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BMC Genomics May 2018Brucellosis is a bacterial zoonotic disease. Annually in the world more than 500,000 new cases of brucellosis in humans are registered. In this study, we propose an...
BACKGROUND
Brucellosis is a bacterial zoonotic disease. Annually in the world more than 500,000 new cases of brucellosis in humans are registered. In this study, we propose an evolutionary model of the historical distribution of B. melitensis based on the full-genomic SNP analysis of 98 strains.
RESULTS
We performed an analysis of the SNP of the complete genomes of 98 B. melitensis strains isolated in different geographical regions of the world to obtain relevant information on the population structure, genetic diversity and the evolution history of the species. Using genomic sequences of 21 strains of B. melitensis isolated in Russia and WGS data from the NCBI database, it was possible to identify five main genotypes and 13 species genotypes for analysis. Data analysis based on the Bayesian Phylogenetics and Phylogeography method allowed to determine the regions of geographical origin and the expected pathways of distribution of the main lines (genotypes and subgenotypes) of the pathogen.
CONCLUSIONS
Within the framework of our study, the model of global evolution and phylogeography of B. melitensis strains isolated in various regions of the planet was proposed for the first time. The sets of unique specific SNPs described in our study, for all identified genotypes and subgenotypes, can be used to develop new bacterial typing and identification systems for B. melitensis.
Topics: Bacterial Typing Techniques; Bayes Theorem; Brucella melitensis; Evolution, Molecular; Genes, Bacterial; Genotype; Phylogeny; Phylogeography; Polymorphism, Single Nucleotide; Species Specificity
PubMed: 29747573
DOI: 10.1186/s12864-018-4762-2 -
American Journal of Physiology. Lung... Jul 2015Acute lung injury (ALI) and the more severe acute respiratory distress syndrome are common responses to a variety of infectious and noninfectious insults. We used a...
Acute lung injury (ALI) and the more severe acute respiratory distress syndrome are common responses to a variety of infectious and noninfectious insults. We used a mouse model of ALI induced by intratracheal administration of sterile bacterial wall lipopolysaccharide (LPS) to investigate the changes in innate lung microbiota and study microbial community reaction to lung inflammation and barrier dysfunction induced by endotoxin insult. One group of C57BL/6J mice received LPS via intratracheal injection (n = 6), and another received sterile water (n = 7). Bronchoalveolar lavage (BAL) was performed at 72 h after treatment. Bacterial DNA was extracted and used for qPCR and 16S rRNA gene-tag (V3-V4) sequencing (Illumina). The bacterial load in BAL from ALI mice was increased fivefold (P = 0.03). The community complexity remained unchanged (Simpson index, P = 0.7); the Shannon diversity index indicated the increase of community evenness in response to ALI (P = 0.07). Principal coordinate analysis and analysis of similarity (ANOSIM) test (P = 0.005) revealed a significant difference between microbiota of control and ALI groups. Bacteria from families Xanthomonadaceae and Brucellaceae increased their abundance in the ALI group as determined by Metastats test (P < 0.02). In concordance with the 16s-tag data, Stenotrohomonas maltophilia (Xanthomonadaceae) and Ochrobactrum anthropi (Brucellaceae) were isolated from lungs of mice from both groups. Metabolic profiling of BAL detected the presence of bacterial substrates suitable for both isolates. Additionally, microbiota from LPS-treated mice intensified IL-6-induced lung inflammation in naive mice. We conclude that the morbid transformation of ALI microbiota was attributed to the set of inborn opportunistic pathogens thriving in the environment of inflamed lung, rather than the external infectious agents.
Topics: Animals; Base Sequence; Bronchoalveolar Lavage Fluid; Brucellaceae; DNA, Bacterial; Disease Models, Animal; Lipopolysaccharides; Lung; Lung Injury; Mice; Mice, Inbred C57BL; Microbiota; RNA, Ribosomal, 16S; Respiratory Distress Syndrome; Sequence Analysis, DNA; Xanthomonadaceae
PubMed: 25957290
DOI: 10.1152/ajplung.00061.2014 -
Revue Scientifique Et Technique... Apr 2013The genus Brucella (Mayer and Shaw, 1920) currently consists often species with validly published names. Within most species further differentiation into biovars exists....
