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Sibele Borsuk - One of the best experts on this subject based on the ideXlab platform.
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the combination of corynebacterium pseudotuberculosis recombinant proteins rpld rcp01850 and rcp09720 for improved detection of Caseous Lymphadenitis in sheep by elisa
Journal of Medical Microbiology, 2019Co-Authors: Mara Thais De Oliveira Silva, Ricardo W Portela, Rodrigo Barros De Pinho, Francisco Silvestre Brilhante Bezerra, Vasco Azevedo, Wanessa Lordelo Pedreira Vivas, Caroline De Santana Ferreira, Sibele BorsukAbstract:Corynebacterium pseudotuberculosis is the causative agent of Caseous Lymphadenitis (CLA), a chronic disease of sheep and goats. Current methods for CLA diagnosis cannot identify all infected animals; therefore, the development of an improved diagnosis is essential. We evaluated recombinant phospholipase D (rPLD) protein individually or combined with rCP01850 or rCP09720 proteins for the detection of CLA in sheep. A total of 40 positive and 25 negative sera samples were analysed by ELISA using the recombinant proteins. ELISA using rPLD (E1), rPLD+rCP01850 (E2) and rPLD+rCP09720 (E3) showed 90, 92.5 and 97.5 % sensitivity and 92, 72 and 92 % specificity, respectively. The area under the receiver operating characteristic curves for E1, E2 and E3 was 0.925, 0.882 and 0.990, respectively. ELISA using rPLD +rCP09720 demonstrated the best sensitivity and specificity. Thus, the combination of these recombinant proteins in indirect ELISA has the potential for the diagnosis of CLA in sheep.
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a panel of recombinant proteins for the serodiagnosis of Caseous Lymphadenitis in goats and sheep
Microbial Biotechnology, 2019Co-Authors: Thiago Doria Barral, Roberto Meyer, Vasco Azevedo, Sibele Borsuk, Dan Loureiro, Ricardo Barros Mariutti, Raghuvir K Arni, Anderson Jesus Santos, Ana Rita Sokolonski, Ricardo Wagner Dias PortelaAbstract:Caseous Lymphadenitis (CLA) is a small ruminant disease characterized by the development of granulomatous lesions in superficial and internal lymph nodes, as well as in some organs, and causes significant economic losses worldwide. The aetiological agent of CLA is the bacterium Corynebacterium pseudotuberculosis; however, the commercially available diagnostic tools present problems with regard to specificity, which can lead to false-negative results. This study aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulins in goats and sheep using recombinant C. pseudotuberculosis PLD, CP40, PknG, DtxR and Grx proteins. For validation of the ELISAs, 130 goat serum samples and 160 sheep serum samples were used. The best ELISA for goats was developed using a combination of PLD and CP40 as antigens at a 1:1 ratio, which presented 96.9% sensitivity and 98.4% specificity. The most effective ELISA for sheep presented 91% sensitivity and 98.7% specificity when recombinant PLD alone was used as the antigen. These ELISAs can be used as highly accurate tools in epidemiological surveys and for the serodiagnosis of C. pseudotuberculosis infection in goats and sheep.
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establishment of an objective endpoint in mice model for Caseous Lymphadenitis vaccine trials
Veterinary Microbiology, 2019Co-Authors: Mara Thais De Oliveira Silva, Rodrigo Barros De Pinho, Francisco Silvestre Brilhante Bezerra, Vinicius Farias Campos, Vasco Azevedo, Sibele BorsukAbstract:Abstract For a long time, the scientific community has described the need for a continued update in practices that ensure the welfare of animals undergoing experimentation. In addition to approaches on principles of care and use of animals, there is a more current emerging concern: defining an appropriate end point in experiments that use animals for research, teaching and testing. The term “endpoint” is defined as the point at which an experimental animal’s pain and/or distress is terminated, minimized, or reduced humanely. In the present study, we established an endpoint in Balb/C mice for Caseous Lymphadenitis vaccine trials, which can be considered as a highly important parameter since several studies are being developed to control the disease efficiently. Mice were monitored daily until the 30th day after infection with pathogenic strain of C. pseudotuberculosis MIC–6 using the most relevant parameters for the appearance of clinical signs of Caseous Lymphadenitis (CLA), such as abscesses, lethargy, and loss of weight and hair. The endpoint was found to be a weight loss of 0.2167 g after five days or 10% weight loss in less than five days. In conclusion, the findings reported here will help improve animal’s well-being during vaccine trials for CLA and consequently represent significant contribution to animal’s welfare.
