RESUMO
Brucella melitensis and Brucella ovis are gram-negative pathogens of sheep that cause severe economic losses and, although B. ovis is non-zoonotic, B. melitensis is the main cause of human brucellosis. B. melitensis carries a smooth (S) lipopolysaccharide (LPS) with an N-formyl-perosamine O-polysaccharide (O-PS) that is absent in the rough LPS of B. ovis. Their control and eradication require vaccination, but B. melitensis Rev 1, the only vaccine available, triggers anti-O-PS antibodies that interfere in the S-brucellae serodiagnosis. Since eradication and serological surveillance of the zoonotic species are priorities, Rev 1 is banned once B. melitensis is eradicated or where it never existed, hampering B. ovis control and eradication. To develop a B. ovis specific vaccine, we investigated three Brucella live vaccine candidates lacking N-formyl-perosamine O-PS: Bov::CAΔwadB (CO2-independent B. ovis with truncated LPS core oligosaccharide); Rev1::wbdRΔwbkC (carrying N-acetylated O-PS); and H38ΔwbkF (B. melitensis rough mutant with intact LPS core). After confirming their attenuation and protection against B. ovis in mice, were tested in rams for efficacy. H38ΔwbkF yielded similar protection to Rev 1 against B. ovis but Bov::CAΔwadB and Rev1::wbdRΔwbkC conferred no or poor protection, respectively. All H38ΔwbkF vaccinated rams developed a protracted antibody response in ELISA and immunoprecipitation B. ovis diagnostic tests. In contrast, all remained negative in Rose Bengal and complement fixation tests used routinely for B. melitensis diagnosis, though some became positive in S-LPS ELISA owing to LPS core epitope reactivity. Thus, H38ΔwbkF is an interesting candidate for the immunoprophylaxis of B. ovis in B. melitensis-free areas.
Assuntos
Vacina contra Brucelose , Brucella melitensis , Brucella ovis , Brucelose , Doenças dos Roedores , Doenças dos Ovinos , Animais , Anticorpos Antibacterianos , Brucella melitensis/genética , Brucella ovis/genética , Brucelose/prevenção & controle , Brucelose/veterinária , Masculino , Camundongos , Ovinos , Doenças dos Ovinos/prevenção & controleRESUMO
PURPOSE.: Plantar fasciitis is the most frequent cause of heel pain. Custom-made plantar supports are a common treatment solution, while the application of kinesiology tape (KT) can be an effective measure to alleviate pain. The objective was to evaluate the effects of KT on the pain of patients with plantar fasciitis. METHODS.: Randomized controlled trial including participants with plantar fasciitis. There was an experimental group (n = 17), whose participants underwent a KT treatment, and a control (sham tape) group (n = 17). The pain, measured with a visual analog scale (VAS), was the primary outcome and was assessed daily until the fifth day of wearing the tape and 24 h after removing it. Inferential statistics looked for time, group, and time per group differences with CI at 95%. RESULTS.: The greatest between-group VAS difference was 3.5 points, and occurred at the 2-day follow-up assessment. Then, pain differences decreased over time until the last assessment point. Statistically significant time, group, and time*group differences were found with p < 0.001. CONCLUSION.: This study supports that KT is effective in reducing pain in the short term in patients with plantar fasciitis, and more effective than a sham intervention with tape.
Kinesiology tape (KT) reduces pain in patients with plantar fasciitis.Pain is reduced from the first day and the tape can be worn up to five days.KT is an effective solution prior to the application of orthopaedic treatment.
RESUMO
The brucellae are Gram-negative pathogens that cause brucellosis, a zoonosis of worldwide importance. The genus Brucella includes smooth and rough species that differ in that they carry smooth and rough lipopolysaccharides, respectively. Brucella abortus, B. melitensis, and B. suis are typical smooth species. However, these smooth brucellae dissociate into rough mutants devoid of the lipopolysaccharide O-polysaccharide, a major antigen and a virulence determinant encoded in regions wbo (included in genomic island-2) and wbk. We demonstrate here the occurrence of spontaneous recombination events in those three Brucella species leading to the deletion of a 5.5-kb fragment carrying the wbkA glycosyltranferase gene and to the appearance of rough mutants. Analysis of the recombination intermediates suggested homologous recombination between the ISBm1 insertion sequences flanking wbkA as the mechanism generating the deletion. Excision of wbkA was reduced but not abrogated in a recA-deficient mutant, showing the existence of both RecA-dependent and -independent processes. Although the involvement of the ISBm1 copies flanking wbkA suggested a transpositional event, the predicted transpositional joint could not be detected. This absence of detectable transposition was consistent with the presence of polymorphism in the inverted repeats of one of the ISBm1 copies. The spontaneous excision of wbkA represents a novel dissociation mechanism of smooth brucellae that adds to the previously described excision of genomic island-2. This ISBm1-mediated wbkA excision and the different %GC levels of the excised fragment and of other wbk genes suggest that the Brucella wbk locus is the result of at least two horizontal acquisition events.
