2.2.4. Studying of conditions of maintenance and storage of cultures M pachydermatis in laboratory conditions

In daily laboratory practice often there is a necessity of conservation of cultures of the originators allocated from a pathological material, for the further studying. It is known, that lipophilic yeast mushrooms of sort Malassezia are very whimsical from the point of view of them kultivirovanija in vitro [57].

Allocation of mushrooms of this sort in culture became possible only after their obligate requirement for sources of lipids (except for kind M pachydermatis) has been opened, that has allowed to develop corresponding nutrient mediums [105].

Safety of cultures M pachydermatis at various regimens of resowings on nutrient mediums

Tab. 9 № the strain Presence of growth of culture p ri various intervals between resowings 7 days 14 days 21 days 1 months 1,5 months 2 months 2,5 months S-a S-a S-a S-a S-a S-a S-a 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 12803 + + + + + + +, with + - +, With - - N/o N/o 15003 + + + + + + +, with + +, with +, With - +, with - - 15503 + + + + + + +, With + +, with +, With - - N/o N/o 16103 + + + + + + + + +, With + - +, With - - 16903 + + + + + + +, With + - +, with - - N/o N/o the Most accessible and widespread method of maintenance of cultures of mushrooms are periodic resowings on agarizovannyh mediums between which cultures are stored in a refrigeration cabinet at 5-8°С. The majority of yeast mushrooms keeps viability at 4-12°С and intervals of resowings 6-8 months [172]. However, at work with cultures M pachydermatis we (as well as other researchers) have faced that at storage they lose viability much faster, rather than other kinds of mushrooms. Thus, there was a necessity for definition of an optimum regimen of maintenance of cultures M pachydermatis. Results of definition of safety of cultures M pachydermatis at various intervals between resowings are presented to tab. 9.

1 2 3 4 5 7 8 9 10 11 12 13 14 15 11903 + + +, with +, with - - N/o N/o N/o N/o N/o N/o N/o N/o 13703 + + + + + + +, with + +, with +, with - - N/o N/o Notes: "s-a" — a mash-agar; — medium Barfatini; «+» — growth of culture in the form of a homogeneous lawn; «+, with» — weak growth of culture in the form of the isolated colonies; «-» - culture growth is not observed.

In work it has been used 7 epizooticheskih strains M pachydermatis, peresevavshihsja on a mash-agar and medium Barfatini in test tubes. Between resowings of a test tube with cultures were stored at 5-8°С in convolutions from a pergament paper. Resowings of cultures spent a smear, distributing an inoculum on all surface agarinic skosa, that provided homogeneous gazonnyj growth. Intervals between resowings made from 7 days to 3 months the Majority of the studied strains kept viability in a current 1,5 monthly periods of storage, however at resowing showed weak growth in the form of separate colonies. Thus one of strains M pachydermatis №6 remained teleorgánic only during 14 days Strains №№ 2 and 4 opposite, remained teleorgánic during 2-month's storage. Obviously, it is bound to individual physiological features of various strains, in particular, their ability to adapt for conditions out of an organism of an animal.

For long storage it is more preferable to use medium Barfatini, providing big viability M pachydermatis. Possibly, presence this medium of an olive oil not only optimum provides nutritious requirements of a mushroom, but also slows down drying of medium.

For short-term storage (7-21 days) cultures M pachydermatis it is quite suitable a mash-agar.

It is possible to conclude, that at maintenance of cultures M pachydermatis intervals between resowings should not exceed 1 month, and the most valuable strains expediently peresevat each 2 weeks. One more widespread method of storage of cultures of mushrooms is cryogenic storage with Glycerinum use in quality kriopro -

tektora [57]. We had been studied viability 7 strains M pachydermatis at storage under Glycerinum layer at temperature-16-18°S. It is established, that all strains remained teleorgánic within 3 months (observation term).

The method of lyophilic drying is widely applied to long long-term storage of cultures of mushrooms, however data on its use with reference to mushrooms of sort Malassezia are limited [70, 57].

Experiment on a lyophilization of cultures M pachydermatis has been spent on the basis of Department of tool methods of researches FGU VGNKI with

vmestno with managing department, JU.G.Oparin

In experiment two strains M pachydermatis №№ 12803 and 15003, of interest for further scientifically - research works have been used. 2 variants of protective medium were applied to a lyophilization: 1) protective medium №4 VGNKI (a zhelatino-peptone-saccharose); 2) milk with an inosite (5 %). The compounding of mediums has been offered JU.G.Oparin on the basis of positive takes of use of the given mediums at a lyophilization of cultures of mushrooms Candida albicans.