The genus Brucella (Mayer and Shaw, 1920) currently consists often species with validly published names. Within most species further differentiation into biovars exists. Genetically, all Brucella species are highly related to each other, exhibiting sequence similarity values of 98% to 100% in aligned regions (core genome). The population structure is clonal. Despite this close genetic relatedness, the various species can be clearly distinguished from each other by application of high-resolution molecular typing tools, in addition to assessment of phenotype and host preference. Accurate species delineation can be achieved by conventional multiplex polymerase chain reaction (PCR), single nucleotide polymorphism (SNP) analysis and multilocus sequence typing (MLST) or multilocus sequence analysis (MLSA). The last is also suitable for phylogenetic reconstructions, owing to the highly clonal evolution of the different species. Highly discriminatory multilocus variable number of tandem repeats (VNTR) analysis (MLVA) allows both species delineation and differentiation of individual isolates and thus represents a perfect first-line toolfor molecular epidemiological studies within outbreak investigations. More recently,whole genome sequencing (WGS)and the resulting global genome-wide SNP analysis have become available. These novel approaches should help in further understanding the evolution, host specificity and pathogenicity of the genus Brucella.
Topics: Brucella; Databases, Factual; Gene Expression Regulation, Bacterial; Genome, Bacterial; Multilocus Sequence Typing; Phylogeny; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Species Specificity; Tandem Repeat Sequences
PubMed: 23837373
DOI: 10.20506/rst.32.1.2189 -
Frontiers in Cellular and Infection... 2021Adhesion to host cells is a key step for successful infection of many bacterial pathogens and may define tropism to different host tissues. To do so, bacteria display...
Adhesion to host cells is a key step for successful infection of many bacterial pathogens and may define tropism to different host tissues. To do so, bacteria display adhesins on their surfaces. is an intracellular pathogen capable of proliferating in a wide variety of cell types. It has been described that BmaC, a large protein that belongs to the classical (type Va) autotransporter family, is required for efficient adhesion of strain 1330 to epithelial cells and fibronectin. Here we show that 1330 harbors two other type Va autotransporters (BmaA and BmaB), which, although much smaller, share significant sequence similarities with BmaC and contain the essential domains to mediate proper protein translocation to the bacterial surface. Gain and loss of function studies indicated that BmaA, BmaB, and BmaC contribute, to a greater or lesser degree, to adhesion of 1330 to different cells such as synovial fibroblasts, osteoblasts, trophoblasts, and polarized epithelial cells as well as to extracellular matrix components. It was previously shown that BmaC localizes to a single bacterial pole. Interestingly, we observed here that, similar to BmaC, the BmaB adhesin is localized mostly at a single cell pole, reinforcing the hypothesis that displays an adhesive pole. Although species have strikingly similar genomes, they clearly differ in their host preferences. Mainly, the differences identified between species appear to be at loci encoding surface proteins. A careful analysis of the putative type Va autotransporter orthologues from several strains showed that the locus from and both, the and loci from are pseudogenes in all strains analyzed. Results reported here evidence that all three autotransporters play a role in the adhesion properties of 1330. However, spp. exhibit extensive variations in the repertoire of functional adhesins of the classical autotransporter family that can be displayed on the bacterial surface, making them an interesting target for future studies on host preference and tropism.
Topics: Adhesins, Bacterial; Adhesives; Brucella abortus; Brucella suis; Type V Secretion Systems
PubMed: 33987105
DOI: 10.3389/fcimb.2021.607610 -
Frontiers in Cellular and Infection... 2021In comparison with the standard monoxenic maintenance in the laboratory, rearing the nematode on its natural microbiota improves its fitness and immunity against...
In comparison with the standard monoxenic maintenance in the laboratory, rearing the nematode on its natural microbiota improves its fitness and immunity against pathogens. Although is known to exhibit choice behavior and pathogen avoidance behavior, little is known about whether actively chooses its (beneficial) microbiota and whether the microbiota influences worm behavior. We examined eleven natural isolates in a multiple-choice experiment for their choice behavior toward four natural microbiota bacteria and found that microbiota choice varied among isolates. The natural isolate MY2079 changed its choice behavior toward microbiota isolate MYb71 in both multiple-choice and binary-choice experiments, in particular on proliferating bacteria: MYb71 was chosen less than other microbiota bacteria or OP50, but only after preconditioning with MYb71. Examining escape behavior and worm fitness on MYb71, we ruled out pathogenicity of MYb71 and consequently learned pathogen avoidance behavior as the main driver of the behavioral change toward MYb71. The change in behavior of MY2079 toward microbiota bacterium MYb71 demonstrates how the microbiota influences the worm's choice. These results might give a baseline for future research on host-microbiota interaction in the model.