Assuntos
Proteínas de Bactérias/metabolismo , Brucella/enzimologia , Deleção de Genes , Regulação Bacteriana da Expressão Gênica/fisiologia , Glicosiltransferases/metabolismo , Antígenos de Bactérias , Proteínas de Bactérias/genética , Brucella/citologia , Brucella/genética , Brucella/metabolismo , Cromossomos Bacterianos , Clonagem Molecular , DNA Bacteriano/química , Ilhas Genômicas , Glicosiltransferases/genética , Dados de Sequência Molecular , Recombinação GenéticaRESUMO
The purpose was to assess the effects of three interventions on bone mineral density (BMD) to prevent the onset or progression of osteoporosis in postmenopausal women. Specifically, thirty-nine postmenopausal women, diagnosed with osteopenia or osteoporosis, implemented either high-impact training (G1), the same training + calcium and vitamin D intake (G2), or walked at an intense pace + calcium and vitamin D (G3). Baseline change (BC) in BMD was estimated using the femoral neck and lumbar spine T-scores. Participants were classified as having suffered fractures and/or falls before (24-month) and during the 2-year intervention. The participants-aged 61.8 years-were allocated into G1 (n = 9), G2 (n = 16), and G3 (n = 14). The groups evolved similarly over time; however, participants in G2 exhibited the largest T-score improvements with BC over 20%. G1 and G3 maintained BMD levels (BC = -7 to 13.3%; p > 0.05). Falls occurred similarly across the interventions, while the participants in G2 had the lowest percentage of fracture events (p = 0.037). Overall, the findings suggest that regular physical exercise may be effective in maintaining or improving BMD in postmenopausal women presenting with osteopenia or osteoporosis. Due to the limited sample size, the results are preliminary and warrant future randomized trials to validate the findings.
Assuntos
Doenças Ósseas Metabólicas , Fraturas Ósseas , Osteoporose Pós-Menopausa , Osteoporose , Densidade Óssea , Doenças Ósseas Metabólicas/terapia , Cálcio/farmacologia , Cálcio da Dieta/uso terapêutico , Feminino , Humanos , Osteoporose Pós-Menopausa/prevenção & controle , Pós-Menopausa , Vitamina D/uso terapêutico , Vitaminas/farmacologia , CaminhadaRESUMO
BACKGROUND: The role of wildlife as a brucellosis reservoir for humans and domestic livestock remains to be properly established. The aim of this work was to determine the aetiology, apparent prevalence, spatial distribution and risk factors for brucellosis transmission in several Iberian wild ungulates. METHODS: A multi-species indirect immunosorbent assay (iELISA) using Brucella S-LPS antigen was developed. In several regions having brucellosis in livestock, individual serum samples were taken between 1999 and 2009 from 2,579 wild bovids, 6,448 wild cervids and4,454 Eurasian wild boar (Sus scrofa), and tested to assess brucellosis apparent prevalence. Strains isolated from wild boar were characterized to identify the presence of markers shared with the strains isolated from domestic pigs. RESULTS: Mean apparent prevalence below 0.5% was identified in chamois (Rupicapra pyrenaica), Iberian wild goat (Capra pyrenaica), and red deer (Cervus elaphus). Roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis aries) and Barbary sheep (Ammotragus lervia) tested were seronegative. Only one red deer and one Iberian wild goat resulted positive in culture, isolating B. abortus biovar 1 and B. melitensis biovar 1, respectively. Apparent prevalence in wild boar ranged from 25% to 46% in the different regions studied, with the highest figures detected in South-Central Spain. The probability of wild boar being positive in the iELISA was also affected by age, age-by-sex interaction, sampling month, and the density of outdoor domestic pigs. A total of 104 bacterial isolates were obtained from wild boar, being all identified as B. suis biovar 2. DNA polymorphisms were similar to those found in domestic pigs. CONCLUSIONS: In conclusion, brucellosis in wild boar is widespread in the Iberian Peninsula, thus representing an important threat for domestic pigs. By contrast, wild ruminants were not identified as a significant brucellosis reservoir for livestock.
