FR2534929A1 - SUPPLEMENT OF NOURISHING LAYERS FOR THE CULTURE OF CAMPYLOBACTER JEJUNI AS OF CEFOPERAZON - Google Patents

Abstract

THE PRESENT INVENTION RELATES TO A NOURISHING LAYER SUPPLEMENT FOR SELECTIVE CULTURE OF CAMPYLOBACTER JEJUNI. ACCORDING TO THE INVENTION, IT CONTAINS, AS AN EFFECTIVE ANTIMICROBIAL SUBSTANCE, CEFOPERAZON. THE INVENTION APPLIES IN PARTICULAR TO THE CULTURE OF BACTERIA.

Description

The present invention relates to a supplement of feeder layers for

  selective cultivation of Campylobacter eujuni, which complement contains at least one substance

effective antimicrobial.

  In recent years, it has been known that the bacterial species Campylobacter plays an important role at the origin of gastroenteritis. In particular, the genus Campylobacter

  faetus Subsp jejuni, which will be called hereinafter more

  Campylobacter jejuni, has been found to be particularly dangerous to humans and to many animals It is therefore necessary to place Campylobacter jejuni in selective culture for the purpose of diagnosis and / or examination of drugs or Resistance O To avoid as far as possible that on a bacteriological feeder layer, other microorganisms, besides Campylobacter jejuni, do not reproduce to a great extent, the effective neat antimicrobial substances have already been mixed with the feeder layers used, against which Campylobacter jejuni has a high resistance to other microorganisms Mixing complements are known to nurse layers which contain, for example, one or more of the following effective antimicrobial substances: Polymycin, Rifampicin, Trimethoprim, Actidion, Colistin, Bacitracin , Novobiocine, Cycloheximide,

Cephazolin, Amphotericin.

  The present invention therefore has the task of creating a complement of the above kind whose effect

  selective to promote Campylobacter jejuni is considerable-

  better than the complements of this kind known

until now.

  The complement according to the invention, which makes it possible to solve the task indicated above, differs from supplements known in the above manner by the fact that, as an effective antimicrobial substance, there is

Cefoperazon.

  Until now, Cefopérazon has never been used as an effective antimicrobial substance in supplements for feeder layers for the selective cultivation of Campylobacter jejuni.

  that Cefoperazon could produce an anti-selective

  microbial particularly valuable in the desired direction.

  In a useful way, the complement according to the invention contains, as effective antimicrobial substances, besides Cefoperazon, Rifampicin and advantageously

  also Amphotericin B and Colistin.

  The complement is used, according to another form of

  the invention, in admixture with a bacterio-alimentary layer

  logic for the selective cultivation of Campylobacter jejuni so that for 1 ml of the total mixture, there is approximately 15 μg of Cefoperazon and approximately 10 μg of Rifampicin and, if appropriate, approximately 2 μg of Amphotericin B and approximately 1 μg of Colistin Such a composition allows a very good growth of Campylobacter jejuni, while the growth of unwanted flora that accompanies it is significantly prevented.

  the pathogenic excitatory Campylobacter jejuni is considerable-

easily facilitated.

  Good solubility in water is another advantage

= complement according to the invention.

  The invention will be better explained with reference to the following examples:

EXAMPLE 1

  In a usual bacteriological feeder layer, consisting of one liter of Columbia agar and 50 ml of ewe's blood, a supplement containing three parts by weight of Cefoperazon and two parts by weight of Rifampicin was mixed. The concentration was chosen so that for about 1 ml of the total mixture, there is 15 Fg Cefoperazon and about 10 μg Rifampicin This mixture for

  feeder layer will be referred to hereinafter as "1-A".

  For comparison with the state of the art, a second feeder layer mixture, hereinafter referred to as "1-B", was prepared having the following composition: 1 liter of Colombian agar, 50 ml of blood of sheep and a supplement particularly recommended to date, containing, for 1 ml of the total mixture, 25 U I. Bacitracin, 50) ig Cycloheximide, 10 IU Colistin, Cephazoline pg and 6 ig Novobiocineo 759 sterile boxes Petri dishes were filled with feeder layer mixture 1-A and the same number of sterile petri dishes were filled with the second feeder layer mixture 1-B. Then, all the dishes were incubated in the usual way in research. bacteriological samples with the same human stool, then all the plates were allowed to rest at 37 ° C in an atmosphere consisting of 85% nitrogen, 10%

carbon dioxide and 5% oxygen.

  After 20 hours, 55 (= 7.24%) boxes containing the 1-A Feeder Mix and 56 (= 7.37%) boxes containing the feed mix were determined.

  feeder layers 1-B, the presence of Campylobacter jejuni.

  Thus, it is possible to obtain, with the complement contained in the feed layer mixture 1-A according to the invention, substantially the same good growth of Campylobacter jejuni as that obtained with the known complement contained in the second feed layer mixture 1-B. However, differences were found in the growth of flora Accompanying only 65% of boxes

  about as much growth in the flora

  In 1-A and 1-B Feeder Mixes 15% of all cans showed increased growth of flora in the 1-A Feeder Mix, while 20% of all cans left to recognize an increased growth of the flora of accompaniment in the second nourishing layer 1-B - The isolations undertaken in the usual way, led, with regard to the flora of accompaniment, with the following results: on the mixture for layer 1-A, it was found, after the first 10% isolation, after two isolations 3% and after three isolations 2% of all the boxes, a stronger growth of the accompanying flora, mainly yeasts and On the other hand, with the mixture for feeder layer 1-B, it was found, after the first isolation 13%, after two isolations 3% and after three isolations 4% of all the boxes a stronger growth. ance of the accompanying flora, mainly

cockles and Pseudomonas.

