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1.
J Immunol Methods ; 500: 113172, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34673003

RESUMEN

A promising strategy for controlling animal brucellosis is vaccination with commercial vaccine strains (Brucella melitensis Rev.1 and Brucella abortus RB51). Owing to safety concerns associated with these vaccines, developing a more effective and safe vaccine is essential. In this study, we examined the capacity of BhuA, 7α-HSDH or FliC antigens in the presence or absence of adjuvant in eliciting immune responses against brucellosis. After cloning, expression and purification, these proteins were used to examine immunologic responses. All immunized mice induced a vigorous IgG, with a predominant IgG2a response. Moreover, splenocytes of immunized mice proliferated and produced IL-2 and IFN-γ, suggesting the induction of cellular immunity. The high IgG2a/IgG1 ratio and IL-2 and IFN-γ indicated a Th1-oriented immune response in test groups. BhuA-, 7α-HSDH- or FliC- poly I:C formulations were the most effective at inducing Th1 immune response compared to groups immunized with naked proteins. Immunization with proteins protected mice against B. melitensis 16M and B. abortus 544. The proteins in adjuvant induced higher levels of protection than proteins only and exhibited similar degree of protection to live attenuated vaccines. Our results, for first time, introduced five potential candidates for subunit vaccine development against B. melitensis and B. abortus infection.


Asunto(s)
Proteínas Bacterianas/inmunología , Vacuna contra la Brucelosis/inmunología , Brucella abortus/fisiología , Brucella melitensis/fisiología , Brucelosis Bovina/inmunología , Flagelina/inmunología , Hidroxiesteroide Deshidrogenasas/inmunología , Proteínas de Transporte de Membrana/inmunología , Células TH1/inmunología , Adyuvantes Inmunológicos , Animales , Anticuerpos Antibacterianos/sangre , Bovinos , Modelos Animales de Enfermedad , Femenino , Inmunidad Humoral , Inmunoglobulina G/sangre , Interferón gamma/metabolismo , Ratones , Poli I-C/inmunología , Vacunas de Subunidad
2.
Front Immunol ; 12: 679560, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34163479

RESUMEN

Brucella abortus is an important zoonotic pathogen that causes severe economic loss to husbandry and poses a threat to human health. The B. abortus A19 live vaccine has been extensively used to prevent bovine brucellosis in China. However, it is difficult to distinguish the serological response induced by A19 from that induced by natural infection. In this study, a novel genetically marked vaccine, A19ΔvirB12, was generated and evaluated. The results indicated that A19ΔvirB12 was able to provide effective protection against B. abortus 2308 (S2308) challenge in mice. Furthermore, the safety and protective efficacy of A19ΔvirB12 have been confirmed in natural host cattle. Additionally, the VirB12 protein allowed for serological differentiation between the S2308 challenge/natural infection and A19ΔvirB12 vaccination. However, previous studies have found that the accuracy of the serological detection based on VirB12 needs to be improved. Therefore, we attempted to identify potential supplementary antigens with differential diagnostic functions by combining label-free quantitative proteomics and protein chip technology. Twenty-six proteins identified only in S2308 were screened; among them, five proteins were considered as potential supplementary antigens. Thus, the accuracy of the differential diagnosis between A19ΔvirB12 immunization and field infection may be improved through multi-antigen detection. In addition, we explored the possible attenuation factors of Brucella vaccine strain. Nine virulence factors were downregulated in A19ΔvirB12. The downregulation pathways of A19ΔvirB12 were significantly enriched in quorum sensing, ATP-binding cassette transporter, and metabolism. Several proteins related to cell division were significantly downregulated, while some proteins involved in transcription were upregulated in S2308. In conclusion, our results contribute to the control and eradication of brucellosis and provide insights into the mechanisms underlying the attenuation of A19ΔvirB12.


