Cell-mediated immune responses in cattle vaccinated with killed Brucella melitensis strain H-38 vaccine or infected with viable Brucella abortus strain 2308 organisms, or both.
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Animals↗
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Titration of a brucella protein allergen in sheep sensitized with Brucella melitensis.
A protein allergen extracted from a rough strain of B. melitensis was titrated in sheep sensitized by vaccination with the attenuated B. melitensis strain Rev. I. Single doses of allergen were given per animal by the intradermal or palpebral routes. Doses over a 100-fold range produced delayed hypersensitivity reactions of similar magnitude by both routes. The most suitable method for application in the field was a dose of 50 mug given by the palpebral route (0.5 ml subcutaneously in the upper eyelid) with readings taken 48 hours after inoculation. Sheep vaccinated with H 38 adjuvant vaccine for 6 to 9 months showed positive reaction to the allergic test, whereas non-vaccinated non-infected sheep showed no reactions. No increase in agglutinations titers following allergic test was observed in either Rev. I or H 38 vaccinated sheep.
Allergens↗
Resistance against a subcutaneous Brucella challenge of mice immunized with living or dead Brucella or by transfer of immune serum.
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Animals↗
Report of the Subcommittee on Taxonomy of the Genus "Brucella". Speciation in the genus "Brucella".
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Amino Acids↗
[Aspects related to the antigenic structure and serological specificity of brucellae phylogenetically related to S- and R-forms and dissociated into R-variants].
The antigenic structures and serological properties of Brucellae S- and R-forms, phylogenetically differentiated, and dissociated R-variants are studied by means of gel-precipitation, immunoelectrophoresis and fixation of the complement. The Brucella suis 1330S, Brucella suis 1330R, Brucella ovis 02 and Brucella abortus 99 are involved in the experiments. Both specific and general antigenic structures are established in all strains studied, but only homologous antibodies from the antigen-antibody complex in the complement-fixation test, induced by the phylogenetically differentiated S- and R-forms (Brucella ovis) or brucellae, diverged into R-variants. Cross complement-fixation tests between the antisera, prepared against S-forms and antigens from natural R-forms (Brucella ovis) or dissociative R-forms (R-variants) have not been observed. Also cross reactions between antisera, induced by R-forms (natural R-forms--Brucella ovis, or dissociative R-variants) and antigens, obtained from S-forms of brucellae are found.
Antigen-Antibody Complex↗
[Carboyydrates localization on ultrathin sections of "Brucella" and "Escherichia" cells in smooth (S) and rough (R) phase (author's transl)].
Carbohydrates localization on ultrathin sections of Brucella abortus, melitensis and Excherichia coli cells has been studied by the periodic acid thiocarbohydrazide-silver proteinate (PATAg) and phosphotungstic acid (PTA) methods. Carbohydrates are mainly localized on the cell envelope of Brucella and Escherichia but there are several differences between these two bacteria. The differences are discussed and a Brucella polysaccharide envelope model is proposed. The PATAg method gives the same silver grain deposits on Brucella S and R and Escherichia S and R. The phosphotungstic acid method differenciates Brucella S and R cells by lack of contrast on the outer leaflet of the outer membrane of the latter, but does not differenciate E. coli S or R cells. The Brucella outer membrane contains less polysaccharides than that of E. coli. There is a seemingly symetric distribution of polysaccharide in the Brucella outer membrane as compared to an asymetric distribution in E. coli. The peptidoglycan of Brucella reacts strongly as compared with that of E. coli. The polysaccharides present a dispersed pattern in Brucella cytoplasm, whereas in E. coli they appear as dense, strongly reactive clusters very close to the cytoplasmic membrane.
Brucella↗
A solid phase assay with radioactively-labelled antibody for the detection of Brucella abortus.
An assay for the detection of Brucella abortus is described. IgG from rabbit antisera against live brucellae or brucella extracts was chemically linked to cellulose to form a solid phase reagent capable of binding brucella antigens present in buffer solutions or serum. After washing away any unbound material the presence of bound antigen was revealed by incubation with radioactively-labelled anti-brucella antibodies. The assay was capable of detecting less than 100 pg brucella antigen in a 20 microliter sample. Experiments in which IgG of non-related specificity was used in place of anti-brucella IgG showed that the test was specific. Normal human serum had only a slight inhibitory effect but anti-brucella antibodies were strongly inhibitory if present in the test sample. The extent of this effect and its relationship to antibody titre was investigated in 12 sera from brucellosis patients.
Antibodies, Bacterial↗
Resistance of normal or immunized guinea pigs against a subcutaneous challenge of Brucella abortus.
