Characterization by restriction endonuclease analysis and DNA hybridization using IS900 of bovine, ovine, caprine and human dependent strains of Mycobacterium paratuberculosis isolated in various localities.

DNA of 90 mycobactin-dependent strains of Mycobacterium paratuberculosis, isolated in 9 countries, was digested with restriction endonuclease PstI and hybridized with a DNA fragment containing insertion sequence IS900. Bovine strains (n = 73) were isolated from 61 animals in 17 herds, ovine strains (n = 15) from 13 animals in 3 herds and the set was completed by 1 caprine and 1 human (Linda) strain. Three types, tentatively designated A (n = 37), B (n = 51) and C (n = 2) were differentiated by restriction fragment length polymorphism (RFLP). Of the bovine strains 27, 45 and 1 were classified as belonging to types A, B and C, respectively; of the 15 ovine strains 10 and 5 belonged to types A and B, respectively; the caprine strain belonged to type C. The human strain Linda, isolated in the U.S.A., from a man with Crohn's disease, belonged to B type. A certain degree of type uniformity was observed among strains isolated within one herd in the course of several years, and the prevalence of a single type was also recorded within individual regions of the Czech Republic and Slovakia. Type A was identified in the course of 2 years in a sheep farm with frequent sales and purchases of animals, and type C was demonstrated in a goat kept in the same farm. Differences between RFLP types of the strains isolated from mesenteric lymph nodes and intestinal mucosa were found in one cow and one sheep. Selected strains of M. paratuberculosis RFLP type A (14 strains) and B (18 strains) were digested with restriction endonuclease BstEII. All the strains of A type were classified into C1 group and all the strains of B type into Cx group.

[Occurrence, economic significance and diagnosis of paratuberculosis]. / Výskyt, ekonomický význam a diagnostika paratuberkulózy.

Paratuberculosis is the most serious chronic bacterial disease of ruminants. Regarding the increasing incidence of the disease in the Czech Republic in connection with changing situation in agriculture and, among other factors, with import of animals we are presenting a summary of the latest data. We assume that high economic losses and expected participation of Mycobacterium paratuberculosis in Crohn's disease in men will evoke a broad interest in the disease among people from various branches of veterinary and human diagnosis.

[Allergic and serologic diagnosis of paratuberculosis in cattle]. / Alergická a sérologická diagnostika paratuberkulózy skotu.

The use of allergenodiagnosis in cattle herds where paratuberculosis is to be eradicated depends on the allergenogenic properties of the causative agent of this disease. An evaluation of the reliability of an intradermal allergic test performed to diagnose paratuberculosis in cattle in the given area revealed that this method is important especially if the result of this test is explicitly negative. A cultivation proof of the causative agent of paratuberculosis is of greatest importance among bacteriological methods for infection diagnosis. The agreement of the bacteriological and serological examination which reached 80% in young cattle pointed out the high specificity of an indirect haemagglutination test. In cows, unlike calves and heifers, the specificity of diagnostic tests was considerably lower. It is recommended to use an elimination method mainly in young cattle: regular examinations of calves from their age of three months will help to form a group of heifers free of infection, observing the basic infection controlling measures and providing conventient diet.

[Biologic characteristics of mycobacterium strains isolated from cattle from herds with clinical paratuberculosis]. / Biologické vlastnosti kmenu mykobaktérií izolovaných u skotu z chovu s klinickou paratuberkulózou.

In the period from 1983 to 1986, bacteriological examination for paratuberculosis was performed in 263 samples of lymph nodes, intestinal mucous membrane and excrements of cattle, kept on a farm where clinical paratuberculosis occurred. Seventy-nine strains of mycobacteria were isolated during the culturing. On selective agar medium with mycobactin as the growth stimulator, 71 strains were isolated which had failed to grow on the conventional mycobacterium-culturing media. In the subculture, the dependence of mycobacteria on the mycobactin declined and the number of mycobacterium strains growing in the subculture on conventional mycobacting-free media doubled. Two thirds of the mycobacteria which did not depend on mycobactin during growth exhibited the same antigenic properties as Mycobacterium avium 1, 2, 3, 8 during serotypification. Ability to induce sensibility to PPD avian tuberculin or paratuberculin was demonstrated during the bioassays of mycobactin. Almost a half of the strains inducing animals' sensitivity to the above-mentioned allergens were found to be virulent to pullets that had tuberculosis in their parenchymatous organs. Of the laboratory animals, the highest virulence of the mycobactin-dependent mycobacterium strains was demonstrated in mice subjected to intravenous infection, accompanied by hyperplasia of the spleen, with reisolation of the mycobacterium culture within six eight weeks after infection.

[Metabolic test in pigs with experimental mycobacteriosis fed different amounts of nutrients]. / Metabolický test u prasat pri experimentální mykobakterióze a odlisném príjmu zivin.

