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Pakistan Journal of Biological Sciences... Jan 2019The economic losses due to brucellosis as well as its potential public health in human worldwide encourage more researches to find novel pathways for effective control...
BACKGROUND AND OBJECTIVE
The economic losses due to brucellosis as well as its potential public health in human worldwide encourage more researches to find novel pathways for effective control methods of the disease. The objective of this study was to investigate the most prevalent Brucella strains obtained from cattle and their virulence genes.
MATERIALS AND METHODS
Three hundred small-holders cows in Menoufia governorate, Egypt, were screened for brucellosis using rose bengal test (RBT) and confirmed by complement fixation test (CFT). Milk samples and supra-mammary lymph nodes of serologically positive cows were collected for bacteriological isolation and identification. The obtained isolates were genotyped using PCR and their virulence genes (omp25, omp31, manA, manB, virB and znuA) were screened.
RESULTS
The prevalence rate of bovine brucellosis was 15 (5%), 11 (3.6%) and 7 (2.33%) by RBT, CFT and bacteriological examination, respectively. The seven isolates were identified and genotyped as Brucella melitensis biotype3. Furthermore, the molecular detection of substantial virulence genes revealed that manA, manB, omp25 and omp31 genes were detected in all tested B. melitensis strains. Meanwhile, the virB genes were detected in 4 strains and the znuA genes were detected in 3 strains among the isolated B. melitensis strains.
CONCLUSION
It was concluded that B. melitensis biotype3 was the pre-dominant Brucella spp. as well as omp25, omp31, manA and manB were the most common related-virulence genes which assumed to play a worthy function in the pathogenesis of brucellosis.
Topics: Animals; Bacterial Outer Membrane Proteins; Bacterial Proteins; Brucella melitensis; Brucellosis; Cattle; Complement System Proteins; DNA, Bacterial; Egypt; Female; Genotype; Mannose-6-Phosphate Isomerase; Multienzyme Complexes; Nucleotidyltransferases; Phenotype; Polymerase Chain Reaction; Rose Bengal; Virulence
PubMed: 31930868
DOI: 10.3923/pjbs.2019.239.246 -
Infection, Genetics and Evolution :... Mar 2018Currently, although the prevalence of brucellosis in Kazakhstan remains high, there are limited data available on the genetic diversity of circulating Brucella strains....
Currently, although the prevalence of brucellosis in Kazakhstan remains high, there are limited data available on the genetic diversity of circulating Brucella strains. Here, MLVA was employed to genotype a panel of 102 Brucella isolates collected from eight Kazakh regions and neighboring countries (Russia, Kyrgyzstan) during the period 1935-2017. MLVA-11 analysis classified 64 B. abortus strains into genotypes 72, 82, 331, 71, 341 and 69, while one genotype was novel, having no correspondence within the MLVA international database. MLVA-11 analysis of 37 B. melitensis strains showed 100% identity with genotypes 116, 114 and 11. One B. suis strain was classified into genotype 33. Phylogeography based on MLVA-15 demonstrated that all B. abortus and B. melitensis strains belonged to "Abortus C" and "East Mediterranean" lineages, respectively. B. abortus strains from Kazakhstan and Russia resulted genetically related to Portuguese, Brazilian and US isolates, suggesting ancient spread of these lineages from Europe westwards to South America and eastwards to Turkey, Russia and Asia. Most of Kazakh B. melitensis isolates were related to strains circulating in China, likely due to long-term trading partnerships between the two countries. In fine-scale MLVA-15 analysis, 17 B. abortus and 12 B. melitensis genotypes were identified; among them 12 are novel. Interestingly, epidemiological information supporting molecular data were retrieved for two clusters within the B. abortus group, thus proving that MLVA is an appropriate tool for effective traceback analyses. Our findings suggest that molecular genotyping should be applied systematically to support control plans for eradication of brucellosis in Kazakhstan.
Topics: Animal Diseases; Animals; Brucella abortus; Brucella melitensis; Cattle; Genetic Variation; Genotype; Geography, Medical; Guinea Pigs; Kazakhstan; Minisatellite Repeats; Multilocus Sequence Typing; Phylogeny; Phylogeography
PubMed: 29278754
DOI: 10.1016/j.meegid.2017.12.022 -
Indian Journal of Medical Microbiology 2023Brucellosis is a bacterial zoonotic disease caused by genus Brucella. The disease is often transmitted to humans by direct or indirect contact with infected livestock or...