Assuntos
Animais Selvagens/microbiologia , Brucelose/veterinária , Reservatórios de Doenças , Animais , Técnicas de Tipagem Bacteriana , Brucella abortus/classificação , Brucella abortus/isolamento & purificação , Brucella melitensis/classificação , Brucella melitensis/isolamento & purificação , Brucella suis/classificação , Brucelose/epidemiologia , Impressões Digitais de DNA , Ensaio de Imunoadsorção Enzimática/métodos , Genótipo , Geografia , Humanos , Polimorfismo Genético , Portugal/epidemiologia , Fatores de Risco , Estudos Soroepidemiológicos , Sorotipagem , Espanha/epidemiologiaRESUMO
OBJECTIVE: To determine the most frequently used outcome measures in total knee replacement rehabilitation trials. LITERATURE SURVEY: Systematic review of randomized trials searched in five databases: Web of Science, MEDical Literature Analysis and Retrieval System, Physiotherapy Evidence Database, Scopus, and Cochrane Library. METHODOLOGY: Trials were included if participants underwent total knee replacement rehabilitation and outcome measures were used to assess rehabilitation outcomes. A descriptive synthesis determined the frequency of using outcome measures and preferred assessment time points. Outcomes were classified into eight categories: patient- and clinician-reported function, performance-based function, balance, anxiety and depressive symptoms, quality of life, and others. SYNTHESIS: Eighty-one trials were included and 102 different outcome measures were classified. The most frequently reported outcome was knee range of motion, used in 54% of trials, followed by a visual analog scale of pain (43%) and Western Ontario and McMaster Universities Arthritis Index (WOMAC; 40%). Patient- and clinician-reported function were the categories most frequently assessed (74%), whereas performance-based measures were implemented by 56% of trials. The most frequent assessment time points were 1 week presurgery (52%) and 3 months postsurgery (39%). CONCLUSIONS: There is consensus regarding the need to evaluate functional outcomes in total knee replacement rehabilitation trials but none regarding the outcome measure that should be used. These findings suggest that most trials include patient- and clinician-reported functional measures, along with pain and performance-based measures in trial designs.
Assuntos
Artroplastia do Joelho , Osteoartrite do Joelho , Avaliação de Resultados em Cuidados de Saúde , Humanos , Osteoartrite do Joelho/cirurgia , Qualidade de Vida , Ensaios Clínicos Controlados Aleatórios como Assunto , Amplitude de Movimento ArticularRESUMO
BACKGROUND: The lipopolysaccharide is a major antigen and virulence factor of Brucella, an important bacterial pathogen. In smooth brucellae, lipopolysaccharide is made of lipid A-core oligosaccharide and N-formylperosamine O-polysaccharide. B. ovis and B. canis (rough species) lack the O-polysaccharide. RESULTS: The polymorphism of O-polysaccharide genes wbkE, manA(O-Ag), manB(O-Ag), manC(O-Ag), wbkF and wbkD) and wbo (wboA and wboB), and core genes manB(core) and wa** was analyzed. Although most genes were highly conserved, species- and biovar-specific restriction patterns were found. There were no significant differences in putative N-formylperosamyl transferase genes, suggesting that Brucella A and M serotypes are not related to specific genes. In B. pinnipedialis and B. ceti (both smooth), manB(O-Ag) carried an IS711, confirming its dispensability for perosamine synthesis. Significant differences between smooth and rough species were found in wbkF and wbkD, two adjacent genes putatively related to bactoprenol priming for O-polysaccharide polymerization. B. ovis wbkF carried a frame-shift and B. canis had a long deletion partially encompassing both genes. In smooth brucellae, this region contains two direct repeats suggesting the deletion mechanism. CONCLUSION: The results define species and biovar markers, confirm the dispensability of manB(O-Ag) for O-polysaccharide synthesis and contribute to explain the lipopolysaccharide structure of rough and smooth Brucella species.