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corynebacterium pseudotuberculosis cp09 mutant and cp40 recombinant protein partially protect mice against Caseous Lymphadenitis
BMC Veterinary Research, 2014Co-Authors: Judson Wallace Rodrigues Silva, Anderson Miyoshi, Ricardo W Portela, Vasco Azevedo, Sibele Borsuk, Fernanda Alves Dorella, Daniela Droppaalmeida, Flavia S Rocha, Wanessa Lordelo Pedreira Vivas, Francine Ferreira PadilhaAbstract:Background: Caseous Lymphadenitis (CLA) is an infectious disease that affects small ruminants and is caused by Corynebacterium pseudotuberculosis. This disease is responsible for high economic losses due to condemnation and trim of infected carcasses, decreased leather and wool yield, loss of sales of breeding stock and deaths from internal involvement. Treatment is costly and ineffective; the most cost-effective strategy is timely immunisation. Various vaccine strategies have been tested, and recombinant vaccines are a promising alternative. Thus, in this study, different vaccine formulations using a recombinant protein (rCP40) and the CP09 live recombinant strain were evaluated. Five groups of 10 mice each were immunised with saline (G1), rCP40 (G2), CP09 (G3), a combination of CP09 and rCP40 (G4) and a heterologous prime-boost strategy (G5). Mice received two immunisations within 15 days. On day 30 after primary immunisation, all groups were challenged with a C. pseudotuberculosis virulent strain. Mice were monitored and mortality was recorded for 30 days after challenge. Results: The G2, G4 and G5 groups showed high levels of IgG1 and IgG2a; G2 presented significant IgG2a production after virulent challenge in the absence of IgG1 and IgG3 induction. Thirty days after challenge, the mice survival rates were 20 (G1), 90 (G2), 50 (G3), 70 (G4) and 60% (G5). Conclusions: rCP40 is a promising target in the development of vaccines against Caseous Lymphadenitis.
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development of an elisa using the recombinant protein cp1957 of corynebacterium pseudotuberculosis for diagnosis of Caseous Lymphadenitis in sheep
BMC Proceedings, 2014Co-Authors: Carlos Guilherme Rosa Reis, Vasco Azevedo, Henrique Ramos Angelo, Andrea De Fatima Silva Rezende, Alexandre Antunes Brum, Sibele BorsukAbstract:Background Caseous Lymphadenitis (CLA) is a disease caused by the bacteria Corynebacterium pseudotuberculosis which affects small ruminants such as sheeps and goats, leading to severe economic losses. The development of more sensitive and specific diagnoses showing effectiveness on asymptomatic animals is essential for disease’s control. This study purposes the use of the recombinant protein CP1957 of C. pseudotuberculosis in indirect ELISA using sheep sera.
Nubia Seyffert - One of the best experts on this subject based on the ideXlab platform.
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serological proteome analysis of corynebacterium pseudotuberculosis isolated from different hosts reveals novel candidates for prophylactics to control Caseous Lymphadenitis
Veterinary Microbiology, 2014Co-Authors: Nubia Seyffert, Renata F Silva, Julien Jardin, Wanderson M Silva, Thiago Luiz De Paula CastroAbstract:Caseous Lymphadenitis (CLA) is a highly prevalent disease in goats and sheep worldwide, which is caused by Corynebacterium pseudotuberculosis. Although several prophylactic methods against CLA have been proposed previously, the identification of new C. pseudotuberculosis proteins that are really produced during the infectious process is still needed to improve efficiency and accuracy in vaccines and diagnostics. In this study, we used optimized conditions for serological proteome analysis (SERPA) in order to identify new immune-reactive proteins in C. pseudotuberculosis culture supernatants of two strains, 1002 and C231, isolated from goats and sheep, respectively. Using a sheep and goat serum pool, 13 novel immune-reactive exoproteins common to the two strains were identified. Four of these proteins present known functions and were already described as immune-reactive proteins in other microorganisms, whereas the other nine are of unknown function and show low similarity with proteins from other bacterial species. These data reveal promising targets for immunoprophylactic methods against CLA.
