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1.
Medicine (Baltimore) ; 102(11): e33182, 2023 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-36930131

RESUMO

Brucellosis is a zoonotic disease caused by Brucella. There is no effective vaccine against human brucellosis. Omp19 and Omp25 are the outer membrane proteins of Brucella. They are widely expressed and highly conserved in Brucella and have high immunogenicity. Herein, we aim to identify multi-epitope vaccine candidates based on Omp19 and Omp25. We analyzed the physicochemical properties and protein structure of Omp19 and Omp25, and predicted the corresponding B cell and T cell epitopes using bioinformatics analysis. Omp19 and Omp25 were composed of 177 amino acids and 213 amino acids, respectively. They were both stable hydrophilic proteins. The instability indices were 44.8 and 23, respectively. The hydrophilicity was -0.1 and -0.317, respectively. In the secondary structure of Omp19 and Omp25 proteins, the α-helix accounted for 12.43% and 23.94%, the ß-sheet was 18.64% and 23.47%, the ß-turn was 6.78% and 4.23%, and the random coil was 62.15% and 48.36%. Finally, 5 B cell epitopes, 3 Th-cell epitopes and 5 CTL cell epitopes of Omp19 protein, and 4 B cell epitopes, 3 Th-cell epitopes, and 5 CTL cell epitopes of Omp25 protein were selected as vaccine candidates. In conclusion, we obtained potential B cell and T cell epitopes of the Brucella outer membrane Omp19 and Omp25 proteins. This lays the foundation for the further design of multi-epitope vaccine of Brucella.


Assuntos
Brucella , Brucelose , Vacinas , Humanos , Epitopos de Linfócito T , Epitopos de Linfócito B , Antígenos de Bactérias , Proteínas da Membrana Bacteriana Externa , Biologia Computacional , Aminoácidos
2.
Int J Mol Sci ; 24(3)2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36768429

RESUMO

Brucella intermedia (formerly Ochrobactrum intermedium), a non-fermentative bacterium, has been isolated from animals and human clinical specimens. It is naturally resistant to polymyxins, including colistin (CO), and may cause opportunistic infections in humans. We isolated six Brucella intermedia strains from Senegalese monkey stool. In order to determine whether an efflux pump mechanism was involved in CO resistance in B. intermedia, we evaluated the effects of verapamil (VRP), reserpine (RSP), phe-arg ß-naphthylamide dihydrochloride (PAßN) and carbonyl cyanide 3-chloro phenyl hydrazone (CCCP), four efflux pump inhibitors, on these colistin-resistant strains. Using the broth microdilution method, a CO and CCCP combination of 2 µg/mL and 10 µg/mL, respectively, significantly reduced the CO minimal inhibitory concentration (MIC) of B. intermedia, supporting an efflux pump mechanism. In contrast, VRP, PAßN and RSP did not restore CO susceptibility. A time kill assay showed a bactericidal effect of the CO-CCCP combination. Genomic analysis revealed a potential implication in the CO resistance mechanism of some conserved efflux pumps, such as YejABEF, NorM and EmrAB, as previously reported in other bacteria. An inhibitory effect of the CO-CCCP combination was observed on biofilm formation using the crystal violet method. These results suggest that the intrinsic CO resistance in Brucella intermedia is linked to an efflux pump mechanism and that the synergistic effect of CO-CCCP may open a new field to identify new treatments to restore antibiotic efficacy in humans.


Assuntos
Brucella , Colistina , Humanos , Colistina/farmacologia , Carbonil Cianeto m-Clorofenil Hidrazona/farmacologia , Hidrazonas , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana
4.
J Equine Vet Sci ; 123: 104227, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36649828

