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1.
Int J Mol Sci ; 22(19)2021 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-34639201

RESUMO

The arsenic acid-resistant (ArsR) family transcriptional regulators are widely distributed in microorganisms, including in the facultative intracellular pathogen Brucella spp. ArsR proteins are implicated in numerous biological processes. However, the specific roles of ArsR family members in Brucella remain obscure. Here, we show that ArsR6 (BSS2_RS07325) is required for Brucella survival both under heat, oxidative, and osmotic stress and in a murine infection model in vivo. RNA-seq and ChIP-seq reveal that 34 potential target genes for ArsR6 protein were identified, among which eight genes were up-regulated and 26 genes were down-regulated, including outer membrane protein 25D (Omp25D). ArsR6 autoregulates its own expression to maintain bacterial intracellular Cu/Ni homeostasis to benefit bacterial survival in hostile environments. Moreover, ArsR6 also regulates the production of virulence factor Omp25D, which is important for the survival of Brucella under stress conditions. Significantly, Omp25D deletion strain attenuated in a murine infection model in vivo. Altogether, our findings reveal a unique mechanism in which the ArsR family member ArsR6 autoregulates its expression and also modulates Omp25D expression to maintain metal ion homeostasis and virulence in Brucella.


Assuntos
Proteínas de Bactérias/metabolismo , Brucella/crescimento & desenvolvimento , Brucelose/microbiologia , Regulação Bacteriana da Expressão Gênica , Transativadores/metabolismo , Fatores de Virulência/metabolismo , Virulência , Animais , Brucella/genética , Brucella/metabolismo , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Transativadores/genética , Fatores de Virulência/genética
2.
BMC Vet Res ; 17(1): 342, 2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-34717610

RESUMO

BACKGROUND: We implemented a longitudinal study to determine the incidence of Brucella infection in cattle, camels, sheep and goats that were being raised in a pastoral area in Isiolo County, Kenya. An initial cross-sectional survey was implemented to identify unexposed animals for follow up; that survey used 141 camels, 216 cattle, 208 sheep and 161 goats. Sera from these animals were screened for Brucella spp. using the Rose Bengal Plate test (RBPT), a modified RBPT, and an indirect multispecies Enzyme Linked Immunosorbent Assay (iELISA). Results of RBPT and iELISA were interpreted in parallel to determine seroprevalence. A total of 30 camels, 31 cattle, 22 sheep and 32 goats that were seronegative by all the above tests were recruited in a subsequent longitudinal study for follow up. These animals were followed for 12 months and tested for anti-Brucella antibodies using iELISA. Seroconversion among these animals was defined by a positive iELISA test following a negative iELISA result in the previous sampling period. All seropositive samples were further tested using real-time PCR-based assays to identify Brucella species. These analyses targeted the alkB and BMEI1162 genes for B. abortus, and B. melitensis, respectively. Data from the longitudinal study were analysed using Cox proportional hazards model that accounted for within-herds clustering of Brucella infections. RESULTS: The overall incidence rate of Brucella infection was 0.024 (95% confidence interval [CI]: 0.014-0.037) cases per animal-months at risk. Brucella infection incidence in camels, cattle, goats and sheep were 0.053 (0.022-0.104), 0.028 (0.010-0.061), 0.013 (0.003-0.036) and 0.006 (0.0002-0.034) cases per animal-month at risk, respectively. The incidence rate of Brucella infection among females and males were 0.020 (0.009-0.036) and 0.016 (0.004-0.091), respectively. Real-time PCR analyses showed that B. abortus was more prevalent than B. melitensis in the area. Results of multivariable Cox regression analysis identified species (camels and cattle) as an important predictor of Brucella spp. exposure in animals. CONCLUSIONS: This study estimated an overall brucellosis incidence of 0.024 cases per animal-months at risk with camels and cattle having higher incidence than sheep and goats. These results will inform surveillance studies in the area.


Assuntos
Brucella/imunologia , Brucelose/veterinária , Camelus/microbiologia , Doenças dos Bovinos/epidemiologia , Doenças das Cabras/epidemiologia , Doenças dos Ovinos/epidemiologia , Animais , Brucelose/epidemiologia , Brucelose/microbiologia , Bovinos , Doenças dos Bovinos/microbiologia , Estudos Transversais , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Doenças das Cabras/microbiologia , Cabras , Incidência , Quênia/epidemiologia , Gado , Estudos Longitudinais , Masculino , Fatores de Risco , Estudos Soroepidemiológicos , Ovinos , Doenças dos Ovinos/microbiologia
3.
Vet Res ; 52(1): 116, 2021 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-34521471