Three trials were conducted with pigs at different levels of nutrition. Seven groups of pigs were perorally infected with a suspension of M. intracellulare. Changes in 12 biochemical parameters of metabolism were studied in blood plasma or serum for 98 to 111 days. In comparison with the group of non-infected animals or with the situation prior to infection, the test pigs showed irregular and temporary changes in a number of metabolites, most frequently an increase in the concentration of urea, cholesterol, iron and vitamin A. In the majority of cases, the results are considered as non-typical of infection processes. It was also in relation to clinical and production parameters that the induced mycobacteriosis was regarded as a condition not disturbing, or just insignificantly influencing, the metabolism of slaughter pigs and the economy of their production. The changes in tocopherolaemia, reduction in alkaline phosphatase activity, increase in the total concentration of protein, albumin and cholesterol were related to the development of the pigs at the weight from 20 to 80 kg, rather than to the infection process. The limited intake of proteins or vitamin supplement allowed for verifying the possibility of using some biochemical parameters for practical diagnosis.

[The epizootiological significance of positive bacteriological findings on Mycobacterium tuberculosis and Mycobacterium bovis in humans]. / Epizootologický význam pozitivního bakteriologického nálezu Mycobacterium tuberculosis a Mycobacterium bovis u lidí.

The relation between the active form of tuberculosis in persons working in agriculture and incidence of tuberculosis in cattle was analyzed in 1974 to 1978, i.e. in the period after the elimination of bovine tuberculosis in Czechoslovakia (in 1968). M. tuberculosis was isolated in 15 cases and M. bovis in four cases of persons employed by the farms on which the Regional Hygienic Station, Brno, was responsible for the microbiological diagnostics of tuberculosis. Direct contact with animals was demonstrated in eight patients; M. tuberculosis was isolated from seven of these patients and M. bovis from one. Seven cattle herds were exposed to spontaneous infection by M. tuberculosis and in one of them tuberculosis was not demonstrated during complex examination. In three herds the examination revealed only a sensitivity of cattle to mammalian tuberculin. In other three herds tuberculosis was detected by allergic tests, patho-anatomic examination and bacteriological examination. M. tuberculosis in cattle was detected in two herds. The occurrence of bovine tuberculosis caused by a cattle tender with a positive finding of M. bovis in sputum was demonstrated in one herd. Virulence for the tested cattle was found in one strain (isolated from a mesenterial lymph node of cattle) of the four strains of M. tuberculosis used for the experimental infection of 17 animals. On the other hand, in three strains of M. tuberculosis, trials with experimental infection demonstrated only allergy to mammalian tuberculin and changes at the sites of subcutaneous inoculation of mycobacteria of regressive nature; these mostly disappeared within 90 days from infection.

[The effect of qualitatively different nutritional intakes on the development of experimentally induced atypical mycobacteriosis and the appearance of indicators in pigs]. / Vliv kvalitativne odlisného príjmu zivin na rozvoj experimentálne vyvolané atypické mykobakteriózy a na produkcní ukazatele u prasat.

A single infection with the culture of M. intracellulare at a rate of 0.1 mg per kg of weight in pigs fed the A 1 and SOL mixtures and the subsequent development of clinically confirmed mycobacteriosis did not exert any significant influence on the performance of the animals, as compared with the non-infected animals. The metabolically determined coefficients of nutrient digestibility in the SOL mixture determined in the animals on the 70th and 91st days from infection were almost the same as those in non-infected animals. The absence of animal protein and biofactor supplement in experimental mixtures adversely affected the performance of the pigs but exerted no unfavourable influence on the development of atypical mycobacteriosis after the administration of 0.01 mg or 0.1 mg of the culture of M. intracellulare per kg of body weight. No elimination of atypical mycobacteria in the faeces was demonstrated in the groups fed mixtures with biofactor supplement. The feeding of infected pigs with the COS 2 and A 1 mixtures, compared with the use of the experimental mixtures with a lower content of nitrogen compounds and vitamins and a higher content of fibre and energy, influenced the parameters of performance but did not result in a different patho-anatomic and bacteriological picture after mass-scale infection (1 milligram of culture per kg of weight).

[The importance of the infective dose, nutritional value of the feed and vitamin content for the susceptibility in pigs to Mycobacterium intracellulare]. / Význam infekcní dávky, nutricní hodnoty krmiva a obsahu vitamínu pro vnímavost prasat k mycobacterium intracellulare.