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subclinical diagnosis of Caseous Lymphadenitis based on elisa in sheep from brazil
Journal of Bacteriology & Parasitology, 2013Co-Authors: Dayana Ribeiro, Nubia Seyffert, Anderson Miyoshi, Ricardo W Portela, Roberto Meyer, Fernanda Alves Dorella, Luis G C Pacheco, Thiago Luiz De Paula Castro, Maria Cecilia Rui Luvizotto, Vasco AzevedoAbstract:Caseous Lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is a chronic contagious disease that affects small ruminants and still remains an important problem for many lamb-producing countries. Animals are considered clinically infected when occurs abscesses in superficial lymph nodes. Visceral or internal form can coexist which no apparent clinical signs of infection are seen. The best procedure to avoid spread of the disease is elimination of infected animals. However, as the chronic and subclinical nature of the infection of CLA alternative methods are required for detection and screening. In this study, we described the performance of indirect Enzyme-Linked Immunosorbent Assay (ELISA) for diagnosis of CLA in asymptomatics sheep. Also, test culture and biochemical identification were achieved to confirm CLA infection. The serological diagnostic was performed in sheep symptomatics (n=50) and asymptomatics (n=374) from nine flocks. Analysis reported high positivity of 71% for ELISA in 85% of asymptomatic animal for CLA with a sensitivity of 88% and specificity of 31%. Results from ELISA test in asymptomatic animals against culture for Caseous Lymphadenitis were more specific (97%) and permitted to exclude healthy animals without symptoms. This study concluded that C. pseudotuberculosis infection could be widely disseminated in sheep flocks in Northwestern region of the state of Sao Paulo, Brazil and only one screening test is not enough. The association with indirect ELISA test and culture could better indicate the real problem of CLA in sheep flocks.
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Caseous Lymphadenitis epidemiology diagnosis and control
The IIOAB Journal, 2011Co-Authors: A S Guimaraes, Filipe Borges, Rebeca Barbosa Pauletti, Nubia Seyffert, Dayana Ribeiro, Andrey Pereira Lage, Marcos Bryan Heinemann, Anderson Miyoshi, A M G GouveiaAbstract:Caseous Lymphadenitis, caused by Corynebacterium pseudotuberculosis, is one of the most important diseases of sheep and goats, causing considerable losses for herd owners. Due to the chronic and generally subclinical nature of infection, control is difficult and prevalence in animals and herds is high. This review describes the principal characteristics of C. pseudotuberculosis, including pathogenesis, epidemiology and principal manifestations of Caseous Lymphadenitis, as well as management practices, diagnostic tests and vaccination as disease control tools.
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high seroprevalence of Caseous Lymphadenitis in brazilian goat herds revealed by corynebacterium pseudotuberculosis secreted proteins based elisa
Research in Veterinary Science, 2010Co-Authors: Nubia Seyffert, A S Guimaraes, Andrey Pereira Lage, Marcos Bryan Heinemann, A M G Gouveia, Bruno Lopes Bastos, Ricardo W Portela, Fernanda Alves Dorella, Luis G C Pacheco, Roberto MeyerAbstract:We conducted a seroepidemiological survey to determine the prevalence of Caseous Lymphadenitis (CLA) in goat herds in Minas Gerais state, Brazil. Serum samples were collected from goats (n = 676) from 108 rural properties in 2001, covering most of the sub-regions of this ca. 586,500 square kilometer state. Antibodies against Corynebacterium pseudotuberculosis secreted proteins were detected by an indirect enzyme-linked immunosorbent assay (ELISA). Most of the animals (78.9%) tested positive for CLA; 98% of flocks presented at least one seropositive animal. Goats managed under an extensive production system had a significantly higher seroprevalence of CLA than those in intensive and semi-intensive operations. The age distribution of the animals in the flocks affected the prevalence of this disease; however, goat breed did not. We found seropositivity against C. pseudotuberculosis to be highly prevalent in these Brazilian goat herds; consequently, appropriate management practices for the control of CLA should be implemented.
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Caseous Lymphadenitis in sheep flocks of the state of minas gerais brazil prevalence and management surveys
Small Ruminant Research, 2009Co-Authors: A S Guimaraes, Nubia Seyffert, Bruno Lopes Bastos, Ricardo W Portela, Roberto Meyer, F B Carmo, J C M Cruz, John A Mcculloch, Andrey Pereira LageAbstract:Abstract Corynebacterium pseudotuberculosis is the etiologic agent of Caseous Lymphadenitis, which is a serious, economically important problem for sheep production. We examined the seroprevalence of infection by C. pseudotuberculosis and possible risk factors associated with Caseous Lymphadenitis in sheep herds of the state of Minas Gerais, Brazil. Samples were collected from 642 sheep from 97 farms. Sera of all of the sheep were tested with ELISA for antibodies against C. pseudotuberculosis. A questionnaire was applied to gather data on the farm, the sheep herd, the farmer, and individual animal data (breed, sex and age). This is the first sero-epidemiological survey for Caseous Lymphadenitis in sheep herds in Minas Gerais. We found a high real prevalence, much higher than that suggested from information obtained with the questionnaire, which points to the scarcity of vaccination against Caseous Lymphadenitis in the sample evaluated. Only a small proportion of the farmers declared that cases of this disease were present in their flocks. The frequency of seropositive sheep varied significantly with breed (χ2 test, P = 0.021). Age group also significantly affected the percentage of seropositivity (χ2 test, P = 0.049), the highest frequency being found in adult animals (more than 12 months old), when compared to the 5–12 months old group (χ2 test, P = 0.021). The prevalence of infection with C. pseudotuberculosis in sheep in the state of Minas Gerais was estimated to be 70.9% (95% confidence interval (CI): 64.7–77.0%) and the prevalence of infected flocks being 95.9% (95% CI: 89.8–98.9%). We concluded that C. pseudotuberculosis infection is widely disseminated in sheep flocks in Minas Gerais and that Caseous Lymphadenitis control and eradication programs are necessary in this state.