RESUMO

The management of infectious diseases in wildlife reservoirs is challenging and faces several limitations. However, detailed knowledge of host-pathogen systems often reveal heterogeneity among the hosts' contribution to transmission. Management strategies targeting specific classes of individuals and/or areas, having a particular role in transmission, could be more effective and more acceptable than population-wide interventions. In the wild population of Alpine ibex (Capra ibex-a protected species) of the Bargy massif (French Alps), females transmit brucellosis (Brucella melitensis) infection in ~90% of cases, and most transmissions occur in the central spatial units ("core area"). Therefore, we expanded an individual-based model, developed in a previous study, to test whether strategies targeting females or the core area, or both, would be more effective. We simulated the relative efficacy of realistic strategies for the studied population, combining test-and-remove (euthanasia of captured animals with seropositive test results) and partial culling of unmarked animals. Targeting females or the core area was more effective than untargeted management options, and strategies targeting both were even more effective. Interestingly, the number of ibex euthanized and culled in targeted strategies were lower than in untargeted ones, thus decreasing the conservation costs while increasing the sanitary benefits. Although there was no silver bullet for the management of brucellosis in the studied population, targeted strategies offered a wide range of promising refinements to classical sanitary measures. We therefore encourage to look for heterogeneity in other wildlife diseases and to evaluate potential strategies for improving management in terms of efficacy but also acceptability.


Assuntos
Brucella melitensis/fisiologia , Brucelose/veterinária , Doenças das Cabras/prevenção & controle , Animais , Animais Selvagens , Brucelose/microbiologia , Brucelose/prevenção & controle , Feminino , França , Doenças das Cabras/microbiologia , Cabras , Masculino
4.
Food Microbiol ; 100: 103873, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34416970

RESUMO

The bulk milk examination is a reliable screening tool for monitoring the quality of milk in the farms. The infection to Neospora caninum, Toxoplasma gondii and Brucella sp. Was evaluated in bulk milk samples of dairy farms in Hamedan province, West part of Iran. All the dairy farms (n = 149) were examined for N. caninum, T. gondii and Brucella infections using milk ring test (MRT), microbiology, serology (Enzyme-linked Immunosorbent Assay), and molecular techniques. Based on molecular methods, Brucella-infection was negative in all farms; while, 55 %, 5.4 % and 2.7 % of samples were positive for N. caninum, T. gondii and mix infection, respectively. The highest Neospora-infection was detected in the farms with history of abortion in fall and winter. There was significant association between Neospora-infection and the presence of dogs and rodents in the farms, herd size, and age of the animals. Also, a significant association was seen between Toxoplasma-infection and the presence of cats and rodents in the farms, as well as age of the animals. Average total bacterial count (TBC) was calculated 1.14 × 106±1.1 × 106. The highest TBC was in the farms from Central locations of studied area (5.7 × 106±2.24 × 106), farms with more than 120 animals (7.9 × 106±2.8 × 106), and farms with ≥50-months age (1.74 × 106±6.3 × 105) in spring and summer (6.9 × 106±3.7 × 106). The number of somatic cells was estimated between 1 × 104 and 2 × 106 (Average = 4.2 × 105±3.39 × 105). The current study was a comprehensive evaluation of Neospora, Toxoplasma and Brucella infections in milk samples of Iranian dairy farms for the first time. Neospora-infection is responsible for economic losses in the region. Health education and milk pasteurization are so helpful for inhibiting the milk borne diseases. To reduce the risk factors, predict and design the appropriate schemes like redundant of heterogeneous animals are recommended.


Assuntos
Brucella/isolamento & purificação , Coccidiose/veterinária , Contaminação de Alimentos/análise , Leite/microbiologia , Leite/parasitologia , Neospora/isolamento & purificação , Toxoplasma/isolamento & purificação , Animais , Animais Domésticos/microbiologia , Animais Domésticos/parasitologia , Brucella/classificação , Brucella/genética , Brucelose/microbiologia , Brucelose/veterinária , Doenças do Gato/microbiologia , Doenças do Gato/parasitologia , Gatos , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/parasitologia , Coccidiose/parasitologia , Doenças do Cão/microbiologia , Doenças do Cão/parasitologia , Cães , Fazendas , Feminino , Masculino , Leite/química , Neospora/classificação , Neospora/genética , Toxoplasma/classificação , Toxoplasma/genética , Toxoplasmose Animal/parasitologia
5.
BMC Vet Res ; 17(1): 289, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34461896

