CABI BIOSIENCE UK Centre (Egham) formerly International Mycological Institute

Contact for additional information: Dr D Smith

Reference Collection, Equipment/Technologies:

Freeze Drying Equipment:
Edwards Modulyo 4K
Edwards Super Modulyo 12K
Edwards Minifast Shelf Freeze-drier

Cryopreservation Equipment:
Controlled rate freezer
Storage in 350l liquid nitrogen vessels with automatic fill system.

Cryogenic Light Microscopy :
A cryogenic light microscope system (CM3 Planer Products Ltd) with an Amstrad PC 1640SD for temperature control of the stage heater. A conduction stage is mounted on a Zeiss Axioskop H-DIC microscope fitted with Plan Neofluar 40/0.75 (PH2) objectives combined with a 1.6x magnification changer. Data are recorded on video (Panasonic AG6200 recorder, JVC TK870E camera) using a video character generator (CM3200-00) and played back for analysis on a video monitor (Sony Trinitron KX14CPI 14").

Additional Services, Equipment/Technologies available from CABI BIOSCIENCE:

TRAINING
Full details available directly from CABI BIOSCIENCE:

IDENTIFICATION and DIAGNOSTIC SERVICE:
Fungal and Bacterial Identification Service utilizing scanning electron microscopy (SEM), chemotaxonomic, molecular, and biochemical facilities as required.
Diseased Plant Diagnostic Service
Environmental and Industrial Diagnostic Service

RESEARCH ADVISORY AND CONSULTANCY SERVICES
Microbial testing of materials: CABI BIOSCIENCE has NAMAS accreditation for mould growth testing of materials, equipment, plastics and biocide challenge testing, allows a wide range of tests to national and international standards.
Culture and extract supply for screening. (Full details available from CABI BIOSCIENCE).
Longer-term Research projects. (Generally funded externally, often in collaboration with overseas institutions).

Biosystematics
CABI BIOSCIENCE can offer expertise in the following areas:

• Advice on sampling protocols
• Assistance in bioprospecting schemes
• Database design and development
• Ex-situ preservation and conservation
• Identification and classification
• Isolation from natural substrata
• Production of nomenclators, bibliographies and checklists
• Repatriation of data
• Selection and monitoring of indicator species
• Selection of species and strains of edible fungi Site surveys and inventories
• Strain and species characterization
• Studies of soil biodiversity
• Sustainable use of ecosystems
• Training and capacity building

Biochemistry and Molecular Biology

Biochemical techniques

• Isoenzyme analysis of intra- and extra-cellular enzymes
• Isoprenoid quinone analysis
• Fatty acid analysis
• Physiological analyses, including enzyme and metabolite production

Biochemical or chemotaxonomic methods have been established at CABI BIOSCIENCE for over 10 years and contribute a significant proportion of all characterization and identification work. Methods include isoenzyme analysis by use of polyacrylamide-gel electrophoresis, analysis of isoprenoid quinones by TLC and HPLC and quantitative analysis of fatty acids by gas chromatography. The latter is based on a dedicated system (MIDI, Delaware, USA) and is one of the best available for rapid and accurate bacterial and yeast characterization.

Techniques have been developed to determine biological activities of compounds of biotechnological importance. Rapid screening methods for mycotoxins (e.g. ochratoxin A, patulin), phytotoxins (e.g. fusaric acid) and antibiotics (e.g. griseofulvin), together with citric acid and industrial enzymes (e.g. cellulase, lipase, pectinase, amylase) have been perfected. A database of TLC characters and UV visible spectra of over 100 secondary metabolites from fungi has been compiled and is updated as further compounds are obtained.

Molecular biology techniques:

• Analysis of mitochondrial DNA by RFLP
• Genomic fingerprinting methods based on nucleic acid amplification techniques (RAPD’s, ITS etc.)
• Genomic fingerprinting methods based on the use of PFGE, including electrophoretic karyotyping
• Probing/hybridization techniques

A wide range of molecular biology techniques are currently in use and include restriction fragment length polymorphism (RFLP) analysis of fungal mitochondrial DNA, electrophoretic karyotyping by use of pulsed field gel electrophoresis (PFGE), genomic fingerprinting by use of rare-cutting restriction endonucleases and PFGE, genomic fingerprinting using nucleic acid amplification techniques such as random amplified polymorphic DNA analyses (RAPD’s) and amplification and subsequent restriction of fungal ITS and IGS regions, and hybridization and probing methods. A recent addition is the DGGE population analysis technique.

The Culture Collection of Algae and Protozoa (CCAP) Oban

Contact for additional information: Mrs Christine Campbell

General Details:

The Collection comprises a gene bank of marine and hypersaline microalgae and cyanobacteria collected from a wide range of geographical locations and ecological niches, ranging from Antarctic seas to soda lakes in Africa. It includes some small multicellular seaweeds and most algal classes are represented. Some material is cryopreserved. CCAP is an IDA for patented strains and can accept valuable strains for confidential safekeeping. The Collection and its associated facilities for cultivating algae under controlled conditions provide an ideal environment for experimental and pilot scale work.

