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1.
Infect Immun ; 88(1)2019 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-31611272

RESUMO

IgA plays an important role in mucosal immunity against infectious pathogens; however, the molecular mechanism of IgA secretion in response to infection remains largely unknown, particularly in Mycoplasma spp. In this study, we found that the levels of IgA in the peripheral blood serum, bronchoalveolar lavage fluid, nasal mucosa, trachea, hilar lymph nodes, and lung tissues of pigs increased significantly after infection with Mycoplasma hyopneumoniae Furthermore, IgA and CD11c were detected in the lungs and hilar lymph nodes by immunohistochemical analysis, and colocalization of these two markers indicates that CD11c+ cells play an important role in IgA mucosal immunity induced by M. hyopneumoniae To investigate the regulatory mechanism of IgA, we separated mouse dendritic cells (DCs) from different tissues and mouse macrophages from the lungs and then cultured mouse B cells together with either DCs or macrophages in vitro In the mouse lung-DC/B (LDC/B) cell coculture, IgA secretion was increased significantly after the addition of whole-cell lysates of M. hyopneumoniae The expression of both Toll-like receptor 2 (TLR2) and TLR4 was also upregulated, as determined by mRNA and protein expression analyses, whereas no obvious change in the expression of TLR3 and TLR7 was detected. Moreover, the IgA level decreased to the same as the control group when TLR2 or TLR4 was inhibited instead of TLR8 or TLR7/9. In conclusion, M. hyopneumoniae can stimulate the response of IgA through TLR2 and TLR4 in a mouse LDC/B cell coculture model, and the coculture model is an ideal tool for studying the IgA response mechanism, particularly that with Mycoplasma spp.


Assuntos
Formação de Anticorpos , Imunoglobulina A/imunologia , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Linfócitos B/imunologia , Células Dendríticas/imunologia , Macrófagos/imunologia , Camundongos , Modelos Teóricos , Suínos
2.
BMC Vet Res ; 15(1): 342, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31619295

RESUMO

BACKGROUND: The objective of this study was to assess the efficacy of a trivalent vaccine mixture and compare it to the respective monovalent vaccines against Mycoplasma hyopneumoniae, porcine circovirus type 2 (PCV2), and porcine reproductive and respiratory syndrome virus (PRRSV). RESULTS: Pigs that were triple challenged with M. hyopneumoniae, PCV2, and PRRSV following vaccination with the trivalent vaccine mixture exhibited a significantly better growth performance when compared to unvaccinated and challenged pigs. A statistical difference was not found when comparing pig populations which were vaccinated with the trivalent vaccine followed by a triple challenge and pigs vaccinated with monovalent M hyopneumoniae vaccine followed by mycoplasmal single challenge in the following areas: M. hyopneumoniae nasal shedding, the number of M. hyopneumoniae-specific interferon-γ secreting cells (IFN-γ-SC), and mycoplasmal lung lesion scores. Pigs vaccinated with the trivalent vaccine mixture followed by a triple challenge resulted in a similar reduction of PCV2 viremia, an increase in the number of PCV2-specific IFN-γ-SC and reduction in interstitial lung lesion scores when compared to pigs vaccinated with a PCV-2 vaccine and challenged with PCV2 only. Lastly, there was a significant difference in the reduction of PRRSV viremia, an increase in PRRSV-specific IFN-γ-SC and a reduction of interstitial lung lesion scores between pigs vaccinated with the trivalent vaccine mixture followed by a triple challenge and pigs vaccinated with a monovalent PRRSV vaccine followed by PRRSV challenge only. CONCLUSION: The trivalent vaccine mixture was efficacious against a triple challenge of M. hyopneumoniae, PCV2, and PRRSV. The trivalent vaccine mixture, however, did not result in equal protection when compared against each respective monovalent vaccine, with the largest vaccine occurring within PRRSV.


Assuntos
Circovirus/imunologia , Mycoplasma hyopneumoniae/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Doenças dos Suínos/prevenção & controle , Vacinação/veterinária , Animais , Vacinas Bacterianas/imunologia , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/prevenção & controle , Feminino , Masculino , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/prevenção & controle , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Sus scrofa , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Doenças dos Suínos/virologia , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/imunologia , Vacinas Virais/imunologia
3.
BMC Vet Res ; 15(1): 327, 2019 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-31511007

RESUMO

BACKGROUND: Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary pathogen of porcine enzootic pneumonia, which has been associated with economic losses due to reduced daily weight gain and feed efficiency. Although it has a small genome and no more than 1000 genes, M. hyopneumoniae can be cultured in cell free media. However, some proteins were not expressed or were only expressed in negligible amounts under culture conditions. Nevertheless, some of these proteins can be expressed at a high level and induce a strong and rapid immune response after M. hyopneumoniae infection. The unexpressed or less expressed proteins may play critical roles in pathogenesis and/or immune response. In order to find the differentially expressed proteins of M. hyopneumoniae between culture condition and infected animals, we established an indirect ELISA for the detection of humoral immunodominant proteins which can discriminate between inactivated bacterin-induced hyperimmune sera and convalescent sera by using Mhp366 protein which did not react with sera from bacterin-immunized pigs, but revealed a strong immunoreaction with porcine convalescent sera. RESULTS: The checkerboard titration method was done by using porcine convalescent sera as positive sera and inactivated bacterin-induced hyperimmune sera as negative sera. The bacterial lysates of fusion proteins and free GST protein without dilution were the optimal coating antigens. The optimal blocking buffer was PBS with 10% FBS and 2.5% skimmed milk. In the checkboard ELISAs, when the sera were diluted at 1:500 and the HRP-labeled rabbit anti-pig IgG were diluted at 1:20000, most positive result was obtained for the assay. CONCLUSIONS: This established indirect ELISA can be used as a tool for the detection of humoral immunodominant proteins of M. hyopneumoniae which can discriminate between inactivated bacterin-induced hyperimmune sera and convalescent sera.