PURPOSE
Brucellosis is a bacterial zoonotic disease caused by genus Brucella. The disease is often transmitted to humans by direct or indirect contact with infected livestock or from laboratory exposure. In this study two clinical isolates of Brucella melitensis were subjected to whole genome sequencing (WGS) using Ion Torrent PGM and Oxford Nanopore MinIon platform.
METHODS
The two hybrid complete genomes were subjected to core gene SNP analysis to identify the relative evolutionary position. To distinguish between the various lineages of B. melitensis, Pangenome analysis was carried out.
RESULTS
Phylogenetic analysis revealed that both the study isolates (ST8) clustered along the other Asian isolates that formed genotype II. Genome wide analyses of 326 B melitensis isolates suggests 2171 gene clusters were shared across all the genomes while 3552 gene clusters were considered as accessory genes.
CONCLUSION
Here we attempted to provide the gain and loss of six unique genes that defined the phylogenetic lineages and complex evolutionary process. As the severity and prevalence of human brucellosis is increasing a better understanding of Brucella genomics and transmission dynamics is needed.
Topics: Humans; Brucella melitensis; Phylogeny; Genome-Wide Association Study; Brucellosis; Genomics; Genotype
PubMed: 37356834
DOI: 10.1016/j.ijmmb.2023.02.003 -
Veterinary Microbiology Dec 2002The genus Brucella contains alpha-Proteobacteria adapted to intracellular life within cells of a variety of mammals. Controversy has arisen concerning Brucella internal... (Review)
Review
The genus Brucella contains alpha-Proteobacteria adapted to intracellular life within cells of a variety of mammals. Controversy has arisen concerning Brucella internal taxonomy, and it has been proposed that the DNA-DNA hybridization-based genomospecies concept be applied to the genus. According to this view, only one species, Brucella melitensis, should be recognized, and the classical species should be considered as biovars (B. melitensis biovar melitensis; B. melitensis biovar abortus; etc.). However, a critical reappraisal of the species concept, a review of the population structure of bacteria and the analysis of Brucella genetic diversity by methods other than DNA-DNA hybridization show that there are no scientific grounds to apply the genomospecies concept to this genus. On the other hand, an enlarged biological species concept allows the definition of Brucella species that are consistent with molecular analyses and support the taxonomical standing of most classical species. Both the host range as a long-recognized biological criterion and the presence of species-specific markers in outer membrane protein genes and in other genes show that B. melitensis, B. abortus, B. ovis, B. canis and B. neotomae are not mere pathovars (or nomenspecies) but biologically meaningful species. The status of B. suis is, however, less clear. These approaches should be useful to define species for the marine mammal Brucella isolates, as illustrated by the grouping of the isolates from pinnipeds or from cetaceans by omp2 gene analysis. It is shown that a correct Brucella species definition is important to understand the evolution of the genus.
Topics: Biological Evolution; Brucella; Brucella melitensis; Classification; DNA, Bacterial; Phylogeny; Species Specificity
PubMed: 12414145
DOI: 10.1016/s0378-1135(02)00210-9 -
International Microbiology : the... Mar 1998The brucellae are Gram-negative bacteria characteristically able to multiply facultatively within phagocytic cells and which cause a zoonosis of world-wide importance.... (Review)
Review
The brucellae are Gram-negative bacteria characteristically able to multiply facultatively within phagocytic cells and which cause a zoonosis of world-wide importance. This article reviews the structure and topology of the main components (lipopolysaccharide, native hapten polysaccharide, free lipids and proteins) of the outer membranes of Brucella abortus and B. melitensis, as well as some distinctive properties (permeability and interactions with cationic peptides) of these membranes. On these data, an outer membrane model is proposed in which, as compared to other Gram-negatives, there is a stronger hydrophobic anchorage for the lipopolysaccharide, free lipids, porin proteins and lipoproteins, and a reduced surface density of anionic groups, which could be partially or totally neutralized by ornithine lipids. This model accounts for the permeability of Brucella to hydrophobic permeants and for its resistance to the bactericidal oxygen-independent systems of phagocytes.
Topics: Bacterial Outer Membrane Proteins; Brucella abortus; Brucella melitensis; Haptens; Lipid A; Lipids; Models, Molecular; Molecular Conformation; Protein Conformation; Structure-Activity Relationship; Surface Properties
PubMed: 10943337
DOI: No ID Found -
Microbiology and Immunology Jul 2013Brucellosis is a globally distributed zoonotic disease that causes animal and human diseases. Although effective, the current Brucella vaccines (Rev.1 and M5-90) have...