Vasco Azevedo - One of the best experts on this subject based on the ideXlab platform.
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the combination of corynebacterium pseudotuberculosis recombinant proteins rpld rcp01850 and rcp09720 for improved detection of Caseous Lymphadenitis in sheep by elisa
Journal of Medical Microbiology, 2019Co-Authors: Mara Thais De Oliveira Silva, Ricardo W Portela, Rodrigo Barros De Pinho, Francisco Silvestre Brilhante Bezerra, Vasco Azevedo, Wanessa Lordelo Pedreira Vivas, Caroline De Santana Ferreira, Sibele BorsukAbstract:Corynebacterium pseudotuberculosis is the causative agent of Caseous Lymphadenitis (CLA), a chronic disease of sheep and goats. Current methods for CLA diagnosis cannot identify all infected animals; therefore, the development of an improved diagnosis is essential. We evaluated recombinant phospholipase D (rPLD) protein individually or combined with rCP01850 or rCP09720 proteins for the detection of CLA in sheep. A total of 40 positive and 25 negative sera samples were analysed by ELISA using the recombinant proteins. ELISA using rPLD (E1), rPLD+rCP01850 (E2) and rPLD+rCP09720 (E3) showed 90, 92.5 and 97.5 % sensitivity and 92, 72 and 92 % specificity, respectively. The area under the receiver operating characteristic curves for E1, E2 and E3 was 0.925, 0.882 and 0.990, respectively. ELISA using rPLD +rCP09720 demonstrated the best sensitivity and specificity. Thus, the combination of these recombinant proteins in indirect ELISA has the potential for the diagnosis of CLA in sheep.
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a panel of recombinant proteins for the serodiagnosis of Caseous Lymphadenitis in goats and sheep
Microbial Biotechnology, 2019Co-Authors: Thiago Doria Barral, Roberto Meyer, Vasco Azevedo, Sibele Borsuk, Dan Loureiro, Ricardo Barros Mariutti, Raghuvir K Arni, Anderson Jesus Santos, Ana Rita Sokolonski, Ricardo Wagner Dias PortelaAbstract:Caseous Lymphadenitis (CLA) is a small ruminant disease characterized by the development of granulomatous lesions in superficial and internal lymph nodes, as well as in some organs, and causes significant economic losses worldwide. The aetiological agent of CLA is the bacterium Corynebacterium pseudotuberculosis; however, the commercially available diagnostic tools present problems with regard to specificity, which can lead to false-negative results. This study aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulins in goats and sheep using recombinant C. pseudotuberculosis PLD, CP40, PknG, DtxR and Grx proteins. For validation of the ELISAs, 130 goat serum samples and 160 sheep serum samples were used. The best ELISA for goats was developed using a combination of PLD and CP40 as antigens at a 1:1 ratio, which presented 96.9% sensitivity and 98.4% specificity. The most effective ELISA for sheep presented 91% sensitivity and 98.7% specificity when recombinant PLD alone was used as the antigen. These ELISAs can be used as highly accurate tools in epidemiological surveys and for the serodiagnosis of C. pseudotuberculosis infection in goats and sheep.
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establishment of an objective endpoint in mice model for Caseous Lymphadenitis vaccine trials
Veterinary Microbiology, 2019Co-Authors: Mara Thais De Oliveira Silva, Rodrigo Barros De Pinho, Francisco Silvestre Brilhante Bezerra, Vinicius Farias Campos, Vasco Azevedo, Sibele BorsukAbstract:Abstract For a long time, the scientific community has described the need for a continued update in practices that ensure the welfare of animals undergoing experimentation. In addition to approaches on principles of care and use of animals, there is a more current emerging concern: defining an appropriate end point in experiments that use animals for research, teaching and testing. The term “endpoint” is defined as the point at which an experimental animal’s pain and/or distress is terminated, minimized, or reduced humanely. In the present study, we established an endpoint in Balb/C mice for Caseous Lymphadenitis vaccine trials, which can be considered as a highly important parameter since several studies are being developed to control the disease efficiently. Mice were monitored daily until the 30th day after infection with pathogenic strain of C. pseudotuberculosis MIC–6 using the most relevant parameters for the appearance of clinical signs of Caseous Lymphadenitis (CLA), such as abscesses, lethargy, and loss of weight and hair. The endpoint was found to be a weight loss of 0.2167 g after five days or 10% weight loss in less than five days. In conclusion, the findings reported here will help improve animal’s well-being during vaccine trials for CLA and consequently represent significant contribution to animal’s welfare.