Assuntos
Vacinas Bacterianas/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Animais , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Mycoplasma hyopneumoniae/química , Pneumonia Suína Micoplasmática/sangue , Suínos , Doenças dos Suínos/microbiologia
4.
Vet Microbiol ; 232: 50-57, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31030844

RESUMO

Mycoplasma (M.) hyopneumoniae is the initiator agent of the porcine respiratory disease complex (PRDC) and the etiological agent of enzootic pneumonia. M. hyorhinis and M. flocculare are also found in extensive gross pneumonia-like lesions, but their role is not known. We investigated the pathogenicity of M. hyorhinis and M. flocculare in specific-pathogen-free pigs pre-infected or not with M. hyopneumoniae. Mono-inoculated pigs with M. flocculare showed no clinical signs, hematological changes or macroscopic lesions upon necropsy. Mono-inoculated pigs with M. hyorhinis showed, overall seven days after inoculation, an increase in mean temperature with increases in white blood cell (monocyte) counts and in concentrations of pig major acute phase protein, whereas the average daily weight gain (ADWG) decreased compared with non-infected animals. M. hyorhinis was detected in serous membranes (polyserositis) but not in bronchi. Co-infected pigs with M. hyopneumoniae and M. hyorhinis or M. flocculare showed lower ADWG during the third week of the experiment and higher haptoglobin concentrations in contrast to pigs only mono-infected with M. hyopneumoniae. In pigs co-infected with M. hyopneumoniae and M. hyorhinis, it was interesting to observe that (i) M. hyorhinis was detected in bronchi of six pigs, (ii) M. hyopneumoniae was detected in polyserositis and (iii) there was a slight delay in the production of anti-M. hyopneumoniae IgG. The extent of pneumonia was not statistically different between groups. These results suggest that mycoplasmal associations appear to induce an additive effect and increase the inflammatory status in pigs, probably involving in the impairment of the immune system.


Assuntos
Coinfecção/veterinária , Mycoplasma hyopneumoniae/imunologia , Mycoplasma hyorhinis/patogenicidade , Mycoplasma/patogenicidade , Pneumonia Suína Micoplasmática/imunologia , Animais , Anticorpos Antibacterianos/sangue , Brônquios/microbiologia , Coinfecção/microbiologia , Ensaio de Imunoadsorção Enzimática , Haptoglobinas , Pneumonia Suína Micoplasmática/patologia , Organismos Livres de Patógenos Específicos , Suínos , Virulência , Ganho de Peso
5.
PLoS One ; 14(4): e0215408, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30986254

RESUMO

Cathepsin L (CTSL) has been proved to help contain leishmaniasis and mycoplasma infection in mice by supporting cellular immune responses, but the regulatory functions of CTSL on mucosal immune responses haven't been tested and remain undefined. Here, we investigated the effects of CTSL on SIgA responses and invariant chain (Ii) degradations in the co-cultured swine dendritic cells (DCs) and B cells system in vitro. When the cells system were transfected with vector CTSL-GFP or incubated with recombinant CTSL (rCTSL) before they were infected with Mycoplasma hyopneumoniae (M.hp), SIgA significantly increased and Ii chain was degraded into smaller intermediates, while SIgA decreased when CTSL was knockdown or inhibited with E64. To confirm the SIgA responses promoted by CTSL contribute to the resistance to mycoplasma pneumonia, pigs injected with rCTSL before they were challenged with M.hp, showed milder clinical symptoms and histopathological damage of lungs, less mycoplasma burden together with higher secretion of SIgA, percentages of CD4+ T cells and level of MHC II molecules comparing with the group without rCTSL. Collectively, these results suggested that rCTSL could provide effective protection for piglets against mycoplasma pneumonia by enhancing M.hp-specific mucosal immune responses through its role in antigen presentation by processing the invariant chain.


Assuntos
Apresentação do Antígeno/efeitos dos fármacos , Catepsina L/farmacologia , Imunidade nas Mucosas/efeitos dos fármacos , Imunoglobulina A/imunologia , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Catepsina L/imunologia , Feminino , Antígenos de Histocompatibilidade Classe II/imunologia , Masculino , Pneumonia Suína Micoplasmática/tratamento farmacológico , Pneumonia Suína Micoplasmática/patologia , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/farmacocinética , Suínos
6.
Vet Microbiol ; 230: 195-201, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30827388