Assuntos
Brucella/genética , Antígenos O/genética , Polimorfismo de Fragmento de Restrição , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Biomarcadores , DNA Bacteriano/genética , Genes Bacterianos , Hexosaminas/genética , Manose-6-Fosfato Isomerase/genética , Dados de Sequência Molecular , Complexos Multienzimáticos/genética , Nucleotidiltransferases/genética , Alinhamento de Sequência , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
Infection of sheep with Brucella ovis results in ovine brucellosis, a disease characterized by infertility in rams, abortion in ewes and increased perinatal mortality in lambs. During the course of the infection both the ovine immune response and host cell gene expression are modified. The objective of this research was to conduct a preliminary characterization of differential gene expression in rams experimentally infected with B. ovis by microarray hybridization and real-time RT-PCR. Of the 600 ruminant inflammatory and immune response genes that were analyzed in the microarray, 20 and 14 genes displayed an expression fold change >1.75 with a P-value <0.05 at 15 and 60 days post-challenge (dpc), respectively. Of these genes, 16 were upregulated and 4 were downregulated in infected rams at 15 dpc. At 60 dpc, 11 and 3 genes were up- and down-regulated in infected rams, respectively. Only four genes, desmoglein, epithelial sodium channel, alpha subunit (ENaC-alpha), interleukin 18 binding protein (IL18BP) and macrophage migration inhibition factor (MIF) were found upregulated in infected rams at both 15 and 60 dpc. The analysis of differentially expressed genes demonstrated activation of inflammatory and innate immune pathways in infected animals. B. ovis infection also resulted in upregulation of genes involved in phagocytosis and downregulation of protective host defense mechanisms, both of which may contribute to the chronicity of B. ovis infection. The gene expression profiles differed between rams with severe and moderate B. ovis infection. This is the first analysis of differential gene expression in rough brucellae and particularly in B. ovis-infected rams. The characterization of the genes and their expression profiles in response to B. ovis infection further contributes to our understanding of the molecular mechanisms of infection and the pathogenesis of brucellosis.
Assuntos
Brucella ovis , Brucelose/veterinária , Regulação da Expressão Gênica/imunologia , Genes MHC da Classe II/imunologia , Doenças dos Ovinos/microbiologia , Animais , Brucella ovis/patogenicidade , Brucelose/imunologia , Brucelose/microbiologia , Genes MHC da Classe II/genética , Inflamação , Masculino , Doenças dos Ovinos/imunologia , VirulênciaRESUMO
BACKGROUND: Brucellosis is a world-wide extended zoonosis that causes a grave problem in developing economies. Animal vaccination and diagnosis are essential to control brucellosis, and the need for accurate but also simple and low-cost tests that can be implemented in low-infrastructure laboratories has been emphasized. METHODOLOGY: We evaluated bovine, sheep, goat and swine lateral flow immunochromatography assay kits (LFA), the Rose Bengal test (RBT) and a well-validated protein G indirect ELISA (iELISA) using sera of Brucella culture-positive and unvaccinated brucellosis free livestock. Sera from cattle vaccinated with S19 and RB51 brucellosis vaccines were also tested. Finally, we compared RBT and LFA using sera of white Fulani cattle of unknown bacteriological status from a brucellosis endemic area of Nigeria. RESULTS AND CONCLUSIONS: Although differences were not statistically significant, RBT showed the highest values for diagnostic sensitivity/specificity in cattle (LFA, 96.6/98.8; RBT, 98.9/100; and iELISA, 96.6/100) and the iELISA yielded highest values in sheep (LFA, 94.0/100; RBT, 92.0/100; iELISA, 100/100), goats (LFA, 95.7/96.2; RBT, 97.8/100; iELISA, 100/100) and pigs (LFA, 92.3/100; RBT, 92.3/100; iELISA, 100/100). Vaccine S19 administered subcutaneously interfered in all tests but conjunctival application minimized the problem. Although designed not to interfere in serodiagnosis, vaccine RB51 interfered in LFA and iELISA but not in the RBT. We found closely similar apparent prevalence results when testing the Nigerian Fulani cattle by RBT and LFA. Although both RBT and LFA (showing similar diagnostic performance) are suitable for small laboratories in resource-limited areas, RBT has the advantage that a single reagent is useful in all animal species. Considering these advantages, its low cost and that it is also useful for human brucellosis diagnosis, RBT might be a good choice for resource-limited laboratories.
Assuntos
Brucelose/veterinária , Cromatografia de Afinidade/métodos , Testes Diagnósticos de Rotina/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Coloração e Rotulagem/métodos , Zoonoses/diagnóstico , Animais , Brucelose/diagnóstico , Bovinos , Corantes Fluorescentes/metabolismo , Cabras , Nigéria , Rosa Bengala/metabolismo , Sensibilidade e Especificidade , Ovinos , SuínosRESUMO
Large discrepancies are usually found when different ELISAs for the diagnosis of pig salmonellosis are compared. Thus, our main goal was to estimate the diagnostic accuracy through Bayesian approaches of two commercial assays (Svanovir "test A" and HerdCheck "test B") for the detection of antibodies to Salmonella spp. in slaughter pigs. Previously, we estimated the agreement between both tests and their relative sensitivity (Se) and specificity (Sp) with respect to bacteriology on caecal content and ileocaecal lymph nodes. Test A, at a cut-off OD%>or=20%, indicated higher prevalence than test B (OD%>or=10%) (14.6% vs. 8.6%). Relative Se with respect to overall bacteriology was low (approximately 30%) and similar for both tests, but the relative Sp was significantly lower for test A compared to B (88% vs. 95%). Both tests failed to detect some pigs infected with Salmonella serogroups B and C1, which they were supposed to identify. In general, tests showed only fair-to-moderate agreement when they were compared (kappa: 0.41). In the Bayesian models, Se of test A varied between 63% and 77%, while Se of test B was 73%. Sp of A was always lower than that of test B (89% vs. 95%). The implications derived from the use of these imperfect serological tests will have to be accounted for in large-scale Salmonella-control programs.
