Proteomic analysis of Brucella melitensis and Brucella ovis for identification of virulence factor using bioinformatics approachs

Paci, Valentina; Krasteva, Ivanka; Orsini, Massimiliano; Febo, Tiziana Di; Luciani, Mirella; Perletta, Fabrizia; Pasquale, Adriano Di; Mattioli, Mauro; Tittarelli, Manuela

doi:10.1016/j.mcp.2020.101581

Cited by 9 publications

(11 citation statements)

References 30 publications

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“…The bacterial strains (BM 16 M), reference antisera and the cultivation of Brucella for proteomic analyses were the same used by Paci et al [7]. To identify BM immunogenic proteins that are not associated with sLPS, the positive serum for BM was treated for 4 h at 37 °C, with sLPS antigen produced according to OIE Manual.…”

Section: Methodsmentioning

confidence: 99%

“…To identify BM immunogenic proteins that are not associated with sLPS, the positive serum for BM was treated for 4 h at 37 °C, with sLPS antigen produced according to OIE Manual. BM total proteins (5 µg/well) were separated by SDS-PAGE using NuPage® 4-12% Bis-Tris pre-cast gels (Life Technologies) at a constant voltage of 200 V [7,8] and transferred onto a nitrocellulose membrane using the iBlot Dry Blotting system (Life Technologies) [9]. After blocking with 5% skim milk in PBS + 0.05% Tween 20 (PBST) for 2 h, the membranes were cut into strips, which were incubated with untreated and sLPS-treated sera diluted 1:100 in PBST + 2.5% skim milk and then with Protein G-peroxidase conjugate (Sigma) diluted 1:100000 for 1 h, at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…Western Blotting (WB) analyses were performed to detect specific immunoreactive bands of BM not associated with sLPS. The immunoreactive bands were identified by nLC-ESI-MS/MS and the exposed Bcell epitopes were predicted through bioinformatics tools, as reported by Paci et al [7]. The bioinformatics pipeline, used to analyze the data obtained by mass spectrometry, was developed to identify potential B-cell target protein candidates for serological diagnosis.…”

Section: Introductionmentioning

confidence: 99%

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Identification of Immunogenic Candidate for New Serological Tests for Brucella melitensis by a Proteomic Approach

Paci¹,

Visciano²,

Krasteva³

et al. 2021

TOMICROJ

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Background: The diagnosis of brucellosis by serological tests is based on antigen suspensions derived from smooth lipopolysaccharide extracts, which can give false positive results linked to cross-reactivity with other Gram-negative microorganisms, especially Yersinia enterocolitica O:9 and Escherichia coli O157:H7. Objective: The objective of the present study was the characterization by proteomic analysis of specific immunogenic proteins not associated with smooth lipopolysaccharide to improve the diagnostic tests used in the ovine brucellosis eradication programs. Methods: The serum from a sheep positive to Brucella melitensis was treated to eliminate all antibodies against such lipopolysaccharide and highlight the reaction towards the immunoreactive proteins in Western Blotting. Results: The immunoreactive bands were identified by nLC-MS/MS and through bioinformatic tools, it was possible to select 12 potential candidates as protein antigens specific for Brucella melitensis. Conclusion: The detection of new antigens not subjected to cross-reactivity with other Gram-negative microorganisms can offer an additional tool for the serological diagnosis of such disease.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of Immunogenic Candidate for New Serological Tests for Brucella melitensis by a Proteomic Approach

Paci¹,

Visciano²,

Krasteva³

et al. 2021

TOMICROJ

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“…Contrarily, Omp25d and Omp22 play critical roles in the entry of B. ovis into mammalian cells, but there is no evidence that they perform this function in B. melitensis ( 82 , 96 ). Many other differences exist between B. melitensis and B. ovis OMPs and their participation in differential pathogenicity and host preference has been predicted in silico or tested in vitro , but not by in vivo challenge of the preferred host ( 43 , 82 , 86 , 97 , 98 ).…”