RESUMO

Pigs harbor several different species of mycoplasmas, of which Mycoplasma hyopneumoniae presents the most significant economic impact on the swine industry. While ELISAs are the predominant diagnostic assay to measure antibody responses during infection with M. hyopneumoniae, the assay itself is only a rough estimate of the total antibody response. It lends little information on pathogen-wide antigen-specific responses. In addition, antibody responses to M. hyopneumoniae as measured by ELISA are slow to develop in infected swine. Our goal was to determine if a protein microarray could be more sensitive and informative of the serological responses of pigs to M. hyopneumoniae infection. The gene sequences of approximately 50 M. hyopneumoniae surface proteins or protein fragments were cloned, mutated to remove UGA codons, expressed in Escherichia coli and purified. The arrays were used to interrogate pig sera from various sources. Sera from naturally-infected swine gave some variability in antigen-specific responses, but, unexpectedly, the responses against the C-terminal portion of the major adhesin P97 was weak in all animals, including those that were experimentally infected. In two of four 118-day experimentally-infected caesarian-derived colostrum-deprived pigs, the strongest antibody responses occurred on days 30 and 54 against members of the P97/P102 paralog families. Our Day 0 results in the other two animals indicate that although thought to be mycoplasma free by all known criteria (serology and PCR), they may have harbored an inapparent Mycoplasma infection. In summary, the protein microarray has the potential to identify new targets for assay development to enhance sensitivity of antibody-based assays.


Assuntos
Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Animais , Reações Cruzadas , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Análise Serial de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Suínos
7.
Artigo em Alemão | MEDLINE | ID: mdl-30340241

RESUMO

OBJECTIVE: The aim of this study was the scientific evaluation of an intradermal vaccination method in comparison to an intramuscular vaccination against Mycoplasma hyopneumoniae in suckling piglets with regard to skin reactions, performance parameters and procedural aspects. Possible effects on animal welfare should be deduced. MATERIAL AND METHODS: Under field conditions, 672 suckling piglets in three batches were vaccinated; 338 intradermally and 334 intramuscularly. In addition to a detailed scoring of the integument, the injection site with the local reaction was evaluated, scoring the swelling size (score 0-5), and rubor and incrustation (score 0-3). Moreover, piglets were weighed individually 1 day before vaccination and 8 days later. In addition, the time required for each vaccination was documented. RESULTS: On the first day after vaccination, 71.3 % of the intramuscularly vaccinated piglets and 2.7 % of the intradermally vaccinated piglets displayed no swelling at the vaccination site. No differences remained by the 7th day after vaccination. Daily weight gain did not differ significantly between the piglets in the intramuscularly (248 g) and intradermally (258 g) vaccinated groups. Intradermal vaccination took a mean of 11 seconds per piglet, while 17 seconds were required for intramuscular vaccination. CONCLUSIONS AND CLINICAL RELEVANCE: In this first study, no negative effects of the intradermal vaccination on performance parameters and no long-standing skin reactions were detected in the suckling piglets. Skin reactions were related to the desired immune reaction of the intradermal vaccination, but were no longer present after 7 days. Moreover, with regard to procedural aspects, the intradermal vaccination offered time saving advantages. To evaluate further possible effects on animal welfare, further analyses via video recordings are required.


Assuntos
Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/efeitos adversos , Erupção por Droga/veterinária , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/prevenção & controle , Doenças dos Suínos/etiologia , Bem-Estar do Animal , Animais , Animais Recém-Nascidos , Erupção por Droga/etiologia , Injeções Intradérmicas , Injeções Intramusculares , Pneumonia Suína Micoplasmática/imunologia , Sus scrofa , Suínos , Doenças dos Suínos/imunologia
8.
Acta Trop ; 187: 214-221, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29949731

RESUMO

BACKGROUND: The productivity of pigs in smallholder systems is affected by high disease burden, most of which might not be obvious, with their epidemiology and impact being poorly understood. This study estimated the seroprevalence and identified the risk factors of a range of bacterial and viral pathogens of potential economic and public health importance in domestic pigs in Uganda. A total of 522 clinically healthy pigs were randomly selected from 276 pig farms in Masaka (142) and Lira (134) districts of Uganda in 2015. RESULTS: Overall the highest animal prevalence was found for Streptococcus suis 73.0% (CI95: 67.0-78.3) in Lira and 68.2% (CI95: 62.7-73.4) in Masaka; followed by Porcine circovirus type 2 with 50.8% (CI95: 44.5-57.2) in Lira and 40.7% (CI95: 35.2-46.5) in Masaka and Actinobacillus pleuro-pneumoniae, 25.6% (CI95: 20.4-31.6) in Lira and 20.5% (CI95: 16.2-25.6) in Masaka. Mycoplasma hyopneumonia prevalence was 20.9% (CI95: 16.2-26.6) in Lira and 10.1% (CI95: 7.1-14.1) in Masaka, while Porcine parvovirus was 6.2% (CI95: 4.0-9.7) in Masaka and 3.4% (CI95: 1.7-6.6) in Lira. Less common pathogens were Influenza A, 8.5% (CI95: 5.6-12.8) in Lira and 2.0% (CI95: 0.9-4.5) in Masaka and Porcine Reproductive and Respiratory Syndrome Virus, 1.7% (CI95: 0.7-4.3) in Lira and 1.3% (CI95: 0.5-3.5) in Masaka. Even less common was Rotavirus A with 0.8% (CI95: 0.2-3.0) in Lira and 0.7% (CI95: 0.2-2.5) in Masaka; the same was for Aujeszky virus with 0.4% (CI95: 0.7-2.4) in Lira and 0.0% (CI95: 0.0-0.1) in Masaka. Co-infection with two pathogens was common and there was a significant association of M. hyo and PCV2 co-occurrence (p = 0.016). Multivariate analysis showed that for S. suis the use of disinfectant reduced odds of sero-positivitey (OR = 0.15; p = 0.017) and pigs less than 6 months were more likely to be infected than older pigs (OR = 3.35; p = 0.047). For M. hyo, crossbred pigs had higher odd of infection compared to local breeds (OR = 1.59; p < 0.001). CONCLUSIONS: The studied pathogens have high prevalences in smallholder pig production systems and might be silent killers, thus affecting productivity and there is a possibility that some pathogens could spread to humans. Given the limited knowledge of veterinary workers and the poor diagnostic capacities and capabilities in these systems, the diseases are potentially usually under-diagnosed. These findings constitute baseline data to measure the impact of future interventions aiming to reduce disease burden in the pig production systems in Uganda.