RESUMO

BACKGROUND: UTP-glucose-1-phosphoryl transferase (UGPase) catalyzes the synthesis of UDP-glucose, which is essential for generating the glycogen needed for the synthesis of bacterial lipopolysaccharide (LPS) and capsular polysaccharide, which play important roles in bacterial virulence. However, the molecular function of UGPase in Brucella is still unknown. RESULTS: In this study, the ubiquitination modification of host immune-related protein in cells infected with UGPase-deleted or wild-type Brucella was analyzed using ubiquitination proteomics technology. The ubiquitination modification level and type of NF-κB Essential Modulator (NEMO or Ikbkg), a molecule necessary for NF-κB signal activation, was evaluated using Coimmunoprecipitation, Western blot, and dual-Luciferase Assay. We found 80 ubiquitin proteins were upregulated and 203 ubiquitin proteins were downregulated in cells infected with B. melitensis 16 M compared with those of B. melitensis UGPase-deleted strain (16 M-UGPase-). Moreover, the ubiquitin-modified proteins were mostly enriched in the categories of regulation of kinase/NF-κB signaling and response to a bacterium, suggesting Brucella UGPase inhibits ubiquitin modification of related proteins in the host NF-κB signaling pathway. Further analysis showed that the ubiquitination levels of NEMO K63 (K63-Ub) and Met1 (Met1-Ub) were significantly increased in the 16 M-UGPase--infected cells compared with that of the 16 M-infected cells, further confirming that the ubiquitination levels of NF-κB signaling-related proteins were regulated by the bacterial UGPase. Besides, the expression level of IκBα was decreased, but the level of p-P65 was significantly increased in the 16 M-UGPase--infected cells compared with that of the 16 M- and mock-infected cells, demonstrating that B. melitensis UGPase can significantly inhibit the degradation of IκBα and the phosphorylation of p65, and thus suppressing the NF-κB pathway. CONCLUSIONS: The results of this study showed that Brucella melitensis UGPase inhibits the activation of NF-κB by modulating the ubiquitination of NEMO, which will provide a new scientific basis for the study of immune mechanisms induced by Brucella.


Assuntos
Brucella melitensis/metabolismo , Quinase I-kappa B/metabolismo , NF-kappa B/metabolismo , UTP-Glucose-1-Fosfato Uridililtransferase/genética , Ubiquitinação , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Brucella melitensis/genética , Brucelose/metabolismo , Brucelose/microbiologia , Regulação da Expressão Gênica , Camundongos , Células RAW 264.7 , Transdução de Sinais , Ubiquitina/genética , Ubiquitina/metabolismo
6.
PLoS One ; 16(7): e0254530, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34283853

RESUMO

Brucellosis and Q fever are neglected zoonoses of global health importance, with unknown true prevalence in occupationally vulnerable settings, partly due to misdiagnosis for other febrile conditions and poor access to primary health care. We examined the seroprevalence of these diseases and associated factors amongst pastoralists and their cattle in Sokoto State, a hub of cattle and pastoral populations in Nigeria. Serum samples randomly collected from 137 pastoralists and 366 cattle from 27 herds in three selected Local Government Areas (LGAs) in the state were analysed for antibodies to Brucella abortus using Rose Bengal Plate Test (RBT) and competitive Enzyme-Linked Immunosorbent Assay (cELISA) as well as antibodies to Coxiella burnetti using indirect ELISA. Consenting pastoralists' knowledge, perception and practices about the diseases were assessed using a semi-structured questionnaire. Data were analysed using descriptive statistics and bivariate analysis at p ≤ 0.05 level of significance. Brucellosis adjusted individual seroprevalence were 0.83% (95%CI: 0.04-4.59%) and 0% among pastoralists; 2.28% (95%CI: 1.16-4.43%) and 5.70% (95%CI: 3.68-8.74%) in cattle by RBT and cELISA, respectively. Adjusted herd-level seroprevalence for brucellosis were 23.20% (95%CI: 11.07-42.54%) and 42.00% (95%CI: 25.27-61.11%) by RBT and cELISA, respectively. For Q fever, higher seroprevalence of 62.57% (95%CI: 54.04-70.46%) and 2.98% (95%CI: 1.57-5.58%) were recorded amongst the pastoralists and their cattle, respectively. with adjusted herd-level seroprevalence of 40.36% (95%CI: 22.57-63.17%). The LGAs of sampling were significantly (OR: 0.2; 95%CI: 0.02-1.00) associated with Q fever infection, though marginal. The majority of the pastoralists had poor knowledge, perception and practices towards the diseases. This is the first study establishing the presence of brucellosis and Q fever at the human-animal interface in Sokoto State, Nigeria. The pastoralists' poor knowledge, perception and practices about these diseases are worrisome and are important factors for consideration in disease control.


Assuntos
Brucelose/sangue , Febre Q/sangue , Estudos Soroepidemiológicos , Zoonoses/sangue , Criação de Animais Domésticos , Animais , Brucella abortus/isolamento & purificação , Brucella abortus/patogenicidade , Brucelose/epidemiologia , Brucelose/microbiologia , Bovinos , Ensaio de Imunoadsorção Enzimática , Cabras/sangue , Cabras/microbiologia , Humanos , Nigéria/epidemiologia , Febre Q/epidemiologia , Febre Q/microbiologia , Fatores de Risco , Zoonoses/epidemiologia , Zoonoses/microbiologia , Zoonoses/transmissão
7.
Infect Immun ; 89(10): e0015621, 2021 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-34125603