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corynebacterium pseudotuberculosis cp09 mutant and cp40 recombinant protein partially protect mice against Caseous Lymphadenitis
BMC Veterinary Research, 2014Co-Authors: Judson Wallace Rodrigues Silva, Anderson Miyoshi, Ricardo W Portela, Vasco Azevedo, Sibele Borsuk, Fernanda Alves Dorella, Daniela Droppaalmeida, Flavia S Rocha, Wanessa Lordelo Pedreira Vivas, Francine Ferreira PadilhaAbstract:Background: Caseous Lymphadenitis (CLA) is an infectious disease that affects small ruminants and is caused by Corynebacterium pseudotuberculosis. This disease is responsible for high economic losses due to condemnation and trim of infected carcasses, decreased leather and wool yield, loss of sales of breeding stock and deaths from internal involvement. Treatment is costly and ineffective; the most cost-effective strategy is timely immunisation. Various vaccine strategies have been tested, and recombinant vaccines are a promising alternative. Thus, in this study, different vaccine formulations using a recombinant protein (rCP40) and the CP09 live recombinant strain were evaluated. Five groups of 10 mice each were immunised with saline (G1), rCP40 (G2), CP09 (G3), a combination of CP09 and rCP40 (G4) and a heterologous prime-boost strategy (G5). Mice received two immunisations within 15 days. On day 30 after primary immunisation, all groups were challenged with a C. pseudotuberculosis virulent strain. Mice were monitored and mortality was recorded for 30 days after challenge. Results: The G2, G4 and G5 groups showed high levels of IgG1 and IgG2a; G2 presented significant IgG2a production after virulent challenge in the absence of IgG1 and IgG3 induction. Thirty days after challenge, the mice survival rates were 20 (G1), 90 (G2), 50 (G3), 70 (G4) and 60% (G5). Conclusions: rCP40 is a promising target in the development of vaccines against Caseous Lymphadenitis.
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development of an elisa using the recombinant protein cp1957 of corynebacterium pseudotuberculosis for diagnosis of Caseous Lymphadenitis in sheep
BMC Proceedings, 2014Co-Authors: Carlos Guilherme Rosa Reis, Vasco Azevedo, Henrique Ramos Angelo, Andrea De Fatima Silva Rezende, Alexandre Antunes Brum, Sibele BorsukAbstract:Background Caseous Lymphadenitis (CLA) is a disease caused by the bacteria Corynebacterium pseudotuberculosis which affects small ruminants such as sheeps and goats, leading to severe economic losses. The development of more sensitive and specific diagnoses showing effectiveness on asymptomatic animals is essential for disease’s control. This study purposes the use of the recombinant protein CP1957 of C. pseudotuberculosis in indirect ELISA using sheep sera.
Ricardo W Portela - One of the best experts on this subject based on the ideXlab platform.
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the combination of corynebacterium pseudotuberculosis recombinant proteins rpld rcp01850 and rcp09720 for improved detection of Caseous Lymphadenitis in sheep by elisa
Journal of Medical Microbiology, 2019Co-Authors: Mara Thais De Oliveira Silva, Ricardo W Portela, Rodrigo Barros De Pinho, Francisco Silvestre Brilhante Bezerra, Vasco Azevedo, Wanessa Lordelo Pedreira Vivas, Caroline De Santana Ferreira, Sibele BorsukAbstract:Corynebacterium pseudotuberculosis is the causative agent of Caseous Lymphadenitis (CLA), a chronic disease of sheep and goats. Current methods for CLA diagnosis cannot identify all infected animals; therefore, the development of an improved diagnosis is essential. We evaluated recombinant phospholipase D (rPLD) protein individually or combined with rCP01850 or rCP09720 proteins for the detection of CLA in sheep. A total of 40 positive and 25 negative sera samples were analysed by ELISA using the recombinant proteins. ELISA using rPLD (E1), rPLD+rCP01850 (E2) and rPLD+rCP09720 (E3) showed 90, 92.5 and 97.5 % sensitivity and 92, 72 and 92 % specificity, respectively. The area under the receiver operating characteristic curves for E1, E2 and E3 was 0.925, 0.882 and 0.990, respectively. ELISA using rPLD +rCP09720 demonstrated the best sensitivity and specificity. Thus, the combination of these recombinant proteins in indirect ELISA has the potential for the diagnosis of CLA in sheep.
