G DETERMINATION OF TRICHOPHYTOCIDAL EFFICIENCY OF SOME DISINFECTION SOLUTIONS USING A SIMPLE ORIENTATION METHOD

Sisak M., A. Ry bnikaf : Determination of Trichophytocidal Efficiency of Some Disinfection Solutions Using a Simple Orientation Method. Acta vet. Brno, 49, 1980: 245--250. The antitrichophytocidal efficiency was tested of some generally used disinfection preparations and of Mykolastanox F a preparation used for local therapy of cattle trichophytosis. Tests were performed on 14 and 22-day-old sporulating cultures of Trichophyton verrucosum Bodin 1902. Of all the disinfection preparations the best results were reached with Persteri1. It was also found that formaldehyde at a concentration of 4 % had a strong fungicidal effect. The effect of the other disinfection preparations tested in the concentrations used was only fungistatic, or that they virtually did not affect the vitality of the T. verrucosum culture. As far as the effect of Mykolastanox F is concerned it was found that older cultures of T. verrucosum were more resistant. A trichophytocidal effect was reached after a 5-minute application of the preparation in a 0.3 % concentration, or after one minute using the preparation in a 0.5 % concentration. Trichophyton oerrucosum, disinfection, Mykolastanox F. Cattle trichophytosis is a dangerous disease contagious to man; its originator is the Trichophyton verrucosum fungus. The germ is very resistant to unfavourable conditions and can survive for a long time in pathological material outside the host organism (L adz ianska and Hal asa 1975). A number of preparations was proposed for therapy and disinfection the majority of which was insufficiently effective and considerably expensive. Hurnp oll ckova et a1. (1978) performed laboratory estimations of the efficiency of veterinary antimycotics and disinfestants. In the preparations only the fungistatic activity was determined whereas the fungicidal effectiveness was not estimated. Our experiments were based on studies of the authors mentioned above. However, tests of the effectiveness of disinfection preparations used commonly for disinfection in microbiological laboratories and of the Mykolastanox F preparation were performed using the immers-ingmethod, thus determining the fungistatic and fungicidal effect. ' Materials and Methods The antitrichophytocidal activity was determined in the following freshly prepared aqueous: solutions: Orthosan BF-12 (2 %, 5% and 10% concentrations), Ajatin (2%, 5% and 10%), Jo-donal B (5%, 10% and 20%), formaldehyde (1%, 2% and 4%), soda lye (1%, 5% and 10%),. and Persteril (1%, 2% and 4%). Segments of 14-day-old and 22-day-old Trichophyton verrUCOSU111 cultures were dipped into these solutions for one and five minutes (segment diameter was 5 mm). The culture of T. verrucosum was obtained through propagation on sweet wort agar at a tempera-ture of 28°C. At a given age the culture represented a group of hyphae with abundant microonidia and sporadically occurring macronidia and chlamydospores.

for a long time in pathological material outside the host organism (L adz ian s k a and Hal a sa 1975).A number of preparations was proposed for therapy and disinfection the majority of which was insufficiently effective and considerably expensive.Hurnp o ll ckova et a1.(1978) performed laboratory estimations of the efficiency of veterinary antimycotics and disinfestants.In the preparations only the fungistatic activity was determined whereas the fungicidal effectiveness was not estimated.Our experiments were based on studies of the authors mentioned above.However, tests of the effectiveness of disinfection preparations used commonly for disinfection in microbiological laboratories and of the Mykolastanox F preparation were performed using the immers-ingmethod, thus determining the fungistatic and fungicidal effect.'
After being exposed to the disinfection solutions the segments were rinsed several times in distilled water and transferred onto Petri dishes with Sabouraud agar.In the control experiment the segments were dipped into and rinsed with water in regular time intervals.Incubation was done in darkness at a temperature of 28°C.
In the second experiment tested were aqueous solutions of Mykolastanox F (in concentrations of 0.05%, 0.1 %, 0.2%, 0.3%, 0.4% and 0.5%) again using segments of 14-day and 22-day-old cultures of T. uerrucosum.The process was the same as in the preceding experiment, only the periods of exposure were different, i.e, one minute, 5 minutes, 10 minutes, 30 minutes and one hour.
Growth changes were assessed in both experiments in the course of 21 days of incubation macroscopically according to the rate of growth of the culture of T. uerrucosum in the experimental variant as compared with the control experiment.
Table 1 Effect of some disinfection solutions on the vitality of a 14-day-old culture of Trichophyton verrucosum Growth changes 5. 2. 21. 12. 9.