RESUMO

Brucellosis is one of the most common global zoonoses and is caused by facultative intracellular bacteria of the genus Brucella. Numerous studies have found that MyD88 signaling contributes to protection against Brucella; however, the underlying mechanism has not been entirely defined. Here, we show that MyD88 signaling in hematopoietic cells contributes both to inflammation and to control of Brucella melitensis infection in vivo. While the protective role of MyD88 in Brucella infection has often been attributed to promotion of gamma interferon (IFN-γ) production, we found that MyD88 signaling restricts host colonization by B. melitensis even in the absence of IFN-γ. In vitro, we show that MyD88 promotes macrophage glycolysis in response to B. melitensis. Interestingly, a B. melitensis mutant lacking the glucose transporter, GluP, was more highly attenuated in MyD88-/- than in wild-type mice, suggesting MyD88 deficiency results in an increased availability of glucose in vivo, which Brucella can exploit via GluP. Metabolite profiling of macrophages identified several metabolites regulated by MyD88 in response to B. melitensis, including itaconate. Subsequently, we found that itaconate has antibacterial effects against Brucella and also regulates the production of proinflammatory cytokines in B. melitensis-infected macrophages. Mice lacking the ability to produce itaconate were also more susceptible to B. melitensis in vivo. Collectively, our findings indicate that MyD88-dependent changes in host metabolism contribute to control of Brucella infection.


Assuntos
Brucelose/metabolismo , Glucose/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Succinatos/metabolismo , Animais , Brucella melitensis/patogenicidade , Brucelose/microbiologia , Citocinas/metabolismo , Glicólise/fisiologia , Interferon gama/metabolismo , Macrófagos/metabolismo , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais/fisiologia
8.
PLoS Pathog ; 17(5): e1009597, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33989349

RESUMO

Macrophages metabolic reprogramming in response to microbial insults is a major determinant of pathogen growth or containment. Here, we reveal a distinct mechanism by which stimulator of interferon genes (STING), a cytosolic sensor that regulates innate immune responses, contributes to an inflammatory M1-like macrophage profile upon Brucella abortus infection. This metabolic reprogramming is induced by STING-dependent stabilization of hypoxia-inducible factor-1 alpha (HIF-1α), a global regulator of cellular metabolism and innate immune cell functions. HIF-1α stabilization reduces oxidative phosphorylation and increases glycolysis during infection with B. abortus and, likewise, enhances nitric oxide production, inflammasome activation and IL-1ß release in infected macrophages. Furthermore, the induction of this inflammatory profile participates in the control of bacterial replication since absence of HIF-1α renders mice more susceptible to B. abortus infection. Mechanistically, activation of STING by B. abortus infection drives the production of mitochondrial reactive oxygen species (mROS) that ultimately influences HIF-1α stabilization. Moreover, STING increases the intracellular succinate concentration in infected macrophages, and succinate pretreatment induces HIF-1α stabilization and IL-1ß release independently of its cognate receptor GPR91. Collectively, these data demonstrate a pivotal mechanism in the immunometabolic regulation of macrophages during B. abortus infection that is orchestrated by STING via HIF-1α pathway and highlight the metabolic reprogramming of macrophages as a potential treatment strategy for bacterial infections.


Assuntos
Brucella abortus/imunologia , Brucelose/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Macrófagos/metabolismo , Proteínas de Membrana/metabolismo , Animais , Brucelose/imunologia , Brucelose/microbiologia , Glicólise , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Inflamassomos/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Óxido Nítrico/metabolismo , Fosforilação Oxidativa , Espécies Reativas de Oxigênio/metabolismo
9.
Vet Med Sci ; 7(4): 1245-1253, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33974356

RESUMO

BACKGROUND: Brucellosis is an infectious zoonotic bacterial disease of humans and other animals. In the Republic of South Africa (RSA), animal brucellosis is widespread and the current available data on the prevalence of this disease rely solely on serological testing. The primary limitation of brucellosis serology is the lack of discriminatory powers to differentiate between Brucella species and biovars as well as the cross-reactivity observed with other Gram-negative bacteria. AIM: The aim of this study was to conduct a retrospective laboratory-based survey on Brucella species and biovars isolated from various animal species in SA between 2008 and 2018. MATERIAL AND METHODS: The isolation of Brucella species and biovar typing was performed using conventional microbiological techniques. RESULTS AND DISCUSSION: A total of 963 strains of Brucella species were included in this study with a frequency of detection for B. abortus (n = 883; 91.6%) followed by B. melitensis (n = 42; 4.4%), B. ovis (n = 29; 3.0%) and B. canis (n = 9; 0.9%). Of the 883 strains of B. abortus, 90.1% were typed as B. abortus biovar-1 while 5.7% as B. abortus biovar-2, and 3.3% and 0.5% were B. abortus S19 and B. abortus RB51 vaccine strains, respectively. Among the 42 B. melitensis strains, 71.4% were reported as B. melitensis biovar-1 and 26.2% as B. melitensis biovar-3 while 2.4% was B. melitensis biovar-2. CONCLUSION: A retrospective study, such as this one, provides useful information that can be critical in formulating policies and strategies for the control and eradication of brucellosis in animal populations in RSA.