Asunto(s)
Vacuna contra la Brucelosis/genética , Vacuna contra la Brucelosis/inmunología , Brucelosis Bovina/diagnóstico , Brucelosis Bovina/prevención & control , Marcadores Genéticos , Vacunas Sintéticas , Animales , Vacuna contra la Brucelosis/administración & dosificación , Brucelosis Bovina/inmunología , Brucelosis Bovina/metabolismo , Bovinos , Cromatografía Líquida de Alta Presión , Citocinas/metabolismo , Diagnóstico Diferencial , Modelos Animales de Enfermedad , Ingeniería Genética , Inmunización , Inmunogenicidad Vacunal , Ratones , Evaluación de Resultado en la Atención de Salud , Proteómica/métodos , Espectrometría de Masas en Tándem , Virulencia
3.
Infect Immun ; 89(7): e0000421, 2021 06 16.
Artículo en Inglés | MEDLINE | ID: mdl-33820813

RESUMEN

Brucella abortus is a facultatively extracellular-intracellular pathogen that encounters a diversity of environments within the host cell. We report that bacteria extracted from infected cells at late stages (48 h postinfection) of the intracellular life cycle significantly increase their ability to multiply in new target cells. This increase depends on early interaction with the cell surface, since the bacteria become more adherent and penetrate more efficiently than in vitro-grown bacteria. At this late stage of infection, the bacterium locates within an autophagosome-like compartment, facing starvation and acidic conditions. At this point, the BvrR/BvrS two-component system becomes activated, and the expression of the transcriptional regulator VjbR and the type IV secretion system component VirB increases. Using bafilomycin to inhibit BvrR/BvrS activation and using specific inhibitors for VjbR and VirB, we showed that the BvrR/BvrS and VjbR systems correlate with increased interaction with new host cells, while the VirB system does not. Bacteria released from infected cells under natural conditions displayed the same phenotype as intracellular bacteria. We propose a model in which the B. abortus BvrR/BvrS system senses the transition from its replicative niche at the endoplasmic reticulum to the autophagosome-like exit compartment. This activation leads to the expression of VirB, which participates in the release of the bacterium from the cells, and an increase in VjbR expression that results in a more efficient interaction with new host cells.


Asunto(s)
Brucella abortus/fisiología , Brucelosis Bovina/microbiología , Interacciones Huésped-Patógeno , Animales , Autofagosomas , Adhesión Bacteriana , Proteínas Bacterianas/genética , Brucelosis Bovina/inmunología , Bovinos , Regulación Bacteriana de la Expresión Génica , Interacciones Huésped-Patógeno/inmunología , Macrófagos/microbiología , Sistemas de Secreción Tipo IV/genética , Sistemas de Secreción Tipo IV/metabolismo , Virulencia/genética
4.
Front Immunol ; 11: 1944, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32983124

RESUMEN

Bovine brucellosis, cause by infection with Brucella abortus, causes reproductive failure in cattle, has a major economic impact to producers, and as a zoonoses, it is a disease of public health concern. Characterization of the protective immune response against Brucella infection is important to our understanding of disease pathogenesis and for the development of diagnostic assays and vaccines. Most of the knowledge regarding protection against Brucella comes from studies in the murine model, but less is known about the immune responses in cattle. Assessment of antigen-specific T cell frequency and functional phenotype are critical to understand the immune status of the host, characterize mechanisms of protective immunity and immunopathology, and to predict immune protection. The frequency of circulating T cells specific for a particular pathogen is often very low, making analysis of such responses difficult. Our goal was to develop a flow-cytometry based approach to better track Brucella-specific T cell responses. Using peripheral blood mononuclear cells (PMBC) from Brucella abortus strain RB51-vaccinated cattle, we optimized an in vitro stimulation protocol based on a combination of antigen and pan-T cell stimulation. We then assessed RB51-specific T cell responses by concurrently measuring proliferation and cytokine production using flow-cytometry. This methodology enhances the detection of peripheral, Brucella-specific responses in cattle following RB51 vaccination. This protocol is versatile in that it can be modified to fit other in vitro stimulation systems and additional functional or phenotypic parameters can be added for flow cytometric detection and characterization of antigen-specific T cells.


Asunto(s)
Vacuna contra la Brucelosis/administración & dosificación , Brucella/patogenicidad , Brucelosis Bovina/prevención & control , Linfocitos T CD4-Positivos/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Inmunogenicidad Vacunal , Activación de Linfocitos/efectos de los fármacos , Animales , Brucella/inmunología , Vacuna contra la Brucelosis/inmunología , Brucelosis Bovina/inmunología , Brucelosis Bovina/metabolismo , Brucelosis Bovina/microbiología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/microbiología , Bovinos , Células Cultivadas , Femenino , Citometría de Flujo , Interacciones Huésped-Patógeno , Inmunoensayo , Interferón gamma/metabolismo , Factores de Tiempo , Vacunación
5.
Braz J Microbiol ; 51(4): 2133-2143, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-32918242