RESUMO

Brucellosis, a bacterial infection caused by Brucella spp., is a widespread zoonosis concerning human and animal health. In equines, brucellosis may occur asymptomatically or with clinical signs such as arthritis, bursitis, and tenosynovitis. This study aims to ascertain the overall seroprevalence of equine brucellosis and its related factors, including geographic region, serological detection method, equine's species, gender, age, and body condition. This is because equine brucellosis is a zoonotic infection with significant epidemiological implications. The systematic literature search was conducted from January 1, 1990, to April 1, 2022, in the following electronic databases: Google Scholar, Scopus, Pub Med, Science Direct, Web of Science, Embase, Springer, and ProQuest. Based on the preferred reporting items for systematic reviews and meta-analyses (PRISMA) and inclusion criteria, 49 qualified studies were acquired from several countries. Because Egger's test result and the asymmetry in the funnel plot indicated significant publication bias, the meta-analysis model was corrected using the trim-and-fill method. After correction, the pooled seroprevalence of equine brucellosis was 1.92% (95% CI, 0.67-3.79; estimated by the random effect model). Based on meta-regression analysis, the probable heterogeneity factors may be geographic areas (continents and countries). The brucellosis seroprevalence in equines was the lowest in Europe (0%, 95% CI, 0-0.1) and the highest in Asia (10.03%, 95% CI, 8-12.9). The highest overall estimate of seroprevalence of Brucella infection in equines based on a subset of serological tests was made using the serum agglutination test (10.2%, 95% CI, 7.7-13.4). The combined seroprevalence of brucellosis in donkeys (7.3%, 95% CI, 4.9-9.7) was higher than in other equine species. The overall seroprevalence of brucellosis in female equines was significantly higher than in males (OR = 1.437; 95% CI, 1.025-2.014); despite this, the overall seroprevalence had no significant difference based on other risk factors such as age and body condition. Understanding the epidemiology of this zoonotic infection in equines is critical for one-health integration. In addition, this study's results may be appropriate for controlling and managing equine brucellosis.


Assuntos
Brucella , Brucelose , Doenças dos Cavalos , Masculino , Animais , Cavalos , Humanos , Feminino , Estudos Soroepidemiológicos , Brucelose/epidemiologia , Brucelose/veterinária , Brucelose/microbiologia , Zoonoses/epidemiologia , Equidae
5.
mBio ; 14(1): e0333822, 2023 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-36622142

RESUMO

Host cell egress is a critical step in the life cycle of intracellular pathogens, especially in microbes capable of establishing chronic infections. The Gram-negative bacterium Brucella belongs to such a group of pathogens. Even though much has been done to understand how Brucella avoids killing and multiplies in its intracellular niche, the mechanism that this bacterium deploys to egress from the cell to complete its cycle has been poorly studied. In the manuscript, we quantify the kinetics of bacterial egress and show that Brucella exploits multivesicular bodies to exit host cells. For the first time, we visualized the process of egress in real time by live video microscopy and showed that a population of intracellular bacteria exit from host cells in vacuoles containing multivesicular body-like features. We observed the colocalization of Brucella with two multivesicular markers, namely, CD63 and LBPA, both during the final stages of the intracellular life cycle and in egressed bacteria. Moreover, drugs that either promote or inhibit multivesicular bodies either increased or decreased the number of extracellular bacteria, respectively. Our results strongly suggest that Brucella hijacks multivesicular bodies to exit the host cells to initiate new infection events. IMPORTANCE How intracellular bacterial pathogens egress from host cells has been poorly studied. This is particularly important because this stage of the infectious cycle can have a strong impact on how the host resolves the infection. Brucella is an intracellular pathogen that infects mammals, including humans, and causes a chronic debilitating illness. The bacterium has evolved a plethora of mechanisms to invade host cells, avoid degradation in the endocytic pathway, and actively multiply within a specialized intracellular compartment. However, how this pathogen exits from infected cells to produce reinfection and complete its life cycle is poorly understood. In the manuscript, we shed some light on the mechanisms that are exploited by Brucella to egress from host cells. We observed for the first time the egress of Brucella from infected cells by time-lapse video microscopy, and we found that the bacterium exits in vesicles containing multivesicular bodies (MVBs) features. Moreover, the drug manipulation of MVBs resulted in the alteration of bacterial egress efficiency. Our results indicate that Brucella hijacks MVBs to exit host cells and that this strongly contributes to the reinfection cycle.