Biomaterial freezing, drying, pumping out and zapajka ampoules were carried out on standard technology. Before and after a lyophilization in cultures defined concentration of yeast cells (the general and teleorgánic) and presence of extraneous microorganisms. In ready ampoules defined residual humidity and vacuum presence. Upon termination of the control of an ampoule with cultures M pachydermatis have been put in pawn on storage at temperature 5-8°С. The general period of storage of ampoules has made 3,5 years. Upon termination of storage in ampoules defined concentration of teleorgánic yeast cells in comparison with initial indicators, studied to a cult - ralnye and morphological properties.

In the course of 3,5-year-old storage in liofilizirovannom a condition there was a sharp depression of viability at both strains M pachyderma - tis. Concentration of teleorgánic cells made only 2,5-5,0 % from the general concentration of yeast cells in ampoules while right after dryings this indicator made 57-69 %. The protective medium №4 VGNKI provided a little big viability of a mushroom, rather than milk about 5 % with an inosite. kulturalnye and morphological properties of strains for a storage time have not undergone visible changes.

Results of studying of viability of cultures M pachydermatis at storage in liofilizirovannom a condition are presented to tab. 10. Tab. 10 the Studied properties the Strain №15003 Strain № 12803 Z.s. №4 VGNKI Z.s. Milk - inosite Z.s. №4

VGNKI Z.s. Milk - an inosite the General concentration of cells, million/smz 1087,0 540,0 590,0 770,0 Initial concentration of teleorgánic cells, million/smz / % to the general 706,5/65,0 372,6 / 69,0 360,0/61,0 439,0 / 57,0 Concentration of teleorgánic cells after storage, million/smz / % to the general 55,0 / 5,0 12,5/2,3 15,0/2,5 19,0 / 2,4 Presence of vacuum in an ampoule Is vacuum There is a vacuum There is a vacuum There is a vacuum Presence of extraneous microorganisms are absent Are absent are absent Are absent kulturalnye and morphological properties Typical for M pachydermatis Typical for M pachydermatis Typical for M pachydermatis Typical for M pachydermatis 41. Changes of properties liofilizirovannyh cultures M pachydermatis at

Storage within 3,5 years

The received results have shown basic possibility is

Usings of lyophilic drying for long storage of cultures M pachydermatis, however for maintenance of larger viability, obviously, optimisation of conditions of a lyophilization is necessary.

<< | >>
A source: Ershov Peter Petrovich Etiologicheskaja. the importance of yeast mushrooms of sort Malassezia at dermal diseases of animals. The dissertation on competition of a scientific degree of the candidate of veterinary sciences Moscow - 2008. 2008

More on topic 2.2.4. Studying of conditions of maintenance and storage of cultures M pachydermatis in laboratory conditions:

  1. studying of ecological conditions and physiological pokazayotelej klarievyh the catfishes containing in akvarialnyh conditions by preparation for reproduction.
  2. Studying of cytomorphological properties clinical izoljatov M pachydermatis
  3. the Appendix № 12 Technique of an establishment of the reasons, conditions, circumstances and the accident mechanism in building, definitions of a circle of persons, in whose duties entered maintenance of safe working conditions
  4. 2.6. Laboratory researches of possibility doizvlechenija micro-residual oil after zavodnenija for conditions of layers of XIII horizon of deposit Uzen
  5. 3.4.2. Studying of a state of health of commodity fishes in conditions astayotichnogo and constant termorezhnmov in UZV.
  6. 2.2. The gear of selection of strategy of control in conditions neoprede - lennosti the future conditions of economic activities
  7. Change of the conditions of the employment contract defined by the parties for the reasons connected with change of organizational or technological working conditions
  8. features of state-legal maintenance of safety of traffic in the conditions of a wartime (1941-1945)
  9. § I Conditions (maintenance) of the international contract (a duty of the parties)
  10. § 2. Maintenance of conditions of correct realisation of the discretion in administrative activity of militia
  11. I. Maintenance of the political rights and freedom of citizens by creation of conditions for their safe realisation.
  12. § 1.1.1. The beginning of studying of medieval settlements in the USSR. Settlements saltovo-majatskoj cultures.
  13. 4.1. Formation of legal and organizational bases of maintenance of safety of traffic in the conditions of new Russian statehood
  14. the chapter I. The economic maintenance of the bank credit in the conditions of the market