Section: Known and Undefined Mechanisms Of B Melitensis ...mentioning

confidence: 99%

Comparative Review of Brucellosis in Small Domestic Ruminants

2022

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Brucella melitensis and Brucella ovis are the primary etiological agents of brucellosis in small domestic ruminants. B. melitensis was first isolated in 1887 by David Bruce in Malta Island from spleens of four soldiers, while B. ovis was originally isolated in Australia and New Zealand in early 1950's from ovine abortion and rams epididymitis. Today, both agents are distributed worldwide: B. melitensis remains endemic and associated with an extensive negative impact on the productivity of flocks in -some regions, and B. ovis is still present in most sheep-raising regions in the world. Despite being species of the same bacterial genus, B. melitensis and B. ovis have extensive differences in their cultural and biochemical characteristics (smooth vs. rough colonial phases, serum and CO2 dependence for in vitro growth, carbohydrate metabolism), host preference (female goat and sheep vs. rams), the outcome of infection (abortion vs. epididymitis), and their zoonotic potential. Some of these differences can be explained at the bacterial genomic level, but the role of the host genome in promoting or preventing interaction with pathogens is largely unknown. Diagnostic techniques and measures to prevent and control brucellosis in small ruminants vary, with B. melitensis having more available tools for detection and prevention than B. ovis. This review summarizes and analyzes current available information on: (1) the similarities and differences between these two etiological agents of brucellosis in small ruminants, (2) the outcomes after their interaction with different preferred hosts and current diagnostic methodologies, (3) the prevention and control measures, and (4) alerting animal producers about the disease and raise awareness in the research community for future innovative activities.

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“…Detection of brucellosis is conducted by various methods, including cultural isolation, serodiagnosis, and molecular diagnosis ( Yagupsky et al, 2019 ). Besides, researchers have used bioinformatics approaches to explore brucellosis based on the molecular biological properties of Brucella , such as by computationally analyzing the surface properties of Brucella outer membrane protein for vaccine design ( Chen et al, 2021 ) or the correlation of the virulence with structural features of Brucella subtypes ( Paci et al, 2020 ). At present, the development of sequencing technologies has obvious advantages in providing digital data in a cost-effective manner ( Mardis, 2017 ) as well as in delivering high-resolution sequencing results, which is a portable precondition for comparative purposes.…”

Section: Introductionmentioning

confidence: 99%

An integrated platform for Brucella with knowledge graph technology: From genomic analysis to epidemiological projection

Xiao

Zhu

et al. 2022

Front. Genet.

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Motivation:Brucella, the causative agent of brucellosis, is a global zoonotic pathogen that threatens both veterinary and human health. The main sources of brucellosis are farm animals. Importantly, the bacteria can be used for biological warfare purposes, requiring source tracking and routine surveillance in an integrated manner. Additionally, brucellosis is classified among group B infectious diseases in China and has been reported in 31 Chinese provinces to varying degrees in urban areas. From a national biosecurity perspective, research on brucellosis surveillance has garnered considerable attention and requires an integrated platform to provide researchers with easy access to genomic analysis and provide policymakers with an improved understanding of both reported patients and detected cases for the purpose of precision public health interventions.Results: For the first time in China, we have developed a comprehensive information platform for Brucella based on dynamic visualization of the incidence (reported patients) and prevalence (detected cases) of brucellosis in mainland China. Especially, our study establishes a knowledge graph for the literature sources of Brucella data so that it can be expanded, queried, and analyzed. When similar “epidemiological comprehensive platforms” are established in the distant future, we can use knowledge graph to share its information. Additionally, we propose a software package for genomic sequence analysis. This platform provides a specialized, dynamic, and visual point-and-click interface for studying brucellosis in mainland China and improving the exploration of Brucella in the fields of bioinformatics and disease prevention for both human and veterinary medicine.

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Proteomic analysis of Brucella melitensis and Brucella ovis for identification of virulence factor using bioinformatics approachs

Cited by 9 publications

References 30 publications

Identification of Immunogenic Candidate for New Serological Tests for Brucella melitensis by a Proteomic Approach

Identification of Immunogenic Candidate for New Serological Tests for Brucella melitensis by a Proteomic Approach

Comparative Review of Brucellosis in Small Domestic Ruminants

An integrated platform for Brucella with knowledge graph technology: From genomic analysis to epidemiological projection

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