Assuntos
Brucella , Humanos , Animais , Corpos Multivesiculares , Reinfecção/metabolismo , Vacúolos/metabolismo , Bactérias , Mamíferos
6.
Emerg Infect Dis ; 29(2): 333-340, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36692350

RESUMO

Human brucellosis is a zoonoses caused by bacteria of the genus Brucella. Infection results in subacute or chronic debilitating disease with nonspecific clinical manifestations and is often associated with consuming unpasteurized dairy products. We report 2 cases of brucellosis in male patients who were hospitalized in distinct towns of French Guiana, an overseas territory of France located on the northeastern shore of South America. Both men were citizens of Brazil working as clandestine goldminers in the deep Amazonian rainforest. Characterization of the 2 bacterial isolates revealed that they represent a potential new species of Brucella. Medical practitioners working in contact with wildlife in this region of the world should be aware of the existence of these pathogens and the potential for human infection.


Assuntos
Brucella , Brucelose , Animais , Humanos , Masculino , Guiana Francesa/epidemiologia , Brucelose/diagnóstico , Brucelose/epidemiologia , Brucelose/microbiologia , Zoonoses/microbiologia , Brasil
7.
J Microbiol Methods ; 205: 106663, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36592896

RESUMO

BACKGROUND: Brucellosis is a very common zoonosis in certain localized areas worldwide, with a high prevalence in most developing countries. The detection of brucellosis still faces many challenges such as the need for more sensitive and specific diagnostic antigens. METHODS: To evaluate the efficacy of Brucella outer membrane proteins (Omps) Omp2b in combination with omp31 and BP26 as diagnostic antigens for the serological detection of human brucellosis, these proteins were prepared by a prokaryotic expression system. Human brucellosis-positive and-negative sera were collected, and the detection effects of the diagnostic antigens were evaluated using an established indirect ELISA (iELISA) method. Receiver operating characteristic (ROC) curves and the area under the ROC curve (AUC), true positives, true negatives, false positives, false negatives, accuracy, positive predictive value, negative predictive value, analytical specificity, and sensitivity were obtained to evaluate the effectiveness of Omp2b and antigen combinations. RESULTS: The iELISA results showed that the AUC of the antigenic proteins was 0.9100, 0.9387, 0.9343, and 0.9448, respectively, and that the combination of Omp31 and BP26 improved the accuracy and was superior to that of Omp2b alone. Analysis at the determined cut-off values showed that the analytical sensitivity of the assay was 0.8739 (95% CI:0.7974-0.9293) and the analytical specificity was 0.8539 (95% CI:0.7632-0.9199) when using Omp2b alone and 0.8649 when using the combination of Omp2b + BP26 (95% CI:0.7869-0.9223) with an analytical specificity of 0.9213 (95% CI:0.8446-0.9678) and 0.8468 (95% CI:0.7662-0.9082) and an analytical sensitivity of 0.9101 (95% CI:0.8305-0.9604). When Omp2b + Omp31 + BP26 was combined, the analytical sensitivity and specificity were 0.8559 (95% CI:0.7765-0.9153) and 0.9326 (95% CI:0.8590-0.9749), respectively. Protein antigens, including antigen combinations, did not cross-react with Yersinia enterocolitica O9 and E. coli O157: H7, indicating that their specificity was better than that of lipopolysaccharide (LPS). CONCLUSIONS: Compared with individual Omp2b, antigen combinations improved the effectiveness in detecting brucellosis, but were still not as effective as LPS antigen. Omp2b, combined with Omp31 and BP26 as diagnostic antigens, can be used to detect human brucellosis.


Assuntos
Brucella , Brucelose , Humanos , Lipopolissacarídeos , Escherichia coli , Brucelose/diagnóstico , Proteínas de Membrana , Ensaio de Imunoadsorção Enzimática/métodos , Sensibilidade e Especificidade , Anticorpos Antibacterianos , Antígenos de Bactérias
9.
J Clin Microbiol ; 61(2): e0028122, 2023 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-36533907

RESUMO

Novel bacterial taxonomy and nomenclature revisions can have significant impacts on clinical practice, disease epidemiology, and veterinary microbiology laboratory operations. Expansion of research on the microbiota of humans, animals, and insects has significant potential impacts on the taxonomy of organisms of clinical interest. Implications of taxonomic changes may be especially important when considering zoonotic diseases. Here, we address novel taxonomy and nomenclature revisions of veterinary significance. Noteworthy discussion centers around descriptions of novel mastitis pathogens in Streptococcaceae, Staphylococcaceae, and Actinomycetaceae; bovine reproductive tract pathogens in Corynebacteriaceae; novel members of Mannheimia spp., Leptospira spp., and Mycobacterium spp.; the transfer of Ochrobactrum spp. to Brucella spp.; and revisions to the genus Mycoplasma.