Guinea pigs were inoculated subcutaneously into a hind footpad with Brucella abortus. Unimmunized guinea pigs had little capacity for barring brucella from spreading to their spleens: the inoculum dose needed to infect 50% of the spleens one month after inoculation was about 10 brucella. When the size of the challenge inoculum reached approximately 5 x 10(3) ID50 a previous immunization with killed or virulent brucella: --did not prevent the challenge bacteria from reaching the lymph nodes near the inoculation site, but reduced the level of infection in these, --barred or reduced the spreading of brucella throughout the remainder of the reticuloendothelial system. Immunization with a small number of virulent brucella was more effective than immunization with a higher number of killed bacteria to produce such modifications of the repartition of challenge brucella.
Animals↗
[Fatty acid makeup of various Brucella species and its relationship to the culture medium].
Gas chromatographic method was applied to the study of the fatty acid composition (in Br. melitensis, Br. abortus, Br. suis, and Br. ovis strains. Fatty acid composition was similar in the mentioned brucellae species, except Br. suis No. 1330 significantly differing by this sign. Methyleneoctadecanoic acid content was considerably elevated, and that of octadecenoic -- reduced in brucellae grown on liver agar with the addition of serum and on meat-peptone agar in comparison with brucellae grown on liver agar; apparently this represents one of the mechanisms of the microorganism adaptation to the less favourable conditions of the nutrient medium. Passage of Br. ovis strain through the guinea pig organism led to the appearance of brucellae forming two types of colonies when grown on liver agar with the addition of serum. The fatty acid composition of brucellae forming small transparent colonies was the same as that of the initial culture with the prevalence of methyleneoctadecanoic acid; as to brucellae with larger colonies with irregular margin and nontransparent centre of the colony--octadecenoic acid prevailed in their fatty acid composition, i.e. their composition was similar to such in brucellae of the melitensis and abortus species grown on liver agar.
Brucella↗
The serological relationship between Brucella spp., Yersinia enterocolitica serotype IX and Salmonella serotypes of Kauffmann-White group N.
The serological relationship between Brucella spp., Yersinia enterocolitica IX, and the group N salmonella serotypes S. godesberg, S. landau, S. morehead, S. neusdorf, S. soerenga and S. urbana was examined using agglutination, antiglobulin, complement fixation, immunodiffusion and fluorescent antibody methods. Antisera to the group N salmonella serotypes all reacted to significant titres in agglutination and complement fixation, but not antiglobulin or immunodiffusion tests with smooth brucella antigens. These antisera also reacted in agglutination, but not antiglobulin, tests with Y. enterocolitica IX. They did not react significantly in any tests with rough brucella antigens. Conversely, antisera to smooth Brucella spp. agglutinated group N salmonellas to low titre and Y. enterocolitica IX to titres similar to those given against the homologous strain. Antiserum to Y. enterocolitica IX on the other hand reacted with smooth brucella antigens to high titre in agglutination, complement fixation and antiglobulin tests, and with the group N salmonella antigens to substantial titres in agglutination tests. In direct fluorescent antibody tests, smooth Brucella strains and Y. enterocolitica IX reacted strongly with FITC-labelled antibody to Br. abortus whereas the group N salmonella strains reacted weakly. In tests with monospecific antisera to the A and M determinants of Br. abortus and Br. melitensis respectively, Y. enterocolitica IX reacted only with the antiserum to the A determinant whereas group N salmonellas reacted to low titre with both A and M antisera. The results of cross-absorption tests confirmed this relationship and suggested that the O30 antigens of group N salmonella serotypes contained antigenic determinants similar to, but not identical with, the antigenic structure shared by smooth Brucella spp. and Y. enterocolitica IX.
Absorption↗
Enzyme-linked immunosorbent assay for bovine immunoglobulin subclass-specific response to Brucella abortus lipopolysaccharides.
An enzyme-linked immunosorbent assay was developed to follow the bovine response, by immunoglobulin class and subclass, to defined smooth and rough lipopolysaccharides (LPS) of Brucella abortus. Binding to smooth LPS of immunoglobulin G1 (IgG1) and IgG2 in sera from Brucella-infected animals was significantly greater than binding in sera from normal uninfected animals. Competition or steric blocking among IgM, IgG1, and IgG2 for binding sites on smooth LPS was shown to occur. Binding of IgM to Brucella smooth LPS with sera from uninfected animals was elevated above the assay control levels, and attempts to eliminate this nonspecific IgM binding were not successful. The same levels of nonspecific IgM binding were also seen with Brucella rough LPS, Escherichia coli LPS, and Pseudomonas solanacearum LPS. Sera from some, but not all, Brucella-infected animals showed elevated binding of IgG1 and IgM to both E. coli LPS and Brucella rough LPS as well as to Brucella smooth LPS. This was interpreted as specific antibody. Cross-reactions between B. abortus smooth or rough LPS and E. coli LPS could not be shown by immunodiffusion.
Animals↗
[Development of spleenic infection in the mouse following intravenous inoculation of various species of Brucella in smooth (S) or rough (R) phases].
The bacterial population in the spleen of mice inoculated intravenously by Brucella abortus 544 (S), Brucella suis 1330 (S), Brucella melitensis 115 (R) or Brucella canis RM 666(R) is studied kinetically. Like the S strains of Brucella, the R strains are able to multiply in the mouse spleen. The consequences for the tests of vaccines against natural infections produced by S or R strains of Brucella are discussed.
