The Nature of pig antibodies againstBrucella suis

1. (1)
   лзучапосъ распрелеление аггплютининов цротив Brucella suis B γ-и β-глобулиновой ?ракпии иммунной свиой сыоротки. лммунохимический анализ показал, что γ-глобулиновый антиген присутствует и в β-, а частью лаже и в α-глобулиновой ?раккпии.     
   
   

Study of antibody formation by the use of metabolic inhibitors

1. (1)
   у взрослых особей реакция на бактериальный антигеп имеет характер преимущественно вторичной реакции, а 6-меркаптопурин не окаэывает на нее влияния даже в больших дозах. Поэтому мы исследовали вопрос, влияет ли 6-меркаптопурип па реакцию обраэоваторых он выэывает первичную реакцию. Мы установили, что дозы в 0,3 и 0,5 мг/100 г веса тела выэывают полное торможение реакции обраэовання антител после влрыскивания антигенов Salmonella paratyphi B и Brucella suis.     
   
   

Isolation of a novel ‘atypical’ <Emphasis Type="Italic">Brucella</Emphasis> strain from a bluespotted ribbontail ray (<Emphasis Type="Italic">Taeniura lymma</Emphasis>)

A pleomorphic Gram-negative, motile coccobacillus was isolated from the gills of a wild-caught bluespotted ribbontail ray after its sudden death during quarantine. Strain 141012304 was observed to grow aerobically, to be clearly positive for cytochrome oxidase, catalase, urease and was initially identified as “Brucella melitensis” or “Ochrobactrum anthropi” by Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and VITEK2-compact^®, respectively. Affiliation to the genus Brucella was confirmed by bcsp31 and IS711 PCR as well as by Brucella species-specific multiplex PCR, therein displaying a characteristic banding pattern recently described for Brucella strains obtained from amphibian hosts. Likewise, based on recA sequencing, strain 141012304 was found to form a separate lineage, within the so called ‘atypical’ Brucella, consisting of genetically more distantly related strains. The closest similarity was detected to brucellae, which have recently been isolated from edible bull frogs. Subsequent next generation genome sequencing and phylogenetic analysis confirmed that the ray strain represents a novel Brucella lineage within the atypical group of Brucella and in vicinity to Brucella inopinata and Brucella strain BO2, both isolated from human patients. This is the first report of a natural Brucella infection in a saltwater fish extending the host range of this medically important genus.     

Cultivation of mesenchymal tissue with reference to antibody formationin vitro

The optimal conditions for the cultivation of rabbit lymphocytes in suspension and antibody formation by these cellsin vitro were studied. The lymphocytes were cultivated in medium in which serum was replaced by serum albumin containing the alpha globulin fraction. Several media were used, with different amounts and proportions of amino acids, vitamins and glucose. The cells survived best in medium in which a component with a high molecular weight was present and in which the glutamine concentration was 0.5–1 mM and the amount of glucose was 0.4–0.5%. Among the other factors determined, the initial pH had a significant influence on survival of the tissue, the optimal limits being 6.9–7.2. Under these conditions 55–60% of the cells survived during 120 hours’ cultivation. The conditions and factors influencing these findings are discussed. Study of the formation of antibodies againstBrucella suis in vitro showed that antibody formation increased only up to 48 hours, after which it remained at the same level.     

The <Emphasis Type="Italic">yajC</Emphasis> gene from <Emphasis Type="Italic">Lactobacillus buchneri</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> and its role in ethanol tolerance

The yajC gene (Lbuc_0921) from Lactobacillus buchneri NRRL B-30929 was identified from previous proteomics analyses in response to ethanol treatment. The YajC protein expression was increased by 15-fold in response to 10 % ethanol vs 0 % ethanol. The yajC gene encodes the smaller subunit of the preprotein translocase complex, which interacts with membrane protein SecD and SecF to coordinate protein transport and secretion across cytoplasmic membrane in Escherichia coli. The YajC protein was linked to sensitivity to growth temperatures in E. coli, involved in translocation of virulence factors during Listeria infection, and stimulating a T cell-mediated response of Brucella abortus. In this study, the L. buchneri yajC gene was over-expressed in E. coli. The strain carrying pET28byajC that produces YajC after isopropyl β-d-1-thiogalactopyranoside induction showed tolerance to 4 % ethanol in growth media, compared to the control carrying pET28b. This is the first report linking YajC to ethanol stress and tolerance.     