RESUMEN

A cross-sectional study was conducted to determine the associated factors of brucellosis in Colombia's preeminent dairy region declared in quarantine. A total of 656 samples were collected from cows ≥ 2-year-old from 40 herds. Samples were screened by the Rose Bengal Plate Test, and the Fluorescence Polarized Assay test and Competitive ELISA were used as confirmatory tests. A cow was classified as positive if the screening and both confirmatory tests were positive. A herd was classified as positive if at least one cow was seropositive. The factors associated to seropositivity were tested using a logistic regression model with explanatory variables regarding cattle management, zootechnical parameters, and sanitary practices. The seroprevalence at the animal level was 6.6% (43/656) and at herd level 27.5% (11/40). In the model, five variables explained the animal cases: purchase or animal transfer between owner's farms (OR = 2.79, 95% CI 1.42, 5.49), history of abortion (OR = 4.22, 95% CI 1.91, 9.33), birth of weak calves (OR = 13.77, 95% CI 2.75, 68.91), use of a bull for mating (OR = 9.69, 95% CI 2.23, 42.18), and the vaccination in adulthood (OR = 3.03, 95% CI 1.04.8.78). In the model at the herd level, two variables explained the cases: birth of weak calves (OR = 9.60, 95% CI 1.54, 59.76) and purchase or animal transfer between owner's farms (OR = 7.22, 95% CI 1.03, 50.62). These results justify the need for a quarantine declaration in the region and the implementation of epidemiological studies as a public health measures used to combat outbreak.


Asunto(s)
Anticuerpos Antibacterianos/sangre , Brucelosis Bovina/epidemiología , Brucelosis Bovina/inmunología , Industria Lechera/estadística & datos numéricos , Salud Pública , Crianza de Animales Domésticos , Animales , Bovinos , Colombia/epidemiología , Estudios Transversales , Femenino , Humanos , Modelos Logísticos , Masculino , Embarazo , Factores de Riesgo , Estudios Seroepidemiológicos
6.
Vet Microbiol ; 242: 108569, 2020 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32122583

RESUMEN

In the present study, we analyzed the immune response of calves to Brucella abortus strain 19 vaccine (S19) and its association with MHC class I (BoLA-A) alleles (exons 2-3 and 4-5). Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for typing of BoLA-A exon 2-3 with DdeI and TaqI restriction enzymes; and exon 4-5 with HinfI in 45 crossbred calves. The PCR-RFLP analysis revealed five BoLA-A alleles each for exon 2-3 (A10/A19, A19, A18/19, A18 and A31) and exon 4-5 (A, B, C, D and E). Immune response against B. abortus S19 was assessed at the 4th week post vaccination; antibody response by standard tube agglutination test (STAT) and cell-mediated immunity by lymphocyte proliferation and lymphocyte-mediated cytotoxicity assays. Further, the macrophage function in terms of nitrite production was also analyzed. The association analysis of various BoLA-A alleles with the elicitation of immune response revealed that calves with certain defined genotypes induced significantly higher cell-mediated immune response in terms of lymphocyte proliferation with higher stimulation indices (S.I.) of 1.59 (BoLA-A19), 1.49 (A18/19) and 1.52 (HinfI-D); lymphocyte mediated cytotoxicity (55.52 % in A19) and nitrite production (43.40 µM in A31). It is assumed that allelic variants of BoLA-A (exons 2-3 and 4-5) were associated with the differential immune response of calves to B. abortus S19 vaccination. Therefore, further studies on association analysis of MHC class-I genes in large number of cattle may generate more information and might be useful for adapting the alternative approach of exploring genetic resistance in the cattle herd against bovine brucellosis.


Asunto(s)
Brucelosis Bovina/inmunología , Antígenos de Histocompatibilidad Clase I/genética , Antígenos de Histocompatibilidad Clase I/inmunología , Inmunidad Celular , Factores de Edad , Alelos , Animales , Vacuna contra la Brucelosis/administración & dosificación , Brucella abortus/genética , Brucelosis Bovina/genética , Bovinos/inmunología , Bovinos/microbiología , Estudios de Asociación Genética , Variación Genética , Genotipo , Activación de Linfocitos , Nitritos/metabolismo
7.
Vet Ital ; 55(2): 149-155, 2019 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-31274176