Topics: Animals; Bacteria; Caenorhabditis elegans; Microbiota; Ochrobactrum; Virulence
PubMed: 34976859
DOI: 10.3389/fcimb.2021.775634 -
Veterinary Microbiology Dec 2002Although Brucella is responsible for one of the major worldwide zoonosis, our understanding of its pathogenesis remains in its infancy. In this paper, we summarize some... (Review)
Review
Although Brucella is responsible for one of the major worldwide zoonosis, our understanding of its pathogenesis remains in its infancy. In this paper, we summarize some of the research in progress in our laboratory that we think could contribute to a better understanding of the Brucella molecular virulence mechanisms and their regulation.
Topics: Animals; Brucella; Brucellosis; Cell Communication; Cell Cycle; Flagella; Humans; Luminescent Measurements; Vibrio; Zoonoses
PubMed: 12414152
DOI: 10.1016/s0378-1135(02)00208-0 -
Microbiology Spectrum Dec 2021Brucellosis is a worldwide zoonosis caused by bacteria from the genus Brucella. Once established, it is very hard to eradicate this disease, since it contaminates...
Brucellosis is a worldwide zoonosis caused by bacteria from the genus Brucella. Once established, it is very hard to eradicate this disease, since it contaminates animals, the environment, and humans, causing problems for veterinary and public health as well as wildlife protection programs. Swabs are used for sampling in bacteriological and/or molecular diagnostics, from seropositive animals with disease symptoms, from genitalia or tissue lesions, as well as from contaminated environments. The aim of this study was to compare main of the commercially used swab types for sampling and diagnostics of Brucella spp. and determine the optimal storage conditions and time frame for testing. To achieve this, we tested bacterial and molecular methods for detection of Brucella abortus, Brucella melitensis, and Brucella suis using nine swab types, all with different tip materials, treated immediately after spiking, after 72 h at +4°C, and after 72 h at -20°C. Flocked swabs showed the highest capacity to preserve bacterial viability and DNA quality, regardless the storage conditions. Flocked swabs immersed in a protective medium provided the best conditions for Brucella survival in all three storage conditions. At the same time, the efficacy of quantitative PCR (qPCR) detection for all swabs, including the positive control, was above 50%, irrespective of the storage conditions, while bacterial survival was significantly lowered when swabs were kept at +4°C or -20°C for 72 h (48.2% and 27.5%, respectively). Compared to the positive control and other types, the flocked swabs maintained higher reproducibility regarding their capacity to preserve live bacteria in all three storage conditions. In order to protect public and veterinary health from highly zoonotic bacteria such as members of the genus Brucella and prevent their dissemination into the environment, direct diagnostics are of utmost importance. However, in addition to the highly specific diagnostic tests, the sampling methods, time necessary for specimens to reach the laboratories, and transport conditions are important factors to consider in order to increase the sensitivity of performed tests, especially bacterial culturing and qPCR. This paper shows how different swab types and storage conditions influence classical bacteriological diagnostics of the most prevalent Brucella species - B. melitensis, B. abortus, and B. suis - but have little impact on molecular methods. The presented results highlight (i) the choice of swab regarding the storage and transport conditions, (ii) the importance of immediate swab treatment upon sampling, and (iii) that molecular methods do not depend on storage conditions, unlike classical bacteriological isolation.
Topics: Animals; Brucella abortus; Brucella melitensis; Brucella suis; Brucellosis; DNA, Bacterial; Humans; Microbial Viability; Polymerase Chain Reaction; Specimen Handling; Zoonoses
PubMed: 34787496
DOI: 10.1128/Spectrum.00728-21 -
Emerging Infectious Diseases Jun 2021We describe the isolation of atypical Brucella inopinata-like species and unique clinicopathologic findings in 2 adult marine toads (Rhinella marina), including...
We describe the isolation of atypical Brucella inopinata-like species and unique clinicopathologic findings in 2 adult marine toads (Rhinella marina), including oophoritis in 1 toad. These findings represent a novel emerging disease in toads and a possible zoonotic pathogen.