Assuntos
Infecções por Circoviridae/veterinária , Fazendas , Infecções por Orthomyxoviridae/veterinária , Pneumonia Suína Micoplasmática/epidemiologia , Síndrome Respiratória e Reprodutiva Suína/epidemiologia , Infecções Estreptocócicas/veterinária , Sus scrofa , Doenças dos Suínos/epidemiologia , Animais , Animais Domésticos/microbiologia , Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/imunologia , Circovirus , Desinfetantes , Vírus da Influenza A , Análise Multivariada , Mycoplasma hyopneumoniae , Razão de Chances , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Vírus da Síndrome Respiratória e Reprodutiva Suína , Prevalência , Fatores de Risco , Estudos Soroepidemiológicos , Infecções Estreptocócicas/epidemiologia , Infecções Estreptocócicas/imunologia , Streptococcus suis , Suínos , Doenças dos Suínos/imunologia , Uganda/epidemiologia
9.
Mem Inst Oswaldo Cruz ; 112(12): 812-816, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29211241

RESUMO

BACKGROUND: The B subunit of Escherichia coli heat-labile enterotoxin (LTB) is a potent mucosal immune adjuvant. However, there is little information about LTB's potential as a parenteral adjuvant. OBJECTIVES: We aimed at evaluating and better understanding rLTB's potential as a parenteral adjuvant using the fused R1 repeat of Mycoplasma hyopneumoniae P97 adhesin as an antigen to characterise the humoral immune response induced by this construct and comparing it to that generated when aluminium hydroxide is used as adjuvant instead. METHODS: BALB/c mice were immunised intraperitoneally with either rLTBR1 or recombinant R1 adsorbed onto aluminium hydroxide. The levels of systemic anti-rR1 antibodies (total Ig, IgG1, IgG2a, and IgA) were assessed by enzyme-linked immunosorbent assay (ELISA). The ratio of IgG1 and IgG2a was used to characterise a Th1, Th2, or mixed Th1/Th2 immune response. FINDINGS: Western blot confirmed rR1, either alone or fused to LTB, remained antigenic; anti-cholera toxin ELISA confirmed that LTB retained its activity when expressed in a heterologous system. Mice immunised with the rLTBR1 fusion protein produced approximately twice as much anti-rR1 immunoglobulins as mice vaccinated with rR1 adsorbed onto aluminium hydroxide. Animals vaccinated with either rLTBR1 or rR1 adsorbed onto aluminium hydroxide presented a mixed Th1/Th2 immune response. We speculate this might be a result of rR1 immune modulation rather than adjuvant modulation. Mice immunised with rLTBR1 produced approximately 1.5-fold more serum IgA than animals immunised with rR1 and aluminium hydroxide. MAIN CONCLUSIONS: The results suggest that rLTB is a more powerful parenteral adjuvant than aluminium hydroxide when administered intraperitoneally as it induced higher antibody titres. Therefore, we recommend that rLTB be considered an alternative adjuvant, even if different administration routes are employed.


Assuntos
Adesinas Bacterianas/imunologia , Adjuvantes Imunológicos/administração & dosagem , Toxinas Bacterianas/administração & dosagem , Enterotoxinas/administração & dosagem , Proteínas de Escherichia coli/administração & dosagem , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/prevenção & controle , Hidróxido de Alumínio , Animais , Toxinas Bacterianas/imunologia , Enterotoxinas/imunologia , Ensaio de Imunoadsorção Enzimática , Proteínas de Escherichia coli/imunologia , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Pneumonia Suína Micoplasmática/imunologia , Suínos
10.
Mem. Inst. Oswaldo Cruz ; 112(12): 812-816, Dec. 2017. graf
Artigo em Inglês | LILACS | ID: biblio-894861

RESUMO

BACKGROUND The B subunit of Escherichia coli heat-labile enterotoxin (LTB) is a potent mucosal immune adjuvant. However, there is little information about LTB's potential as a parenteral adjuvant. OBJECTIVES We aimed at evaluating and better understanding rLTB's potential as a parenteral adjuvant using the fused R1 repeat of Mycoplasma hyopneumoniae P97 adhesin as an antigen to characterise the humoral immune response induced by this construct and comparing it to that generated when aluminium hydroxide is used as adjuvant instead. METHODS BALB/c mice were immunised intraperitoneally with either rLTBR1 or recombinant R1 adsorbed onto aluminium hydroxide. The levels of systemic anti-rR1 antibodies (total Ig, IgG1, IgG2a, and IgA) were assessed by enzyme-linked immunosorbent assay (ELISA). The ratio of IgG1 and IgG2a was used to characterise a Th1, Th2, or mixed Th1/Th2 immune response. FINDINGS Western blot confirmed rR1, either alone or fused to LTB, remained antigenic; anti-cholera toxin ELISA confirmed that LTB retained its activity when expressed in a heterologous system. Mice immunised with the rLTBR1 fusion protein produced approximately twice as much anti-rR1 immunoglobulins as mice vaccinated with rR1 adsorbed onto aluminium hydroxide. Animals vaccinated with either rLTBR1 or rR1 adsorbed onto aluminium hydroxide presented a mixed Th1/Th2 immune response. We speculate this might be a result of rR1 immune modulation rather than adjuvant modulation. Mice immunised with rLTBR1 produced approximately 1.5-fold more serum IgA than animals immunised with rR1 and aluminium hydroxide. MAIN CONCLUSIONS The results suggest that rLTB is a more powerful parenteral adjuvant than aluminium hydroxide when administered intraperitoneally as it induced higher antibody titres. Therefore, we recommend that rLTB be considered an alternative adjuvant, even if different administration routes are employed.