Assuntos
Anticorpos Antibacterianos/sangue , Ensaio de Imunoadsorção Enzimática/veterinária , Salmonelose Animal/diagnóstico , Salmonella/imunologia , Doenças dos Suínos/diagnóstico , Criação de Animais Domésticos , Animais , Teorema de Bayes , Canadá , Distribuição de Qui-Quadrado , Ensaio de Imunoadsorção Enzimática/normas , Salmonella/isolamento & purificação , Salmonelose Animal/sangue , Salmonelose Animal/microbiologia , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/sangue , Doenças dos Suínos/microbiologiaRESUMO
OBJECTIVE: The aim of the study was to analyze the effects of endurance and high-impact training oriented toward preventing osteoporosis in postmenopausal women with calcium and vitamin D supplementation. METHODS: This study was a randomized clinical trial. Thirty-six postmenopausal women were randomized to the control and experimental groups. Thirty-four women completed the 2-year interventions. The control group training involved walking at an intense pace. The experimental group conducted high-impact training specifically oriented to prevent osteoporosis. Dual-energy x-ray absorptiometry was used to estimate the T-scores of the lumbar spine and femoral neck. RESULTS: The fast-walking group showed constant T-scores in the femoral neck and improved T-scores in the lumbar spine. High-impact exercises produced improvements in both anatomical levels. Significant differences were found in the femoral neck (ΔControlâ=â-0.04, ΔExperimentalâ=â0.28). The differences were not significant in the lumbar spine (ΔControlâ=â0.27, ΔExperimentalâ=â0.47). Cohen's effect size (dâ=â0.52) suggested a medium practical significance of the trial. The power was 51%. CONCLUSIONS: Calcium plus vitamin D supplementation combined with specifically oriented exercises had a higher impact in the femoral neck than walking at an intense pace. As there were no differences at the lumbar spine level, the results were, however, inconclusive concerning which type of exercise was the most convenient. Importantly, the fact that the T-scores did not decrease after 2 years supports the belief that both proposed interventions can be conveniently used to prevent osteoporosis in postmenopausal women. A trial with a larger sample size would provide consistency to the findings and is warranted given the possible effects and benefits.
Assuntos
Densidade Óssea , Treino Aeróbico/métodos , Osteoporose Pós-Menopausa/prevenção & controle , Caminhada/fisiologia , Absorciometria de Fóton , Conservadores da Densidade Óssea/administração & dosagem , Cálcio da Dieta/administração & dosagem , Cálcio da Dieta/farmacologia , Feminino , Colo do Fêmur/diagnóstico por imagem , Colo do Fêmur/efeitos dos fármacos , Colo do Fêmur/patologia , Humanos , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/efeitos dos fármacos , Vértebras Lombares/patologia , Pessoa de Meia-Idade , Osteoporose Pós-Menopausa/diagnóstico por imagem , Pós-Menopausa , Vitamina D/administração & dosagem , Vitamina D/farmacologiaRESUMO
Brucellosis is a worldwide extended zoonosis with a heavy economic and public health impact. Cattle, sheep and goats are infected by smooth Brucella abortus and Brucella melitensis, and represent a common source of the human disease. Brucellosis diagnosis in these animals is largely based on detection of a specific immunoresponse. We review here the immunological tests used for the diagnosis of cattle brucellosis. First, we discuss how the diagnostic sensitivity (DSe) and specificity (DSp), balance should be adjusted for brucellosis diagnosis, and the difficulties that brucellosis tests specifically present for the estimation of DSe/DSp in frequentistic (gold standard) and Bayesian analyses. Then, we present a systematic review (PubMed, GoogleScholar and CABdirect) of works (154 out of 991; years 1960-August 2017) identified (by title and Abstract content) as DSe and DSp studies of smooth lipopolysaccharide, O-polysaccharide-core, native hapten and protein diagnostic tests. We summarize data of gold standard studies (nâ¯=â¯23) complying with strict inclusion and exclusion criteria with regards to test methodology and definition of the animals studied (infected and S19 or RB51 vaccinated cattle, and Brucella-free cattle affected or not by false positive serological reactions). We also discuss some studies (smooth lipopolysaccharide tests, protein antibody and delayed type hypersensitivity [skin] tests) that do not meet the criteria and yet fill some of the gaps in information. We review Bayesian studies (nâ¯=â¯5) and report that in most cases priors and assumptions on conditional dependence/independence are not coherent with the variable serological picture of the disease in different epidemiological scenarios and the bases (antigen, isotype and immunoglobulin properties involved) of brucellosis tests, practical experience and the results of gold standard studies. We conclude that very useful lipopolysaccharide (buffered plate antigen and indirect ELISA) and native hapten polysaccharide and soluble protein tests exist, provided they are applied taking into account the means available and the epidemiological contexts of this disease: i) mass vaccination; ii) elimination based on vaccination combined with test-and-slaughter; and iii) surveillance and existence of false positive serological reactions. We also conclude that the insistence in recent literature on the lack of usefulness of all smooth lipopolysaccharide or native hapten polysaccharide tests in areas where S19 vaccination is implemented is a misinterpretation that overlooks scientific and practical evidence.