RESUMEN

One of the major constraints in the diagnosis of animal brucellosis is the cross-reactivity that occurs between Brucella and Yersinia surface antigens. With the aim to find a method to distinguish Brucella from Yersinia infection, the expansion of interferon gamma producing (IFN-γ+) T cell subsets obtained from peripheral blood mononuclear cells (PBMC) isolated from cattle either infected by Brucella abortus or experimentally immunized with Yersinia enterocolitica O:9 were compared. The lymphocytes were analyzed by flow cytometry after PBMC were in vitro re-exposed to Yersinia or Brucella antigens. The results highlighted a statistically significant difference in the expansion of the CD4+ and CD8+ IFN-γ+ T cells occurring when PBMC of animals immunized with Yersinia are in vitro exposed to Y. enterocolitica O:9 antigen but not to Brucella antigen. This method could thus be suggested in those cases where results obtained by serodiagnosis need to be further clarified.


Asunto(s)
Brucella abortus/fisiología , Brucelosis Bovina/inmunología , Interferón gamma/inmunología , Yersiniosis/inmunología , Yersinia enterocolitica/fisiología , Animales , Bovinos , Citometría de Flujo/veterinaria , Leucocitos Mononucleares/inmunología
8.
Hig. aliment ; 33(288/289): 1918-1922, abr.-maio 2019.
Artículo en Portugués | LILACS, VETINDEX | ID: biblio-1482432

RESUMEN

A equipe técnica do Centro Estadual de Pesquisa em Sanidade Animal Geraldo Manhães Carneiro (CEPGM), da Empresa de Pesquisa Agropecuária do Estado do Rio de Janeiro (PESAGRO-RIO), realizou em 2009, levantamento epidemiológico para Brucelose através do teste de triagem Teste do Anel do Leite (TAL) no município de Carmo. Foram testados 30 tanques de expansão, entre eles, comunitários, da Cooperativa e Laticínio local e tanques individuais. Como resultado do teste realizado no CEPGM, observou-se a prevalência de 38% para Brucelose bovina no município, indicando expressiva necessidade de aplicação de medidas de profilaxia e controle. Assim, foi instituída vacinação gratuita no município, em março de 2009. Durante 9 anos e 6 meses ininterruptos, foram vacinadas 18.777 bezerras. Em agosto de 2018, um novo levantamento epidemiológico foi realizado, procurando atingir os mesmos pontos de coleta do levantamento de 2009. Foi observada a prevalência de 6,6% para Brucelose, representando a expressiva redução de 83% na prevalência da doença nos rebanhos do município em questão.


Asunto(s)
Femenino , Animales , Recién Nacido , Bovinos , Brucelosis Bovina/epidemiología , Brucelosis Bovina/inmunología , Brucelosis Bovina/prevención & control , Vacuna contra la Brucelosis , Prevalencia
9.
Microbiol Immunol ; 62(11): 694-701, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30350322

RESUMEN

Brucellosis caused by the bacteria of the genus Brucella is an important zoonosis and constitutes a serious public health hazard. Brucellosis is diagnosed mainly by the Rose Bengal plate test and indirect ELISA, both of which have poor specificity because false positive serological reactions occur when screening animals that have been vaccinated with B. abortus S19. Fluorescence polarization assay (FPA) was evaluated for screening samples from cattle and buffalo calves with persistent antibody titers induced by vaccination. The standardized FPA exhibited relative sensitivity and specificity of 0.94 and 0.95, respectively, and the area under the curve, kappa and accuracy were 0.98, 0.87 and 0.95, respectively. Comparison of competitive ELISA and FPA revealed that, FPA is more specific than competitive ELISA. The high specificity, sensitivity and 95% accuracy of FPA indicate that, it is suitable for testing vaccinated animals because it can distinguish between infected from vaccinated animals.