Brucella suis vaccine strain S2-infected immortalized caprine endometrial epithelial cell lines induce non-apoptotic ER-stress

Brucella, which is regarded as an intracellular pathogen responsible for a zoonotic disease called brucellosis, survives and proliferates within several types of phagocytic and non-phagocytic cells. Brucella infects not only their preferred hosts but also other domestic and wild animal species, inducing abortion and infertility. Therefore, the interaction between uterine cells and Brucella is important for understanding the pathogenesis of this disease. In this study, we describe the Brucella suis vaccine strain S2 (B.suis.S2) infection and replication in the immortalized caprine endometrial epithelial cell line hTERT-EECs and the induced cellular and molecular response modulation in vitro. We found that B.suis S2 was able to infect and replicate to high titers and inhibit the proliferation of EECs and induce non-apoptotic pathways, as determined by B.suis.S2 detection using MTT and acridine orange/ethidium bromide (AO/EB) staining and flow cytometry. We explored the evidence of non-apoptotic pathways using real-time quantitative RT-PCR and by western blot analysis. Finally, we discovered the over-expression of GRP78, ATF4, ATF6, PERK, eIF2α, CHOP, and cytochrome c (Cyt-c) but not IRE1, xbp-1, and caspase-3 in B.suis.S2 (HK)-attacked and B.suis.S2-infected cells, suggesting that the molecular mechanism of ER stress sensor activation by B.suis.S2 is basically concomitant with that by B.suis.S2 (HK) and that ER stress, especially the PERK pathway, plays an important role in the process of B.suis.S2 infecting EEC, which may, in part, explain the role of the uterus in the pathogenesis of B.suis.S2.

Electronic supplementary material

The online version of this article (doi:10.1007/s12192-014-0564-x) contains supplementary material, which is available to authorized users.     

A multicenter point-prevalence study: antimicrobial prescription frequencies in hospitalized patients in turkey

Background

Of the diverse presentation of neurobrucellosis, intra-medullary spinal cord abscess is extremely rare. Only four other cases have been reported so far. We present a case of spinal cord intra-medullary abscess due to Brucella melitensis.

Case presentation

A forty-year-old female presented with progressive weakness of both lower limb with urinary incontinence of 6 months duration. She was febrile. Neurological examination revealed flaccid areflexic paraplegia with T₁₀ below sensory impairment including perianal region. An intramedullary mass was diagnosed on Magnetic Resonance Image (MRI) scan extending from T₁₂ to L₂. At surgery, a large abscess was encountered at the conus medullaris, from which Brucella melitensis was grown on culture. She was started on streptomycin and doxycycline for 1 month, followed by rifampicin and doxycycline for 1 month. At 2-year follow-up, she had recovered only partially and continued to have impaired bladder function.

Conclusion

Neurobrucellosis, if not treated early, can result in severe neurological morbidity and sequale, which may be irreversible. Hence it is important to consider the possibility of neurobrucellosis in endemic region and treat aggressively.     

Brucellosis caused by the wood rat pathogen <Emphasis Type="Italic">Brucella neotomae:</Emphasis> two case reports

Background

Brucellosis is a chronic bacterial disease caused by members of the genus Brucella. Among the classical species stands Brucella neotomae, until now, a pathogen limited to wood rats. However, we have identified two brucellosis human cases caused by B. neotomae, demonstrating that this species has zoonotic potential.

Cases presentation

Within almost 4 years of each other, a 64-year-old Costa Rican white Hispanic man and a 51-year-old Costa Rican white Hispanic man required medical care at public hospitals of Costa Rica. Their hematological and biochemical parameters were within normal limits. No adenopathies or visceral abnormalities were found. Both patients showed intermittent fever, disorientation, and general malaise and a positive Rose Bengal test compatible with Brucella infection. Blood and cerebrospinal fluid cultures rendered Gram-negative coccobacilli identified by genomic analysis as B. neotomae. After antibiotic treatment, the patients recovered with normal mental activities.

Conclusions

This is the first report describing in detail the clinical disease caused by B. neotomae in two unrelated patients. In spite of previous claims, this bacterium keeps zoonotic potential. Proposals to generate vaccines by using B. neotomae as an immunogen must be reexamined and countries housing the natural reservoir must consider the zoonotic risk.

