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M Królak

Publications and source records attributed to M Królak.

18 recordsLinked to original sources
PubMed1987

[Activity of porcine anti-Brucella abortus immunoglobulins in the acid plate agglutination test (APAT)].

Serological activity of swine IgM and IgG against Brucella abortus in RBPT was determined in relation to four other reactions used in Poland for diagnosing brucellosis standard agglutination test, complement fixation test, antiglobulin test, 2-mercaptoethanol test). Isolation of IgG was performed by the method of filtration on Sephadex gel G-200 of swine sera raised against Brucella abortus S19 by double immunization with suspension of killed bacteria. The presence of a certain Ig class in the fractions thus obtained was confirmed by immunoelectrophoresis and immunodiffusion tests. RBPT revealed the reaction of antibodies of IgM and IgG class which proves usability of this reaction diagnosis both early (IgM) and chronic (IgG) infection with brucellosis. Both classes of antibodies mentioned above were active also in SAT and CTT. Also the results obtained in AGT and MET were found interesting. In one of the sera, the absence of incomplete antibodies was observed, whereas positive reaction in antiglobulin test was found in its fractions containing IgG. This phenomenon was determined as concealment of incomplete agglutinins through higher level of complete antibodies in normal serum. In swine (the results were different from those obtained for cattle), apart from incomplete antibodies in IgG class, the presence of these agglutinins in IgM class was noted. On the other hand, the results obtained in MET proved that IgM antibodies of swine were not totally reduced when affected by 2-mercaptoethanol.

Agglutination Tests↗
PubMed1986

Distinct fixation of defined amounts of complement by the Brucella abortus antigen and bovine antibodies under different incubation conditions in the CFT.

Different times and temperatures of the 1st phase incubation of the complement fixation test (CFT) were used. It was found that a similar decrease in complement (C') activity occurred after 2 h at 37 degrees C, after 8 h at 20 degrees C and after 18 h at 4 degrees C. Simultaneously, in the same periods a double increase in standard serum titre at 4 and 20 degrees C was noted in comparison with titre at 37 degrees C. This increase of sensitivity of the CFT at 4 and 20 degrees C appeared to be a simulating one and resulted from the different pattern of reaction in each temperature in the block titration of antigen. The positive reactions in the range from the lowest to the highest dilutions of antigen were appearing and increasing and then disappearing in sequence at 4, 20 and 37 degrees C, creating the typical C' fixation area for each temperature. Similar results were obtained in four different C' fixation techniques, most frequently used in diagnosis of animal brucellosis. On the basis of our own examination results and literature data the warm and short C' fixation (37 degrees C, 30 min) was chosen in the standard technique of the CFT used in routine diagnostic survey for animal brucellosis in Poland.

Animals↗
PubMed1986

[Standardization of the complement fixation test (CFT) in brucellosis. I. The Polish standard of anti-Brucella abortus serum].

The purpose of this paper was to prepare an indigenous standard of anti-Brucella abortus serum for the complement fixation test (CFT) as a homologue of the secon International Standard of anti-Brucella abortus Serum (ISABS-II), which contains 1000 international units of complement-fixing antibodies in 1 cm3. The indigenous Standard of anti-Brucella Abortus Serum (ISABAS) was prepared in 1975. The material used was the serum of a cow infected with Br.abortus, biotype 1, under natural conditions. The cow was beheaded on the 54th day after abortion. The serum obtained was filtered through a Seitz EK filter and appropriately diluted with normal bovine serum. The preparation was lyophilized in 8500 ampules, each of which contained 1 cm3 of serum. They were filled with nitrogen before closing them. It was shown that ISABAS activity in CFT was approximate to that of ISABS-II. The studies in CVL, Weybridge showed that one ampule contained 1115 international units of complement-fixing antibodies. ISABAS titres in the agglutination test (AT) and antiglobulin test (AGT) were 320+ + and 2560+ + + respectively. Reduction with 2-mercaptoethanol and separation on a column with Sephadex G-200 gel showed that anti-Brucella antibodies occurred in ISABAS mainly in IgG class. ISABAS lyophilizate is characterized by stability of CFT titre, good solubility and a low anticomplementary activity. The average weight of the lyophilizate in an ampule is 79.57 mg, standard deviation - 0.36%. ISABS may serve as the basis of antigen standardization and other elements of indigenous CFT technique in brucellosis.

Animals↗
PubMed1986

[Standardization of the complement fixation test (CFT) in brucellosis. II. The antigen].

The activity of cellular antigens and those as aqueous extracts prepared from 4 laboratory strains of Brucella abortus: 99, S-19, 119-3 and "Krotz" has been compared in this paper. The purpose of the studies was to select an antigen of the highest activity and to test it by the Polish Standard of anti-Brucella abortus serum. It was shown that cellular antigens are more active in CFT than aqueous extract antigens of the same brucella strain. Cellular suspension of Br. abortus strain S-19 was chosen for production of the domestic antigen. A chessboard system was used in standardization of this antigen. In the working dilution of the antigen in CF such an amount of it was used that gave the titre 250++ according to the Polish standard of anti-Brucella abortus serum. The multiplier 4 (1000:250) was obtained which serves to calculate CFT titres in terms of international units of the complement fixing antibodies (icftu) in 1 cm3 of the serum studied. In examinations of all species of farm animals by the standard CFT technique, 10-20 icftu were assumed as a doubtful result and greater than or equal to 20 icftu as a positive result. A definite excess of the antigen used in working solution limits the prozone range and permits to carry out CFT in mass diagnostic examinations initially using one lowest serum dilution.

