompanying flora be determined on the culture medium mixture I-A as compared to culture medium I-B. 15% of all dishes showed an increased growth of accompanying flora on the culture medium mixture I-A, whereas 20% of all dishes showed an increased growth of accompanying flora on the second culture medium I-B.
Isolations carried out in the normal manner led to the following results in relation to the accompanying flora: 10% of all dishes after the first isolation 3% after two isolations and 2% after three isolations showed a stronger growth of accompanying flora, predominantly yeast and E.Coli bacteria, on the culture medium mixture I-A. In contrast, 13% of all dishes after the first isolation, 3% after two isolations and 4% after three isolations showed a stronger growth of accompanying flora, predominantly cocci and pseudomonas, on the culture medium mixture l-B.
It can be seen that on the culture medium mixture I-A the pathogenic germs campylobacter jejuni can be better separated from the accompanying flora than on the culture medium mixture I-B containing the known supplement, because the supplement containing cefoperazon exercises a stronger antimicrobial action on the accompanying flora than the known supplement, without impairing the growth of campylobacter.
Example 2 A supplement containing cefoperazon, rifampicin, amphotericin B and colistin as antimicrobially active substances was mixed with a bacteriological culture medium consisting of 1 litre of columbia agar and 50 ml of sheep's blood. The concentration was such that each ml of total mixture contained about 15 ug of cefoperazon, 10 g of rifampicin, 2 ug of amphotericin B and 10 I.E. of colistin. This culture medium mixture is indicated hereinafter as "2-A".
For comparison with the state of the art, the known culture medium mixture 1-B described in Example 1 was again used.
1570 sterile petri dishes were covered with the culture medium mixture 2-A and the same number of sterile petri dishes were covered with the known culture medium mixture 1-B. All dishes were then incubated in the normal manner with samples of human faeces, after which all dishes were allowed to stand in an atmosphere of 85% nitrogen, 10% carbon dioxide and 5% oxygen at 37 C.
After 18 hours, an increase of campylobacterjejuni could be determined in 148 dishes (ie 9.42%) containing the culture medium mixture 2-A and in 146 dishes (ie 9.29%) containing the known culture medium 1-B. Thus, the supplement contained in the culture medium mixture 2-A has no impeding influence on the growth of campylobacter jejuni, compared with the known supplement in the culture medium mixture I-B.
Differences in the growth of the accompanying flora were even clearer than in the case of Example 1. No growth of accompanying flora could be determined in 71.6% of the dishes containing the culture medium mixture 2-A, whereas this was the case only in 45.7% of the dishes comprising the known culture medium mixture 1-B. An accompanying flora was observed in the remaining 28.4% of dishes containing the culture medium mixture 2-A, namely in the first quadrant in the case of 23.1%, in the second quadrant in the case of 3.6%, in the third quadrant in the case of 1.0% and in the fourth quadrant in the case of 0.7% of all dishes containing the culture medium mixture 2-A.In contrast, a growth of accompanying flora was determined in the case of 54.3% of the dishes containing the known culture medium mixture 1-B, namely in the first quadrant in the case of 37.6%, in the second quadrant in the case of 10.0%, in the third quadrant in the case of 2.9% and in the fourth quadrant in the case of 3.8% of all dishes containing the culture medium mixture 1-B.
It is clearthatthe required selective growth of campylobacter jejuni with the culture medium mixture 2-A is considerably more efficient than with the known culture medium mixture 1-B, because the described supplement is more suitable for suppressing the growth of the accompanying flora than the known supplement.
The supplement described in Example 2 comprising the antimicrobially active substances cefoperazon, rifampicin, amphotericin B and colistin has also proved more effective in the required sense than the supplement described in Example 1,which contains only cefoperazon and rifampicin. Thus the supplement described in Example 2 is preferred.
It will be appreciated that the use of the described culture media supplement considerably simplifies the isolation of the pathogenic agent campylobacter jejuni. The supplement containing cefoperazon is further advantageous in that it has good water solu bility.