Assuntos
Brucelose Bovina/diagnóstico , Testes Imunológicos/veterinária , Animais , Teorema de Bayes , Brucelose Bovina/imunologia , Brucelose Bovina/microbiologia , Bovinos , Testes Imunológicos/métodos , Sensibilidade e EspecificidadeRESUMO
The properties of drug-vaccine delivery systems based on the use of biodegradable polymers, and its application in the control of experimental infection by Salmonella enterica serovar. Abortusovis (SAO), are described in this manuscript. Micelles of major membrane antigens from SAO (HSao extract) can be encapsulated in microparticles of poly(epsilon-caprolactone) or in nanoparticles of Gantrez polymer. The encapsulation process was optimized by the combined use of cyclodextrins. The resulting particles contained unaltered significant amounts of the antigenic complex. To establish the protective value of these subunit vaccines, particles were injected in one single dose (20 microg of HSao) subcutaneously in BALB/c mice in order to observe the protection conferred against experimental infection with the virulent strains S. Abortusovis 15/5. Control non-immunized animals resulted infected, as well as the group that received unloaded or HSao loaded into microparticles. In contrast, nanoparticles conferred a significant protection when compared to unvaccinated controls, similar to that induced by the attenuated commercial vaccine Rv6. In conclusion, protection against experimental infection in mice after one single shoot, and its potential for mucosal vaccination suggest that HSao-nanoparticles may represent a serious alternative to the conventional attenuated vaccines against S. Abortusovis.
Assuntos
Antígenos de Bactérias/imunologia , Salmonelose Animal/prevenção & controle , Vacinas contra Salmonella/administração & dosagem , Vacinas contra Salmonella/imunologia , Salmonella enterica/imunologia , Animais , Composição de Medicamentos , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Microesferas , Tamanho da Partícula , Distribuição Aleatória , Salmonelose Animal/microbiologia , Salmonella enterica/patogenicidade , Vacinas de Subunidades Antigênicas/administração & dosagem , Vacinas de Subunidades Antigênicas/imunologiaRESUMO
When brucellosis false positive serological reactions happen in cattle, the serial use of pairs of specificity-correlated serological tests (rose bengal, complement fixation, competitive ELISA) results in specificities lower than expected. In this situation, highly specific tests, such as the indirect ELISA used alone, may be more adequate than serial testing.