Asunto(s)
Brucella abortus/inmunología , Brucelosis Bovina/diagnóstico , Enfermedades de los Bovinos/diagnóstico , Polarización de Fluorescencia/métodos , Polarización de Fluorescencia/veterinaria , Animales , Anticuerpos Antibacterianos/sangre , Vacuna contra la Brucelosis/inmunología , Brucella abortus/genética , Brucelosis Bovina/sangre , Brucelosis Bovina/inmunología , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/prevención & control , ADN Bacteriano/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Genes Bacterianos/genética , Sensibilidad y Especificidad , Pruebas Serológicas/veterinaria , Vocalización Animal
10.
J S Afr Vet Assoc ; 89(0): e1-e7, 2018 Mar 29.
Artículo en Inglés | MEDLINE | ID: mdl-29781672

RESUMEN

Brucellosis is of worldwide economic and public health importance. Heifer vaccination with live attenuated Brucella abortus strain 19 (S19) is the cornerstone of control in low- and middle-income countries. Antibody persistence induced by S19 is directly correlated with the number of colony-forming units (CFU) per dose. There are two vaccination methods: a 'high' dose (5-8 × 1010 CFU) subcutaneously injected or one or two 'low' doses (5 × 109 CFU) through the conjunctival route. This study aimed to evaluate serological reactions to the 'high' dose and possible implications of the serological findings on disease control. This study included 58 female cases, vaccinated at Day 0, and 29 male controls. Serum was drawn repeatedly and tested for Brucella antibodies using the Rose Bengal Test (RBT) and an indirect enzyme-linked immunosorbent assay (iELISA). The cases showed a rapid antibody response with peak RBT positivity (98%) at 2 weeks and iELISA (95%) at 8 weeks, then decreased in an inverse logistic curve to 14% RBT and 32% iELISA positive at 59 weeks and at 4.5 years 57% (4/7 cases) demonstrated a persistent immune response (RBT, iELISA or Brucellin skin test) to Brucella spp. Our study is the first of its kind documenting the persistence of antibodies in an African communal farming setting for over a year to years after 'high' dose S19 vaccination, which can be difficult to differentiate from a response to infection with wild-type B. abortus. A recommendation could be using a 'low' dose or different route of vaccination.


Asunto(s)
Vacuna contra la Brucelosis/inmunología , Brucella abortus/inmunología , Brucelosis Bovina/inmunología , Crianza de Animales Domésticos , Animales , Anticuerpos Antibacterianos/sangre , Vacuna contra la Brucelosis/administración & dosificación , Brucelosis Bovina/sangre , Brucelosis Bovina/microbiología , Brucelosis Bovina/prevención & control , Estudios de Casos y Controles , Bovinos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Estudios Longitudinales , Masculino , Sudáfrica
11.
Trop Anim Health Prod ; 50(5): 957-963, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29492808

RESUMEN

Brucella poses a great threat to animal and human health. Vaccination is the most promising strategy in the effort to control Brucella abortus (B. abortus) infection, but the currently used live vaccines interfere with diagnostic tests and could potentially result in disease outbreak. Therefore, new subunit vaccines and combined immunization strategies are currently under investigation. In this study, immunogenicity and protection ability of a recombinant adenovirus and plasmid DNA vaccine co-expressing P39 and lumazine synthase proteins of B. abortus were evaluated based on the construction of the two molecular vaccines. Four immunization strategies (single adenovirus, single DNA, adenovirus/DNA, DNA/adenovirus) were investigated. The results showed that the immunization strategy of DNA priming followed by adenovirus boosting induced robust humoral and cellular immune responses, and it significantly reduced the numbers of B. abortus in a mouse model. These results suggest that it could be a potential antigen candidate for development of a new subunit vaccine against B. abortus infection.


Asunto(s)
Vacuna contra la Brucelosis/inmunología , Brucelosis/inmunología , Complejos Multienzimáticos/inmunología , Vacunas de ADN/inmunología , Adenoviridae , Animales , Anticuerpos Antibacterianos/sangre , Proteínas Bacterianas/inmunología , Brucella abortus/inmunología , Brucelosis/prevención & control , Brucelosis Bovina/inmunología , Bovinos , Proliferación Celular , Citocinas/inmunología , Femenino , Ratones , Ratones Endogámicos BALB C
12.
Prev Vet Med ; 151: 57-72, 2018 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-29496108

RESUMEN

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 (n = 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 (n = 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.