Equipment /Techniques:

There are several illuminated incubaters with light/dark cycling, capable of maintaining constant temperatures (±1 C) between 0°C and 40°C. There are two large walk-in constant temperature (10-30°C) rooms with lighting and robust shelving for larger scale (200-300 litres in total) cultivation. Gases can be supplied. Single axenic cultures of up to 20 litres can be handled if necessary. There are two laminar flow cabinets for conducting aseptic operations and two top loading autoclaves. There is a useful range of light microscopes available, one equipped with a colour TV camera.

Routine cryopreservation techniques and techniques of algal lipid analyses are employed by CCAP staff.

Access to the facilities of DML:

Associated with the core activities of CCAP are the general facilities of DML, including EM, SEM, centrifuges, HPLC, GC, Fluorometer, AA photospectrometer, X-ray Fluorescence Spectrometer, Inductively Coupled Plasma Mass Spectrometer (ICP-MS), scintillation counter and autoanalyser. There is a radiation suite. There are seagoing boats and extensive experimental aquarium facilities supplied with running seawater. There are good library facilities.

The Culture Collection of Algae and Protozoa (CCAP) Windermere.

Contact for additional information: Dr J.G. Day

Equipment /Techniques:

Fully equipped purpose built laboratory suites, including separate, dedicated, media-prep and wash-up facility.
Four temperature controlled rooms/ walk in incubators.
Various small incubators both illuminated and non-illuminated.
Class III containment room
Various laminar flow cabinets.
Cryopreservation facilities allowing controlled-rate, uncontrolled two-step cooling and vitrification.
Cryostorage facility.
Various microscopes: Including bright field, phase contrast and fluorescence.
Molecular biology equipment: PCR machine, gel electrophoresis apparatus, pulse-field gel electrophoresis apparatus, DNA sequencing apparatus

Core research:

Cryopreservation: Protocol development and improvement; mechanisms of lethal and sublethal cryoinjury.
Taxonomic: Using both traditional and modern molecular techniques.

Contract research:

Wide range of areas and techniques including biochemical, biotechnological, ecological and microbiological approaches have been used to fulfil CR project including discrete CCAP projects and input into larger IFE projects.

Access to the broader facilities of IFE.

This includes:
Electron microscopy (Both SEM & TEM).
Flow cytometry.
Fritsch collection of algal illustrations.
LAN, Internet connection and Computing support.
Analytical chemistry facilities.

The European Collection of Cell Cultures (ECACC)

Contact for additional information: Dr P Packer /The CAMR Business Development Department

Equipment/Technologies available at ECACC:

Contract Master and Working Cell Bank storage of Animal cells, Human cells, bacteria and DNA Probes.

Safe and Patent Deposit Facilities.
Tissue Culture.
Contract Mycoplasma, bacteria and fungi testing of Cell Lines.
Cell Culture Scale-up.
EBV Transformations.
DNA Extraction.
Serum Testing Facs.
Immortalisation of animal cells.
Genetic "fingerprinting" of animal cells.
Primary Cell Culture.
Tissue modelling.
In-vitro animal cell modelling.
Trans-Epithelial Electrical Resistance Measurement.
Simulated-Microgravity (Rotary wall Vessels).
3D Culturing using poly-hema coated Cell Culture Surfaces.
Plasmid Amplification.
Preparation and supply of purified nucleic acid or inactivated Pathogens for laboratories which cannot handle live micro-organisms.
Provision for characterisation of pathogens and their products.
Provision for raising hydridomas/purified antibody against pathogens (Automass).
Provision of bulk production of antibody Recombinant Protein Technology.
Diagnosis of exotic viral isolates.

Equipment/Technologies at CAMR and available to ECACC:

DNA Sequencer
Mass Spectrometer
Scanning Electron Microscope
Transmission Electron Microscope
Protein Modeller
Pathogen containment facilities-ACDP Class II, III and IV
WHO Reference site for Anthrax and Ebola
Research into a variety of pathogens including HIV, E.coli 0157, Yersinia pestis, Neisseria menigitidis etc.
Production of Toxins, e.g. Botulinum Toxin.
Contract Research and Development
Pharmaceutical Manufacturing Facilities (to GMP and including, freeze-drying, packaging etc.).
Vaccine Manufacture Contract.
Manufacturing of Biopharmaceuticals Pilot Fermentation Capacity.
Large Scale Animal Cell Culture.
Diagnostics and detection.
Research and Development Production Monitoring, analysis and control of micro-organisms in natural and man-made environments.
Equipment testing and validation for microbial integrity.
Bioremediation and waste management.
Aerobiology Investigation

The National Collection of Industrial, Food and Marine Bacteria (NCIMB)

Contact for additional information: Dr P Green

Preservation equipment
Edwards EF6 centrifugal and shelf freeze-dryers.
Edwards Supermodulyo freeze-dryer.
Cryotechnics Biosystems 20 liquid nitrogen storage system with autofill

Ancillary equipment
Range of incubators, ovens, centrifuges, water baths, steam sterilisers, autoclaves, water purification systems, media dispensers, safety and sterile airflow cabinets, anaerobic cabinets, microscopes and photomicroscopes, vacuum pouching devices, phastsystem electropheresis, BIOLOG automated ID system, Gas gramatograph for FAME analysis.