Brucellosis is a globally distributed zoonotic disease that causes animal and human diseases. Although effective, the current Brucella vaccines (Rev.1 and M5-90) have several drawbacks. The first involves residual virulence for animals and humans and the second is the inability to differentiate natural infection from that caused by vaccination. Therefore, Brucella melitensis 16M hfq mutant (16MΔhfq) was constructed to overcome these drawbacks. Similarly to Rev.1 and M5-90, 16MΔhfq reduces survival in macrophages and mice and induces strong protective immunity in BALB/c mice. Moreover, these vaccines elicit anti-Brucella-specific IgG1 and IgG2a subtype responses and induce secretion of gamma interferon and interleukin-4. The Hfq antigen also allows serological differentiation between infected and vaccinated animals. These results show that 16MΔhfq is an ideal live attenuated vaccine candidate against virulent Brucella melitensis 16M infection. It will be further evaluated in sheep.
Topics: Animals; Antibodies, Bacterial; Brucella Vaccine; Brucella melitensis; Brucellosis; Cell Survival; Disease Models, Animal; Female; Gene Deletion; Host Factor 1 Protein; Immunoglobulin G; Interferon-gamma; Interleukin-4; Macrophages; Mice; Mice, Inbred BALB C; Survival Analysis; Vaccines, Attenuated; Virulence Factors
PubMed: 23647412
DOI: 10.1111/1348-0421.12065 -
Veterinary Microbiology Mar 2021Brucellosis is an infectious disease of several terrestrial and marine animals and humans caused by bacteria of the genus Brucella. This study aimed to identify Brucella...
Brucellosis is an infectious disease of several terrestrial and marine animals and humans caused by bacteria of the genus Brucella. This study aimed to identify Brucella species and biovars circulating in cattle and to analyze their geographic distribution across Algeria. Two hundred ninety eight milk and lymph node samples from 161 seropositive cattle of different local and foreign breeds were collected from 97 dairy farms in 56 towns of 13 wilayas (states/ provinces) of the central, eastern, western and southern regions. The samples were cultured on selective media and the obtained isolates were identified using bacteriological and molecular tests. Eighty-five Brucella isolates (72 B. abortus and 13 B. melitensis) were recovered from 63 animals in 37 dairy farms. In total, 71 (83.5 %) B. abortus bv 3, 11 (12.9 %) B. melitensis bv 2, 2 (2.4 %) B. melitensis bv 3 and 1 (1.2 %) unidentified B. abortus biovar were detected. The identification of B. abortus biovar 3 and B. melitensis biovar 2 is a new finding for Algeria and the Maghreb, respectively. B. abortus (84.7 %) was the main etiological agent of brucellosis. B. abortus showed a scattered distribution across Algeria. The fact that 60 % of the seropositive cattle showed no clinical signs, but 36 % were culture positive is an alarming observation. These data will rise awareness for the current epidemiological situation of bovine brucellosis in Algeria. To the best of our knowledge, this is the first representative countrywide bacteriological investigation of Brucella species and biovars in cattle across Algeria, which is a developing country where resources might be limited and the working conditions might not be very friendly.
Topics: Aborted Fetus; Algeria; Animals; Bacterial Typing Techniques; Brucella abortus; Brucella melitensis; Brucellosis; Cattle; Cattle Diseases; Dairying; Female; Genotype; Geography; Multilocus Sequence Typing; Phylogeny; Risk Factors
PubMed: 33571821
DOI: 10.1016/j.vetmic.2021.109004 -
Journal of Microbiology and... Apr 2020For control of brucellosis in small ruminants, attenuated Rev1 is used but it can be virulent for animals and human. Based on these aspects, it is essential to identify...
For control of brucellosis in small ruminants, attenuated Rev1 is used but it can be virulent for animals and human. Based on these aspects, it is essential to identify potential immunogens to avoid these problems in prevention of brucellosis. The majority of OMPs in the Omp25/31 family have been studied because these proteins are relevant in maintaining the integrity of the outer membrane but their implication in the virulence of the different species of this genus is not clearly described. Therefore, in this work we studied the role of Omp31 on virulence by determining the residual virulence and detecting lesions in spleen and testis of mice inoculated with the LVM31 mutant strain. In addition, we evaluated the conferred protection in mice immunized with the mutant strain against the challenge with the Bm133 virulent strain. Our results showed that the mutation of 31 caused a decrease in splenic colonization without generating apparent lesions or histopathological changes apparent in both organs in comparison with the control strains and that the mutant strain conferred similar protection as the Rev1 vaccine strain against the challenge with Bm133 virulent strain. These results allow us to conclude that Omp31 plays an important role on the virulence of in the murine model, and due to the attenuation shown by the strain, it could be considered a vaccine candidate for the prevention of goat brucellosis.