     

Construction of an expression plasmid (vector) encoding Brucella melitensis outer membrane protein,a candidate for DNA vaccine

Background:

DNA vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. Bacteria of the genus Brucella are facultative intracellular pathogens for which new and efficient vaccines are needed.

Methods:

To evaluate the use of a DNA immunization strategy for protection against brucellosis, a plasmid containing the DNA encoding the Brucella melitensis (B. melitensis) 31 kDa outer membrane protein, as a potent immunogenic target, was constructed.

Results:

The constructed plasmid, pcDNA3.1+omp31, was injected intramuscularly into mice and the expression of omp31 RNA was assessed by RT-PCR. The integrity of the pcDNA3.1+omp31 construct was confirmed with restriction analysis and sequencing. Omp31 mRNA expression was verified by RT-PCR.

Conclusion:

Our results indicate that the pcDNA3.1+omp31 eukaryotic expression vector expresses omp31 mRNA and could be useful as a vaccine candidate.Key Words: Brucella melitensis, omp31, DNA Vaccine, pcDNA3.1     

Deficiency of the BMP Type I receptor ALK3 partly protects mice from anemia of inflammation

Background

Inflammatory stimuli induce the hepatic iron regulatory hormone hepcidin, which contributes to anaemia of inflammation (AI). Hepcidin expression is regulated by the bone morphogenetic protein (BMP) and the interleukin-6 (IL-6) signalling pathways. Prior results indicate that the BMP type I receptor ALK3 is mainly involved in the acute inflammatory hepcidin induction four and 72 h after IL-6 administration. In this study, the role of ALK3 in a chronic model of inflammation was investigated. The intact, heat-killed bacterium Brucella abortus (BA) was used to analyse its effect on the development of inflammation and hypoferremia in mice with hepatocyte-specific Alk3-deficiency (Alk3^fl/fl; Alb-Cre) compared to control (Alk3^fl/fl) mice.

Results

An iron restricted diet prevented development of the iron overload phenotype in mice with hepatocyte-specific Alk3 deficiency. Regular diet leads to iron overload and increased haemoglobin levels in these mice, which protects from the development of AI per se. Fourteen days after BA injection Alk3^fl/fl; Alb-Cre mice presented milder anaemia (Hb 16.7 g/dl to 11.6 g/dl) compared to Alk3^fl/fl control mice (Hb 14.9 g/dl to 8.6 g/dl). BA injection led to an intact inflammatory response in all groups of mice. In Alk3^fl/fl; Alb-Cre mice, SMAD1/5/8 phosphorylation was reduced after BA as well as after infection with Staphylococcus aureus. The reduction of the SMAD1/5/8 signalling pathway due to hepatocyte-specific Alk3 deficiency partly suppressed the induction of STAT3 signalling.

Conclusion

The results reveal in vivo, that 1) hepatocyte-specific Alk3 deficiency partly protects from AI, 2) the development of hypoferremia is partly dependent on ALK3, and 3) the ALK3/BMP/hepcidin axis may serve as a possible therapeutic target to attenuate AI.

     

Determination of the cell adhesion specificity of Streptococcus suis with the complete set of monodeoxy analogues of globotriose

Streptococcus suis causes meningitis and other serious infections in pigs and humans, and binds to host cell globotriosylceramide. In order to determine the essential hydroxyls involved in binding, the complete set of monodeoxy derivatives of the receptor trisaccharide Gal1-Gal1-4Glc were tested as inhibitors of bacterial hemagglutination. Removal of the 4-, 6, 2 or 3-hydroxyls abolished inhibitory activity, which indicated that they were critically involved in binding. The same results were obtained using synthetic lipid-linked monodeoxy derivatives of the trisaccharides in a thin-layer overlay assay. The P_N and P_O subtypes of the S. suis adhesin showed similar binding patterns. The hydroxyls of the glucose moiety were not critical for binding, although the adhesin binds better to the trisaccharide Gal1-4Gal1-4Glc than the disaccharide Gal1-4Gal.     