The influence of infectious dose and nutrition on the pig susceptibility to Mycobacterium intracellulare, serotype 4, 8, was studied with 48 pigs--crossbreds LW X X L--divided into four equal groups. All groups of pigs were fed ad libitum by the mixture consisting of maize, wheat, barley, soybean meal, salt lick and MKP 1 mineral feed supplement. Individual groups were given different levels of meals of animal origin and biofactor supplements containing vitamins A, D2, B2, B12 and niacin. Single per os administration of the infectious dose of Mycobacterium intracellulare amounted to 0.01 or 0.1 mg of mycobacterial culture per one kg of live weight. The extent of patho-anatomical findings depended on the infection intensity. The infectious dose of 0.01 mg of the culture resulted in the changes in the form of exudative lymphadenitis with characteristic caseous degeneration in lymphatic glands only in three out of 24 pigs, i. e. in 12.5%. A tenfold dose of mycobacteria caused caseous lesions in the mandibular and intestinal glands in 50% out of 22 infected animals. The result of complex examination of the lymph glands proves the favourable effect of biofactor supplement on the resistance in pigs, particularly those infected with lower mycobacterial doses. Mycobacteriosis was proved in all experimental pigs fed the mixture containing no biofactor supplement. In the group of animals fed the mixture with biofactor supplement the result was positive in 66.6% of infected animals. It was observed that the biofactor supplement also had a positive influence on active excretion of M. intracellulare by faeces. In pigs infected with high dose of mycobacteria this excretion was proved in 27.2% of animals. In the pigs fed the mixture containing biofactor supplement, regardless of the infectious dose and amount of animal proteins, no elimination of mycobacteria by faeces was observed. Out of 14 pigs with the microscopic finding of tuberculosis in the lymph glands, the reaction with an induration diameter larger than 10 mm was found in eight animals, i. e. 57.2%, after intradermal tuberculinization with purified avian tuberculin after 74 days from infection.

[Use of allergological diagnosis in the elimination of tuberculosis from poultry kept in a small flock]. / Uplatnení alergenodiagnostiky pri ozdravování tuberkulózou nakazené drubeze z malochovu.

The use of intradermal tuberculinization during the sanitation of TBC-infected small flocks of poultry with the application of the elimination method depends on the extent to which the flock is infected. The results suggest that the elimination method can be recommended for practical use in cases of 5% to 10% incidence of reactions to avian tuberculin. Thorough sanitation of the environment after the elimination of the infected birds is an essential part of the elimination method. When the incidence of reagents to tuberculin was up to 5%, two thirds of the small flocks were successfully sanitated by the elimination of positive reagents in a single operation. In flocks with a positive-reaction incidence from 5.1 to 10%, the elimination method was successful in 84.7% of the treated stocks; in some cases the tuberculinization had to be repeated three times before an infection-free state was obtained. When the rate of positive reactions was above 20%, better results were obtained from a radical method based on the elimination of the poultry and sanitation of the environment.

[Preventive and therapeutic efficacy of the vaccine against trychophytosis in cattle]. / Preventivní a lécebná úcinnost vakcíny proti trichofytóze u skotu.

The effectiveness of vaccine against dermatomycosis, caused by Trichophyton verrucosum, was studied in young cattle in three herds. The vaccines of Czechoslovak and Soviet production had almost the same effectiveness in prevention. In healthy calves without clinical signs of trichophytosis vaccinated at an age of one to three months with a dose of 2.5 to 5.0 ml, applied intramuscularly, and revaccinated with due respect to the rules of hygiene in the calf-houses, an incidence of 0 to 13% in the three- to five-month period of study was observed. In the non-vaccinated control groups, the incidence under the same infection conditions and in the same period was 36.3 to 100.0%. At the early stage of vaccination--during the inoculation of the animals suspected of being infected--the incidence of the disease was 50.0 to 68.8% and mostly showed no difference from incidence observed in the nonvaccinated control cattle. The therapeutic effect of the vaccine and Mykolastanox F is characterized by the fact that zero prevalence was obtained in the animals ten weeks after vaccination. Animals treated locally with Mykolastanox F showed zero prevalence 12 weeks after the beginning of treatment.

[Virulence of Mycobacterium avium and Mycobacterium intracellulare in pigs during experimental and spontaneous infection]. / Virulence Mycobacterium avium a Mycobacterium intracellulare pro prasata pri experimentální a spontánní infekci

The virulence of M. avium and M. intracellulare was studied in 16 experimentally infected pigs and on 174 animals coming from herds spontaneously infected with the mentioned Mycobacterium species. The result of the patho-anatomic examination and cultivation from samples of lymph nodes, organs, muscles, and excrements proved a higher virulence of M. avium to experimentally and spontaneously infected pigs, as compared with M. intracellulare. The strains of M. avium and M. intracellulare sensibilized all pigs to aviary tuberculin within 80 days from peroral and intranasal infection. Patho-anatomic changes in pigs spontaneously infected with M. avium were proved in 10% of the animals that showed a negative reaction to aviary tuberculin.