Assuntos
Brucella , Leptospira , Feminino , Animais , Bovinos , Humanos , Animais Domésticos , Bactérias , Zoonoses/microbiologia
10.
Microb Pathog ; 174: 105930, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36496059

RESUMO

Brucellosis is a zoonotic disease caused by Gram-negative bacteria of the genus Brucella. These pathogens cause long-lasting infections, a process in which Brucella modifications in the lipopolysaccharide (LPS) and envelope lipids reduce pathogen-associated molecular pattern (PAMP) recognition, thus hampering innate immunity activation. In vivo models are essential to investigate bacterial virulence, mice being the most used model. However, ethical and practical considerations impede their use in high-throughput screening studies. Although lacking the complexity of the mammalian immune system, insects share key-aspects of innate immunity with mammals, and Galleria mellonella has been used increasingly as a model. G. mellonella larvae have been shown useful in virulence analyses, including Gram-negative pathogens like Klebsiella pneumoniae and Legionella pneumophila. To assess its potential to study Brucella virulence, we first evaluated larva survival upon infection with representative Brucella species (i.e.B. abortus 2308W, B. microti CCM4915 and B. suis biovar 2) and mutants in the VirB type-IV secretion system (T4SS) or in the LPS-O-polysaccharide (O-PS). As compared to K.pneumoniae, the Brucella spp. tested induced a delayed and less severe mortality profile consistent with an escape of innate immunity detection. Brucella replication within larvae was affected by the lack of O-PS, which is reminiscent of their attenuation in natural hosts. On the contrary, replication was not affected by T4SS dysfunction and the mutant induced only slightly less mortality (not statistically significant) than its parental strain. We also evaluated G. mellonella to efficiently recognise Brucella and their LPS by quantification of the pro-phenoloxidase system and melanisation activation, using Pseudomonas LPS as a positive control. Among the brucellae, only B. microti LPS triggered an early-melanisation response consistent with the slightly increased endotoxicity of this species in mice. Therefore, G. mellonella represents a tool to screen for potential Brucella factors modulating innate immunity, but its usefulness to investigate other mechanisms relevant in Brucella intracellular life is limited.


Assuntos
Brucella , Mariposas , Animais , Camundongos , Mariposas/microbiologia , Lipopolissacarídeos , Larva/microbiologia , Interações Hospedeiro-Patógeno , Mamíferos
11.
Vet Med Sci ; 9(1): 523-534, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36583994

RESUMO

BACKGROUND: Brucellosis, caused by Brucella spp., is a major zoonotic public health threat. Although several Brucella vaccines have been demonstrated for use in animals, Brucella spp. can cause human infection and to date, there are no human-use vaccines licensed by any agency. Recently, methods in vaccine informatics have made major breakthroughs in peptide-based epitopes, opening up a new avenue of vaccine development. OBJECTIVES: The purpose of this article was to identify potential antigenic peptides in Brucella by proteome and peptidome analyses. METHODS: Mouse infection models were first established by injection with Brucella and spleen protein profiles were then analysed. Subsequently, the major histocompatibility complex class I or II (major histocompatibility complex [MHC]-I/II)-binding peptides in blood samples were collected by immunoprecipitation and peptides derived from Brucella proteins were identified through liquid chromatography-mass spectrometry (LC-MS/MS). These peptides were then evaluated in a variety of ways, such as in terms of conservation in Brucella and synchronicity in predicted peptides (similarity and coverage), which allowed us to more effectively measure their antigenic potential. RESULTS: The expression of the inflammatory cytokines IL1B and IFN-γ was significantly altered in the spleen of infected mice and some Brucella proteins, such as Muri, AcpP and GroES, were also detected. Meanwhile, in blood, 35 peptides were identified and most showed high conservation, highlighting their potential as antigen epitopes for vaccine development. In particular, we identified four proteins containing both MHC-I- and MHC-II-binding peptides including AtpA, AtpD, DnaK and BAbS19_II02030. They were also compared with the predicted peptides to estimate their reliability. CONCLUSIONS: The peptides we screened could bind to MHC molecules. After being stimulated with antigen T epitopes, Memory T cells can stimulate T cell activation and promote immune responses. Our results indicated that the peptides we identified may be good candidate targets for the design of subunit vaccines and these results pave the way for the study of safer vaccines against Brucella.