Asunto(s)
Brucelosis Bovina/diagnóstico , Pruebas Inmunológicas/veterinaria , Animales , Teorema de Bayes , Brucelosis Bovina/inmunología , Brucelosis Bovina/microbiología , Bovinos , Pruebas Inmunológicas/métodos , Sensibilidad y Especificidad
13.
Microb Pathog ; 110: 196-201, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28669636

RESUMEN

Bovine brucellosis is an important zoonotic disease caused by the bacterium Brucella abortus that leads to economic losses due to animal discard and commercial restrictions. Since positive animals for brucellosis are culled, little is known about the pathogenesis of this disease. Therefore, the aims of this study were to evaluate possible changes in the activity of deaminase adenosine (ADA) and the oxidative stress in cows seropositives for brucellosis (Experiment I), and to evaluate the seroprevalence of B. abortus in dairy cows from the Western state of Santa Catarina, Southern Brazil (Experiment II). The Experiment I evaluated 20 pregnant cows: ten seropositives for B. abortus and ten seronegatives that were used as controls. The ADA activity and markers of oxidative stress (TBARS, catalase (CAT) and superoxide dismutase (SOD)) were evaluated in these animals. A reduction in the activity of ADA and catalase enzymes in seropositive animals was observed (p < 0.001). Conversely, there was an increase in TBARS levels and superoxide dismutase activity in cows infected by B. abortus (p < 0.001). The presence of oxidative stress and a reduction of ADA might be related to the modulation of the inflammatory response. The experiment II was performed due to a high number of herds with restrictions imposed by cases of brucellosis in the state of Santa Catarina in the last two years, and thus, the seroprevalence for B. abortus was evaluated in 1242 serum samples of cows of 69 herds. The serodiagnosis was performed using two tests: buffered acidified antigen and 2-mercaptoethanol. However, none of the serum samples were positive for B. abortus. Although we did not find seropositive animals for brucellosis in our study, the disease still requires continued surveillance, due to its economic impact, and to the oxidative stress caused by it, which may have contributed to cases of abortion in three seropositive cows (Experiment I) in the final third of the gestation.


Asunto(s)
Brucella abortus/patogenicidad , Brucelosis Bovina/sangre , Brucelosis Bovina/inmunología , Enfermedades de los Bovinos/sangre , Enfermedades de los Bovinos/inmunología , Estrés Oxidativo , Adenosina , Animales , Anticuerpos Antibacterianos/sangre , Biomarcadores/sangre , Brasil/epidemiología , Brucelosis Bovina/diagnóstico , Brucelosis Bovina/epidemiología , Catalasa/sangre , Bovinos , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Femenino , Inflamación , Embarazo , Estudios Seroepidemiológicos , Pruebas Serológicas , Superóxido Dismutasa/sangre
14.
Vet Res ; 48(1): 17, 2017 04 04.
Artículo en Inglés | MEDLINE | ID: mdl-28376905

RESUMEN

Brucellosis, caused by Brucella spp., is an important zoonosis worldwide. Vaccination is an effective strategy for protection against Brucella infection in livestock in developing countries and in wildlife in developed countries. However, current vaccine strains including S19 and RB51 are pathogenic to humans and pregnant animals, limiting their use. In this study, we constructed the Brucella abortus (B. abortus) S2308 mutant strain Δ22915, in which the putative lytic transglycosylase gene BAB_RS22915 was deleted. The biological properties of mutant strain Δ22915 were characterized and protection of mice against virulent S2308 challenge was evaluated. The mutant strain Δ22915 showed reduced survival within RAW264.7 cells and survival in vivo in mice. In addition, the mutant strain Δ22915 failed to escape fusion with lysosomes within host cells, and caused no observable pathological damage. RNA-seq analysis indicated that four genes associated with amino acid/nucleotide transport and metabolism were significantly upregulated in mutant strain Δ22915. Furthermore, inoculation of ∆22915 at 105 colony forming units induced effective host immune responses and long-term protection of BALB/c mice. Therefore, mutant strain ∆22915 could be used as a novel vaccine candidate in the future to protect animals against B. abortus infection.


Asunto(s)
Vacuna contra la Brucelosis/inmunología , Brucella abortus/genética , Brucelosis Bovina/prevención & control , Animales , Adhesión Bacteriana/inmunología , Vacuna contra la Brucelosis/uso terapéutico , Brucella abortus/inmunología , Brucelosis Bovina/inmunología , Brucelosis Bovina/microbiología , Bovinos , Femenino , Técnica del Anticuerpo Fluorescente/veterinaria , Ratones , Ratones Endogámicos BALB C , Células RAW 264.7 , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Transcriptoma/genética
15.
Vet J ; 216: 133-5, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27687940

RESUMEN

Intestinal infection by Yersinia enterocolitica serotype O:9 (YeO9) in cattle has been linked to false positive serological reactivity (FPSR) in diagnostic tests for brucellosis. Although eradicated in Ireland, brucellosis monitoring still identifies seropositive animals, usually one or two (termed singletons) per herd, which are classed as FPSR. To investigate a link between FPSR and YeO9, faeces and blood were collected from singleton FPSR cattle, and from companion animals, in eight selected herds with more than one FPSR animal, for YeO9 culture and Brucella serology. YeO9 was isolated from 76/474 (16%) FPSR singletons in 309 herds, but not from any of 621 animals in 122 control non-FPSR herds. In the FPSR herds 52/187 (27.8%) animals were culture positive, and 17% of the isolates were from seronegative animals. Seropositive animals were more likely to have a rising antibody titre when culture positive.