Results
The results of antitrichophytocidal efficiency of disinfection means are given in Tabs. 1 and 2. It was evident that the most effective of all the preparations tested was Persteril.After a five-minute application of this preparation all the concentrations showed a fungicidal effect.Even a 45 % concentration of formaldehyde had a fungicidal effect after a five-minute exposure.The effect of the other preparations tested was only fungistatic; the growth of the culture segments in the variants Jodonal 5 % and 10 %, Orthosan 2 0/0' and soda lye 1 % was approximately the same as in the control.When comparing the resistance of cultures of various age no considerable differences were found.
Tests of the Mykolastanox F solutions showed that older cultures (22 days) of T. verrucosum were more resistant.At a concentration of 0.5 % this substance had a trichophytocidal effect as soon as after one minute.When the concentration was reduced to 0.3 % and 0.2 % a fungicidal effect occurred within 5 minutes
-----iime of incubation--------- (days)   -----------'------------'--'---- and one hour, respectively.The effect of Mykolastanox F in concentrations of 0.1 % and 0.05 % was only fungistatic, there was no fungicidal effect even after one hour, as can be seen in Tabs. 3 and 4. Discussion Tkacik (1969) studied the effect of disinfection solutions on the Trichophyton verrucosum fungus in vitro.He reached the best results when using formalin and peroacetic acid.A 1 % concentration of formalin was effective for the fungus from the pathological material after a five-minute exposure.In the authors' experiments, using a cultural fungus form, a 1 %concentration of formalin caused only growth inhibition, whereas, macroscopically, growth was observed after 9 days of incubation.However, it is necessary to note that the authors studied the growth changes only up to 7 days.The same results were obtained when testing peroacetic acid; however, on the contrary, in the variants with soda lye in a concentration of 10% no fungicidal effects were observed in the authors' experiments as compared with those of Tkacik, As far as Persteril is concerned, very good results were obtained also in practice, Krupicka and Svec (1965) recommended a once-over local treatment of the affected cattle with peroacetic acid in concentrations from 1 0;0 to 4 %.A 1 % concentration of this substance was suitable for the disinfection of barns and equipment.Similar good results were obtained by the German authors Arculari us et al. (1976) who used peroacetic acid against cattle trichophytosis in the form of sprays.For therapeutic and prophylactic effects the most suitable was a 2 % concentration of this substance.The disadvantage of Persteril is its strong corrosive effect and its irritating effect on the eyes of animals and the eyes and hands of operators.
The results obtained 'with Mykolastanox F proved that its trichophytocidal properties were good.During the tests it was found than in 0.3 % and 0.5 % concentrations its effect was strongly fungicidal even when exposed for a very short time.The results of field estimations of the efficiency of Mykolastanox F were very good.With once-over applications of this substance Kovac et al. (1974) managed to cure cattle affected with trichophytosis in 90 %, with further treatment they reached 100 %.For prophylactic treatment the authors recommend the whole body of the animal to be sprayed with a 0.25 % solution of MykolastanoxF.Sisak et al. (1977) treated animals with Mykolastanox Fat a 0.1 % concentration applied in the form of a spray and with Mykolastanox F at a 0.5 % concentration applied on one third" of the animal's body.Application of this substance at a 0.5 0/0 concentration gave very good results, however, the application of 0.1 % Mykolastanox F was not effective enough.These results are in accordance with the results of the present study with tests of Mykolastanox F in vitro.
The disinfections preparations Orthosan, jodonal, and Aiatin, commonly used for disinfection in microbiology, proved to be insufficiently effective in the concentrations investigated.That is why the authors do not recommend them for disinfection against Trichophyton verrucosum germs.
F on the vitality of a 22-day-old culture of Trichophyton oerrucosum ---~-

Table 2 Effect of some disinfection solutions on the vitality of a 22-day..old culture of Trichophyton onrrucosum
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