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PubMed1978

Brucellosis in reindeer, Rangifer tarandus L., inoculated experimentally with Brucella suis, type 4.

An investigation of brucellosis caused by Brucella suis, type 4, in reindeer, Rangifer tarandus L., and other ungulates inoculated experimentally with virulent isolates was undertaken to observe the course of infection, follow titres of serum agglutins, and determine the extent to which intraspecific and interspecific transmission might occur among confined animals. Titres rose to maximum levels within 1 to 2 months following inoculation, decreased during the next 4 months, and persisted at low levels thereafter. Titres in uninoculated associates of these animals followed the same pattern, but at lower levels. Abortion occurred in reindeer that received large numbers of organisms; females receiving smaller numbers at an earlier stage of gestation produced normal fawns. Possible explanations for the holarctic distribution of B. suis, type 4, in reindeer are considered.

Animals↗
PubMed2026

Comprehensive genomic and computational insights into Brucella suis: pan-genome analysis, evolutionary perspectives, and in-silico vaccine design.

BACKGROUND: Brucella suis is a zoonotic intracellular pathogen responsible for brucellosis, mainly in swine and humans. Although numerous genome sequences are publicly available, an integrative genomic analysis combining pan-genome architecture, structural organization, evolutionary relationships, and vaccine-associated targets remains limited. RESULTS: In this study, we analyzed 91 publicly available B.suis genomes to characterize their pan-genome composition and genomic structure. The pan-genome exhibited an open configuration, indicating continued genomic diversification. A total of 2,146 core genes were identified, representing conserved functions essential for species maintenance, while the accessory genome reflected strain-level variability. Phylogenetic reconstruction based on single-copy orthologs revealed distinct evolutionary clades among the strains. A complementary phylogenetic analysis of pan-genome gene presence-absence patterns further supported clade differentiation and highlighted variation in accessory gene repertoires. Comparative synteny and genome structural analyses demonstrated largely conserved chromosomal organization with localized rearrangements across strains. Screening of the core proteome identified 64 putative antigenic proteins with predicted surface localization and immunogenic properties. Additionally, resistance-associated determinants related to tetracycline and doxycycline were detected in one genome within the dataset. CONCLUSIONS: This comprehensive genomic analysis defines the pan-genome structure, evolutionary relationships, and genome organization of B.suis. The integration of core and pan-genome-based phylogenies provides complementary insights into strain diversification, while the identified conserved antigenic candidates offer a foundation for future experimental validation and rational vaccine development strategies.

Genome, Bacterial↗
PubMed1978

Comparative studies on the ultrastructure of L forms obtained from S and R variants of Brucella suis 1330.

Electron-microscopic studies were carried out on S and R variants of Brucella suis 1330 and the production of L forms induced with the use of penicillin. No variations were established in the ultrastructure of the initial forms. However, essential differences were observed in the ultrastructure between the latter and the L-transformed Brucellae as well as changes that differentiated the ultrastructure of L forms obtained from S variants from that of L forms obtained from R variants. In both cases the L-transformed Brucella organisms have suffered substantial changes in their electron-microscopic structure. There was no cell wall, the cytoplasm was profoundly altered - chiefly the ribosomal apparatus, which, from a polyribosomal complex located peripherally in the cell, adjacent to the cytoplasmic membrane in the initial forms, has disintegrated into ribosomal units parts of which have often penetrated deeply in the nucleotide mass. Individual ribosomes remained connected with each other through DNA filaments, but there were equally cases in which links of such types were not present. The structure of the cytoplasmic membrane invariably showed three layers. The L forms obtained from R variants occasionally showed the presence of microtubules similar to those established with other microorganisms. Such structures were missing in the L forms obtained from S variants.

Brucella↗
PubMed1979

Brucella suis infection in pregnant cattle.

Six pregnant, Bos taurus cows with stages of gestation ranging from 11 to 33 weeks were each inoculated into the right conjunctival sac with 0.2 ml of a smooth culture of Brucella suis type I containing 27 x 10(6) viable organisms. The 6 cows produced 7 calves of which one single calf and one twin calf were stillborn, the cause of which was not determined. Br. suis was not isolated from any of the cows or calves using either special media or guinea pig inoculation. No abnormality was found in any of the cows or calves at autopsy. Microscopic examination of placentas and tissues from stillborn calves revealed no abnormality. Serologically, 2 weeks after inoculation all 6 cows had positive reactions to the Rose Bengal Test (RBT) and serum agglutination (SAT) titres of 25 iu to 116 iu. However, these reactions disappeared within 11 weeks. Only 2 cows had a complement fixation (CFT) titre which lasted a maximum of 5 weeks and reached a titre of 4/4. Following the anamnestic use of Br. abortus strain 45/20 vaccine on 3 of the cows, positive RBT reactions, SAT titres of 33 iu, 29 iu and 58 iu and CFT titres of 4/16, 1/8 and 3/8 respectively were recorded 6 weeks after vaccination.