Assuntos
Brucella , Animais , Camundongos , Proteoma , Cromatografia Líquida/veterinária , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem/veterinária , Epitopos , Peptídeos
12.
J Int Med Res ; 50(12): 3000605221139668, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36482666

RESUMO

Brucellosis is a common zoonotic infectious disease worldwide that can affect almost all organs in the human body. Osteonecrosis of the femoral head has numerous possible causes. Patients usually experience movement disorders, severe pain, and an extremely high disability rate. We herein describe a 32-year-old male farmer living in Shanxi, China who accidentally injured his right hip while working. After 1 month of rest, the pain in the right hip was relieved, and the patient began to walk again. However, after 2 months of walking, he developed aggravated pain and claudication in the right lower limb. Examination revealed shortening of the right lower limb. We carried out a comprehensive and systematic examination and performed total hip arthroplasty. Hip trauma with Brucella infection may progress to osteonecrosis of the femoral head in a short period of time without typical symptoms of Brucella infection. Systematic examination and treatment should be performed at an early stage to prevent the eventual deterioration of the disease and serious complications that are difficult to treat.


Assuntos
Brucella , Brucelose , Necrose da Cabeça do Fêmur , Humanos , Adulto , Cabeça do Fêmur , Necrose da Cabeça do Fêmur/diagnóstico por imagem , Necrose da Cabeça do Fêmur/etiologia , Necrose da Cabeça do Fêmur/cirurgia , Caminhada , Brucelose/complicações
13.
J Med Case Rep ; 16(1): 492, 2022 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-36585719

RESUMO

BACKGROUND: Brucellosis is one of the most common zoonotic diseases in the world. Osteoarticular complications, especially vertebral system involvement, are most commonly reported. However, reports and coreports of pulmonary complications and thoracal spondylodiscitis and epidural abscess are rare. CASE PRESENTATION: Spondylodiscitis was detected at the T11-12 vertebral level, followed by epidural and paravertebral abscess, and then empyema was detected in a 17-year-old Asian female patient without any additional disease. The patient had used various antibiotics and the disease could not be proven bacteriologically. Also, the Rose Bengal test was negative. However, serologically high titer Brucella positivity was detected in the blood and pleural fluid sample. Drainage was required for bilateral empyema. Disease duration prolonged due to multiple complications. The patient was cured with combined long-term treatment for brucellosis. CONCLUSIONS: Although some are rare, brucellosis is a zoonotic disease that can cause many complications. The gold standard for diagnosis is the growth of bacteria in blood culture or tissue culture. However, isolation of the microorganism can be very difficult. Clinical suspicion and serological tests are important guides.


Assuntos
Brucella , Brucelose , Discite , Animais , Humanos , Feminino , Adolescente , Discite/tratamento farmacológico , Brucelose/complicações , Brucelose/diagnóstico , Brucelose/tratamento farmacológico , Antibacterianos/uso terapêutico , Coluna Vertebral , Zoonoses/tratamento farmacológico
14.
J Med Case Rep ; 16(1): 418, 2022 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-36329543