Equipment available via NCIMB Environmental
HPLC, Gas chromatograph with FID detection
AA spectrometer and graphite furnace
Dionex anion chromatograph, IR Spectrophotometer
Gamma Spectrometer, X-ray diffraction electron microscope (for inorganic solids analysis) with EDX

Identification and analytical services
Identification of non-pathogenic bacterial cultures using phentotypic characterisation, biochemical, chemotaxonomic and molecular techniques. "Traditional" identification techniques have been used at NCIMB for the last 50 years and are still in demand by many clients who wish either a simple identification based on phenotype or detailed strain characterisation. Over the years these techniques have been supplemented with other methodologies including API, BIOLOG chemical oxidation spectral analysis, MIDI fatty acid analyses and cell wall analysis.

Molecular techniques include DNA sequencing facilities which allow culture confirmation or authentication, possible strain differentiation, as well as routine bacterial identification.

Analytical services include tailored chemical and microbiological analyses of a range of freshwater and marine systems, products and wastes.

R+D and consultancy services
Contract or collaborative R+D or consultancy is possible in a number of areas. Current projects are associated with molecular probe design, PCR based bacterial analytical kit development and user friendly bacterial transportation and delivery systems.

The National Collection of Pathogenic Fungi (NCPF)

Contact for additional information: Dr DW Warnock or Dr Colin Campbell

Equipment /Techniques available "in house" or via the PHLS:

Liquid Nitrogen storage facility.
Freeze-drying equipment.
Molecular biology, esp. PCR, RFLP.
Serological techniques.
Immunofluorescence microscopy.
Stereomicroscopy.
Photography, Macro and Micro.
Antifungal drug susceptibility testing.
Dedicated Laboratory for Hazard group 3 fungi.
PC based record maintenance.
PC identification keys for some groups of fungi.
Comprehensive library of medical mycological literature.
Representative collection of histological preparations of mycotic diseases.

The National Collection of Plant Pathogenic Bacteria (NCPPB)

Contact for additional information: Dr D Stead

Equipment /Techniques

(Available via the Central Science Laboratory of which NCPPB is a part):
Bacterial identification by fatty acid profiling-MIDI MIS system.
We have a range of commercial libraries as well as self generated libraries based largely on the NCPPB strains- in total we have entries for over 1000 taxa
Identification by Protein profiling. This and the genetic fingerprinting are enhance by use of Gelcompar software( perhaps the best available)
Equipment and skills for almost all genetic fingerprinting technologies including:
RFLP by pulsed field gel electrophoresis
Amplified fragment length polymorphism
PCR - RFLP
REP - PCR(including rep,eric and BOX elements)
RAPDs
Detection of bacteria by PCR, serological techniques including:
Immunofluorescence
ELISA
Immunofluorescent colony staining
Immunomagnetic capture PCR
Traditional methods for identification including:
BIOLOG
Culture maintenance by lyophilisation and -80C storage. We also have a -150C freezer and liquid Nitrogen facilities.
State of the art Quarantine Glasshouses with full environmental control for use with alien pathogens. We advise MAFF on UK licensing for handling plant pathogens and are fully licenced ourselves.

Access to a range of electron microscope and mass spectrometer facilities within CSL.

The National Collection of Type Cultures (NCTC)

Contact for additional information: DR M. Costas or Dr B. Holmes

Equipment /Techniques:

Standard microbiolgical facilities.
Centrifugal Freeze-driers.
Shelf Freeze-drier
Liquid nitrogen storage facility.
Category III facility including freeze-drier.
e-mail and Internet links.

Additional relevant Equipment /Techniques available within CPHL:

Molecular genetics unit..
Electron microscopes.
Full range of chemical analytical equipment.
Within CPHL and PHLS access to specialist reference laboratories equipped to deal with specific groups of bacteria.

The National Collection of Wood Rotting Fungi (NCWRF)

Contact for additional information: Dr J.K. Carey

Equipment /Techniques:

Liquid Nitrogen storage facility.

Additional relevant Equipment /Techniques at the Buildings Research Establishment (BRE):

ELISA equipment.
Electron microscopes.
Full range of chemical analytical equipment.

The National Collection of Yeast Cultures (NCYC)

Contact for additional information: Dr I Roberts

Liquid Nitrogen storage facility.
Freeze drying equipment.
Computers + accounts software + database software + catalogue software + web server.
Microscopy - light and electron.
Microtitre plate reader
PCR equipment and general molecular biology equipment.
Automated DNA sequencer.
Florescence Activated Cell Sorter (less automated sorting).
Automated micromanipulator for spore dissection.