Assuntos
Animais , Feminino , Camundongos , Toxinas Bacterianas/toxicidade , Adjuvantes Imunológicos/administração & dosagem , Adesinas Bacterianas/imunologia , Proteínas de Escherichia coli/administração & dosagem , Proteínas de Escherichia coli/imunologia , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/prevenção & controle , Enterotoxinas/administração & dosagem , Suínos , Ensaio de Imunoadsorção Enzimática , Mycoplasma hyopneumoniae , Hidróxido de Alumínio
11.
PLoS One ; 12(9): e0185387, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28945819

RESUMO

Pneumocystis carinii f. sp. suis (PCS) nucleic acid and antibody profiles on two Austrian-farrow-to-finish farms were investigated. Furthermore, associations with other respiratory pathogens were evaluated. Respiratory specimen and sera from pigs of five age classes between the 1st week and the 3rd month of life as well as samples from sows were analyzed. On Farm A, PCS infection occurred early in life. The suckling piglets were already infected in the 1st week of life and the pigs remained positive until the 3rd month of life. On Farm B, pigs were infected later, between 3 and 4 months of age. The maximum PCS nucleic acid load on Farm A was 8.3 log10 genome copies/mL BALF, whereas on Farm B the PCS burden was significantly lower, with 4.0 log10 genome copies/mL BALF. Anti-PCS antibodies were detected in sows, as maternal antibodies in suckling piglets and as an immunological reaction to infection. On both farms, PCS infection was accompanied by several co-infections. On Farm A, there were concurrent infections with PRRSV, a virulent strain of Haemophilus parasuis, and Mycoplasma hyopneumoniae. On Farm B, PCS was accompanied by infections with swine influenza virus, Mycoplasma hyopneumoniae, and a non-virulent strain of Haemophilus parasuis. The results clearly show that the PCS profiles can vary between farms. Younger pigs may be more susceptible as they had higher PCS burdens. It is possible that PCS may contribute to a respiratory disease in pigs and further investigation of its potential role is warranted.


Assuntos
Pneumocystis carinii/patogenicidade , Pneumonia por Pneumocystis/veterinária , Doenças dos Suínos/microbiologia , Fatores Etários , Animais , Animais Recém-Nascidos , Anticorpos Antifúngicos/análise , Anticorpos Antifúngicos/sangue , Áustria , Coinfecção/imunologia , Coinfecção/microbiologia , Coinfecção/veterinária , Estudos Transversais , DNA Bacteriano/análise , DNA Bacteriano/sangue , DNA Bacteriano/genética , DNA Fúngico/análise , DNA Fúngico/sangue , DNA Fúngico/genética , DNA Viral/análise , DNA Viral/sangue , DNA Viral/genética , Feminino , Infecções por Haemophilus/imunologia , Infecções por Haemophilus/microbiologia , Infecções por Haemophilus/veterinária , Haemophilus parasuis/genética , Haemophilus parasuis/isolamento & purificação , Masculino , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/microbiologia , Infecções por Orthomyxoviridae/veterinária , Pneumocystis carinii/genética , Pneumocystis carinii/imunologia , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/microbiologia , Pneumonia por Pneumocystis/imunologia , Pneumonia por Pneumocystis/microbiologia , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/microbiologia , Síndrome Respiratória e Reprodutiva Suína/virologia , Sus scrofa , Suínos , Doenças dos Suínos/imunologia
12.
BMC Vet Res ; 13(1): 274, 2017 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-28851359

