Immune response of BALB/c mice infected with two strains of Corynebacterium pseudotuberculosis.

This work evaluated aspects of the immune response of BALB/c mice infected with Corynebacterium pseudotuberculosis (T1 and C57). The fifteen BALB/c mice were euthanized after 70 days of infection and morphologically evaluated, also analyzing the innate and adaptive immune responses. The C57 strain induced more pronounced morphological changes than the T1 strain. There was an increase in CD4+ and CD8+ T cells identified during infection with the C57 strain. Cytokines of the inflammatory profile IL-1α and IL-6 and regulatory IL-13 and IL-10 presented significant differences. Cytokines IL-2, IL-4, INF-γ, IL-22, IL-21, and IL-27 did not differ significantly between groups. The obtained results contribute to a better understanding of the type of response and the immunological mechanisms involved during infection with different strains of C. pseudotuberculosis.

Bioinformatic Approach of B and T Cell Epitopes of PLD and CP40 Proteins of Corynebacterium pseudotuberculosis ovis Mexican Isolate 2J-L towards a Peptide-Based Vaccine.

Mapping B and T cell epitopes constitutes an important action for peptide vaccine design. PLD and CP40 virulence factors of Corynebacterium pseudotuberculosis biovar ovis, a causal agent of Caseous Lymphadenitis, have been evaluated in a murine model as good candidates for vaccine development. Therefore, the goal of this work was to in silico analyze B and T cell epitopes of the PLD and CP40 proteins of a Mexican isolate of Corynebacterium pseudotuberculosis ovis. The Immune Epitope Data Base and Resource website was employed to predict the linear and conformational B-cell, T CD4+, and T CD8+ epitopes of PLD and CP40 proteins of Corynebacterium pseudotuberculosis ovis Mexican strain 2J-L. Fifty B cell epitopes for PLD 2J-L and forty-seven for CP40 2J-L were estimated. In addition, T CD4+ and CD8+ cell epitopes were predicted for PLD 2J-L (MHC I:16 epitopes, MHC II:10 epitopes) and CP40 2J-L (MHC I: 15 epitopes, MHC II: 13 epitopes). This study provides epitopes, paying particular attention to sequences selected by different predictor programs and overlap sequences as B and T cell epitopes. PLD 2J-L and CP40 2J-L protein epitopes may aid in the design of a promising peptide-based vaccine against Caseous Lymphadenitis in Mexico.

In silico approach to identify microsatellite candidate biomarkers to differentiate the biovar of Corynebacterium pseudotuberculosis genomes.

Corynebacterium pseudotuberculosis is the causative bacterial agent of the zoonotic disease known as caseous lymphadenitis, and it presents several mechanisms of response to host defenses, including the presence of virulence factors (VFs). The genomes of these bacteria have several polymorphic markers known as microsatellites, or simple sequence repeats (SSRs), that can be used to characterize the genome, to study possible polymorphisms existing among strains, and to verify the effects of such polymorphic markers in coding regions and regions associated with VFs. In this study, several SSRs were identified within coding regions throughout the 54 genomes of this species, revealing possible polymorphisms associated with coding regions that could be used as strain-specific or serotype-specific identifiers of C. pseudotuberculosis. The similarities associated with SSRs amongst the different serum variants of C. pseudotuberculosis, biovars equi and ovis, were also evaluated, and it was possible to identify SSRs located in coding regions responsible for a VF enrolled in pathogenesis known to mediate bacterial adherence (SpaH-type pili virulence factor). Phylogenetic analyses revealed that strains sharing SSR patterns, including the possible polymorphisms identified in the same position of gene-coding regions, were displayed by strains with a common ancestor, corroborating with the Genome Tree Report of the NCBI. Statistical analysis showed that the microsatellite groups belonging to equi and ovis biovars have a significance of 0.006 (p-value) in similarity, thus indicating them as good biomarker candidates for C. pseudotuberculosis.

The Association of Bacterin and Recombinant Proteins Induces a Humoral Response in Sheep against Caseous Lymphadenitis.

In this study, we investigated the capacity of the recombinant proteins SpaC, NanH, SodC, and PLD of C. pseudotuberculosis to trigger protective humoral and cellular immune responses against experimentally induced C. pseudotuberculosis infection in sheep. The antigens were produced in a heterologous system and were purified by affinity chromatography. Nine sheep were randomly divided into three groups, which were immunized as follows: Group 1 (control)-a mix of adjuvants composed of the inactivated T1 strain of C. pseudotuberculosis and commercial Montanide™ISA 61 VG (T1M); Group 2-rSpaC, rSodC, rPLD, and T1M; Group 3-rNanH, rSodC, rPLD, and T1M. All groups were immunized twice (on days 0 and 30) and challenged on day 90 of the experiment. Humoral and cellular immune responses were evaluated by Enzyme-Linked Immunosorbent Assay (ELISA) to quantify the IgG antibodies and interferon-gamma (IFN-y). Both vaccine formulations with recombinant proteins (groups 2 and 3) could induce a significant humoral IgG immune response in sheep. The proteins rSodC, rPLD, and rNanH were more immunogenic, inducing significant levels of IgG antibodies after the first dose of the vaccine or after the challenge, maintaining constant levels until the end of the experiment. However, it was not possible to differentiate between the cellular responses induced by the vaccines. This lack of effectiveness points toward the need for further studies to improve the efficacy of this subunit-based vaccine approach.