Animals↗
PubMed1986

[Standardization of the complement fixation test (CFT) in brucellosis. IV. Incubation conditions].

A differentiated time and temperature of incubation were used in the first CFT phase. It was found that the activity of complement fixation (C') decreased in the reaction medium. This was observed in the control system in which at 1 C'H50 hemolysis inhibition increase from 50 to 90% was found after 2 hr at 37 degrees C, after 8 hr at 20 degrees C and after 18 hr at 4 degrees C. Simultaneously, a double increase of the standard serum titre was obtained at 4 and 20 degrees C as compared with that at 37 degrees C. At stable concentration of the antigen and complement the effect of C' fixation was the strongest at 4 degrees C, slightly weaker at 20 degrees C and the weakest at 37 degrees C. The above dependence resulted from a different reaction distribution in the chessboard system at each temperature. In the order from the lowest to the highest antigen dilutions positive reactions were increasing and then subsequently expired at 4, 20 and 37 degrees C, forming an optimal antigen dilution typical for each temperature. In our own standard CFT technique a heat and short C' fixation (37 degrees C for 30 min) were chosen which provided the required reaction sensitivity and permitted to make mass diagnostic examinations. It was also shown that the temperature and time of initial incubation of the antigen-antibody complex had no influence on the effect of C' fixation by this complex. This is supported by joint introduction of the antigen and complement into sera.

Animals↗
PubMed1986

[Standardization of the complement fixation test (CFT) in brucellosis. III. The hemolytic system].

The purpose of this paper was to developed simple and accurate methods for standardization of the particular components of a hemolytic system and to choose optimal amounts of these components in CFT. On the basis of comparative studies and the data from literature the following procedure was chosen: 1. Sheep's blood was conserved with Alsever liquid and used after 5 days of stabilization. A 2% suspension of erythrocytes was standardized by centrifuging in calibrated test tubes at 1000 r for 10 min and spectrophotometrically, assuming the following criterion: the suspension of erythrocytes dissolved in distilled water at 1:10 ratio should show 0.500 extinction (E541), which corresponds to 5 X 10(8) erythrocytes in 1 cm3. 2. Lyophilized hemolysin was dissolved, conserved with glycerol and used after 5 days of stabilization. Hemolysin was titrated with 100% hemolysis in the presence of the excess of complement. The suspension of erythrocytes was sensitized with 5 units of hemolysin directly before using it. 3. Lyophilized complement (C) was dissolved, conserved with boric acid +K2SO4 containing liquid and used after 5 days of stabilization. Titration of C was done by 50% hemolysis; 3C'H50 was used in CFT. 4. A barbital buffer was used as diluent at pH 7.3-7.4, alternatively 0.85% NaCl with an addition of Ca and Mg at optimal concentration. The hemolytic system was incubated at 37 degrees C for 30 min.

Animals↗
PubMed1982

[Barbital buffer in tablets as a solvent for complement fixation test (CFT)].

The purpose of this paper was to develop a tableting technology of barbital buffer as a standard diluent easy in use for complement fixation test (CFT). As starting point, barbital (veronal) buffer after Alton et al. [1] was assumed. Among 4 solubilizers polyvinyl pyrrolidone (PVP 10 000) was chosen, which provided a good solubility of the barbital in distilled water at 20 degrees C and it affected favourably the complement activity (C'). PVP 10 000 added to the components of barbital buffer at an amount of 0.275 per 10 tablets, in the form of solution for granulation, provided the required physical properties of the tablets. As compared with the barbital buffer after Alton et al. [1], the tablets produced affected favourably C' activity, decreased the range of spontaneous haemocyte lysis, and they did not effect CFT titers of two anti-Brucella abortus standard sera as well as 77 sera of cattle infected with brucellosis under natural conditions. The described physical and biological properties of the barbital buffer tablets with PVP addition indicate their usability as diluent for CFT. A standard diluent used in the form of tablets in all Poland's laboratories should provide a better reproducibility of CFT results in diagnostic examinations towards brucellosis as well as other infectious diseases in men and animals.

Animals↗
PubMed1979

[Comparison of the values of selected reagents for the diagnosis of swine brucellosis. I. Preparation and standardization of antiglobulin serum].

The purpose of these studies was to make and to evaluate serums (SAG) helpful to the carrying out of OAG in diagnosis of pig brucellosis. Immunological serums directed against globulin fraction of pig serum were produced on rabbits immunizing them with "Suiglobin" preparation or with own preparation--"purified globulins" obtained from pig serum. Królak immunization method was used and in parallel that of Ressang's for the sake of comparison. Obtained SAG was titrated in the quick and slow OAG according to Hajdu. It was shown that rabbit immunization according to Ressang gives better results than that of Królak's. With the help of the immunoelectrophoresis reaction, in all SAG, the presence of the antibodies precipitating subfractions IgG and IgM of pig serum was found. Moreover, it was shown that to determine the final, working dilution of SAG, its titration was necessary in the slow, sample variant of OAG according to Hajdu.

Animals↗