Unit: Institut Pasteur Collection
Director: BIZET Chantal
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 for the creation of the Biological Resources Centres.
1. Strain stock management
7.800 strains are available on stock. Every year, new bacterial strains are added to the collection (about 400) 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).
2. Check up of the CIP strains
a) Total protein electrophoresis and chimiotaxonomy
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 (about 3000 strains).
b) A database with 888 profiles obtained after ribotyping has been performed.
This laboratory is necessary for the Collection strain check and strain expertise for industrial laboratories
3. Quality System
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.
The CIP has renewed its certification in November 2003 according to the new referential ISO 9001 version 2000.
4. Biological Resources Centre of the Institut Pasteur
The Institut Pasteur is the French institution having on one site, the most important collections specialized in microorganisms (bacteria, fungi, virus ).
In most of cases, these collections are kept inside research laboratories. Only four of these collections are "opened" i.e. they send strains to laboratories, pharmaceutical industries, research laboratories, education when receiving an order form. These four collections are recorded in the World Directory of Collections of Cultures of Microorganisms. It includes the collection of the Institut Pasteur (CIP), the cyanobacteria collection, the fungi collection (UMIP) and the collection of sporulated entomopathogenical bacteria (IBEC).
Two goals were in focus :
1) to increase the number of microorganisms strains available for the scientific researchers and industrials for which including, production, check (including molecular), preservation, stocking and distribution are made according to Quality Insurance and legal demandings, specifically regarding safety,
2) to create et run the software for the management of the collection, used by all the collections into the CRBIP. Such a software should be able to manage all data for different kind of microorganisms. These data concerning taxonomy, keywords references for bibliography, morphological, physiological, physiopathological and molecular characters, pratical information (such as used medium, temperature for growing ), management of the production, stockage and distribution.
Presently 350 strains of Leptospira (coming from Leptospirose CNR) have been included into the CRBIP, after having check their viability and identity and having a minimum stock. A hundred of Mycobacterium strains (coming from Mycobacterium CNR) are being to be included.
So, the first step regarding the creation of the Biological Resources Centre of the Institut Pasteur is completed. The main goal was to deal with the creation of a software for running all the collections (ARPAS). The database has been created for adding more collections. Now, we have to improve our software and also to implement Quality Insurance.
Connections to ARPAS are done through our Internet, so a web site must be created. In providing information to scientists, teachers, industrials..., regarding strains it will be a positive action for the CRBIP.
5. Identification of strains of Acinetobacter by a phenotypic and molecular technic
Acinetobacter specy was created in 1954 by Brisou and Prévot. It is made of short not mobile bacilla, stric anaerobic, Gram negative. Phylogenetical analysis based upon ARN sequences ribosomal 16S enable us to file Acinetobacter in the subspecy gamma of Proteobacteria. Indide it, 21 genomic groups have been got through hybridation AND-AND studies. However, for a long time, Acinetobacter specy gets only 7 species. Recently, 9 new species of Acinetobacter have been described. It shows that the diversity for this specy is very important and that a lot of strains are not classified.
22 strains of Acinetobacter inside the collection of the Institut Pasteur have been caracterised in order to define the taxonomic status of these strains by different methods including phenotipical analysis, electrophoresis of proteins and ARN sequences ribosomal 16S.
Results of ARNr 16S sequences show that 4 strains can be bound to genomospecies 9, 4 strains to Acinetobacter baumannii, 1 strain to genomospecies 10, 1 strains to genomospecies 13. 6 strains are an independent cluster.
A good relation with the results of other used methods for identification of these strains has not yet been got.
6. Gene sequencing of rRNA 16S for bacterial type strains of the Collection
Within the framework of a proposal submission and in order to contribute to the enrichment of the database of the CIP, the gene coding for the 16S rRNA of type strains is sequenced.
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. Bacterial strains from external collections are preserved in the CIP. It's submitted to a maintenance contract. CIP provides industrial laboratories with spore suspensions.
This activity is going to be submitted to Quality Insurance.
8. National relationships
The Ministry of Research created Biological Resources Centers (BRC) at a national level which is managed by a Committee of Biological Resources. The CIP belongs to this Committee. Its missions are to create a national network for BRC and to implement a Quality Insurance and to check the launch of quotations for creation and finance of accredited BRC,
9. International relationships
The CIP is a member of different official organizations : l'ECCO (European Culture Collections' Organization - www.eccosite.org), EBRCN (European Biological Resource Centres Network - www.ebrcn.org), CABRI (Common Access to Biotechnological Resources and Information - 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 :
The Presidence has been given to France and a study group is in charge of determining if it's possible to create and implement biological resources centers. The CIP belongs to the French delegation and helps in writing documents regarding the creation of the Insurance Quality implementation.
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 European Collections.
Aims 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.
Keywords: taxonomy, sequencing, biodiversity, collection, bacterial strains, studies, preservation