Application of biotechnology in the diagnosis and control of brucellosis in the Near East Region

Although much information is available on the diagnosis and control of brucellosis, most of it is concerned with Brucella abortus in cattle. In contrast, no specific serological test for Brucella melitensis infection in small ruminants has been developed. Antigens prepared from Brucella abortus, as well as tests used for diagnosis of Brucella abortus infection in cattle, are also used for diagnosis of Brucella melitensis in small ruminants, buffaloes, camels, swine and other animals. Control policy for Brucella melitensis has not yet been established in all countries of the region, probably due to lack of information on the best methods. Moreover, little information exists on the efficacy of a mass vaccination strategy in small ruminants, regarding vaccine strain persistence in the host and its excretion in the milk of adult animals, or horizontal transfer in the field. In addition, recent data have suggested that Brucella melitensis may have evolved to more virulent strains which represent a higher risk for humans. It is suggested that a project should be financed to establish or develop a feasible regional brucellosis control programme, whose objectives will be: (i) to survey different animal species and evaluate the applicability of several serological and molecular biological techniques using specific diagnostics prepared by genetic engineering; (ii) to characterize bacteriological and genetic properties of Brucella isolates as well as the Brucella melitensis Rev. 1 vaccinal isolates recovered from different animals in various countries of the region and compare them to the standard Elberg strain. This will provide an insight into the mechanisms involved in the attenuation of the strain and its immune properties. This knowledge will be used to boost the immune protection conferred by conventional Brucella melitensis Rev. 1 vaccination, to develop new effective attenuated vaccine and/or to design subunit or DNA vaccines for whole flock vaccination.     

Detection of a specifically amplified DNAfragment in Brucella abortus by arbitrarilyprimed polymerase chain reaction

Randomly amplified polymorphic DNA (RAPD) profiles of Brucella and non-Brucella DNA were established after polymerase chain reaction (PCR) amplification. Five arbitrary oligonucleotide primers were screened to generate Brucella-specific DNA fingerprints. The arbitrary primer OPB-01 (5-GTTTCGCTCC-3) produced DNA bands specific to Brucella. Amplification conditions must be optimized for reproductibility. Accordingly, we optimized and established the conditions, which included Mg²⁺, enzyme (DNA polymerase), primer, template and deoxyribonucleoside triphosphate (dNTP) concentrations as well as the optimum number of thermal cycles to produce OPB-01 directed Brucella DNA fingerprints.The optimized RAPD method can produce a 1.3 kb DNA fragment specific to Brucella. This DNA fragment was common to eight biovars of B. abortus and one biovar of B. melitensis. The fragment was not detected in genetically related species such as Ochrobactrum anthropi and other non-Brucella organisms associated with farm animals. We anticipate the use of this fragment as a possible probe for the detection of Brucella organisms.     

Characterization of Novel Brucella Strains Originating from Wild Native Rodent Species in North Queensland,Australia