Assuntos
Antígenos de Bactérias/imunologia , Brucella/imunologia , Brucelose Bovina/diagnóstico , Testes Sorológicos/veterinária , Yersinia enterocolitica/imunologia , Testes de Aglutinação/veterinária , Animais , Bovinos , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Ensaio de Imunoadsorção Enzimática/veterinária , Reações Falso-Positivas , Reprodutibilidade dos Testes , Rosa Bengala/análise , Sensibilidade e Especificidade , Testes Sorológicos/métodos , Testes Sorológicos/normasRESUMO
Swine brucellosis due to Brucella suis is considered an emerging zoonotic disease whose control is based on serological testing and the subsequent culling of seropositive animals or the full depopulation of affected flocks. Here we assessed the performance of several serological tests (Rose Bengal Test [RBT], indirect ELISA [i-ELISA], blocking ELISA [b-ELISA], and two competitive ELISAs [c-ELISA]) for diagnosing swine brucellosis caused by B. suis biovar 2. Both frequentistic and Bayesian statistical inference were used. A frequentistic analysis, using sera from known gold standard (GS) populations (i.e., from truly infected or brucellosis free animals), resulted in maximum (100%) diagnostic sensitivity (Se) and specificity (Sp) in the RBT, i-ELISA and b-ELISA tests. However, c-ELISAs resulted in lower diagnostic Se (ranging from 68.5% to 92.6%, according to the different cut-offs selected). A Bayesian analysis of tests yielding the best diagnostic performance with GS sera (RBT, i-ELISA and b-ELISA), but using a large collection of field sera, resulted in similar Se among tests but markedly lower (≈ 80%) than that resulting from the frequentistic analysis using the GS serum populations. By contrast, the estimated Sp in the Bayesian analysis was only slightly lower than 100%, thus similar to that obtained frequentistically. Our results show that adequate diagnostic tests for brucellosis in swine are available, but also emphasize the need for more extensive validation studies before applying these tests under field conditions.
Assuntos
Brucella suis/isolamento & purificação , Brucelose/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Doenças dos Suínos/microbiologia , Zoonoses/microbiologia , Animais , Teorema de Bayes , Brucelose/sangue , Brucelose/diagnóstico , Brucelose/microbiologia , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/normas , Feminino , Rosa Bengala/química , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/sangue , Doenças dos Suínos/diagnósticoRESUMO
Brucella melitensis is the main etiological agent of brucellosis in sheep and goats, and is also the main agent responsible for human brucellosis, a predominantly occupational disease related to professions in direct contact with livestock. As there is currently no viable method of preventing human brucellosis to safeguard people attention must be directed toward effectively controlling the disease in sheep and goats. This review focuses on the different strategies in different socioeconomic and epidemiologic situations that can be applied to either control or eradicate brucellosis in sheep and goats.
Assuntos
Vacina contra Brucelose/administração & dosagem , Brucella melitensis , Brucelose/veterinária , Doenças das Cabras/prevenção & controle , Doenças dos Ovinos/prevenção & controle , Animais , Brucelose/diagnóstico , Brucelose/prevenção & controle , Brucelose/transmissão , Doenças das Cabras/diagnóstico , Doenças das Cabras/transmissão , Cabras , Humanos , Doenças Profissionais/prevenção & controle , Ovinos , Doenças dos Ovinos/diagnóstico , Doenças dos Ovinos/transmissão , ZoonosesRESUMO
Brucella suis is responsible for swine brucellosis worldwide. Of the five different B. suis biovars (bv.), bv. 2 appears restricted to Europe where it is frequently isolated from wild boar and hares, can infect pigs and can cause human brucellosis. In this study, the differential gene expression profile was characterized in spleens of Eurasian wild boar naturally infected with B. suis bv. 2. Of the 20,201 genes analyzed in the microarray, 633 and 1,373 were significantly (fold change > 1.8; P < 0.01) upregulated and downregulated, respectively, in infected wild boar. The analysis was focused on genes that were over represented after conditional test for biological process gene ontology. Upregulated genes suggested that B. suis bv. 2 infection induced cell maturation, migration and/or proliferation in infected animals. The genes downregulated in infected wild boar impaired the activity of several important cellular metabolic pathways such as metabolism, cytoskeleton organization and biogenesis, immune response and lysosomal function and vesicle-mediated transport. In addition, the response to stress, sperm fertility, muscle development and apoptosis seemed to be also impaired in infected animals. These results suggested that B. suis bv. 2 may use strategies similar to other smooth brucellae to facilitate intracellular multiplication and the development of chronic infections. To our knowledge, this is the first report of the analysis of gene expression profile in hosts infected with B. suis bv. 2, which is important to understand the molecular mechanisms at the host-pathogen interface in the main reservoir species with possible implications in the zoonotic cycle of the pathogen.