RESUMO

BACKGROUND: Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary agent of enzootic pneumonia in pigs. Pigs are often infected with different M. hyopneumoniae strains. This study assessed the efficacy of vaccination against experimental infection with two genetically different M. hyopneumoniae strains in weaned piglets. At 33 days of age (D0), 45 M. hyopneumoniae-free piglets were randomly assigned to three different groups: 1) negative control group (NCG; n = 5): not vaccinated, not infected, 2) positive control group (PCG; n = 20): not vaccinated, infected, and 3) vaccination group (VG; n = 20): single vaccination with an inactivated whole-cell M. hyopneumoniae vaccine (Hyogen®, Ceva) (D1), infected. The PCG and VG were endotracheally inoculated with 7 × 107 CCU in 7 ml of the highly virulent M. hyopneumoniae strain F7.2C (D24) and 7 × 107 CCU in 7 ml low virulent strain F1.12A (D25). A respiratory disease score (RDS) was assessed from D24 until D53. At D53 (euthanasia), macroscopic lung lesions (MLL) were scored, log copies of M. hyopneumoniae DNA (qPCR) and IL-1 and IL-6-concentrations (ELISA) on bronchoalveolar lavage fluid were determined. RESULTS: The RDS and MLL at euthanasia were respectively 0, 1.20 and 0.55 (P < 0.001) and 0, 7.56 and 0.68 (P < 0.001) for NCG, PCG and VG, respectively. The qPCR results for PCG and VG were 3.99 and 1.78 log copies (P < 0.001), respectively, with a significant difference between PCG and VG. The IL-1 and IL-6 results at euthanasia for NCG, PCG and VG were 17.61, 1283.39 and 53.04 pg/ml (P < 0.001) and 148.10, 493.35 and 259.80 pg/ml (P = 0.004), respectively with a significant difference between PCG and VG. CONCLUSIONS: Vaccination with Hyogen® in pigs was efficacious against an experimental challenge with both a low and highly virulent M. hyopneumoniae strain as the vaccinated pigs coughed significantly less, and showed significantly less lung lesions compared to the non-vaccinated challenged pigs: the vaccinated animals showed a 52.9% lower RDS and 91.0% lower MLL compared to the PCG. In the bronchoalveolar lavage fluid collected at the necropsy of the vaccinated pigs, a significantly lower amount of M. hyopneumoniae-DNA and a significantly lower IL-1 and IL-6 concentration was found compared to the pigs of the PCG.


Assuntos
Vacinas Bacterianas/administração & dosagem , Mycoplasma hyopneumoniae , Pneumonia Suína Micoplasmática/prevenção & controle , Animais , Anticorpos Antibacterianos/análise , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/análise , DNA Bacteriano/análise , Relação Dose-Resposta Imunológica , Pulmão/patologia , Mycoplasma hyopneumoniae/imunologia , Mycoplasma hyopneumoniae/isolamento & purificação , Mycoplasma hyopneumoniae/patogenicidade , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/patologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Especificidade da Espécie , Suínos
13.
PLoS One ; 12(4): e0175034, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28380065

RESUMO

Immunopathological events are key for the development of enzootic pneumonia (EP), which is macroscopically observed as cranioventral pulmonary consolidation (CVPC). This study aimed to investigate the putative association between the humoral immune response against Mycoplasma hyopneumoniae (M. hyopneumoniae) and prevalence and extension of CVPC in 1) experimentally infected pigs, 2) slaughtered pigs and 3) sequentially necropsied pigs in a longitudinal study. CVPC was scored by means of the European Pharmacopoeia recommended methodology. Specific IgG, IgG1 and IgG2 antibodies were assessed in serum. In addition, mucosal IgG and IgA antibodies were analyzed in broncho-alveolar lavage fluid (BALF) from experimentally challenged pigs. The systemic humoral immune response in experimentally infected pigs was delayed in onset whereas humoral respiratory mucosal immune response appeared more rapidly but declined earlier. Although low, BALF IgG antibodies showed the highest correlation with CVPC scores (r = 0.49, p<0.05). In slaughter-aged pigs, both percentage of lungs with CVPC and mean lung lesion score were significantly higher in M. hyopneumoniae seropositive farms compared to the seronegative ones (p<0.001). Similarly, seropositive sequentially necropsied pigs showed more severe CVPC than seronegative ones. Overall, mean serological values might help to forecast prevalence and severity of EP-like lung lesions using a population based approach. Remarkably, the specific systemic humoral immune response was found to be predominated by the IgG2 subclass, suggesting a dominant Th1-mediated immune response to M. hyopneumoniae.


Assuntos
Imunidade Humoral , Pulmão/patologia , Mycoplasma hyopneumoniae , Pneumonia Suína Micoplasmática/patologia , Animais , Imunidade Humoral/imunologia , Imunoglobulina G/imunologia , Pulmão/imunologia , Pulmão/microbiologia , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Suínos
14.
BMC Vet Res ; 13(1): 91, 2017 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-28388953

RESUMO

BACKGROUND: Mycoplasma hyopneumoniae (M. hyo) and Porcine Circovirus Type 2 (PCV2) are major pathogens that cause significant health problems in swine worldwide. Maternal derived immunity (MDI) has been suggested as a significant immediate defence factor for newborn piglets and may interfere with piglet's vaccination-induced immunity. The study aimed to assess the efficacy of a novel combination vaccine (consisting of PCV2 subunits and inactivated M. hyo strain J), against PCV2 and M. hyo natural infection [Porcilis® PCV M Hyo (MSD Animal Health, Boxmeer, the Netherlands)], in the presence of strong maternally derived PCV2 immunity (antibody titre averaged 11.08 log2), under field conditions. The study was performed according to a controlled, randomized and blinded design in a Greek swine unit with Enzootic Pneumonia (EP) and subclinical PCV2 infection. In total, 600 healthy three-week-old suckling piglets were allocated randomly, either to treatment (vaccinated with the test product) or control group (injected with sterile buffered saline). RESULTS: Vaccination significantly reduced the severity of lung lesions at slaughter (lesions of cranio-ventral pulmonary consolidation) (P < 0.001). The overall mean lung lesion score (LLS) was 9.6 in the vaccinated group and 12.2 in controls. The level of PCV2 viraemia was significantly reduced in vaccinated pigs. Furthermore, 25 g higher average daily weight gain (ADWG) was observed during the finishing phase (P < 0.001) and 18 g greater ADWG overall (P < 0.001). CONCLUSIONS: Results of LLS, PCV2 viremia and ADWG support the test product's efficacy in the face of strong maternally derived PCV2 immunity.


