R

Technical Data
Hoyle Medium Base M015
Intended use
A highly selective medium used for the isolation and differentiation of Corynebacterium diphtheriae types from clinical
and non-clinical specimens.
Composition**
Ingredients g/L
Peptone 10.000
HM peptone B# 10.000
Sodium chloride 5.000
Agar 15.000
Final pH ( at 25°C) 7.8±0.2
**Formula adjusted, standardized to suit performance parameters
# - Equivalent to Beef extract
Directions
Suspend 40.0 grams in 940 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 and aseptically add 50 ml of laked blood
and 10 ml of PTe 3.5% Selective Supplement (1 ml per vial) (FD047). Mix well and pour into sterile Petri plates.
Principle And Interpretation
The most common disease caused by Corynebacterium diphtheriae is diphtheria, an acute communicable disease
manifested by both local infection of the upper respiratory tract and the systemic effects of the toxin, which are most
notable in the heart and peripheral nerves (1). Hoyle Medium Base, formulated by Hoyle (2), is the modification of the
original formulation of Neill, for the isolation and differentiation of C. diphtheriae. This medium is not inhibitory to some
mitis types of Corynebacterium , as the original formulation.
Peptone and HM peptone B supply carbon, nitrogen substances, amino acids, vitamins and other essential growth nutrients.
Potassium tellurite is a selective agent, which inhibits most of the normal flora of the upper respiratory tract except
Corynebacterium. Hoyle's Medium is a highly selective medium and should be used in conjunction with a non-selective
media such as Loeffler Serum Medium (M537) and Blood Agar Base (M089) with 10% horse blood (3). C.diphtheriae are
usually present in small numbers permitting the formation of well isolated colonies. So, inoculation is done by directly
rubbing the swab over the entire surface of the medium. Incubation should be carried out till 72 hours if the results are
negative. To study the morphology, gentian violet staining is done. To demonstrate the characteristic morphology and
staining reactions of C.diphtheriae by Neissers or Alberts method, it is advisable to use colonies from Loeffler Medium.
The toxigenicity of C.diphtheriae strains can be determined by Eleks (4) method.
Type of specimen
Clinical samples - Throat swab, nasal swab, wound swab
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,5).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions :
In Vitro diagnostic Use only. For professional use only. 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 clinical specimens. Safety guidelines may be referred in individual safety data sheets.
Limitations :
1.Each lot of the medium has been tested for the standard organisms specified on the COA. However, hospital isolates may
show variable growth depending on nutritional requirement.
2.Biochemical and serological tests must be carried out for confirmation.

Please refer disclaimer Overleaf.

 HiMedia Laboratories Technical Data

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
Firm, comparable with 1.5% Agar gel
Colour and Clarity of prepared medium
Basal Medium: Amber coloured, clear to slightly opalescent gel . After Addition of blood & Tellurite : Brownish
red coloured opaque gel forms in Petri plates
Reaction
Reaction of 4.0% w/v aqueous solution at 25°C. pH : 7.8±0.2
pH
7.60-8.00
Cultural Response
Cultural characteristics observed with added 50 ml of laked blood and tellurite solution after an incubation at 35-37°C for
18-24 hours .
Organism Inoculum Growth Recovery Colony
(CFU) characteristics

** Bacillus spizizenii >=104 inhibited 0%

ATCC 6633 (00003*)

C. diphtheriae type
50-100 good-luxuriant >=50% grey colonies
intermedius 14779 with darker
centers
Corynebacterium 50-100 good-luxuriant >=50% grey colonies
diphtheriae type mitis with shining
surface
Escherichia coli ATCC >=104 inhibited 0%
25922 (00013*)
Enterococcus faecalis 50-100 good-luxuriant >=50% black minute
ATCC 29212 (00087*) colonies
Key : (*) Corresponding WDCM numbers. **Formerly known as Bacillus subtilis subsp. spizizenii
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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (3,5).
Reference
1.Murray P. R., Baron J. H., Pfaller M. A., Jorgensen J. H. and Yolken R. H., 8th Ed., American Society for Microbiology,
Washington, D.C. (Ed.), 2003, Manual of Clinical Microbiology.
2.Hoyle I., 1941, Lancet., 1:175.
3.Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.
4.Elek S. D., 1948, Brit. Med. A1:493.Isenberg, H.D. Clinical Microbiology Procedures Handbook 2nd Edition.

Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data

5 Jorgensen, J.H., Pfaller, M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)
Manual of Clinical Microbiology, 11th Edition. Vol. 1.

Revision : 05/2024

HiMedia Laboratories Pvt. Limited, IVD In vitro diagnostic 30°C Storage temperature
Plot No.C-40, Road No.21Y, medical device
MIDC, Wagle Industrial Area,
Thane (W) -400604, MS, India 10°C

EC REP CEpartner4U, Esdoornlaan 13, Do not use if

CE Marking
3951DB Maarn, NL package is damaged
www.cepartner4u.eu

Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in this and
other related HiMedia™ publications. The information contained in this publication is based on our research and development work and is to the best
of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to specifications and information related
to the products at any time. Products are not intended for human or animal or therapeutic use but for laboratory,diagnostic, research or further
manufacturing use only, unless otherwise specified. Statements contained herein should not be considered as a warranty of any kind, expressed or
implied, and no liability is accepted for infringement of any patents.

HiMedia Laboratories Pvt. Ltd. Corporate Office : Plot No.C-40, Road No.21Y, MIDC, Wagle Industrial Area, Thane (W) - 400604, India.
Customer care No.: 022-6147 1919 Email: [email protected] Website: www.himedialabs.com