Topics: Animals; Brucella; Brucellosis; Bufo marinus; Female
PubMed: 34014155
DOI: 10.3201/eid2706.204001 -
Lipids in Health and Disease Sep 2017Alkaline thermostable lipase and biosurfactant producing bacteria are very interested at detergent applications, not only because of their eco-friendly characterize, but...
BACKGROUND
Alkaline thermostable lipase and biosurfactant producing bacteria are very interested at detergent applications, not only because of their eco-friendly characterize, but alsoproduction lipase and biosurfactant by using cheap materials. Ochrobactrum intermedium strain MZV101 was isolated as washing powder resistant, alkaline thermostable lipase and biosurfactant producing bacterium in order to use at detergent applications.
METHODS
O. intermedium strain MZV101 produces was lipase and biosurfactant in the same media with pH 10 and temperature of 60 °C. Washing test and some detergent compatibility character of lipase enzyme and biosurfactant were assayed. The antimicrobial activity evaluated against various bacteria and fungi.
RESULTS
Lipase and biosurfactant produced by O. intermedium strain MZV101 exhibited high stability at pH 10-13 and temperature of 70-90 °C, biosurfactant exhibits good stability at pH 9-13 and thermostability in all range. Both lipase and biosurfactant were found to be stable in the presence of different metal ions, detergents and organic solvents. The lipase enzyme extracted using isopropanol with yield of 69.2% and biosurfactant with ethanol emulsification index value of 70.99% and yield of 9.32 (g/l). The single band protein after through from G-50 Sephadex column on SDS-PAGE was calculated to be 99.42 kDa. Biosurfactant O. intermedium strain MZV101 exhibited good antimicrobial activity against Gram-negative bacteria and against various bacterial pathogens. Based upon washing test biosurfactant and lipase O. intermedium strain MZV101considered being strong oil removal.
CONCLUSION
The results of this study indicate that isolated lipase and biosurfactant with strong oil removal, antimicrobial activity and good stability could be useful for detergent applications.
Topics: Anti-Infective Agents; Bacterial Proteins; Detergents; Hydrogen-Ion Concentration; Lipase; Ochrobactrum; Polymerase Chain Reaction; Solvents; Surface-Active Agents; Temperature
PubMed: 28923075
DOI: 10.1186/s12944-017-0565-8 -
Veterinary Medicine and Science Aug 2020In this study, Multiple Locus Variable Number Tandem Repeat Analysis (MLVA-16) was performed on 18 Brucella isolates identified bacteriologically and molecularly...
In this study, Multiple Locus Variable Number Tandem Repeat Analysis (MLVA-16) was performed on 18 Brucella isolates identified bacteriologically and molecularly (AMOS-PCR) as Brucella abortus (n = 6) and Brucella melitensis (n = 12). This was aimed to study the genetic association among some Egyptian Brucella genotypes isolated during the period from 2002 to 2013 along with the global genotypes database. MLVA-16 analysis for B. melitensis and B. abortus strains illustrates a total of 11, and 3 genotypes with 10 and 1 singleton genotypes, respectively. B. melitensis strains displayed greater markers diversity by VNTRs analysis of the 16 loci than B. abortus and this was attributed mainly to the diverging in panel 2B markers. B. melitensis genotype M4_Fayoum_Giza (3,5,3,13,1,1,3,3,8,21,8,7,5,9,5,3) was the only predominated genotype circulating between two different governorates. The most common B. abortus genotype, GT A3_Dakahlia (4,5,4,12,2,2,3,3,6,21,8,4,4,3,4,4), was present in three identical isolates. In phylogeny, Egyptian B. abortus bv1 genotypes were closely related to East Asian strain (for the first time), Western Mediterranean and Americas clonal lineages. B. melitensis local genotypes exhibit a genetic relatedness mostly to Western Mediterranean clonal lineage and one strain of Eastern Mediterranean clonal lineage. In conclusion, the geographic location is not the only factor stands behind the high genetic similarity of the Egyptian Brucella genotypes. These low variations may be a result of a stepwise mutational event of the most variable loci from a very limited number of ancestors especially during the transmission through non-preference hosts. The authors encourage the authorities in charge to establish pre-movement testing to reduce the risk of brucellosis spread.
Topics: Brucella abortus; Brucella melitensis; Egypt; Genetic Variation
PubMed: 32266769
DOI: 10.1002/vms3.260