Assuntos
Infecções por Circoviridae/veterinária , Imunidade Materno-Adquirida , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/prevenção & controle , Doenças dos Suínos/prevenção & controle , Vacinação/veterinária , Animais , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/prevenção & controle , Circovirus/imunologia , Feminino , Grécia , Pulmão/patologia , Masculino , Pneumonia Suína Micoplasmática/imunologia , Suínos , Doenças dos Suínos/imunologia , Vacinas Combinadas/imunologia , Viremia , Ganho de Peso
15.
Vet Microbiol ; 198: 1-8, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28061998

RESUMO

Mycoplasma hyopneumoniae (Mhp) is the primary etiological agent responsible for swine enzootic pneumonia (EP), a disease that cause tremendous economic losses all over the swine industry. Dendritic cells (DCs), the most effective antigen-presenting cells, are widely distributed beneath respiratory epithelium. DCs uptake and present antigens to T cells, to initiate protective immune responses or generate immune-mediated pathology in different infections. In this study, we investigated the changes in the different DCs subpopulations, T cells and SIgA positive cells counts in porcine nasal cavity after long time Mhp infection. We further evaluated the role of porcine DCs in Mhp exposure. Our results showed that the number of SLA-II-DR+SWC3a+DCs, SLA-II-DR+CD11b+ DCs, T cells, SIgA positive cells in nasal cavity were decreased after Mhp 28 days infection in vivo experiment. The antigen presenting ability of DCs were inhibited by Mhp exposure. DCs couldn't activate T-cell proliferation by down-regulating the antigen presenting molecule CD1a expression and promoting high level of IL-10 production. Further more, the expression levels of IL-12 and IFN-γ in DCs were decreased, suggesting that DCs favour for Th2 immune response development after Mhp exposure in vitro. Taken together, Mhp infection impairs the immune function which allows the persistence of Mhp and cause predispose pigs to secondary infections. The decline of DCs presentation ability is the reason why dysfunction and persistence in Mhp infection. These findings are benefit for exploring the pathogenic mechanisms of Mhp in pigs.


Assuntos
Células Dendríticas/imunologia , Células Dendríticas/microbiologia , Interações Hospedeiro-Patógeno/imunologia , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/imunologia , Animais , Contagem de Células , Proliferação de Células , Células Cultivadas , Citocinas/genética , Células Dendríticas/patologia , Regulação da Expressão Gênica/imunologia , Imunoglobulina A Secretora/metabolismo , Macrófagos/citologia , Cavidade Nasal/imunologia , Cavidade Nasal/microbiologia , Cavidade Nasal/patologia , Pneumonia Suína Micoplasmática/patologia , Organismos Livres de Patógenos Específicos , Suínos , Linfócitos T/citologia , Linfócitos T/imunologia
16.
Anim Sci J ; 88(4): 575-585, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27612216

RESUMO

To clarify the genetic influence of mycoplasmal pneumonia of swine (MPS) lesion-selected Landrace (La) on MPS resistance and immune characteristics in three-way crossbred pigs (LaWaDa), the LaWaDa pigs were compared with the non-selected crossbred (LbWbDb) and purebred (La) pigs. The MPS lesion score in the three lines was as follows: La line < LaWaDa line < LbWbDb line, with significant differences among the lines. The proportions of myeloid cells and T cells were lower and higher, respectively, in the LaWaDa pigs compared with those in the other two lines. Messenger RNA (mRNA) expression of interleukin (IL)-6, IL-10, transforming growth factor-ß, and interferon-γ in peripheral blood was significantly increased after vaccination in the La and LaWaDa lines. IL-4 mRNA expression in the LaWaDa line was intermediate to the La and LbWbDb lines. Furthermore, principal component analysis for immune traits and MPS lesions was executed to clarify the characteristics of each pig line. These findings suggest that the immune responses in the three pig lines are genetically distinct and that MPS resistance and some immunity characteristics from the La line were transmitted to the three-way crossbred pigs.


Assuntos
Resistência à Doença/genética , Resistência à Doença/imunologia , Imunidade Inata/genética , Imunidade Inata/imunologia , Imunocompetência/genética , Imunocompetência/imunologia , Mycoplasma pneumoniae/imunologia , Pneumonia Suína Micoplasmática/genética , Pneumonia Suína Micoplasmática/imunologia , Seleção Artificial/genética , Animais , Vacinas Bacterianas/imunologia , Interferon gama/sangue , Interleucina-10/sangue , Interleucina-6/sangue , Suínos , Fator de Crescimento Transformador beta/sangue
17.
Artigo em Inglês | MEDLINE | ID: mdl-27638117

RESUMO

By selective breeding for five generations, a Landrace line has been recently established to improve resistance to mycoplasmal pneumonia of swine (MPS), daily gain (DG), back fat thickness (BF), and plasma cortisol concentrations (COR). To clarify the involvement of swine leukocyte antigen (SLA) polymorphisms in the selection process, we investigated possible associations of 11 SLA-class II haplotypes with selected traits or immune parameters. Pigs with the low-resolution SLA haplotype Lr-0.23 or Lr-0.13, which increased in frequency with the passage of generations, had less severe pathological lesions of MPS, increased leukocyte phagocytic activity, and higher white blood cell counts. In contrast, Lr-0.12 and Lr-0.2, which decreased in subsequent generations, were weakly associated with more severe pathological lesions of MPS. Therefore, in the studied Landrace line, the Lr-0.23 and Lr-0.13 haplotypes are potentially useful genetic markers for selecting and breeding animals with less severe pathological lesions of MPS.