Topics: Animals; Bacterial Outer Membrane Proteins; Brucella Vaccine; Brucella melitensis; Brucellosis; Disease Models, Animal; Female; Immunization; Male; Mice; Mice, Inbred BALB C; Mutation; Spleen; Vaccines, Attenuated; Virulence
PubMed: 31986561
DOI: 10.4014/jmb.1908.08056 -
PLoS Neglected Tropical Diseases Mar 2022Brucellosis is an infectious disease caused by bacteria of the genus Brucella. Although it is the most common zoonosis worldwide, there are increasing reports of drug...
BACKGROUND
Brucellosis is an infectious disease caused by bacteria of the genus Brucella. Although it is the most common zoonosis worldwide, there are increasing reports of drug resistance and cases of relapse after long term treatment with the existing drugs of choice. This study therefore aims at identifying possible natural inhibitors of Brucella melitensis Methionyl-tRNA synthetase through an in-silico approach.
METHODS
Using PyRx 0.8 virtual screening software, the target was docked against a library of natural compounds obtained from edible African plants. The compound, 2-({3-[(3,5-dichlorobenzyl) amino] propyl} amino) quinolin-4(1H)-one (OOU) which is a co-crystallized ligand with the target was used as the reference compound. Screening of the molecular descriptors of the compounds for bioavailability, pharmacokinetic properties, and bioactivity was performed using the SWISSADME, pkCSM, and Molinspiration web servers respectively. The Fpocket and PLIP webservers were used to perform the analyses of the binding pockets and the protein ligand interactions. Analysis of the time-resolved trajectories of the Apo and Holo forms of the target was performed using the Galaxy and MDWeb servers.
RESULTS
The lead compounds, Strophanthidin and Isopteropodin are present in Corchorus olitorius and Uncaria tomentosa (Cat's-claw) plants respectively. Isopteropodin had a binding affinity score of -8.9 kcal / ml with the target and had 17 anti-correlating residues in Pocket 1 after molecular dynamics simulation. The complex formed by Isopteropodin and the target had a total RMSD of 4.408 and a total RMSF of 9.8067. However, Strophanthidin formed 3 hydrogen bonds with the target at ILE21, GLY262 and LEU294, and induced a total RMSF of 5.4541 at Pocket 1.
CONCLUSION
Overall, Isopteropodin and Strophanthidin were found to be better drug candidates than OOU and they showed potentials to inhibit the Brucella melitensis Methionyl-tRNA synthetase at Pocket 1, hence abilities to treat brucellosis. In-vivo and in-vitro investigations are needed to further evaluate the efficacy and toxicity of the lead compounds.
Topics: Anti-Bacterial Agents; Brucella melitensis; Ligands; Methionine-tRNA Ligase; Molecular Dynamics Simulation
PubMed: 35312681
DOI: 10.1371/journal.pntd.0009799 -
Veterinary Research Nov 2019Epidemiological investigations implemented in wild and domestic ruminants evidenced a reservoir for Brucella in Capra ibex in the French Alps. Vaccination was considered...
Epidemiological investigations implemented in wild and domestic ruminants evidenced a reservoir for Brucella in Capra ibex in the French Alps. Vaccination was considered as a possible way to control Brucella infection in this wildlife population. Twelve ibexes and twelve goats were allocated into four groups housed separately, each including six males or six non-pregnant females. Four to five animals were vaccinated and one or two animals were contact animals. Half of the animals were necropsied 45 days post-vaccination (pv), and the remaining ones at 90 days pv. Additional samples were collected 20 and 68 days pv to explore bacterial distribution in organs and humoral immunity. Neither clinical signs nor Brucella-specific lesions were observed and all vaccinated animals seroconverted. Brucella distribution and antibody profiles were highly contrasted between both species. Proportion of infected samples was significantly higher in ibex compared to goats and decreased between 45 and 90 days pv. Two male ibex presented urogenital excretion at 20 or 45 days pv. The bacterial load was higher 45 days in ibexes compared to goats, whereas it remained moderate to low 90 days pv in both species with large variability between animals. In this experiment, differences between species remained the main source of variation, with low impact of other individual factors. To conclude, multiplicative and shedding capacity of Rev.1 was much higher in ibex compared to goats within 90 days. These results provide initial information on the potential use in natura of a commercial vaccine.
Topics: Animals; Bacterial Shedding; Brucella Vaccine; Brucella melitensis; Brucellosis; Goat Diseases; Goats; Species Specificity; Vaccination
PubMed: 31775863
DOI: 10.1186/s13567-019-0717-0