We report on the characterization of a group of seven novel Brucella strains isolated in 1964 from three native rodent species in North Queensland, Australia, during a survey of wild animals. The strains were initially reported to be Brucella suis biovar 3 on the basis of microbiological test results. Our results indicated that the rodent strains had microbiological traits distinct from those of B. suis biovar 3 and all other Brucella spp. To reinvestigate these rodent strains, we sequenced the 16S rRNA, recA, and rpoB genes and nine housekeeping genes and also performed multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA). The rodent strains have a unique 16S rRNA gene sequence compared to the sequences of the classical Brucella spp. Sequence analysis of the recA, rpoB, and nine housekeeping genes reveals that the rodent strains are genetically identical to each other at these loci and divergent from any of the currently described Brucella sequence types. However, all seven of the rodent strains do exhibit distinctive allelic MLVA profiles, although none demonstrated an amplicon for VNTR 07, whereas the other Brucella spp. did. Phylogenetic analysis of the MLVA data reveals that the rodent strains form a distinct clade separate from the classical Brucella spp. Furthermore, whole-genome sequence comparison using the maximal unique exact matches index (MUMi) demonstrated a high degree of relatedness of one of the seven rodent Brucella strains (strain NF 2653) to another Australian rodent Brucella strain (strain 83-13). Our findings strongly suggest that this group of Brucella strains isolated from wild Australian rodents defines a new species in the Brucella genus.Brucella species are facultative intracellular Gram-negative members of the Alphaproteobacteria class capable of causing brucellosis in a range of animal hosts, including domesticated livestock, wildlife, marine mammals, and humans (1, 5, 7, 29, 32, 33, 36, 47). Brucellosis is the most prevalent zoonotic disease worldwide, causing spontaneous abortion and fetal death in animals and severe flu-like symptoms, focal complications, and often, chronic disease in humans (7, 11, 22, 27, 40, 41, 49, 50). Brucella species are typically transmitted to humans through consumption of unpasteurized dairy products or exposure to fluids or tissues from infected animals (45, 49). Animals are primary hosts of all Brucella spp., which include Brucella abortus (cattle), B. canis (dogs), B. melitensis (goats, cows, and sheep), B. suis (swine), B. ovis (rams), and B. neotomae (desert rats) (3, 7, 8). Recently, three additional Brucella species have been recognized: B. pinnipedialis (seals), B. ceti (dolphins) (5), and B. microti. B. microti was initially isolated from the common vole in the Czech Republic (33, 35). In the mid-1980s, DNA-DNA hybridization studies demonstrated a very high level of genetic similarity (98.5%) among the Brucella spp., which led to the adoption of a monospecies concept for the Brucella genus, with all the species at that time renamed as biovars of B. melitensis (46). However, 20 years later, the use of a phylogenetic-evolutionary approach to Brucella taxonomy was accepted. By that approach, host preferences, virulence, and pathogenicity were considered important criteria in the delineation of Brucella species, and consequently, the multispecies taxonomy was restored to the Brucella genus (28). With the development of more advanced molecular typing methodologies such as multilocus sequence analysis (MLSA) (48), multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) (23), and techniques interrogating single nucleotide polymorphisms (SNPs) (17, 47), Brucella spp. can be quickly genotyped and strains can be readily examined for their phylogenetic and evolutionary relationships (15).Recently, we reported on two unusual human brucellosis cases, one of which led to the identification of a novel species, Brucella inopinata, whose type strain is strain BO1 and which was associated with a breast implant infection in a patient in Oregon (12, 36). The second brucellosis case involved an atypical Brucella strain (strain BO2) isolated from the lung biopsy fluid of a patient with chronic destructive pneumonia in Australia (44). Because both patients denied common risk factors associated with human brucellosis, the primary hosts of these strains remain unknown. However, nucleotide sequence analysis of the outer membrane proteins (omp2a and omp2b) of both strain BO2 and strain BO1^T demonstrated close clustering to an atypical B. suis strain (strain 83-210) isolated from a rodent in Australia (30, 44). Further genetic analysis of the 16S rRNA genes from strains BO1^T and BO2 and the 16S rRNA gene from atypical Brucella strain 83-13 (available at http://www.broadinstitute.org), which was isolated from a rodent in Australia and briefly described by Corbel and Brinley-Morgan in 1984 (9), showed that strain 83-13 yielded notable genetic similarity to the novel human isolates, which led us to speculate that atypical human Brucella strains BO1^T and BO2 may have an animal reservoir in rodents from Australia.Rodent brucellosis is self-limiting and is mostly associated with wild rodents that cohabitate among domestic livestock presumably infected with classical Brucella spp. (27). Over 22 different wild rodent species worldwide have been reported to be susceptible to Brucella infection, as demonstrated by serology and/or culture (41). Earlier field studies in Argentina, Venezuela, and Denmark reported on the prevalence of B. suis in hares, opossums, and rats and B. abortus in ferrets and capybaras (10, 11, 27). However, two rodent-specific Brucella spp. have been identified, including B. neotomae, isolated from the desert wood rat (Neotomae lepida) in Utah (39), and most recently, B. microti, isolated from the common vole (Microtus arvalis) in the Czech Republic (35). In the early 1960s, Cook et al. reported on the isolation and biochemical identification of seven Brucella suis biovar 3 strains from three known species of wild native rats from Australia (6). We reinvestigated the microbiological characteristics of these seven rodent B. suis biovar 3 strains and performed genetic analyses with respect to the microbiological characteristics and genetics of the classical and atypical Brucella species. In this report, we describe this group of Brucella strains isolated from wild rodents in Australia, which confer unique microbiological and molecular characteristics distinct from those of any of the currently described species.     