Agglutination Tests↗
PubMed2026

First case of Brucella suis biovar 1 infection in a dog in Switzerland.

As one of the most common zoonoses globally, brucellosis threatens not only animals but also humans with Brucella (B.) melitensis, B. abortus and B. suis biovar 1&3, being more virulent for humans than B. canis, B. ovis and B. suis biovar 2. Canine brucellosis is caused mainly by B. canis, however, B. suis infections in dogs have been reported sporadically in Europe and more frequently in Australia. B. suis infection in dogs is mainly associated with hunting or wild-animal exposure and rarely with consumption of commercial raw meat products. In Switzerland, B. canis is sporadically diagnosed in dogs and B. suis biovar 2 is present in wild boar and brown hare. We report a case of brucellosis in a dog from Switzerland, neither having a history of hunting nor travel-associated risk of exposure. The intact male dog showed clinical signs (fever and epididymitis) consistent with brucellosis, which was confirmed by culture and molecular methods in urine and by serological methods. Culture and subsequent whole genome sequencing revealed the isolate as B. suis biovar 1, ST-14. The most closely related strain was shown to be a strain isolated in 2021 from a dog in Germany. Based on the lack of any previous report, infection by B. suis in dogs has not previously been diagnosed in Switzerland. This case highlights the need for vigilance regarding this less expected pathogen of high zoonotic importance in dogs.

Animals↗
PubMed1978

Identification of amino sugars from bacterial lipopolysaccharides by gas chromatography electron impact and chemical ionization mass spectrometry.

Amino sugars isolated from lipopolysaccharides of Brucella suis, Brucella abortus and Neisseria gonorrhoeae colony types 1 and 4 were identified using gas chromatography electron impact and chemical ionization mass spectrometry. Lipopolysaccharides were obtained by aqueous ether or aqueous phenol extraction. Isolated lipopolysaccharides were hydrolyzed in 1% acetic acid followed by hydrolysis of the polysaccharide moiety in 2 NHCl for 6 h at 100 degrees C. Amino sugars were first isolated by elution from Dowex 50 H+ and then N-acetylated, followed by trimethylsilylation. Trimethylsilyl ethers of 2-acetamido-2-deoxysugars; N-acetylglucosamine, N-acetylmannosamine, N-acetylgalactosamine, and a 2-acetamido-2.6-dideoxysugar, N-acetylquinovosamine, were identified by their fragmentation patterns. In the electron impact mode, N-acetylglucosamine and N-acetyl-galactosamine were distinguished from one another by comparing peak intensities at m/e 233 and 305. However, N-acetylglucosamine and N-acetylmannosamine could not be differentiated by electron impact mass spectrometry. In the chemical ionization mode, N-acetylglucosamine and N-acetylmannosamine both with base peaks at m/e 494, could be distinguished from N-acetylgalactosamine and N-acetylquinovosamine by their base peaks at m/e 420 and 332, respectively. N-Acetylglucosamine and N-acetylmannosamine were differentiated from one another by comparing peak intensities at m/e 330, 404, 420, and 510 [MH]+. This is the first report of chemical ionization mass spectrometry applied to the identification of amino sugars in bacterial lipopolysaccharides and shows that some 2-amino-2-deoxysugars can be differentiated by both electron impact and chemical ionization mass spectrometry.

Amino Sugars↗
PubMed1978

[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↗
PubMed1979

Immunologic properties of soluble materials from Brucella species.

The purpose in this study was to examine the immunogenic properties of various preparations of aqueous ether extracts of Brucella suis and Brucella canis. The B suis strain 3b and B canis strain RM-6-66 were grown on tryptose agar, and aqueous ether extracts were prepared from the cells. The ether was removed, and the extracts were clarified by centrifugation for 10 hours at 144,000 X g and fractionated by gel chromatography. The B suis endotoxin-containing precipitate, obtained from aqueous ether extracted material by ethanol precipitation, and fraction 1, prepared from ultracentrifugal supernate by column chromatography, protected mice against homologous infection. The B canis aqueous ether-extracted material also protected mice against B suis infections.

Animals↗
PubMed1979

Brucella-induced cholecystitis.

Acute cholecystitis is a rare complication of systemic brucellosis. This report details the occurrence of acute noncalculous cholecystitis in which Brucella suis was cultured from both the blood and the gallbladder. A discussion of the various diagnostic tests and the recommended therapy for brucellosis is included.

Acute Disease↗
PubMedDate not supplied

[Adherent cell and T lymphocyte cooperation for the "in vitro" production of Brucella-induced interferon by murine spleen cells (author's transl)].

Although Brucella is a good in vivo inducer of interferon, in vitro infection of murine spleen cells by Brucella suis has not, to the present time, led to in vitro synthesis. In the present work we show that normal spleen cells can however synthetize interferon in vitro when cultured together with adherent cells obtained from the spleens of syngeneic mice 45 min after in vivo inoculation. Induction and synthesis are thus shown to be distinct phenomena. Moreover soluble factors are shown to be involved in the induction phenomenon and T cells to be essential for synthesis. This in vitro brucella-induced interferon differs from in vivo brucella-induced interferon: its acid lability and its antigenic properties are characteristic of type II "immune" interferon.

