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  Collec


  Director : Chantal Bizet (bizet@pasteur.fr)


  abstract

 

The role of the Collection de l'Institut Pasteur (CIP) is to ensure the maintenance and the enrichment of its bacterial collection, the production of a catalogue (the 7th edition was published in 1998), the diffusion of information on the distributed strains and finally the development of research on identification, taxonomy and preservation of bacteria. The CIP is the first French ISO 9001 certificated collection and participates to several international and national projects.



  report

cale

Strain stock management (F. Bimet)

7.000 strains are available on stock (1.850 different species). Every year, new bacterial strains are added to the collection coming either from researchers of the Institut Pasteur, or from foreign collections or from newly deposited strains after having connection with foreign or French researchers. We've got also plasmids, transposons, host strains, probes. Titrated spore suspensions (species Bacillus and Clostridium) are available. Strains are preserved in two different places and by two different ways (freeze dried and deep-frozen at - 80°C, or deep-frozen in nitrogen liquid and at - 80°C for strains which can't be freeze-dried).

Computer system (F. Bimet)

During the whole year, an update of the database is performed regularly in order to enter strain biochemical properties, information regarding the catalogue and the stock. The catalogue can be viewed on the Institut Pasteur Web site at : http://CIP.pasteur.fr

A software for the Collection management has been implemented.

Studies (D. Clermont, F. Bimet, C. Bizet)

Studies are made according to industrial laboratory requests, regarding bacterial identification by the study of the cell wall components and protein electrophoresis and also by the study of the strain susceptibility to different antibiotics.

The CIP organizes quality controls for public or private laboratories. An external quality control was organized thanks to a partnership with the Comité de Lutte contre les Infections Nosocomiales located in the Centre of France (CLIN). 21 laboratories received 6 multiresistant strains for testing. Differences were discovered, so this kind of quality control will continue.

Bacterial strains from external collections are preserved in the CIP. It's submitted to a maintenance contract.

National relationships (C. Bizet)

1. The Ministry of Research created Biological Ressources Centers (BRC) at a national level which will be managed by a Committee of Biological Resources. The CIP belongs to this Committee. Its missions are the following :

- to create a national network for BRC and to implement a Quality Insurance,

- to check the launch of quotations for creation and finance of accredited BRC,

Three working groups were set up :

- "animal vegetal" group,

- "human" group,

- "microbiology" group.

The CIP is responsible for the last one.

2. Secrétariat Général de la Défense Nationale (SGDN)

The CIP is in the relation with the SGDN for the sending of the strains on the French territory and abroad.

3. Tranportation Ministry

The CIP attends a transportation training for dangerous goods. This training is for technical officers for logistic who are responsible for checking technical logistics on air freight for the French territory.

International relationships (C. Bizet)

The CIP is a member of different official organizations : l'ECCO (European Culture Collections' Organization — www.cba.unige.it), CABRI (Common Access to Biotechnological Resources and Information — www.cabri.org and WFCC (World Federation for Culture Collection).

1. In 1999, OECD (Organization for Economic Cooperation and Development) initialized an international meeting in Tokyo in order to look at the necessary conditions to set up Biological Resource Centers (BRC). It is requested that :

- governments help the creation of BRC and an accreditation system must be implement,

- links must be created between BRC so that an international BRC network can exist,

The Presidence has been given to France and a study group is in charge of determining if it's possible to implement it. An international Secretary Office has been created and the CIP is a member.

2. European Biological Resource Centers Network (EBRCN)

The CIP participates to EBRCN.

This European project enables Europe to :

. show its study skills in Biological Resource Centers (CRB) in computing

. create global policy

. maintain a high level of quality for the services provided by Collections.

Goals of this project are the following :

. to create a biological resource centers network including alive organisms, components and associated data bases, and to answer OECD new decisions in an international context.

. to develop European standards for BRC, based upon management quality system

. to create a framework in order to increase complementarity and to decrease redundancy among European BRC.

. to introduce new techniques in Information Technology in order to bring added value to the catalogue and to give a better access.

. to get and provide information on BRC.