Claims (13)

1. A supplement for use in culture media, for the selective growth of campylobacter jejuni, said suppiement comprising cefoperazon as an antimicrobially active substance.
2. A supplement as claimed in claim 1,further comprising rifampicin as an antimicrobially active substance in addition to cefoperazon.
3. A supplement as claimed in claim 2, which contains substantially 2 parts by weight of rifampicin to 3 parts by weight of cefoperazon.
4. A supplement as claimed in claim 1, further comprising rifampicin and amphotericin B as antimicrobially active substances.
5. A supplement as claimed in claim 4, which contains substantially 10 parts by weight of rifampicin and substantially 2 parts by weight of amphotericin B to 15 parts by weight of cefoperazon.
6. A supplement as claimed in claim 1, further comprising rifampicin, amphotericin B and colistin as antimicrobially active substances.
7. A supplement as claimed in claim 6, in which the antimicrobially active substances are present in substantially the following proportions: cefoperazon 10 - 20 pg rifampicin 7 - 13 ijg amphotericin B 1 - 3 > 9 colistin 5-15 I.E.
8. A supplement according to claim 6, in which the antimicrobially active substances are present in substantiallythefollowing proportions: cefoperazon 15 iSug rifampicin 10 pug amphotericin B 2,ag colistin 1OI.E.
9. A bacteriological culture medium for the selective growth of campylobacter jejuni, containing a supplement as claimed in claim 1, in which substantially 15 9 of cefoperazon are used per ml of total medium.
10. A bacteriological culture medium for the selective growth of campylobacter jejuni, containing a supplement as claimed in claim 3, in which substantially 15 clog of cefoperazon and about 10 ijg of rifampicin are used per ml of total medium.
11. A bacteriological culture medium for the selective growth of campylobacter jejuni, containing a supplement as claimed in claim 6, in which substantially 15 of cefoperazon, about 10 g of rifampicin, about 2 CLg of amphotericin and about 10 I.E. ofcolistin are used per ml of total medium.
12. A supplement for use in culture media for the selective growth of campylobacter jejuni, substan tially as herein described with reference to the accompanying examples of the invention.
13. A bacteriological culture medium for the selective growth of campylobacter jejuni containing a supplement as claimed in any one of claims 1 to 8 or 12.
GB08325721A 1982-10-20 1983-09-26 Supplement for culture media for the growth of campylobacter jejuni Expired GB2129013B (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE19823238819 DE3238819A1 (en) 1982-10-20 1982-10-20 ADDITIONAL SOURCES FOR BREEDING CAMPYLOBACTER JEJUNI

Publications (3)

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GB8325721D0 GB8325721D0 (en) 1983-10-26
GB2129013A true GB2129013A (en) 1984-05-10
GB2129013B GB2129013B (en) 1985-09-25

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JP (1) JPH062048B2 (en)
BE (1) BE897927A (en)
DE (1) DE3238819A1 (en)
FR (1) FR2534929B1 (en)
GB (1) GB2129013B (en)
IT (1) IT1164439B (en)
NL (1) NL8303506A (en)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2301211C3 (en) * 1973-01-11 1978-06-22 C.H. Boehringer Sohn, 6507 Ingelheim Synthetic nutrient medium for microorganisms
US4229530A (en) * 1978-08-29 1980-10-21 Board Of Regents, University Of Texas System Methods and media for rapid detection of pathogenic neisseria

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IT8322931A0 (en) 1983-09-20
FR2534929B1 (en) 1985-11-22
GB8325721D0 (en) 1983-10-26
DE3238819C2 (en) 1990-04-12
IT8322931A1 (en) 1985-03-20
JPH062048B2 (en) 1994-01-12
GB2129013B (en) 1985-09-25
IT1164439B (en) 1987-04-08
BE897927A (en) 1984-01-30
NL8303506A (en) 1984-05-16
DE3238819A1 (en) 1984-04-26
FR2534929A1 (en) 1984-04-27
JPS6037978A (en) 1985-02-27

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PCNP Patent ceased through non-payment of renewal fee

Effective date: 19940926