Animals↗
PubMed1978

[Brucella titers in subclinical infections due to Yersinia enterocolitica serotype O:9 in a pig-breeding farm (author's transl)].

According to the Federal German Veterinary Public Health Regulations part of the stock of a pig-breeding farm in Lower Franconia (South Germany) had to be destroyed due to the findings of positive serological titers against Brucella. B. suis was not isolated from various specimens examined by the local veterinary public health authorities, but the authors found Yersinia enterocolitica serotype O:9 in 15 of 117 pigs investigated (12.8%). The organisms were cultured from 6 out of 117 specimens of fecal material (5.1%), and from 10 out of 55 pharyngeal tonsils (18.2%). The examination of 45 mesenteric and 2 submandibular lymphnodes remained negative. In 9 out of 93 sera titers against B. abortus between 1:20 and 1:40 were detected in tube agglutination but not in complement fixation tests. These sera and an additional serum specimen gave simultaneous reactions against Y. enterocolitica serotype O:9 ranging from 1:40 to 1:640 (Table 2). By cross-absorption antibodies to Brucella were completely removed, but to Yersinia titers between 1:10 and 1:320 remained detectable in 7 sera. It may be concluded that the Brucella titers observed in the animals were caused by subclinical infections with Y. enterocolitica serotype O:9 which is known to give rise to marked serological cross-reactions with various Brucella antigens. The importance of the above findings will be discussed in view of the current veterinary public health regulations.

Animals↗
PubMed1977

[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↗
PubMed1979

Use of an enzyme immunoassay test for characterizing the A and M antigens of Brucella.

An enzyme immunoassay test was developed for detecting the A and M antigens of brucellae. One hundred fifteen isolates, including 96 strains of Brucella abortus, 5 strains of B. suis, 1 strain of B. melitensis and 1 strain of B. neotomae, were accurately serotyped with the enzyme immunoassay test. No reactions were observed with 11 isolates of B. canis and 1 isolate of B. ovis. This rapid serological test provides for considerable savings in time and materials as compared with the standard tube agglutination test.

Agglutination Tests↗
PubMed1977

[Differentiation of Brucella by acrylamide-gel electrophoresis (author's transl)].

The strains of Brucella (Tab. 1) were grown on Tryptose-blood-agar at 37 degrees C, and the strains of Bordetella, Pasteurella and Actinobacillus on blood-agar. After 24 or 48 h they were harvested and extracted in phenol-acetic acid-water solution (4:2:1) (1 ml/50 mg bacterial wet weight) at 4 degrees C over 48 h. After centrifugation (8000 X g, 1 h) 2 volumes of the supernatant were mixed with 1 volume of a 40% sucrose solution in 35% acetic acid. Different volumes (0.20, 0.15 or 0.10 ml) of this were added to 7.5% acrylamide gel (2 ml in tubes 6 X 100 mm), containing 5 M urea and 12% acetic acid. The solution in both electrode chambers was 10% acetic acid. During the first 15 min it was focused with 2 mA/tube, then it was separated for 3 or 5 h with 4 mA/tube. The protein bands were stained with amido black 10B. All species could be exactly differentiated from each other, but the protein bands of biotypes of Bruc. suis, Bruc. melitensis and Bruc. abortus were identical in each case. All investigated strains of Bruc. canis were identical too. A small relationship was noticed between Bruc. canis and Bruc. suis and between Bordetella bronchiseptica and Bruc. canis respectively Bruc. suis strains.

Bacterial Proteins↗
PubMed1978

Brucella cultures typed by the Who Brucellosis Centre at the Commonwealth Serum Laboratories, Melbourne.

Results of the typing of Brucella cultures received at the WHO Brucellosis Centre, CSL, Melbourne, from 1968-1976 are presented. The distribution of the biotypes of cultures recovered throughout Australia is shown on a host and State basis and atypical cultures are discussed. Cultures identified from Australia were Br. abortus, biotypes 1, 2 and 4 and Strain 19; Br. suis, biotype 1 and Br. ovis. Br. melitensis biotypes 1, 2 and 3 were recorded only as exotic human infections from the Mediterranean area and from laboratory infections. Br. abortus, biotype 1 was the most common bovine and human isolate and was found in horses, a goat and a sheep. There was a low incidence of Br. abortus, biotype 2 in cattle and it was found in 1 horse. Br. abortus, biotype 3 was not found in Australia but was submitted from Tanzania. Br. abortus, biotype 4 was rare in cattle and was found once in a horse. Br. abortus, Strain 19 was found occasionally in cattle and once from a test guinea pig. Br. suis, biotype 1 was found in both man and pigs in Queensland and New South Wales whereas biotype 3 was isolated only in New Guinea from pigs and cattle. Br. ovis was submitted from 3 States. Br. canis has not been found in Australia.

Animals↗