Assuntos
Haplótipos , Antígenos de Histocompatibilidade Classe II/imunologia , Pneumonia Suína Micoplasmática/genética , Pneumonia Suína Micoplasmática/imunologia , Suínos/imunologia , Animais , Cruzamento , Resistência à Doença/genética , Marcadores Genéticos , Genótipo , Antígenos de Histocompatibilidade Classe I , Antígenos de Histocompatibilidade Classe II/genética , Hidrocortisona/sangue , Contagem de Leucócitos , Fagocitose , Fenótipo , Pneumonia Suína Micoplasmática/microbiologia , Pneumonia Suína Micoplasmática/fisiopatologia , Polimorfismo Genético
18.
J Gen Virol ; 97(10): 2501-2515, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27498789

RESUMO

In pigs, influenza A viruses and Mycoplasma hyopneumoniae (Mhp) are major contributors to the porcine respiratory disease complex. Pre-infection with Mhp was previously shown experimentally to exacerbate the clinical outcomes of H1N1 infection during the first week after virus inoculation. In order to better understand the interactions between these pathogens, we aimed to assess very early responses (at 5, 24 and 48 h) after H1N1 infection in pigs pre-infected or not with Mhp. Clinical signs and macroscopic lung lesions were similar in both infected groups at early times post-H1N1 infection; and Mhp pre-infection affected neither the influenza virus replication nor the IFN-induced antiviral responses in the lung. However, it predisposed the animals to a higher inflammatory response to H1N1 infection, as revealed by the massive infiltration of neutrophils and macrophages into the lungs and the increased production of pro-inflammatory cytokines (IL-6, IL-1ß and TNF-α). Thus, it seems it is this marked inflammatory state that would play a role in exacerbating the clinical signs subsequent to H1N1 infection.


Assuntos
Vírus da Influenza A Subtipo H1N1/fisiologia , Interferons/imunologia , Mycoplasma hyopneumoniae/fisiologia , Infecções por Orthomyxoviridae/veterinária , Pneumonia Suína Micoplasmática/microbiologia , Doenças dos Suínos/microbiologia , Doenças dos Suínos/virologia , Animais , Suscetibilidade a Doenças , Vírus da Influenza A Subtipo H1N1/genética , Interferons/genética , Interleucina-6/imunologia , Pulmão/imunologia , Pulmão/microbiologia , Pulmão/virologia , Macrófagos/imunologia , Mycoplasma hyopneumoniae/genética , Infiltração de Neutrófilos , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , Pneumonia Suína Micoplasmática/imunologia , Suínos , Doenças dos Suínos/imunologia , Fator de Necrose Tumoral alfa/imunologia
19.
Vet Res Commun ; 40(2): 81-8, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27142053

RESUMO

In this study, we investigated the dynamics of Mycoplasma hyopneumoniae infections in 66 pig farms, with different production systems (one-, two-, and three-site systems), and considered different risk factors. Serological assay was used to detect serum antibodies against M. hyopneumoniae and real time polymerase chain reaction (RT-PCR) was performed to detect M. hyopneumoniae DNA in tracheobronchial swabs. Results demonstrated that M. hyopneumoniae infection status was predominantly influenced by the age of the animals and the type of production system. Infection rates were higher in older animals and the prevalence was higher in the one- and two-site systems than in the three-site systems. Dynamics of infection by RT-PCR showed that earlier M. hyopneumoniae infection on one-site farms occurs earlier, while on two- and three-site farms occurs later but spreads faster, suggesting that contact between animals of different age favors the transmission.


Assuntos
Criação de Animais Domésticos , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/patologia , Pneumonia Suína Micoplasmática/transmissão , Fatores Etários , Animais , Anticorpos Antibacterianos/sangue , Mycoplasma hyopneumoniae/genética , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/sangue , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Soroconversão , Suínos
20.
J Vet Med Sci ; 78(8): 1319-22, 2016 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-27075114

RESUMO

Mycoplasma hyopneumoniae causes porcine enzootic pneumonia, an economically important disease of swine. A more sensitive and reliable method for detection of serum antibodies is needed for epidemiological investigations and to evaluate the effect of immunization. We expressed the M. hyopneumoniae protein P65 in Escherichia coli and produced a monoclonal antibody (mAb) that bound specifically to recombinant P65. Using this mAb, a blocking enzyme linked immunosorbent assay (ELISA) was developed. The blocking ELISA had similar specificity to and sensitivity with the commercial ELISA produced by IDEXX. Thus, this blocking ELISA is a useful test for serological confirmation of M. hyopneumoniae infection.


Assuntos
Proteínas de Bactérias/imunologia , Ensaio de Imunoadsorção Enzimática/veterinária , Mycoplasma hyopneumoniae/imunologia , Pneumonia Suína Micoplasmática/diagnóstico , Animais , Anticorpos Monoclonais/imunologia , Western Blotting/veterinária , Reações Cruzadas/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Pneumonia Suína Micoplasmática/imunologia , Pneumonia Suína Micoplasmática/microbiologia , Sensibilidade e Especificidade , Suínos
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