Histolytic and other intestinal amoebae can be grown in Book and Drbohlav’s diphasic medium for amoeba. It contains an egg slant with an overlay of sterile serum or liver extract. A lapful of sterile rice powder is added to this before inoculation with feces. The culture is incubated at 37° and subculture at 48 hour intervals. Amoeba can be demonstrated in the liquid phase of the culture.
Leis mania and Trypanosomes
Nosy, McNeal and Nicolle (NNN) medium is used for that cultivation of leis mania and trypanosomes. It consists of 2 parts of salt agar and 1 part of defibrinated rabbits blood. Selective isolation of either leishmanina or trypanosomes can be done with NNN medium.
ISOLATION OF FUNGI
Fungi can be isolated by growing in Seaboard’s glucose agar. It contains:
Peptone 40 g
Dextrose 20 g
in 100 ml of water, pH 5 .4
Addition of antibiotics to this medium prevents bacterial contamination. The other media used for growth and isolation of fungi are Czapek – Dox medium and Cornmeal agar.
ISOLATION OF VIRUSES
Viruses can be isolated by:
1. Animal inoculation
2. Inoculation into embrocated eggs.
3. Tissue culture.
1. Animal inoculation: Viruses can be isolated by inoculating them in animals like monkeys, mice, guinea pigs and rabbits. In the mice, inoculation can be done through several routes such as intracerebral, subcutaneous and intraperitoneal.
2. Embrocated eggs: In embrocated eggs, viruses can be inoculated in:
1. Chorioallantoic membrane (CAM)
2. Yolk sac
3. Amniotic sac
3. Tissue culture: Viruses can be cultivated in bits of tissues, organs or cell culture. The various types of cell cultures used are:
1. Primary cell culture
2. Diploid cell culture
3. Continuous cell lines