Differences in chromosome number and genome rearrangements in the genus Brucella

We have studied the genomic structure and constructed the Spe I, Pac I and I- Ceu I restriction maps of the four biovars of the pathogenic bacterium Brucella suis . B . suis biovar 1 has two chromosomes of 2.1 Mb and 1.15 Mb, similar to those of the other Brucella species: B . melitensis , B . abortus , B . ovis and B . neotomae . Two chromosomes were also observed in the genome of B . suis biovars 2 and 4, but with sizes of 1.85 Mb and 1.35 Mb, whereas only one chromosome with a size of 3.1 Mb was found in B . suis biovar 3. We show that the differences in chromosome size and number can be explained by rearrangements at chromosomal regions containing the three rrn genes. The location and orientation of these genes confirmed that these rearrangements are due to homologous recombination at the rrn loci. This observation allows us to propose a scheme for the evolution of the genus Brucella in which the two chromosome-containing strains can emerge from an hypothetical ancestor with a single chromosome, which is probably similar to that of B . suis biovar 3. As the genus Brucella is certainly monospecific, this is the first time that differences in chromosome number have been observed in strains of the same bacterial species.     

Comparative assessment of passive surveillance in disease-free and endemic situation: Example of <Emphasis Type="Italic">Brucella melitensis</Emphasis> surveillance in Switzerland and in Bosnia and Herzegovina

Background

Globalization and subsequent growth in international trade in animals and animal products has increased the importance of international disease reporting. Efficient and reliable surveillance systems are needed in order to document the disease status of a population at a given time. In this context, passive surveillance plays an important role in early warning systems. However, it is not yet routinely integrated in the assessment of disease surveillance systems because different factors like the disease awareness (DA) of people reporting suspect cases influence the detection performance of passive surveillance. In this paper, we used scenario tree methodology in order to evaluate and compare the quality and benefit of abortion testing (ABT) for Brucella melitensis (Bm) between the disease free situation in Switzerland (CH) and a hypothetical disease free situation in Bosnia and Herzegovina (BH), taking into account DA levels assumed for the current endemic situation in BH.

Results

The structure and input parameters of the scenario tree were identical for CH and BH with the exception of population data in small ruminants and the DA in farmers and veterinarians. The sensitivity analysis of the stochastic scenario tree model showed that the small ruminant population structure and the DA of farmers were important influential parameters with regard to the unit sensitivity of ABT in both CH and BH. The DA of both farmers and veterinarians was assumed to be higher in BH than in CH due to the current endemic situation in BH. Although the same DA cannot necessarily be assumed for the modelled hypothetical disease free situation as for the actual endemic situation, it shows the importance of the higher vigilance of people reporting suspect cases on the probability that an average unit processed in the ABT-component would test positive.

Conclusion

The actual sensitivity of passive surveillance approaches heavily depends on the context in which they are applied. Scenario tree modelling allows for the evaluation of such passive surveillance system components under assumed disease free situation. Despite data gaps, this is a real opportunity to compare different situations and to explore consequences of changes that could be made.

     

Outer membrane vesicle of<Emphasis Type="Italic">Neisseria meningitidis</Emphasis> serogroup B as an adjuvant to induce specific antibody response against the lipopolysaccharide of<Emphasis Type="Italic">Brucella abortus</Emphasis> S99

Brucellosis is a worldwide zoonosis and represents one of the most important public health problems in many countries, especially around the Mediterranean basin, Middle East, India and Central and South America. Currently subunit vaccines are being considered to develop effective vaccines for human which has been evidenced by vaccines currently available against the infections such as meningococcal diseases and influenza. The application of new adjuvants of microbial origins is also underway to design subunit vaccines promoting the immune responses to the antigenic determinant(s) of the vaccine. In order to explore the efficacy ofBrucella abortus lipopolysaccharide (LPS) combined with different adjuvants and proteins (as a vaccine candidate) in the induction of immune response as an effective and long-lasting immunity againstBrucella, we evaluated the outer membrane vesicle ofNeisseria meningitidis serogroup B (GBMOMV) as a subcutaneous adjuvant and a part of a brucellosis candidate vaccine to induce high titres of specific anti-Brucella abortus S99 LPS in animal model (mice). The obtained results were compared with complete and incomplete Freund’s adjuvants (CFA and IFA). LPS+GBMOMV was the most immunogenic compound that stimulated following the first injection and increase in IgG titre of about 3.90, 3.18 and 1.58 fold higher than that produced against LPS, LPS+IFA and LPS+CFA, respectively. The highest anti-LPS IgG titre was detected two weeks after the third injection (the day 42) of LPS+GBMOMV. Purified GBMOMV can be considered as a safe subcutaneous adjuvant and a part of a candidate vaccine when combined with lipopolysaccharide ofBrucella abortus S99.