Assuntos
Brucella abortus/isolamento & purificação , Brucella canis/isolamento & purificação , Brucella melitensis/isolamento & purificação , Brucella ovis/isolamento & purificação , Brucelose/veterinária , Animais , Animais Selvagens , Brucelose/epidemiologia , Brucelose/microbiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/microbiologia , Doenças do Cão/epidemiologia , Doenças do Cão/microbiologia , Cães , Doenças das Cabras/epidemiologia , Doenças das Cabras/microbiologia , Cabras , Prevalência , Estudos Retrospectivos , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/microbiologia , Carneiro Doméstico , África do Sul/epidemiologia
10.
BMC Infect Dis ; 21(1): 460, 2021 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-34016047

RESUMO

BACKGROUND: This case report describes the clinical process of a shepherd who suffered brucellosis-related endocarditis (BE) and spondylitis (BS) and was infected with Brucella melitensis biovar 3 (B. melitensis biovar 3). CASE PRESENTATION: A 55-year-old male patient was admitted to The First Affiliated Hospital of Shihezi University on October 11, 2018, due to over 3 months of intermittent fever, back pain, and heart trouble. The Rose Bengal Plate test was positive, the standard agglutination test titer for brucellosis was 1/800, and the blood culture was positive for B. melitensis biovar 3. Three instances of transthoracic echocardiography examination at days 1, 25, and 376 after admission to the hospital and magnetic resonance imaging (MRI) and computed tomography (CT) checks at days 5 and 38 revealed that the size of the vegetation on the posterior leaflet of the mitral valve increased from 0.7 × 1.4 cm to 1.2 × 1.5 cm and that the left atrium and ventricle were enlarged. The MRI and CT results showed hyperplasia of the second and third vertebra, a cold abscess formed on both sides of the psoas major muscles, and the vertebra hyperplasia became aggravated at a later time point. The patient's situation deteriorated, and heart failure was discovered on October 22, 2019. At the moment of submission of this manuscript, the patient remains in bed at home because of severe debility caused by brucellosis. CONCLUSIONS: This is the first reported case of endocarditis combined with spondylitis caused by B. melitensis biovar 3 in a shepherd. Brucellosis infection can cause work-power losses because of misdiagnosis or a lack of proper treatment. Early diagnosis and treatment are essential for a successful outcome.


Assuntos
Brucella melitensis , Brucelose/microbiologia , Endocardite Bacteriana/microbiologia , Espondilite/microbiologia , Testes de Aglutinação , Brucelose/diagnóstico , Brucelose/patologia , Endocardite Bacteriana/diagnóstico , Endocardite Bacteriana/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Valva Mitral/microbiologia , Valva Mitral/patologia , Espondilite/diagnóstico
11.
Medicine (Baltimore) ; 100(21): e26076, 2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34032738

RESUMO

ABSTRACT: There has been no ideal surgical approach for lumbar brucella spondylitis (LBS). This study aims to compare clinical efficacy and safety of posterior versus anterior approaches for the treatment of LBS.From April 2005 to January 2015, a total of 27 adult patients with lumbar brucella spondylitis were recruited in this study. The patients were divided into 2 groups according to surgical approaches. Thirteen cases in group A underwent 1-stage anterior debridement, fusion, and fixation, and 14 cases in group B underwent posterior debridement, bone graft, and fixation. The clinical and surgical outcomes were compared in terms of operative time, intraoperative blood loss, hospitalizations, bony fusion time, complications, visual analog scale score, recovery of neurological function, deformity correction.Lumbar brucella spondylitis was cured, and the grafted bones were fused within 11 months in all cases. It was obviously that the operative time and intraoperative blood loss of group A were more than those of group B (P = .045, P = .009, respectively). Kyphotic deformity was signifcantly corrected in both groups after surgery; however, the correction rate was higher in group B than in group A (P = .043). There were no significant differences between the two groups in hospitalizations, bony fusion time, and visual analog scale score in the last follow-up (P = .055, P = .364, P = .125, respectively).Our results suggested that both anterior and posterior approaches can effectively cure lumbar brucella spondylitis. Nevertheless, posterior approach gives better kyphotic deformity correction, less surgical invasiveness, and less complications.