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brazilian green propolis as a therapeutic agent for the post surgical treatment of Caseous Lymphadenitis in sheep
Frontiers in Veterinary Science, 2019Co-Authors: Mauricio Alcântara Kalil, Roberto Meyer, Laerte Marlon Santos, Thiago Doria Barral, Daniela Meria Rodrigues, Neila De Paula Pereira, Maria Da Conceicao Aquino Sa, Marcelo Andres Umszaguez, Bruna Aparecida Souza Machado, Ricardo W PortelaAbstract:: As antibiotics are ineffective when used to treat Caseous Lymphadenitis, the surgical excision of lesions is often required. Iodine solution (10%) is currently the choice for the post-surgical treatment; however, it may cause histotoxicity. Propolis are resinous substances composed by a mixture of different plants parts and molecules secreted by bees. As green propolis has already proven to possess anti-bacterial and wound healing properties, this study aimed to evaluate the use of a green propolis-based ointment as a therapeutic agent for the post-surgical treatment of Caseous Lymphadenitis. The Caseous lesions of 28 sheep were surgically excised before dividing animals into two groups: (1) iodine-treated animals and (2) sheep treated with an ointment made with a previously characterized green propolis extract. Clinical data of animals, size of the scar area, the presence of moisture and secretion in the surgical wound, the humoral immune response against the bacterium and the susceptibility of C. pseudotuberculosis clinical isolates to the green propolis extract were analyzed. The green propolis-treated group presented complete healing of the surgical wound 1 week before the iodine-treated group. Additionally, animals treated with the green propolis ointment had fewer cases of wound secretion, but it was not statistically different from the iodine-treated group. No clinical signs indicating green propolis toxicity or other side effects were found, associated with a faster and more organized hair recovery by propolis use. The green propolis extract was able to inhibit the growth of 23 from the 27 C. pseudotuberculosis clinical isolates, with minimum inhibitory and minimum bactericide concentrations ranging from 01 to 08 mg/mL, and did not interfere with the humoral immune response against the bacterium. In addition, green propolis was able to inhibit biofilm formation by four of the C. pseudotuberculosis clinical isolates. We concluded that green propolis is a promising therapeutic agent to be used in the post-surgical treatment of Caseous Lymphadenitis in small ruminants due to its effects on surgical wound healing, hair recovery, inhibition of wound contamination and bacterial growth.
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corynebacterium pseudotuberculosis cp09 mutant and cp40 recombinant protein partially protect mice against Caseous Lymphadenitis
BMC Veterinary Research, 2014Co-Authors: Judson Wallace Rodrigues Silva, Anderson Miyoshi, Ricardo W Portela, Vasco Azevedo, Sibele Borsuk, Fernanda Alves Dorella, Daniela Droppaalmeida, Flavia S Rocha, Wanessa Lordelo Pedreira Vivas, Francine Ferreira PadilhaAbstract:Background: Caseous Lymphadenitis (CLA) is an infectious disease that affects small ruminants and is caused by Corynebacterium pseudotuberculosis. This disease is responsible for high economic losses due to condemnation and trim of infected carcasses, decreased leather and wool yield, loss of sales of breeding stock and deaths from internal involvement. Treatment is costly and ineffective; the most cost-effective strategy is timely immunisation. Various vaccine strategies have been tested, and recombinant vaccines are a promising alternative. Thus, in this study, different vaccine formulations using a recombinant protein (rCP40) and the CP09 live recombinant strain were evaluated. Five groups of 10 mice each were immunised with saline (G1), rCP40 (G2), CP09 (G3), a combination of CP09 and rCP40 (G4) and a heterologous prime-boost strategy (G5). Mice received two immunisations within 15 days. On day 30 after primary immunisation, all groups were challenged with a C. pseudotuberculosis virulent strain. Mice were monitored and mortality was recorded for 30 days after challenge. Results: The G2, G4 and G5 groups showed high levels of IgG1 and IgG2a; G2 presented significant IgG2a production after virulent challenge in the absence of IgG1 and IgG3 induction. Thirty days after challenge, the mice survival rates were 20 (G1), 90 (G2), 50 (G3), 70 (G4) and 60% (G5). Conclusions: rCP40 is a promising target in the development of vaccines against Caseous Lymphadenitis.
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association between haptoglobin and igm levels and the clinical progression of Caseous Lymphadenitis in sheep
BMC Veterinary Research, 2013Co-Authors: Bruno Lopes Bastos, Ricardo W Portela, Vasco Azevedo, Dan Loureiro, Jose Tadeu Raynal, Maria T Guedes, Vera Vale, Lilia F Mouracosta, Jose Eugenio Guimaraes, Roberto MeyerAbstract:Background Sheep Caseous Lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis (Cp), is associated with direct economic losses and presents significant zoonotic potential. Despite the importance of the disease, a satisfactory vaccine model has not been developed. Thus, this study aimed to investigate the association between haptoglobin (Hp) and IgM levels and the clinical progression of CLA in primarily infected sheep and in sheep immunized with Cp- secreted antigens adjuvanted with Quillaja saponaria saponins. These animals were kept with CLA-positive sheep to simulate natural exposure that occurs in field conditions. During the experiment, the Hp and IgM levels were monitored for 21 days, and the development of internal CLA lesions was investigated through necropsies on day182 post-immunization.