Assuntos
Brucella suis/fisiologia , Brucelose/veterinária , Reservatórios de Doenças/microbiologia , Perfilação da Expressão Gênica , Baço/microbiologia , Doenças dos Suínos/genética , Doenças dos Suínos/microbiologia , Animais , Animais Selvagens/genética , Animais Selvagens/metabolismo , Animais Selvagens/microbiologia , Brucella suis/classificação , Brucella suis/isolamento & purificação , Brucelose/genética , Brucelose/metabolismo , Brucelose/microbiologia , Regulação da Expressão Gênica , Masculino , Dados de Sequência Molecular , Baço/metabolismo , Sus scrofa/microbiologia , Suínos , Doenças dos Suínos/metabolismoRESUMO
The attenuated Brucella melitensis Rev 1 vaccine, used against brucellosis infection, interferes with serological diagnosis tests, may induce abortions in pregnant animals, and may infect humans. In order to overcome these drawbacks, we developed acellular vaccines based on a Brucella ovis antigenic complex (HS) containing outer membrane proteins and R-LPS entrapped in poly(anhydride) conventional and mannosylated nanoparticles (NP-HS and MAN-NP-HS) or in poly(epsilon-caprolactone) microparticles (HS-PEC) as antigen delivery systems and immunoadjuvants. Brucellosis free rams were vaccinated subcutaneously with a single dose of particles containing 3mg of HS, and challenged 6 months thereafter. Protection was evaluated by clinical, bacteriological and serological examinations, in comparison with non-vaccinated control rams. HS-PEC vaccine induced protection (7 out of 13 animals were infected) equivalent to that induced by the reference Rev 1 vaccine (8/14). In contrast, animals immunized with NP-HS were not protected, showing similar results to that obtained in the control unvaccinated rams. Furthermore HS-PEC vaccine did not interfere against B. melitensis serodiagnostic tests. In summary, HS-PEC microparticles could be used as a safe and effective vaccine against brucellosis in rams.
Assuntos
Adjuvantes Imunológicos/farmacologia , Vacina contra Brucelose/imunologia , Brucella ovis/imunologia , Brucelose/veterinária , Portadores de Fármacos , Nanopartículas , Doenças dos Ovinos/prevenção & controle , Animais , Vacina contra Brucelose/administração & dosagem , Brucelose/microbiologia , Brucelose/patologia , Brucelose/prevenção & controle , Feminino , Injeções Subcutâneas , Lipossomos/farmacologia , Gravidez , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/patologia , Vacinação/métodos , Vacinas Acelulares/administração & dosagem , Vacinas Acelulares/imunologiaRESUMO
Vaccination with the live attenuated Brucella melitensis Rev 1 vaccine is used to control ovine brucellosis caused by Brucella ovis in sheep. The objective of this study was to identify possible correlates of protective response to B. ovis infection through the characterization by microarray hybridization and real-time RT-PCR of inflammatory and immune response genes differentially expressed in rams previously immunized with B. melitensis Rev 1 and experimentally challenged with B. ovis. Gene expression profiles were compared before and after challenge with B. ovis between rams protected and those vaccinated but found infected after challenge. The TLR10, Bak and ANXI genes were expressed at higher levels in vaccinated and protected rams. These genes provide possible correlates of protective response to B. ovis infection in rams immunized with the B. melitensis Rev 1 vaccine.
Assuntos
Vacinas Bacterianas/uso terapêutico , Biomarcadores/análise , Brucella ovis/imunologia , Brucelose/veterinária , Doenças dos Ovinos/genética , Doenças dos Ovinos/prevenção & controle , Animais , Anexina A1/biossíntese , Anexina A1/genética , Biomarcadores/metabolismo , Brucelose/genética , Brucelose/imunologia , Brucelose/prevenção & controle , Expressão Gênica , Perfilação da Expressão Gênica , Masculino , RNA/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ovinos , Doenças dos Ovinos/imunologia , Receptor 10 Toll-Like/biossíntese , Receptor 10 Toll-Like/genética , Vacinas Atenuadas/uso terapêutico , Proteína Killer-Antagonista Homóloga a bcl-2/biossíntese , Proteína Killer-Antagonista Homóloga a bcl-2/genéticaRESUMO
Classical brucellosis vaccines induce antibodies to the O-polysaccharide section of the lipopolysaccharide that interfere in serodiagnosis. Brucella rough (R) mutants lack the O-polysaccharide but their usefulness as vaccines is controversial. Here, Brucella melitensis R mutants in all main lipopolysaccharide biosynthetic pathways were evaluated in sheep in comparison with the reference B. melitensis Rev 1 vaccine. In a first experiment, these mutants were tested for ability to induce anti-O-polysaccharide antibodies, persistence and spread through target organs, and innocuousness. Using the data obtained and those of genetic studies, three candidates were selected and tested for efficacy as vaccines against a challenge infecting 100% of unvaccinated ewes. Protection by R vaccines was 54% or less whereas Rev 1 afforded 100% protection. One-third of R mutant vaccinated ewes became positive in an enzyme-linked immunosorbent assay with smooth lipopolysaccharide due to the core epitopes remaining in the mutated lipopolysaccharide. We conclude that R vaccines interfere in lipopolysaccharide immunosorbent assays and are less effective than Rev 1 against B. melitensis infection of sheep.