Animals↗
Response of high and low antibody producer to Brucella.
Biozzi's high and low lines of antibody producers have been applied to an analysis of murine in vivo responses to Brucella. High responders (H/Ab) are better producers of anti-Brucella antibodies and of Brucella induced interferon than low responders (L/Ab). There is no interstrain difference between H/Ab and L/Ab mice as regards to cutaneous hypersensitivity reactions to melitin. Non-immunized L/Ab mice are more resistant to infection with live Brucella than H/Ab. Immunization with formalin killed Brucella leads to a specific protection of both H/Ab and L/Ab mice but magnifies their inter-strain differences: L/Ab are much better protected by preimmunization than H/Ab. The difference between H/Ab and L/Ab mice is not related to an earlier clearance of intravenously inoculated bacteria from the blood stream but to later events affecting the balance between multiplication and digestion of the injected bacteria in the spleen. The lower resistance of H/Ab mice is not due to negative, facilitating-like, activities of immune antibodies: on the contrary, these are shown to have protective properties. These results are discussed in terms of their contribution to present knowledge on Biozzi's mice and their macrophagic functions, on the relations between interferon synthesis and the immune system, and on the mechanism of natural and post-vaccinal protection toward Brucella.
Animals↗
On the growth of Brucella species in presence of erythritol in defined and undefined media and amniotic fluid of human, cow and sheep.
Different concentrations of erythritol (25-1000 microng/ml) in defined medium were shown to enhance the growth of 75.00 to 91.66 per cent of Brucella abortus and 69.23 to 100.00 per cent of Brucella melitensis strains studied. Compared to these figures only 13.04 to 60.86% (in tryptose broth), and 41.40 to 62.50% (in glycerol dextrose broth), of Brucella abortus strains, 25.00 to 83.30 per cent (in tryptose broth) and 83.30 to 100.00 per cent (in glycerol dextrose broth) of Brucella melitensis strains showed increased growth over the control. 82.20 per cent of amniotic fluid samples from humans showed anti-brucella activity with lytic zones varying from 7-19 mm. Amniotic fluid from cows and sheep had no antibrucella activity.
Amniotic Fluid↗
Evolution and taxonomy in the genus Brucella: steroid hormone induction of filterable forms with altered characteristics after reversion.
Several strains of Brucella abortus, Brucella suis, and Brucella melitensis were exposed to physiologic concentrations of testosterone, progesterone, and diethylstilbestrol by incorporating them into the growth medium. The hormones induced Brucella to form cell wall-defective organisms, including filterable forms. The filterable forms from 1 strain of B melitensis had altered characteristics when it reverted to growth as an intact cell. This is the 1st reported instance of induction of filterable forms of Brucella by progesterone, testosterone, and diethylstilbestrol.
Bacteriophages↗
Isolation and properties of a phage lytic for non-smooth Brucella organisms.
A phage for non-smooth cells of Brucella abortus was isolated from a mixture of three brucella-phages incubated with rough brucella cells in the presence of N-methyl-N'-nitro-N-nitrosoguanidine. It did not lyse smooth cells of Br. abortus strains nor those of other Brucella species, but during the course of its replication in rough organisms a small proportion of phage mutants were produced which were similar in properties to smooth-specific phages. The rough-specific phage, R, itself resembled Weybridge phage in its morphological and serological properties. Phage R was found to contain DNA and segregated into high and low density fractions, both with plaque-forming activity, on ultra-centrifugation in CsSO4 gradients. Unlike the smooth-specific phages it attached to heat-labile receptors present on non-smooth brucella cells.
Adsorption↗
An immunological enhancement phenomenon in experimental brucella infection of the chicks.
The passive transfer of chicken anti-brucella immune globulins promote the survival of Brucella abortus in the spleen of chickens infected with this bacteria. This enhancing effect was reduced significantly when the immune globulins were absorbed with heat-killed brucella cells to remove all the anti-brucella agglutinins and immune adherence antibodies. Therefore, it is most probable that the enhancing effect is related to the presence of brucella antibodies.
Animals↗
Experimental studies on the serological relationships between Yersinia enterocolitica and Brucella abortus.
Yersinia enterocolitica O9 was shown to provoke O, OH and H antibody response in calves. Brucella abortus failed to generate Yersinia H antibody response and generated Brucella O antibodies that cross-reacted with Yersinia O and OH antigens. The presence of Yersinia H agglutinins along with a higher titre of Yersinia OH antibody than Brucella O antibody is suggestive of subclinical infection with yersiniosis rather than brucellosis. Cross-absorption of sera from calves with established Yersinia infections indicated that absorption of sera with Brucella O antigens, although completely removing Brucella O antibodies, failed to remove completely Yersinia O, OH and H antibodies, and thus provides an additional method of distinguishing between the two infections.
Agglutination Tests↗