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subclinical diagnosis of Caseous Lymphadenitis based on elisa in sheep from brazil
Journal of Bacteriology & Parasitology, 2013Co-Authors: Dayana Ribeiro, Nubia Seyffert, Anderson Miyoshi, Ricardo W Portela, Roberto Meyer, Fernanda Alves Dorella, Luis G C Pacheco, Thiago Luiz De Paula Castro, Maria Cecilia Rui Luvizotto, Vasco AzevedoAbstract:Caseous Lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is a chronic contagious disease that affects small ruminants and still remains an important problem for many lamb-producing countries. Animals are considered clinically infected when occurs abscesses in superficial lymph nodes. Visceral or internal form can coexist which no apparent clinical signs of infection are seen. The best procedure to avoid spread of the disease is elimination of infected animals. However, as the chronic and subclinical nature of the infection of CLA alternative methods are required for detection and screening. In this study, we described the performance of indirect Enzyme-Linked Immunosorbent Assay (ELISA) for diagnosis of CLA in asymptomatics sheep. Also, test culture and biochemical identification were achieved to confirm CLA infection. The serological diagnostic was performed in sheep symptomatics (n=50) and asymptomatics (n=374) from nine flocks. Analysis reported high positivity of 71% for ELISA in 85% of asymptomatic animal for CLA with a sensitivity of 88% and specificity of 31%. Results from ELISA test in asymptomatic animals against culture for Caseous Lymphadenitis were more specific (97%) and permitted to exclude healthy animals without symptoms. This study concluded that C. pseudotuberculosis infection could be widely disseminated in sheep flocks in Northwestern region of the state of Sao Paulo, Brazil and only one screening test is not enough. The association with indirect ELISA test and culture could better indicate the real problem of CLA in sheep flocks.
Roberto Meyer - One of the best experts on this subject based on the ideXlab platform.
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a panel of recombinant proteins for the serodiagnosis of Caseous Lymphadenitis in goats and sheep
Microbial Biotechnology, 2019Co-Authors: Thiago Doria Barral, Roberto Meyer, Vasco Azevedo, Sibele Borsuk, Dan Loureiro, Ricardo Barros Mariutti, Raghuvir K Arni, Anderson Jesus Santos, Ana Rita Sokolonski, Ricardo Wagner Dias PortelaAbstract:Caseous Lymphadenitis (CLA) is a small ruminant disease characterized by the development of granulomatous lesions in superficial and internal lymph nodes, as well as in some organs, and causes significant economic losses worldwide. The aetiological agent of CLA is the bacterium Corynebacterium pseudotuberculosis; however, the commercially available diagnostic tools present problems with regard to specificity, which can lead to false-negative results. This study aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulins in goats and sheep using recombinant C. pseudotuberculosis PLD, CP40, PknG, DtxR and Grx proteins. For validation of the ELISAs, 130 goat serum samples and 160 sheep serum samples were used. The best ELISA for goats was developed using a combination of PLD and CP40 as antigens at a 1:1 ratio, which presented 96.9% sensitivity and 98.4% specificity. The most effective ELISA for sheep presented 91% sensitivity and 98.7% specificity when recombinant PLD alone was used as the antigen. These ELISAs can be used as highly accurate tools in epidemiological surveys and for the serodiagnosis of C. pseudotuberculosis infection in goats and sheep.