RESUMO

BACKGROUND: Human brucellosis is the most frequently contracted zoonotic infection worldwide. Although being an old disease that carries minimal risks of mortality, it remains a source of considerable sequelae and disability. However, noncontiguous multifocal spinal involvement is an exceptional presentation of brucellosis; additionally, an associated paravertebral abscess is extremely rare. CASE PRESENTATION: This paper focuses on a 67-year-old Lebanese woman with noncontiguous multifocal Brucella spondylodiscitis, involving the T12-L1 and L3-L4 segments, with paravertebral abscess formation. She presented with a 3-week history of acute severe lumbar back pain, radiating to the lower extremities and associated with impaired mobility and lower extremity weakness. The patient complained of night sweating but had no fever. No lymphadenopathy, hepatomegaly, or splenomegaly could be observed. She had painful percussion of the lumbar spine, painful passive mobilization, and paravertebral tenderness, yet her neurological examination was completely normal. BrucellaCapt test was positive at a titer of 1/5120 (reference range 1/180). The patient was treated with an inpatient regimen for 2 weeks, which was followed by an outpatient oral antibiotic regimen with doxycycline, rifampin, and ciprofloxacin to complete a total treatment duration of 3 months. Magnetic resonance imaging was performed at the end of the treatment and showed a complete resolution of the paravertebral abscess. CONCLUSION: Noncontiguous multifocal Brucella spondylodiscitis with paravertebral abscess is an extremely rare presentation. It may be effectively managed by antibiotic therapy, without surgery or drainage, in the absence of neurological complications. Nonetheless, the principal challenge to an efficient management is establishing the diagnosis of Brucella in the first place. In endemic countries, a strong suspicion of spinal involvement of brucellosis should be elicited in front of back pain presentations-even in the absence of fever and other related symptoms.


Assuntos
Brucella , Brucelose , Discite , Dor Lombar , Feminino , Humanos , Idoso , Discite/diagnóstico por imagem , Discite/tratamento farmacológico , Discite/complicações , Abscesso/diagnóstico por imagem , Abscesso/terapia , Abscesso/complicações , Brucelose/complicações , Brucelose/diagnóstico , Brucelose/tratamento farmacológico , Antibacterianos/uso terapêutico , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/patologia , Imageamento por Ressonância Magnética , Dor nas Costas/etiologia , Dor nas Costas/tratamento farmacológico , Dor Lombar/tratamento farmacológico
15.
Curr Microbiol ; 80(1): 2, 2022 Nov 22.
Artigo em Inglês | MEDLINE | ID: mdl-36418790

RESUMO

The purpose of this study was to elucidate the roles of peptidoglycan-associated lipoprotein (Pal protein) in the proliferation of Brucella in macrophage and bacterial virulence, and to evaluate the immune effect of Pal protein to Salmonella enteritidis. Murine macrophage-like cell line Raw264.7 was stimulated by recombinant Pal protein, and the expression of TNF-α and IFN-γ were up-regulated, but not it of IL-1ß and IL-6. The macrophages infection and in vitro simulated stress assays showed that deletion of pal gene reduced the proliferation of Brucella in macrophages, the survival in acidic, oxidative and polymyxin B-contained environment. The mice infection assay showed that mice challenged with the pal mutant strain were found to have more severe splenomegaly, but less bacterial load. After oral immunization of mice, Pal protein induced a higher titer of mucosal and humoral antibody (IgA and IgG) against heat-killed Salmonella enteritidis, and a stronger Th1 cellular immune response. The challengte experiments showed Pal protein elevated the survival rate and reduced the bacterial load of spleens in immunized mice. In conclusion, our results revealed the important roles of pal gene in Brucella virulence, and Pal protein was a potentially valuable adjuvant against mucosal pathogens, such as Salmonella enteritidis.


Assuntos
Brucella , Camundongos , Animais , Salmonella enteritidis/genética , Virulência , Macrófagos , Proliferação de Células
16.
PLoS One ; 17(11): e0261595, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36413520