Asunto(s)
Brucella/inmunología , Brucelosis Bovina/inmunología , Enfermedades de los Bovinos/epidemiología , Yersiniosis/veterinaria , Yersinia enterocolitica/aislamiento & purificación , Animales , Anticuerpos Antibacterianos/sangre , Brucelosis Bovina/diagnóstico , Bovinos , Enfermedades de los Bovinos/microbiología , Pruebas de Fijación del Complemento/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Reacciones Falso Positivas , Heces/microbiología , Irlanda/epidemiología , Prevalencia , Serogrupo , Yersiniosis/epidemiología , Yersiniosis/microbiología , Yersinia enterocolitica/genética
16.
Vet Immunol Immunopathol ; 177: 1-6, 2016 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-27436438

RESUMEN

Brucellosis is a globally important zoonotic infectious disease caused by gram negative bacteria of the genus Brucella. While many species of Brucella exist, Brucella melitensis, Brucella abortus, and Brucella suis are the most common pathogens of humans and livestock. The virulence of Brucella is largely influenced by its ability to evade host factors, including phagocytic killing mechanisms, which are critical for the host response to infection. The aim of this study was to characterize the bovine neutrophil response to virulent Brucella spp. Here, we found that virulent strains of smooth B. abortus, B. melitensis, B. suis, and virulent, rough, strains of Brucella canis possess similar abilities to resist killing by resting, or IFN-γ-activated, bovine neutrophils. Bovine neutrophils responded to infection with a time-dependent oxidative burst that varied little between Brucella spp. Inhibition of TAK1, or SYK kinase blunted the oxidative burst of neutrophils in response to Brucella infection. Interestingly, Brucella spp. did not induce robust death of bovine neutrophils. These results indicate that bovine neutrophils respond similarly to virulent Brucella spp. In addition, virulent Brucella spp., including naturally rough strains of B. canis, have a conserved ability to resist killing by bovine neutrophils.


Asunto(s)
Brucella/inmunología , Brucella/patogenicidad , Brucelosis Bovina/inmunología , Bovinos/inmunología , Bovinos/microbiología , Neutrófilos/inmunología , Neutrófilos/microbiología , Animales , Brucella abortus/inmunología , Brucella abortus/patogenicidad , Brucella canis/inmunología , Brucella canis/patogenicidad , Brucella melitensis/inmunología , Brucella melitensis/patogenicidad , Brucella suis/inmunología , Brucella suis/patogenicidad , Brucelosis Bovina/microbiología , Muerte Celular/inmunología , Femenino , Interacciones Huésped-Patógeno/inmunología , Humanos , Técnicas In Vitro , Quinasas Quinasa Quinasa PAM/metabolismo , Neutrófilos/metabolismo , Estallido Respiratorio , Especificidad de la Especie , Quinasa Syk/metabolismo , Virulencia/inmunología , Zoonosis/inmunología , Zoonosis/microbiología
18.
Microb Pathog ; 95: 175-185, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-27057678

RESUMEN

Brucella abortus RB51 is an attenuated vaccine strain that has been most frequently used for bovine brucellosis. Although it is known to provide good protection in cattle, it still has some drawbacks including resistance to rifampicin, residual virulence and pathogenicity in humans. Thus, there has been a continuous interest on new safe and effective bovine vaccine candidates. In the present study, we have constructed unmarked mutants by deleting singly cydD and cydC genes, which encode ATP-binding cassette transporter proteins, from the chromosome of the virulent Brucella abortus isolate from Korean cow (referred to as IVK15). Both IVK15ΔcydD and ΔcydC mutants showed increased sensitivity to metal ions, hydrogen peroxide and acidic pH, which are mimic to intracellular environment during host infection. Additionally, the mutants exhibited a significant growth defect in RAW264.7 cells and greatly attenuated in mice. Vaccination of mice with either IVK15ΔcydC or IVK15ΔcydD mutant could elicit an anti-Brucella specific immunoglobulin G (IgG) and IgG subclass responses as well as enhance the secretion of interferon-gamma, and provided better protection against challenge with B. abortus strain 2308 than with the commercial B. abortus strain RB51 vaccine. Collectively, these results suggest that both IVK15ΔcydC and IVK15ΔcydD mutants could be an attenuated vaccine candidate against B. abortus.


