Campylo Thioglycollate HiVeg™ Medium Base

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SKU:
MV908
For isolation, maintenance and transport of cultures of Campylobacter species.


Intended Use

Recommended for maintenance, transport and storage of cultures of Campylobacter species.

Composition**

Ingredients g/L
HiVeg™ hydrolysate 20.000
Sodium chloride 2.500
Dipotassium hydrogen phosphate 1.500
Sodium thioglycollate 0.600
L-Cystine 0.400
Sodium sulphite 0.200
Agar 1.600
Final pH (at 25°C) 7.0±0.2

**Formula adjusted, standardized to suit performance parameters

Directions

Suspend 26.8 grams in 1000 ml purified/distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. To make the medium selective for Campylobacter species, add reconstituted contents of two vials of Campylobacter supplement-I (Blaser-Wang, FD006). Mix well and pour into sterile test tubes or dispense as desired.

Principle And Interpretation

Campylobacter infections occur sporadically in the summer months and usually follow ingestion of improperly handled or cooked food, primarily poultry products (1). Dekeyser et al (2) reported that Campylobacter jejuni could be isolated on a selective media supplemented with antimicrobials from the faeces of patients having diarrhea and gastroenteritis (by the filtration technique). The antimicrobials help to inhibit the normal enteric flora of faeces. Skirrow used a selective medium with three antimicrobials i. e. vancomycin, polymyxin B and trimethoprim. (3). Later on, Blaser et al isolated C.jejuni by direct inoculation of faeces sample on an agar medium containing four antibiotics (4,5). They also reported that C.jejuni could be isolated from faeces sample held at refrigeration temperature for duration of 8-10 hours in Thioglycollate Broth, incorporated with the four antibiotics (5). Blaser et al later included the fifth antibiotic cephalothin to inhibit non-pathogenic Campylobacter fetus (5). Campylo Thioglycollate Medium Base (with antibiotics) is generally used as a holding medium when immediate examination and testing of samples is not possible (6). Campylo Thioglycollate Medium Base is also recommended by APHA for maintenance, transport and storage of cultures of Campylobacter species (7). It is also used for enrichment of Campylobacter species from stool samples (1).

The medium contains necessary nutrients to promote growth of Campylobacter species. Moreover the supplement FD006 (Blaser-Wang) consists of five antibiotics viz. amphotericin B, cephalothin, polymyxin B, trimethoprim and vancomycin which inhibits multiplication of normal microbial flora in faecal specimens thus facilitating isolation of C.jejuni. Cephalothin may not always inhibit C.fetus species and some strains may grow at 42°C. Further tests should be performed to confirm C. jejuni. Campylo Thioglycollate HiVeg™ Medium Base is prepared by using vegetable peptones in place of animal based peptones which make the media free of BSE/ TSE risks.

Rectal swabs can be directly inoculated into the medium in tubes. About 5 drops of stool sample (prepare a saline suspension if stool is solid) can be placed on the medium about 1cm below the surface. Inoculated Campylo Thioglycollate Medium Base can be refrigerated and subcultured on Campylobacter Agar Base (M994) with Campylobacter Supplement-I (Blaser-Wang, FD006).

Type of specimen

Food and dairy samples

Specimen Collection and Handling:

For food and dairy samples, follow appropriate techniques for sample collection and processing as per guidelines (6,7). After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions :

Read the label before opening the container. Wear protective gloves/ protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations :

  • 1. Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
  • 2. It is generally used as a holding medium when immediate examination and testing of samples is not possible (9).

Performance and Evaluation

Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.

Quality Control

Appearance
Cream to yellow homogeneous free flowing powder

Gelling
Highly viscous solution comparable with 0.16% Agar gel.

Colour and Clarity of prepared medium
Light to medium amber coloured, very slightly opalescent solution

Reaction
Reaction of 2.68% w/v aqueous solution at 25°C. pH: 7.0±0.2

pH
6.80-7.20

Cultural Response

Cultural characteristics observed with added Blaser-Wang Selective Supplement (FD006) in an atmosphere of 5-15% O2 and 5-12% CO2 after an incubation at 42°C for 18-24 hours.

Organism Growth
Campylobacter coli ATCC 33559 good-luxuriant
Campylobacter jejuni ATCC 33291 (00005*) good-luxuriant
Escherichia coli ATCC 25922 (00013*) none-poor
Helicobacter pylori ATCC 43504 good-luxuriant

Key: *Corresponding WDCM numbers.

Storage and Shelf Life

Store between 10-30°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition. Seal the container tightly after use. Product performance is best if used within stated expiry period.

Disposal

User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow established laboratory procedures in disposing of infectious materials and material that comes into contact with sample must be decontaminated and disposed of in accordance with current laboratory techniques (8,9).

More Information
Product Name Campylo Thioglycollate HiVeg™ Medium Base
SKU MV908
Product Type HiVeg™
Physical Form Powder
Origin Animal Free (Veg)
Packaging type HDPE
References 1. Murray P. R., Baron E. J., Pfaller M. A., Tenover F. C., Yolken R. H ., (Eds.), 1995, Manual of Clinical Microbiology,6th Ed, ASM Press.2.Dekeyser, Gossuin-Detrain, Butzler and Sternan, 1972, J. Infect. Dis ., 125:390.3.Skirrow M. B., 1977, Br. Med. J., 2:9.4.Blaser, Cravens, Powers and Wang, 1978, Lancet, 2:979.5.Blaser et al, 1979, Ann. Intern. Med., 91:179.6.Reller, Wang and Blaser, 1979, ASCP check sample, Microbiology No.MB -99.Commission of Continuing Education,ASCP, Chicago.7.Speck M. L., (Ed.), 1984, Compendium of Methods for The Microbiological Examination of Foods, 2nd Ed., APHA,Washington D.C.
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