RESUMO

Bovine brucellosis is endemic in Rwanda, although, there is a paucity of documented evidence about the disease in slaughtered cattle. A cross-sectional study was conducted in slaughtered cattle (n = 300) to determine the seroprevalence of anti-Brucella antibodies using the Rose Bengal Test (RBT), and indirect enzyme-linked immunosorbent assay (i-ELISA). Corresponding tissues were cultured onto a modified Centro de Investigación y Tecnología Agroalimentaria (CITA) selective medium and analysed for Brucella spp. using the 16S-23S ribosomal interspacer region (ITS), AMOS, and Bruce-ladder PCR assays. The seroprevalence was 20.7% (62/300) with RBT, 2.9% (8/300) with i-ELISA, and 2.9% (8/300) using both tests in series. Brucella-specific 16S-23S ribosomal DNA interspace region (ITS) PCR detected Brucella DNA in 5.6% (17/300; Brucella culture prevalence). AMOS-PCR assay identified mixed B. abortus and B. melitensis (n = 3), B. abortus (n = 3) and B. melitensis (n = 5) while Bruce-ladder PCR also identified B. abortus (n = 5) and B. melitensis (n = 6). The gold standard culture method combined with PCR confirmation identified 5.6% Brucella cultures and this culture prevalence is higher than the more sensitive seroprevalence of 2.9%. This emphasizes the need to validate the serological tests in Rwanda. The mixed infection caused by B. abortus and B. melitensis in slaughtered cattle indicates cross-infection and poses a risk of exposure potential to abattoir workers. It is essential to urgently strengthen a coordinated national bovine brucellosis vaccination and initiate a test-and-slaughter program that is not presently applicable in Rwanda.


Assuntos
Brucella , Brucelose Bovina , Brucelose , Bovinos , Animais , Brucella/genética , Estudos Soroepidemiológicos , Ruanda/epidemiologia , Brucelose Bovina/epidemiologia , Estudos Transversais , Brucelose/epidemiologia , Brucelose/veterinária , Rosa Bengala
17.
World J Microbiol Biotechnol ; 39(1): 23, 2022 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-36422675

RESUMO

Brucellosis is a chronic infectious disease caused by Brucella, which is characterized by inflammation of reproductive organs and fetal membranes, abortion, infertility, and local inflammatory lesions of various tissues. Due to the widespread prevalence and spread of brucellosis, it has not only caused huge losses to animal husbandry, but also brought serious impacts on human health and safety. Therefore, rapid and accurate diagnosis is of great significance for the effective control of brucellosis. Therefore, we have developed a rapid vertical flow technique (RVFT) using Prussian blue nanoparticles (PBNPs) as a marker material for the detection of brucellosis antibodies. Lipopolysaccharide (LPS) was purified and used to detect brucellosis antibodies to improve the sensitivity of this technique. To enhance the sensitivity of serum antibody detection, a single multifunctional compound buffer was created using whole blood as a biological sample while retaining the advantages of typical lateral flow immunoassays. After signal amplification, standard Brucella-positive serum (containing Brucella antibody at 4000 IU mL-1) could be detected in this system even at a dilution factor of 1 × 10-2. The detection limit was 40 IU mL-1, which is ten times that before signal amplification. This RVFT displayed good specificity and no cross-reactivity. This RVFT effectively avoided the false negative phenomenon of lateral flow immunoassays, was easy to operate, had a short reaction time, has good repeatability, and could elicit results that were visible to the naked eye for 2 ~ 3 min without any equipment. Since this method is very important for controlling the prevalence of brucellosis, it holds great promise for application in primary medical units and veterinary brucellosis detection.


Assuntos
Brucella , Brucelose , Nanopartículas , Animais , Humanos , Brucelose/diagnóstico , Brucelose/veterinária , Anticorpos Antibacterianos , Catálise
18.
Int J Mol Sci ; 23(22)2022 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-36430916

RESUMO

Brucella transfers effectors into host cells, manipulating cellular processes to its advantage; however, the mechanism by which effectors regulate cellular processes during infection is poorly understood. A growing number of studies have shown that apoptosis and autophagy are critical mechanisms for target cells to cope with pathogens and maintain cellular homeostasis. BtpB is a Brucella type IV secretion system effector with a complex mechanism for manipulating host infection. Here, we show that the ectopic expression of BtpB promoted DNA fragmentation. In contrast, an isogenic mutant strain, ΔbtpB, inhibited apoptosis compared to the wild-type strain B. suis S2 in RAW264.7 cells. In addition, BtpB inhibited autophagy, as determined by LC3-II protein levels, the number of LC3 puncta, and p62 degradation. We also found that BtpB reduced autophagolysosome formation and blocked the complete autophagic flux. Moreover, our results revealed that the autophagy inhibitor, chloroquine, reduces Brucella's intracellular survival. Overall, our data unveil new mechanisms of virulence implicating the effector BtpB in regulating host intracellular infection.


