THE HISTORY OF SEARCH ING METHODS OF SELECTIVE NUTRIENT MEDIA PREPARATION FOR MYCOPLASMAS CULTIVATION

Abstract

The results’ analysis obtained in searching of selective nutrient mycoplasmal media (SNMM) for the
period of 120 years and published by several generations of scientists from different countries of the world,
including the authors of this article, is presented. Mycoplasma, the causative  agent of contagious cattle
pleuropneumonia, was first cultivated by E. Nocard, E. R. Roux et al. (1898) in beef – extract broth (BEB)
that was embedded in a collodion bag and sewn into the rabbit peritoneal cavity. The first variants of the
medium were based on BEB or on Marten’s broth and the blood  serum of animals, mainly horses.
D. G. Edward (1947) suggested to add yeast extract to the media. R. M. Chanock et al. (1962) introduced the
horse blood serum, yeast extract, lactalbumin hydrolyzate and DNA into the medium. It was the first time
that they had cultivated the causative agent of the so –called human atypical pneumonia, Eaton agent, which
was considered a virus. It was a powerful impetus to further studying mycoplasma properties and features of
the diseases caused by them −  mycoplasmosis of humans, animals, birds, reptiles, fish, arthropods,  and
plants.  A number of  methods have been suggested to diagnoze mycoplasmoses: from microbiological
(mycoplasmological, cultural) to genetic and molecular types of polymerase chain reaction (PCR). But only
mycoplasmic  is  the basic  method  –“gold standard”, all the rest are additional. Only with its help it is
possible to obtain mycoplasma  cultures, to study their biological properties, to make diagnostics and
vaccines from them. With this purpose, it is necessary to have SNMM, that would provide mycoplasmas with
plastic material and energy, and to know the conditions for their successful cultivation. M. penetrans culture
was isolated by S.C.Lo et al. (1991, 1992) from human urine samples infected with the immunodeficiency
virus, 29 years after the identification of the Eaton agent. It has led to the thought that, although more than
200 species of Mollicutes  class have been cultivated at present, they are only part of the total number
existing in nature. Therefore, their search in different hosts should be continued, but using more improved
SNMM. Several hundreds have been tested with this aim and it has been concluded that it is impossible to
produce a universal medium for all the species described, because they have different nutritional and
cultivation conditions  and requirements. Certain media meet the requirements  of a few species only.
Phytoplasma for Haemobartonella and Eperythrozoon cannot be cultivated in vitro so far.