  It is recognized that on the complement-containing feeder layer mixture according to the invention, the Campylobacter jejuni pathogen could be better separated from the accompanying flora than on the feeder mixture containing the known complement 1-B, whereas obviously the complement according to the invention exerts a stronger antimicrobial effect on the accompanying flora than the known complement, without influencing the growth of Campylobacter.

EXAMPLE 2

  In a bacteriological feeder layer consisting of one liter of Colombian agar and 50 ml of sheep's blood, a supplement was mixed which contained the effective antimicrobial substances according to the invention.

  Cefoperazon, Rifampicin, Amphotericin B and Colistin.

  The concentration was such that for 1 ml of the total mixture there was about 15 μg Cefoperazon, 10 μg Rifampicin, 2 μg Amphotericin B and 10 μl Colistin.

  for feeder layer will be referred to hereinafter as "2-A".

  For a comparison with the state of the art, the feed layer mixture was again used

1-B indicated in Example 1.

  1570 sterile petri dishes were filled with feeder layer mixture 2-A and the same number of sterile petri dishes were filled with the known feeder layer mixture 1-B Then all the dishes were incubated in the usual way, with samples of a human stool, and then all the dishes were allowed to sit in an atmosphere consisting of 85% nitrogen,

  % carbon dioxide and 5% oxygen at 37 C.

  After 18 hours, in 148 dishes (= 9.42%) could be established with the feeder layer mixture 2-A and in 146 dishes (= 9.29%) with the known feed layer mixture. Thus, the complement according to the invention contained in the mixture for feeder layer 2-A has no detrimental influence on the growth of Campylobacter jejuni, in comparison with the complement known in the mixture for layer

feeder 1-B.

  However, differences in growth of the accompanying flora are even more remarkable than in the case of Example 1 In 71.6% of the dishes with the feed layer mixture 2-A, it was not possible to establish no growth of flora accompanying, while this was the case only for 47.5% of the boxes with the known feeder mixture mixture 1-B In the 28.4% of the remaining boxes with the nourishing layer mixture 2-A, it showed flora of accompaniment, in the first quadrant to 23,1% in the second quadrant to 3,6% / in the third quadrant to 1,0% and in the fourth quadrant to 0, 7% of all dishes with the 2-A feeder mix, however, 54.3% of the dits

  known feeder layer i-B, a growth of flora -

  37.6% in the first quadrant at 10.0% in the second quadrant at 2.9% in the third quadrant and 3.8% in the fourth quadrant for all

  with feeder layer mix 1-B.

  It is clear that the desired selective cultivation of Campylobacter jejuni is considerably more effective with the complement-containing 2-A feeder layer mixture of the present invention than with the known feeder-layer mixture 1-B, while the complement according to the invention. invention is better adapted to decrease the growth of the

  accompanying flora that the complement known.

  The complement described in Example 2 containing the effective antimicrobial substances Cefoperazon, Rifampicin, Amphotericin B and Colistin was also found to be more effective, in the desired sense than the complement described in

  Example 1, which contained only Cefoperazon and Rifampicin.

  Therefore, the complement according to the invention indicated in

Example 2 is preferred.

  However, the present invention relates to all the supplements for feeder layers allowing the selective cultivation of Campylobacter jejuni, which contain

  Cefoperazone and, where appropriate, other antimicrobial

effective.

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Claims (4)

CLAIMS CATIONS   1.Complement for feeder layers for the selective cultivation of Campylobacter jejuni, said supplement containing at least one effective antimicrobial substance characterized in that as an effective antimicrobial substance there is Cefoperazon. 2 supplement according to claim 1, characterized in that it contains, as antimicrobienneseeffectives substances,   besides Cefoperazon, rifampicin.   3 supplement according to claim 2, characterized in that it contains, for three parts by weight of Cefoperazon,   about two parts by weight of Rifampicin.   4 supplement according to claim 1, characterized in that it contains, as antimicrobial substances   effective, besides Cefoperazon, Rifampicin and 11 Amphopera-   Tericin B. Supplement according to Claim 4, characterized   in that it contains, for fifteen parts by weight of Cefopera-   about 10 parts by weight of Rifampicin and about 2 parts by weight of Amphotericin B. 6 Supplement according to Claim 1, characterized in that it contains, as effective antimicrobial substances, besides Cefoperazon, Rifampicin, Amphotericin B and Colistine.   7 A supplement according to claim 6, characterized in that the effective antimicrobial substances are present in the following proportions: Cefoperazone 10 20 Fg Rifampicin 7 13 pg Amphotericin B 1 3 pg Colistin 5 15 U I. 8 Supplement according to Claim 6, characterized in that the effective antimicrobial substances are present somewhat in the following proportions: Cefoperazon 15 μg Rifampicin 10, u Opg Amphotericin B 2 μg Colistin 10 μ I. 9 Addition according to claim 1, used in admixture with a bacteriological feeder layer for culture selective Campilobacter characterized in that, for 1 ml of the total mixture, about 15 μg of cefoperazon. A supplement according to claim 2, in admixture with a bacteriological feeder layer for the selective cultivation of Campylobacter, characterized in that about 15 μg of the total mixture is used for 1 ml of the total mixture.   Cefoperazone and about 10 pug of Rifampicin.   11 A supplement according to claim 6 used in a mixture with a bacteriological feeder layer for the selective cultivation of Campylobacter, characterized in that approximately 15 μg of Cefoperazonzon, about 10 μg of Rifampicin, about 2 μg of Rifampicin are used in 1 ml of the total mixture. pg   Amphotericin and about 10 IU Colistin.