Assuntos
Brucella , Camundongos , Animais , Brucella/genética , Autofagia/genética , Apoptose , Células RAW 264.7 , Sistemas de Secreção Tipo IV/genética
19.
BMC Surg ; 22(1): 394, 2022 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-36401260

RESUMO

BACKGROUND: This study aimed to assess the clinical efficacy of one-stage posterior surgery combined with anti-Brucella therapy in the treatment of lumbosacral brucellosis spondylitis (LBS). METHODS: From June 2010 to June 2020, the clinical and radiographic data of patients with LBS treated by one-stage posterior surgery combined with anti-Brucella therapy were retrospectively analyzed. The visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) and Oswestry Disability Index scores (ODI) were used to evaluate the clinical outcomes. Frankel's classification system was employed to access the initial and final neurologic function. Fusion of the bone grafting was classified by Bridwell's grading system. RESULTS: A total of 55 patients were included in this study with a mean postoperative follow-up time of 2.6 ± 0.8 years (range, 2 to 5). There were 40 males and 15 females with a mean age of 39.8 ± 14.7 years (range, 27 to 57). The Brucella agglutination test was ≥ 1:160 in all patients, but the blood culture was positive in 43 patients (78.1%). A statistical difference was observed in ESR, CRP, VAS, ODI, and JOA between preoperative and final follow-up (P < 0.05). Neurological function was significantly improved in 20 patients with preoperative neurological dysfunction after surgery. According to Bridwell's grading system, the fusion of bone grafting in 48 cases (87.2%) was defined as grade I, and grade II in 7 cases (12.7%). None of the infestation recurrences was observed. CONCLUSION: One-stage posterior surgery combined with anti-Brucella therapy was a practical method in the treatment of LBS with severe neurological compression and spinal sagittal imbalance.


Assuntos
Brucella , Brucelose , Fusão Vertebral , Espondilite , Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Estudos Retrospectivos , Fusão Vertebral/métodos , Vértebras Lombares/cirurgia , Desbridamento/métodos , Espondilite/cirurgia , Brucelose/complicações , Brucelose/tratamento farmacológico , Brucelose/cirurgia
20.
Sci Rep ; 12(1): 19459, 2022 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-36376442

RESUMO

We aimed to evaluate the clinical efficacy of the single-stage posterior surgical treatment for patients of lumbar brucella spondylitis combined with spondylolisthesis. In this study, we performed a retrospective analysis of 16 patients with lumbar brucellosis spondylitis combined with spondylolisthesis from January 2015 to January 2019. All patients underwent single-stage posterior lumbar debridement, reduction, interbody fusion, and instrumentation. Preoperative and postoperative of the visual analog scale (VAS), the Oswestry disability index (ODI), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) were compared. In addition, the spondylolisthesis reduction rate, reduction loss rate, interbody fusion rate, and complication rate were recorded. VAS, ODI, ESR, and CRP were conducted with repeated analysis of variance data at different follow-ups. The postoperative follow-up was 12-36 months, with an average of (25.0 ± 8.1) months. VAS, ODI, ESR, and CRP were significantly better at 2-week and 1-year follow-up than preoperative results (P = 0.000, respectively). In addition, 1 year after the operation, VAS, ODI, ESR, and CRP showed a significant improvement (P = 0.000, respectively). The average spondylolisthesis reduction in 2 weeks after operation was (91.2 ± 6.7)%, and the median reduction loss rate in 1 year after operation was 8.0 (5.0, 9.8)%. At the last follow-up, all patients achieved interbody fusion, no loosening and fracture of instrumentation were found, and no recurrence happened. Single-stage posterior operation for lumbar debridement, reduction, interbody fusion, and instrumentation is beneficial for treating lumbar brucellosis spondylitis combined with spondylolisthesis. Furthermore, the reconstruction of spinal stability may relieve pain, heal lesions, and improve patients' living.


Assuntos
Brucella , Brucelose , Fusão Vertebral , Espondilite , Espondilolistese , Humanos , Espondilolistese/cirurgia , Fusão Vertebral/métodos , Estudos Retrospectivos , Vértebras Lombares/cirurgia , Espondilite/cirurgia , Brucelose/cirurgia , Resultado do Tratamento
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