Assuntos
Transplante Ósseo/métodos , Brucelose/cirurgia , Vértebras Lombares/cirurgia , Dor Pós-Operatória/diagnóstico , Espondilite/cirurgia , Adulto , Perda Sanguínea Cirúrgica/estatística & dados numéricos , Transplante Ósseo/efeitos adversos , Brucella/isolamento & purificação , Brucelose/diagnóstico , Brucelose/microbiologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Duração da Cirurgia , Medição da Dor/estatística & dados numéricos , Dor Pós-Operatória/etiologia , Espondilite/diagnóstico , Espondilite/microbiologia , Resultado do Tratamento
12.
Int J Mol Sci ; 22(7)2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33916050

RESUMO

Brucellosis is a highly prevalent zoonotic disease characterized by abortion and reproductive dysfunction in pregnant animals. Although the mortality rate of Brucellosis is low, it is harmful to human health, and also seriously affects the development of animal husbandry, tourism and international trade. Brucellosis is caused by Brucella, which is a facultative intracellular parasitic bacteria. It mainly forms Brucella-containing vacuoles (BCV) in the host cell to avoid the combination with lysosome (Lys), so as to avoid the elimination of it by the host immune system. Brucella not only has the ability to resist the phagocytic bactericidal effect, but also can make the host cells form a microenvironment which is conducive to its survival, reproduction and replication, and survive in the host cells for a long time, which eventually leads to the formation of chronic persistent infection. Brucella can proliferate and replicate in cells, evade host immune response and induce persistent infection, which are difficult problems in the treatment and prevention of Brucellosis. Therefore, the paper provides a preliminary overview of the facultative intracellular parasitic and immune escape mechanisms of Brucella, which provides a theoretical basis for the later study on the pathogenesis of Brucella.


Assuntos
Brucella/fisiologia , Brucelose/microbiologia , Interações Hospedeiro-Patógeno , Animais , Doença Crônica , Humanos
13.
Sci Rep ; 11(1): 8881, 2021 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-33893352

RESUMO

Brucellosis, caused by several species of the genus Brucella, is a zoonotic disease that affects humans and animal species worldwide. Information on the Brucella species circulating in different hosts in Kenya is largely unknown, thus limiting the adoption of targeted control strategies. This study was conducted in multi-host livestock populations in Kenya to detect the circulating Brucella species and assess evidence of host-pathogen associations. Serum samples were collected from 228 cattle, 162 goats, 158 sheep, 49 camels, and 257 humans from Narok and Marsabit counties in Kenya. Information on age, location and history of abortion or retained placenta were obtained for sampled livestock. Data on age, gender and location of residence were also collected for human participants. All samples were tested using genus level real-time PCR assays with primers specific for IS711 and bcsp31 targets for the detection of Brucella. All genus positive samples (positive for both targets) were further tested with a speciation assay for AlkB and BMEI1162 targets, specific for B. abortus and B. melitensis, respectively. Samples with adequate quantities aggregating to 577 were also tested with the Rose Bengal Test (RBT). A total of 199 (33.3%) livestock and 99 (38.5%) human samples tested positive for genus Brucella. Animal Brucella PCR positive status was positively predicted by RBT positive results (OR = 8.3, 95% CI 4.0-17.1). Humans aged 21-40 years had higher odds (OR = 2.8, 95% CI 1.2-6.6) of being Brucella PCR positive compared to the other age categories. The data on detection of different Brucella species indicates that B. abortus was detected more often in cattle (OR = 2.3, 95% CI 1.1-4.6) and camels (OR = 2.9, 95% CI 1.3-6.3), while B. melitensis was detected more in sheep (OR = 3.6, 95% CI 2.0-6.7) and goats (OR = 1.7, 95% CI 1.0-3.1). Both B. abortus and B. melitensis DNA were detected in humans and in multiple livestock host species, suggesting cross-transmission of these species among the different hosts. The detection of these two zoonotic Brucella species in humans further underpins the importance of One Health prevention strategies that target multiple host species, especially in the multi-host livestock populations.


Assuntos
Brucella/genética , Brucelose/epidemiologia , Interações Hospedeiro-Patógeno , Gado , Adulto , Animais , Brucelose/microbiologia , Ecossistema , Feminino , Humanos , Quênia/epidemiologia , Masculino , Epidemiologia Molecular , Adulto Jovem
14.
Microbes Infect ; 23(4-5): 104809, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33753207

RESUMO

The objective of this project was to conduct a feasibility study to determine whether the Brucella abortus S19 vaccine infects and persists in mice and determine whether S19 can be used as a challenge strain for vaccine trial studies. Groups of BALB/c mice were inoculated (intraperitoneally, subcutaneously, intranasally) and euthanized to determine colonization titers in the spleens and lungs. This study showed that S19 does infect and persist in the tissues of mice for 8 weeks and demonstrates that S19 can be used, safely and economically under BSL2 containment, as the challenge strain for future trials to evaluate vaccine efficacy.


