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A Rybnikár

Publications and source records attributed to A Rybnikár.

14 recordsLinked to original sources

Isolation of microbial DNA by newly designed magnetic particles.

Carboxyl group-containing magnetic nonporous poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) (P(HEMA-co-GMA)) microspheres and cobalt ferrite nanoparticles modified with alginic acid (natural carboxylic polysaccharide) were used for isolation of microbial DNA of lactic acid bacteria (LAB) from dairy products, lyophilised cell cultures, and bacterial colonies grown on hard media, and Trichophyton fungi DNA from lyophilised cells. DNA from the samples with lysed cells was reversibly adsorbed to the particles in the presence of high poly(ethylene glycol) (PEG 6000) and sodium chloride concentrations. The optimal final PEG and NaCl concentrations were 9.1 wt.% and 2.0 M, respectively. The adsorbed DNA was released from the particles in low ionic strength TE buffer. The quality of isolated DNA was checked by PCR amplification. Moreover, PCR amplicons were isolated on cobalt ferrite nanoparticles modified with alginic acid and checked by restriction analysis.

Bifidobacterium↗

Experimental dermatophytosis in calves caused by Trichophyton verrucosum culture.

Microconidia suspension of Trichophyton verrucosum virulent culture isolated originally from the mycotic lesions of cattle was inoculated epicutaneously to calves in various doses. Four application methods were selected, namely, onto the sheared and gently scarified skin, onto the sheared and non-scarified skin, onto the non-sheared and gently scarified skin, and onto the non-sheared and non-scarified skin. The inoculated area was 10 x 10 cm for all cases mentioned. When the application was performed onto the sheared and scarified skin, at least 18,000 microconidia of T. verrucosum per one animal had to be used to induce the mycotic disease in 100% of animals. This value was 10 times higher when the application was performed onto the non-sheared and scarified skin, 100 times higher in the case of the sheared and non-scarified skin and 1,000 times higher in the case of the non-sheared and non-scarified skin. The minimum infectious dose still capable of inducing dermatophytosis was 180 microconidia per 100 cm(2) of the scarified skin, 18,000 microconidia per 100 cm(2) of the sheared and non-scarified skin, and 180,000 microconidia per 100 cm(2) of the non-sheared and non-scarified skin.

Animals↗

[The Varix Set--instruments for varices surgery].

The authors describe new, modern instruments for the operation of varicosities, a VARIX set of their own design. The complex of instruments makes sparing and cosmetically effective strisping of the trunk of the large and small saphenous vein possible, as well as extirpation of collateral branches and epifascial severing of perforator veins. Operation with these instruments is easy, safe and sparing for surrounding tissues.

Humans↗

[The minimal effective dose of vaccine against trichophytosis in heifers].

Calves at the age of one month were vaccinated with a lyophilized vaccine against bovine trichophytosis, or with an avirulent vaccine against bovine trichophytosis (mfd by Bioveta, Ivanovice in Haná). Prophylactic doses of the vaccines (15 mil. CFU of production strain Trichophyton verrucosum per calf) were used for immunization, and doses 10 times, 100 times and 1000 times lower. The calves were revaccinated with the same doses in 12 days after the first vaccination. Twenty-eight days later since revaccination, the vaccinated calves and a group of control nonvaccinated calves was challenged epicutaneously with a virulent strain of T. verrucosum. The protectiveness of both vaccines implanted at doses of 2 x 15 mil. and 2 x 1.5 mil. CFU per test animal was very good. No dermal lesion were observed in the challenged calves of these groups, or if any, they were not clear and could be observed for a short time. If the vaccines were used diluted at a ratio 10(-2) (150 thousand CFU of production strain), trichophytic lesions persisting for the whole period of observation were found in four of the seven calves vaccinated with a lyophilized vaccine against bovine trichophytosis and in two of the eight calves implanted an avirulent vaccine after challenge. Mycotic lesions were formed after challenge in all test animals in the groups vaccinated with doses of 2 x 15 thousand CFU of production strain per calf. The extent of these lesions was practically the same as in all nonvaccinated controls--on the surface of infected skin the hair was shed and scales and crusts were formed. A challenge strain of T. verrucosum was cultivated from these lesions.

Animals↗

[Experimental trichophytosis in calves caused by cultures of Trichophyton verrucosum and Trichophyton equinum].

Tests were performed on 130 one-month-old calves. In 16 animals, treatment with a suspension of Trichophyton verrucosum culture at the dosage of 5 mil. conidia for an area of 10 x 10 of the clipped skin caused clinical trichophytic changes. The length of the incubation time was shorter in the calves scarified before inoculation of the infecting culture (10 to 13 days), as compared with the group of calves without scarification of the skin (11 to 19 days). Only 3 in 6 calves, infected by the same dose into non clipped skin, did contract trichophytosis, manifesting themselves as separate mycotic deposits, which appeared 26 to 35 days after inoculation. Trichophytic changes lasted longest in the group with scarification of the infected area. The ID50 in T. verrucosum and T. equinum cultures was about 1500 conidia per one calf in the case of the method of infecting into clipped scarified skin (area 100 sq cm). A 10 times lower dose (150 conidia per one calf) of the T. verrucosum suspension did not induce visible mycotic changes, but the T. equinum inoculation produced infection in one of 10 calves.

