Fastidious organisms having exacting nutritional requirement can be cultivated on infusion media.
Meat infusions were included in the first media used for the cultivation of bacteria.
Huntoon prepared a medium using fresh beef heart and peptone which became known as Heart Infusion Agar.
He demonstrated it could be used to support the growth of nutritionally fastidious microorganisms without the addition of enrichment, such as animal blood.
Heart Infusion Agar is a general-purpose growth medium. It is recommended for the cultivation of nutritionally fastidious microorganisms and as a basal medium with a variety of applications.
Tryptose and beef heart infusion provide nutritional requirements for the pathogenic bacteria.
Beef heart infusion supply nitrogenous compounds and amino acids necessary for the growth of nutritionally fastidious bacteria.
Sodium chloride provides essential electrolytes and maintains osmotic equilibrium.
Agar is a solidifying agent.
Preparation and Method of Use of Heart Infusion Agar
Suspend 40 grams in 1000 ml of distilled water.
Heat to boiling to dissolve the medium completely.
Sterilize by autoclaving at 15 lbs pressure (121ยฐC) for 15 minutes.
If desired 5% v/v sterile defibrinated blood may be added.
Mix well and dispense as desired.
Inoculate and streak the specimen as soon as possible after it is received in the laboratory.
If the material is being cultured directly from a swab, roll the swab over a small area of the agar surface and streak for isolation.
Incubate aerobically or in 5-10% CO2 at 33-37ยฐC for 18-24 hours.
Examine for typical colony morphology
Result Interpretation on Heart Infusion Agar
Organisms
Growth
Escherichia coli
luxuriant growth; beta hemolysis
Staphylococcus aureus
Good-luxuriant growth; beta hemolysis
Neisseria meningitides
Luxuriant growth; no hemolysis
Streptococcus pneumoniae
Good growth; alpha hemolysis
Streptococcus pyogenes
Good growth; alpha hemolysis; beta hemolysis
Uses of Heart Infusion Agar
It is recommended for use in qualitative procedures for isolation of a wide variety of fastidious microorganisms.
It can also be used for the cultivation of Vibrio and Streptococci species.
It is used for mass cultivation of bacteria required in vaccine preparation.
On the supplementation of blood, Heart Infusion Agar can be used to study hemolytic reactions.
It can also be supplemented with glucose, horse serum and antibiotics for the cultivation a wide variety of organisms.
This medium was used for isolation and enumeration of hemolytic Streptococci in milk.
The deep agar fill can be used as an oxygen gradient to determine the oxygen requirements of the inoculum by observing wherein the tube the growth is prominent.
Limitations
It is recommended that biochemical, immunological, molecular, or mass spectrometry testing be performed on colonies from pure culture for complete identification.
Due to the various nutritional requirements of some organisms, occasional isolates may be encountered which fail to grow or grow poorly on this medium.
Sagar Aryal is a microbiologist and a scientific blogger. He is doing his Ph.D. at the Central Department of Microbiology, Tribhuvan University, Kathmandu, Nepal. He was awarded the DAAD Research Grant to conduct part of his Ph.D. research work for two years (2019-2021) at Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Saarbrucken, Germany. Sagar is interested in research on actinobacteria, myxobacteria, and natural products. He is the Research Head of the Department of Natural Products, Kathmandu Research Institute for Biological Sciences (KRIBS), Lalitpur, Nepal. Sagar has more than ten years of experience in blogging, content writing, and SEO. Sagar was awarded the SfAM Communications Award 2015: Professional Communicator Category from the Society for Applied Microbiology (Now: Applied Microbiology International), Cambridge, United Kingdom (UK). Sagar is also the ASM Young Ambassador to Nepal for the American Society for Microbiology since 2023 onwards.