Assuntos
Brucella abortus/classificação , Brucella abortus/fisiologia , Brucelose/microbiologia , Modelos Animais de Doenças , Animais , Brucelose/patologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Organismos Livres de Patógenos Específicos
15.
J Microbiol Methods ; 183: 106182, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33647359

RESUMO

BACKGROUND: Clinical diagnosis of human brucellosis (HB) is often difficult due to non-specific symptoms. Immunological tests have been the most common method used in HB diagnosis, but molecular methods based on quantitative polymerase chain reaction (qPCR) have largely replaced these diagnostic methods. The aim of this study was to validate a HB diagnostic qPCR method; assessing different target Brucella genes, and the influence of biological matrices (serum vs. whole blood) on analytical parameters. MATERIAL AND METHODS: Two target genes, IS711 and bcsp31, for HB molecular diagnosis were evaluated, together with biological matrix type (whole blood and serum) using samples spiked with Brucella abortus. In addition, diagnostic parameters of this qPCR method were evaluated in paired whole blood and serum samples from patients with suspected HB. RESULTS: Both genes could be potential diagnostic targets, but IS711 showed a lower limit of detection. In spiked matrix experiments, whole blood showed a lower limit of detection than serum after probit regression (224 vs. 3681 CFU/mL) and ANOVA analysis showed a significant (p < 0.001) difference between the Cq of whole blood at all dilutions and that of serum. In 12 paired clinical samples, no serum samples and only one whole blood sample tested positive for Brucella using this qPCR detection method. CONCLUSIONS: This standardized qPCR-based Brucella detection method could improve diagnosis of HB, serving as a rapid, highly sensitive, and specific test. Whole blood is better suited to qPCR-based HB diagnosis due to the presence of higher target DNA loads in this matrix, compared to serum.


Assuntos
Proteínas de Bactérias/genética , Sangue/microbiologia , Brucella/isolamento & purificação , Brucelose/microbiologia , Reação em Cadeia da Polimerase/métodos , Soro/microbiologia , Brucella/classificação , Brucella/genética , Brucelose/sangue , Brucelose/diagnóstico , DNA Bacteriano/sangue , DNA Bacteriano/genética , Humanos
16.
J Microbiol Methods ; 184: 106185, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33684449

RESUMO

The widely used serodiagnostic test (RBPT, CFT, I-ELISA and FPA) for diagnosis of brucellosis cannot detect vertically infected or carrier animals that are seronegative, a persistent source of infection to other susceptible animals in the herd. For reducing transmission of disease within the herd, these animals must be detected using a rapid, sensitive, user friendly penside diagnostic test. In the present study, Lateral Flow immunoassay (LFA) strip test was developed for detection of Brucellaspp. from clinical samples (bovine aborted foetal stomach contents). The LFA strip was fabricated by printing anti-Brucella polyclonal antibodies (PAb) and anti-bovine antibodies IgG on test and control line, respectively. For conjugation, colloidal gold nanoparticles (30 nm GNP, Sigma, USA) were conjugated with anti-brucella PAb. The LFA strip test was able to detect 107 cfu/ml B.abortus S99 inactivated organism in PBS and it did not exhibit any cross reactivity with selected non Brucella pathogens. To further validate, 115 clinical specimens were tested using LFA strip test. The relative sensitivity (DSn) and relative specificity (DSp) of LFA strip test was determined by ROC curve analysis using PCR and culture method as reference standard. DSn and DSp of LFA strip test was observed as 78.57% (95%CI: 49.2-95.3); 93.07% (95%CI: 86.2-97.2) and 80.0% (95%CI:51.9-95.7); 94.0% (95%CI:0.795-0.925) using culture and PCR as reference diagnostic tests, respectively. It may be concluded that, the LFA strip test can be used as a rapid penside diagnostic test for screening of brucellosis. To the best of our knowledge, this is the first report on development of GNP based LFA strip test for detection of Brucella spp. from bovine aborted fetal content samples.


Assuntos
Brucella/isolamento & purificação , Brucelose/veterinária , Doenças dos Bovinos/diagnóstico , Imunoensaio/métodos , Animais , Anticorpos Antibacterianos/análise , Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/análise , Antígenos de Bactérias/imunologia , Brucella/genética , Brucella/imunologia , Brucelose/diagnóstico , Brucelose/microbiologia , Bovinos , Doenças dos Bovinos/microbiologia , Imunoensaio/instrumentação , Nanopartículas Metálicas/química , Sensibilidade e Especificidade
17.
BMC Vet Res ; 17(1): 126, 2021 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-33743687

RESUMO

BACKGROUND: A novel Brucella strain closely related to Brucella (B.) melitensis biovar (bv) 3 was found in Croatian cattle during testing within a brucellosis eradication programme. CASE PRESENTATION: Standardised serological, brucellin skin test, bacteriological and molecular diagnostic screening for Brucella infection led to positive detection in one dairy cattle herd. Three isolates from that herd were identified to species level using the Bruce ladder method. Initially, two strains were typed as B. melitensis and one as B. abortus, but multiplex PCR based on IS711 and the Suis ladder showed that all of them to belong to B. melitensis, and the combination of whole-genome and multi-locus sequencing as well as Multi-Locus Variable numbers of tandem repeats Analysis (MLVA) highlighted a strong proximity within the phylogenetic branch of B. melitensis strains previously isolated from Croatia, Albania, Kosovo and Bosnia and Herzegovina. Two isolates were determined to be B. melitensis bv. 3, while the third showed a unique phylogenetic profile, growth profile on dyes and bacteriophage typing results. This isolate contained the 609-bp omp31 sequence, but not the 723-bp omp31 sequence present in the two isolates of B. melitensis bv. 3. CONCLUSIONS: Identification of a novel Brucella variant in this geographic region is predictable given the historic endemicity of brucellosis. The emergence of a new variant may reflect a combination of high prevalence among domestic ruminants and humans as well as weak eradication strategies. The zoonotic potential, reservoirs and transmission pathways of this and other Brucella variants should be explored.