Animals↗

[The development of immunity after vaccination of cattle against trichophytosis].

Trials were conducted with 262 one-month-old calves. Good immunity was found to have developed within the period of 21 to 28 days after the administration of the Czechoslovak live vaccine against bovine trichophytosis. After epicutaneous inoculation of a highly virulent culture of Trichophyton verrucosum, performed 28 and 35 days after re-vaccination, no clinical form of trichophytosis was observed to rise in the calves. The same doses of challenge inoculum induced profound trichophytic changes. In the groups of calves artificially infected on the day of re-vaccination and seven days after re-vaccination, the proportions of experimental animals that fell ill were 88.2% and 44.4%, respectively; in the majority of cases, the course of the disease was not so serious as in the controls. Calves challenged 11 to 21 days after the second administration of the vaccines were mostly resistant to experimental infection: slight changes occurred in 15.4 to 7.0% of the tested animals.

Animals↗

[Lyophilization of dermatophytes].

When 16 dermatophyte strains were lyophilized, the highest survival was recorded in cultures where microconidia constituted a prevalent part. The viability was considerably reduced by lyophilization in the strains producing mainly macroconidia, chlamydospores and hyphae. A 2.5% solution of gelatine + 3.75% solution of saccharose proved to be an effective protecting lyophilization medium for the cultures of Trichophyton mentagrophytes and T. verrucosum to be lyophilized and maintained for two years. Very good results were also obtained when saccharose solutions (3.75, 7.5 and 15%) were used alone; however, an increased relative residual moisture content of the lyophilizates was recorded in some samples. Lyophilized cultures of dermatophytes were stored with success at refrigerator temperatures. When the lyophilizates were kept at laboratory temperature, the viability of the strains was reduced substantially irrespective of their being kept in the dark or exposed to light. The nutrient medium for the reconstitution of the lyophilized strains should be chosen on the basis of the growth requirements of individual cultures. In our trials the best results were obtained with the sweet wort media and the Sabouraud agar; the best media imported from foreign countries were the Mycological agar and Mycobiotic agar.

Freeze Drying↗

[Testing the protective effectiveness of avirulent vaccines against trichophytosis in cattle].

The protective action of live avirulent vaccine against bovine trichophytosis, produced by the Bioveta National Corporation at Ivanovice na Hané, was very good and fully comparable with the earlier vaccine produced from the virulent live culture of Trichophyton verrucosum. Experimental challenge with a massive dose of a virulent strain of T. verrucosum produced only minor surface dermal changes in the calves vaccinated with these vaccines, but these changes spontaneously disappeared in a short time. In the control non-vaccinated calves, large and deep lesions were induced by the challenge, spreading to the neighbouring tissues. Compared with both live biopreparations, the protective effect of inactivated vaccine prepared by the irradiation of virulent vaccine with Co60 was much lower.

Animals↗

[Disinfection for dermatomycoses in veterinary practice].

Sporulating cultures of two strains of Trichophyton equinum, two strains of Trich. mentagrophytes and one strain of Microsporum canis were tested by a simple dipping technique for their sensitivity to ten currently used disinfectants. Antifungal effects on all cultures of dermatophytes were observed already after one-minute treatment with 0.5%-4% peracetic acid, 4% formaldehyde solution and concentrated solution of Lastanox super. The above strains were inactivated after five-minute treatment with 1% and 2% formaldehyde solution, 20% Iodonal B solution and Ajatin solution with 10% active substance. Fungistatic effects of various intensity were observed in the other disinfectants.

Animals↗

Immunity in cattle vaccinated against ringworm.

Protective properties of a live, freeze-dried vaccine against ringworm, produced by Bioveta, Ivanovice na Hané, Czechoslovakia, were tested in a group of 422 calves. Vaccinated and control calves were challenged by epicutaneous inoculation of a virulent culture of Trichophyton verrucosum. Between 4.4 and 9.5% of calves challenged between days 14 and 25 after revaccination showed only mild clinical signs of ringworm and 99-100% were fully protected from day 28, the immunity persisting for at least one year. All control calves showed signs of ringworm after challenge. In most cases, extensive mycotic lesions, also penetrating into deep skin layers, were observed. Protective properties of the Czechoslovak vaccine and a USSR vaccine were high and comparable.

Animals↗

Long-term maintenance of lyophilized fungal cultures of the genera Epidermophyton, Microsporum, Paecilomyces and Trichophyton.

Twenty-six sporulating strains of lyophilized microscopic fungi of the genera Epidermophyton, Microsporum, Paecilomyces and Trichophyton have been maintained at 2-8 degrees C for 8-14 years. In 23 strains the number of viable particles reduced less than 10-fold during this period compared with immediately after lyophilization. The viability of other three investigated strains ranged from 3.4% to 7.1% in comparison with the original values.

Epidermophyton↗

Protective efficacy of vaccines against bovine dermatophytosis after double and single vaccination.

The protective efficacy of two Czech vaccines against bovine Trichophyton verrucosum infection after double vaccination of calves in prophylactic doses, with an interval of 13 days between vaccination and revaccination, was very good. When these preparations were applied in a double dose only once, less protective effect was produced. The use of 0.5% aluminium hydroxide to dilute the vaccines had no influence on the effect of single vaccination.

Animals↗