Quality System (D. Clermont)

Process regarding activities and tasks are written in an organized process system which suits to the CIP organization. Tasks are recorded. Implemented processes are always followed up and improved. Regularly, internal audits are made in order to check and measure differences between what is written and what is done. Quality indices are provided with laboratory problems and requests from customers. If necessary, corrective actions are undertaken. Certification (ISO 9001) has been granted for 3 years. But, every year, a follow up audit must be made in order to keep it.

After having written a new Quality Manual and implementing new processes, the CIP has renewed its certification in July 2001 according to the new referential ISO 9001 version 2000.

Creation of a ribotype database (D. Clermont)

Ribotyping is carried out by using the Riboprinter microbial characterization system (Qualicon Inc. Wilmington, Del.) in order to create a database and to satisfy customers' studies requests and identify the strains of the CIP.

Re-identification of bacterial strains after description of new species (F. Bimet, D. Clermont)

119 Bacillus subtilis strains have been studied by ribotyping and whole cell protein electrophoresis in order to identify 2 subspecies described recently : subtilis and spizizeni.

The results obtained show that the tested strains are divided into two groups by ribotyping, each of them including a subspecies type strain. No distinct similar group was identified by whole cell protein electrophoresis. Ribotyping allows a better distinction of the Bacillus subtilis subspecies than whole cell protein electrophoresis.

Spore sale (D. Clermont)

Spores of 6 different strains are available as European sporicid standards. Spore antiseptic activity is compared after having preserved them at + 4°C and at - 80°C. It's done thanks to a collaboration with MICRAAM laboratory of Marseille.

The 4°C preservation of spore suspensions has been chosen after comparing the results obtained. The sensitivity of the fresh spore suspensions against the two antiseptic tested products was more important than the sensivity of the preserved spore suspensions. Contradictory results were obtained with the spores of Clostridium sporogenes. In order to define a range of use for Bacillus cereus and Bacillus subtilis subsp. subtilis spore suspensions, their antiseptic activities are going to be performed at regular intervals during about one year.

Total protein electrophoresis and chimiotaxonomy (F. Bimet

This laboratory helps in developing bacterial identification methods by analysis of bacterial cell wall components : lipids, peptidoglycane, sugars... and total proteins. Main techniques used are chromatography

and derivated techniques and also electrophoresis in discontinued gel. Results are used thanks to a

computerized process with specific softwares.

We are still working on feeding several databanks with information regarding protein profiles and nature of cell wall components. This laboratory is necessary for the Collection strain check and strain expertise for industrial laboratories

Gene sequencing of rRNA 16S for negative Gram bacterial strains of the Collection (D. Clermont)

Within the framework of a bid and in order to contribute to the enrichment of the database of the CIP, the gene coding for the 16S rRNA of Gram negative strains is going to be sequenced. The first strains concerned are the type strains. The search of a simple accurate and universal (if possible) method for DNA extraction is under way. About fourty strains are ready to be sequenced.

Bacterial identification by phlogenetic analysis (C. Dauga)

Phylogenetic trees based on 16S rRNA genes are a good help for bacterial strain identfication. The study of a large sample of bacteria with several strains by species increases the accuracy of this tool. More variable genes (gyrB, recA) allow to refine identification for short phylogenetic distances. At last, the choice of phylogenetic method is important to assess the results.

More than 200 sequences of genes from gram-positive bacteria i.e. Corynebacterium were included in phylogenetic trees. They allowed to identify 80 unknown strains at species level.



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  publications

puce Publications of the unit on Pasteur's references database


  personnel

  Office staff Researchers Scientific trainees Other personnel
 

COUTELLIER Sabrina
ROPARS Sandrine

BIZET Chantal
DAUGA Catherine

BIZET Chantal
DAUGA Catherine

BIMET François
BREMONT Sylvie
CANDREA Adina
CLERMONT Dominique
Carole FEURER
GRANDET Patricia
HARMANT Christine
NOWAKOWSKI Mireille
RIVERA-MELLADO Teofilo
VIVIER Catherine


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