Assuntos
Brucella/isolamento & purificação , Brucelose/veterinária , Doenças dos Bovinos/microbiologia , Animais , Brucella/classificação , Brucelose/microbiologia , Bovinos , Croácia , Feminino , Variação Genética , Genoma Bacteriano , Tipagem de Sequências Multilocus/veterinária , Reação em Cadeia da Polimerase Multiplex/veterinária , Filogenia
19.
Proc Natl Acad Sci U S A ; 118(11)2021 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-33688053

RESUMO

Cattle are natural hosts of the intracellular pathogen Brucella abortus, which inflicts a significant burden on the health and reproduction of these important livestock. The primary routes of infection in field settings have been described, but it is not known how the bovine host shapes the structure of B. abortus populations during infection. We utilized a library of uniquely barcoded B. abortus strains to temporally and spatially quantify population structure during colonization of cattle through a natural route of infection. Introducing 108 bacteria from this barcoded library to the conjunctival mucosa resulted in expected levels of local lymph node colonization at a 1-wk time point. We leveraged variance in strain abundance in the library to demonstrate that only 1 in 10,000 brucellae introduced at the site of infection reached a parotid lymph node. Thus, cattle restrict the overwhelming majority of B. abortus introduced via the ocular conjunctiva at this dose. Individual strains were spatially restricted within the host tissue, and the total B. abortus census was dominated by a small number of distinct strains in each lymph node. These results define a bottleneck that B. abortus must traverse to colonize local lymph nodes from the conjunctival mucosa. The data further support a model in which a small number of spatially isolated granulomas founded by unique strains are present at 1 wk postinfection. These experiments demonstrate the power of barcoded transposon tools to quantify infection bottlenecks and to define pathogen population structure in host tissues.


Assuntos
Brucella abortus/fisiologia , Brucelose/veterinária , Doenças dos Bovinos/microbiologia , Animais , Brucella abortus/genética , Brucella abortus/crescimento & desenvolvimento , Brucella abortus/patogenicidade , Brucelose/microbiologia , Bovinos , Feminino , Linfonodos/microbiologia , Virulência
20.
Mol Immunol ; 133: 44-52, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33631554

RESUMO

Brucella is an intracellular zoonotic pathogen that can affect many hosts. Brucella melitensis Rev.1 is a live attenuated, is one of the most effective vaccine strain against brucellosis. It can be used safely in sheep, goats, and even cattle. Although many studies are available on this topic, there is no effective vaccine strain for sheep and goats that distinguishes the antibody titer produced between the field infections and vaccinations. Outer membrane protein 19 (Omp 19) is both virulent and a protective antigen found on the cell-wall of the Brucella strain. In this study, used the suicide plasmid pJQ200KS, which contained homologous region without Omp19 Open Reading Frame (ORF) that was transferred to B. melitensis Rev.1 and further transformed into spheroplasts along with penicillin, ampicillin, and glycine by electroporation. To obtain a mutant vector from Escherichia coli, we used the heat shock transformation method along with the blue-white colony screening using X-gal media, whereas for the gene transfer in Brucella, we used electroporation. A scanning electron microscope (S.E.M) was used to observe the spheroplast transformation while the mutant vector and deletion mutants were confirmed through PCR and sequence analysis. In the mouse model efficacy trials, three commercial vaccines were found to comply with the OIE standards. Although the deletion mutants 19 and 44/10 had similar efficiency as the commercial vaccines in terms of stimulation power, the ELISA test with Omp19 protein showed the same results as the negative control. The Rev.1 Omp19 deletion mutants obtained in this study contained sufficient residual virulence, and their protective immunity was similar to the commercial vaccines. The study showed that a vaccine prepared using a B. melitensis Rev.1 ΔOmp19 can act as a marker vaccine or differentiate infected from vaccinated animals (DIVA) through the ELISA test that detects the Omp19 protein.


Assuntos
Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Vacina contra Brucelose/imunologia , Brucella melitensis/genética , Brucella melitensis/imunologia , Brucelose/prevenção & controle , Lipoproteínas/genética , Animais , Brucella abortus/genética , Brucelose/microbiologia , Modelos Animais de Doenças , Eletroporação/métodos , Feminino , Camundongos , Plasmídeos/genética , Vacinação , Vacinas Atenuadas/imunologia , Virulência/genética , Virulência/imunologia
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