Asunto(s)
Transportadoras de Casetes de Unión a ATP/deficiencia , Vacunas Bacterianas/inmunología , Brucella abortus/inmunología , Brucella abortus/patogenicidad , Brucelosis Bovina/prevención & control , Factores de Virulencia/deficiencia , Animales , Anticuerpos Antibacterianos/sangre , Carga Bacteriana , Vacunas Bacterianas/administración & dosificación , Vacunas Bacterianas/genética , Brucella abortus/genética , Brucella abortus/aislamiento & purificación , Brucelosis Bovina/inmunología , Bovinos , Modelos Animales de Enfermedad , Eliminación de Gen , Inmunoglobulina G/sangre , Interferón gamma/metabolismo , Leucocitos Mononucleares/inmunología , Macrófagos/inmunología , Macrófagos/microbiología , Ratones , Ratones Endogámicos BALB C , Células RAW 264.7 , Bazo/microbiología , Bazo/patología , Vacunas Atenuadas/administración & dosificación , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunología , Virulencia
19.
Vet Immunol Immunopathol ; 167(3-4): 166-70, 2015 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-26188737

RESUMEN

Transcription of non-classical major histocompatibility complex class I (MHC-I) was assessed in the bovine placenta throughout gestation. Additionally, the effect of Brucella abortus infection on expression of non-classical MHC-I was also evaluated using a chorioallantoic membrane explant model of infection. The non-classical MHC-I genes MICB and NC3 had higher levels of transcription in the intercotyledonary region when compared to the placentome, which had higher levels of transcription at the second trimester of gestation. NC1 and classical MHC-I had very low levels of transcription throughout gestation. Trophoblastic cells of B. abortus-infected chorioallantoic membrane explants had an increase in transcription of non-classical MHC-I at 4h post infection. Therefore, this study provides an analysis of non-classical MHC-I transcription at different stages of gestation and different placental tissues, and during B. abortus infection. These findings provide additional knowledge on immune regulation in placental tissues, a known immune-privileged site.


Asunto(s)
Brucelosis Bovina/genética , Brucelosis Bovina/inmunología , Genes MHC Clase I , Placenta/inmunología , Complicaciones Infecciosas del Embarazo/veterinaria , Animales , Brucelosis Bovina/complicaciones , Bovinos , Femenino , Antígenos de Histocompatibilidad Clase I/genética , Histocompatibilidad Materno-Fetal/genética , Embarazo , Complicaciones Infecciosas del Embarazo/genética , Complicaciones Infecciosas del Embarazo/inmunología , Transcripción Genética , Trofoblastos/inmunología
20.
Vaccine ; 33(31): 3659-66, 2015 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-26048781

RESUMEN

Brucella abortus live vaccines have been used successfully to control bovine brucellosis worldwide for decades. However, due to some limitations of these live vaccines, efforts are being made for the development of new safer and more effective vaccines that could also be used in other susceptible species. In this context, understanding the protective immune responses triggered by B. abortus is critical for the development of new vaccines. Such understandings will enhance our knowledge of the host/pathogen interactions and enable to develop methods to evaluate potential vaccines and innovative treatments for animals or humans. At present, almost all the knowledge regarding B. abortus specific immunological responses comes from studies in mice. Active participation of macrophages, dendritic cells, IFN-γ producing CD4(+) T-cells and cytotoxic CD8(+) T-cells are vital to overcome the infection. In this review, we discuss the characteristics of the immune responses triggered by vaccination versus infection by B. abortus, in different hosts.


Asunto(s)
Vacuna contra la Brucelosis/inmunología , Brucella abortus/inmunología , Brucelosis Bovina/inmunología , Brucelosis Bovina/prevención & control , Inmunidad Celular , Inmunidad Humoral , Animales , Vacuna contra la Brucelosis/administración & dosificación , Bovinos , Ratones
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