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brazilian green propolis as a therapeutic agent for the post surgical treatment of Caseous Lymphadenitis in sheep
Frontiers in Veterinary Science, 2019Co-Authors: Mauricio Alcântara Kalil, Roberto Meyer, Laerte Marlon Santos, Thiago Doria Barral, Daniela Meria Rodrigues, Neila De Paula Pereira, Maria Da Conceicao Aquino Sa, Marcelo Andres Umszaguez, Bruna Aparecida Souza Machado, Ricardo W PortelaAbstract:: As antibiotics are ineffective when used to treat Caseous Lymphadenitis, the surgical excision of lesions is often required. Iodine solution (10%) is currently the choice for the post-surgical treatment; however, it may cause histotoxicity. Propolis are resinous substances composed by a mixture of different plants parts and molecules secreted by bees. As green propolis has already proven to possess anti-bacterial and wound healing properties, this study aimed to evaluate the use of a green propolis-based ointment as a therapeutic agent for the post-surgical treatment of Caseous Lymphadenitis. The Caseous lesions of 28 sheep were surgically excised before dividing animals into two groups: (1) iodine-treated animals and (2) sheep treated with an ointment made with a previously characterized green propolis extract. Clinical data of animals, size of the scar area, the presence of moisture and secretion in the surgical wound, the humoral immune response against the bacterium and the susceptibility of C. pseudotuberculosis clinical isolates to the green propolis extract were analyzed. The green propolis-treated group presented complete healing of the surgical wound 1 week before the iodine-treated group. Additionally, animals treated with the green propolis ointment had fewer cases of wound secretion, but it was not statistically different from the iodine-treated group. No clinical signs indicating green propolis toxicity or other side effects were found, associated with a faster and more organized hair recovery by propolis use. The green propolis extract was able to inhibit the growth of 23 from the 27 C. pseudotuberculosis clinical isolates, with minimum inhibitory and minimum bactericide concentrations ranging from 01 to 08 mg/mL, and did not interfere with the humoral immune response against the bacterium. In addition, green propolis was able to inhibit biofilm formation by four of the C. pseudotuberculosis clinical isolates. We concluded that green propolis is a promising therapeutic agent to be used in the post-surgical treatment of Caseous Lymphadenitis in small ruminants due to its effects on surgical wound healing, hair recovery, inhibition of wound contamination and bacterial growth.
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association between haptoglobin and igm levels and the clinical progression of Caseous Lymphadenitis in sheep
BMC Veterinary Research, 2013Co-Authors: Bruno Lopes Bastos, Ricardo W Portela, Vasco Azevedo, Dan Loureiro, Jose Tadeu Raynal, Maria T Guedes, Vera Vale, Lilia F Mouracosta, Jose Eugenio Guimaraes, Roberto MeyerAbstract:Background Sheep Caseous Lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis (Cp), is associated with direct economic losses and presents significant zoonotic potential. Despite the importance of the disease, a satisfactory vaccine model has not been developed. Thus, this study aimed to investigate the association between haptoglobin (Hp) and IgM levels and the clinical progression of CLA in primarily infected sheep and in sheep immunized with Cp- secreted antigens adjuvanted with Quillaja saponaria saponins. These animals were kept with CLA-positive sheep to simulate natural exposure that occurs in field conditions. During the experiment, the Hp and IgM levels were monitored for 21 days, and the development of internal CLA lesions was investigated through necropsies on day182 post-immunization.
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subclinical diagnosis of Caseous Lymphadenitis based on elisa in sheep from brazil
Journal of Bacteriology & Parasitology, 2013Co-Authors: Dayana Ribeiro, Nubia Seyffert, Anderson Miyoshi, Ricardo W Portela, Roberto Meyer, Fernanda Alves Dorella, Luis G C Pacheco, Thiago Luiz De Paula Castro, Maria Cecilia Rui Luvizotto, Vasco AzevedoAbstract:Caseous Lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is a chronic contagious disease that affects small ruminants and still remains an important problem for many lamb-producing countries. Animals are considered clinically infected when occurs abscesses in superficial lymph nodes. Visceral or internal form can coexist which no apparent clinical signs of infection are seen. The best procedure to avoid spread of the disease is elimination of infected animals. However, as the chronic and subclinical nature of the infection of CLA alternative methods are required for detection and screening. In this study, we described the performance of indirect Enzyme-Linked Immunosorbent Assay (ELISA) for diagnosis of CLA in asymptomatics sheep. Also, test culture and biochemical identification were achieved to confirm CLA infection. The serological diagnostic was performed in sheep symptomatics (n=50) and asymptomatics (n=374) from nine flocks. Analysis reported high positivity of 71% for ELISA in 85% of asymptomatic animal for CLA with a sensitivity of 88% and specificity of 31%. Results from ELISA test in asymptomatic animals against culture for Caseous Lymphadenitis were more specific (97%) and permitted to exclude healthy animals without symptoms. This study concluded that C. pseudotuberculosis infection could be widely disseminated in sheep flocks in Northwestern region of the state of Sao Paulo, Brazil and only one screening test is not enough. The association with indirect ELISA test and culture could better indicate the real problem of CLA in sheep flocks.
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high sero prevalence of Caseous Lymphadenitis identified in slaughterhouse samples as a consequence of deficiencies in sheep farm management in the state of minas gerais brazil
BMC Veterinary Research, 2011Co-Authors: A S Guimaraes, Marcos Bryan Heinemann, Ricardo W Portela, Roberto Meyer, F B Carmo, Andrey Pereira LageAbstract:Background Caseous Lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is one of the most important diseases of sheep and goats, causing considerable economic losses for herd owners.