Vaccines for caseous lymphadenitis: up-to-date and forward-looking strategies.

Caseous lymphadenitis (CLA) is an infectious chronic disease responsible for economic losses in sheep and goat breeding worldwide. CLA has no effective treatment, evidencing the vaccination schedule as the best control strategy. Although some commercial vaccines have been available, none of them provides total protection, which is sometimes insufficient and does not reach the same efficiency when compared in sheep and goats. They also have questionable safety levels and side effects. In light of this, several experimental vaccines are in development in order to improve safety, reproducibility, and protective immune response against the etiologic agent of CLA, Corynebacterium pseudotuberculosis. In this review, we discussed aspects as antigen, adjuvant, routes of administration, protection level, and animal models used in CLA vaccine development, as well the challenges and future perspectives. KEY POINTS: Caseous lymphadenitis (CLA) does not have an appropriate commercial vaccine. Different experimental vaccines are in development aiming to protect against Corynebacterium pseudotuberculosis. An ideal vaccine for CLA is necessary for the disease control.

A journey through the Corynebacterium pseudotuberculosis proteome promotes insights into its functional genome.

BACKGROUND: Corynebacterium pseudotuberculosis is a Gram-positive facultative intracellular pathogen and the etiologic agent of illnesses like caseous lymphadenitis in small ruminants, mastitis in dairy cattle, ulcerative lymphangitis in equines, and oedematous skin disease in buffalos. With the growing advance in high-throughput technologies, genomic studies have been carried out to explore the molecular basis of its virulence and pathogenicity. However, data large-scale functional genomics studies are necessary to complement genomics data and better understating the molecular basis of a given organism. Here we summarize, MS-based proteomics techniques and bioinformatics tools incorporated in genomic functional studies of C. pseudotuberculosis to discover the different patterns of protein modulation under distinct environmental conditions, and antigenic and drugs targets. METHODOLOGY: In this study we performed an extensive search in Web of Science of original and relevant articles related to methods, strategy, technology, approaches, and bioinformatics tools focused on the functional study of the genome of C. pseudotuberculosis at the protein level. RESULTS: Here, we highlight the use of proteomics for understating several aspects of the physiology and pathogenesis of C. pseudotuberculosis at the protein level. The implementation and use of protocols, strategies, and proteomics approach to characterize the different subcellular fractions of the proteome of this pathogen. In addition, we have discussed the immunoproteomics, immunoinformatics and genetic tools employed to identify targets for immunoassays, drugs, and vaccines against C. pseudotuberculosis infection. CONCLUSION: In this review, we showed that the combination of proteomics and bioinformatics studies is a suitable strategy to elucidate the functional aspects of the C. pseudotuberculosis genome. Together, all information generated from these proteomics studies allowed expanding our knowledge about factors related to the pathophysiology of this pathogen.

Assessment of caseous lymphadenitis in goats in a slaughterhouse in the Brazilian semi-arid region and estimates of economic losses due to carcass condemnation / Avaliação da linfadenite caseosa em caprinos em um abatedouro na região do semiárido brasileiro e estimativas de perdas econômicas por condenação de carcaças

Caseous lymphadenitis (CLA) caused by the pathogenic bacterium, Corynebacterium pseudotuberculosis, results in economic loss in goat farming. Its prevalence in Brazilian herds varies from 5 to 50%, generating expenses with treatment, loss in production, carcass and organ condemnation, and reduced price of the hide. This study aimed to describe the distribution of lesions and quantify the loss associated with CLA in goat breeding due to condemnation in a slaughterhouse located in a semiarid region. The study was conducted at the municipal slaughterhouse of the city of Patos, State of Paraíba, Northeastern Brazil, where goats from this intermediate geographical region are slaughtered. In 2017, 3,662 animals were slaughtered, an average of 305 per month. During the study period, from March to August 2017, 304 goats of both sexes and different ages of undefined breed were assessed. In the ante-mortem examination, inspection and palpation of the superficial lymph nodes was done; in the post-mortem examination, organ and viscera were assessed. The caseous material collected was sent for microbiological analysis. Of the 304 individuals, 227 [74.67% (95% CI: 69.50-79.23)] did not have any lesions, while 77 [25.33% (95% CI: 20.77-30.50)] showed abscesses suggestive of CLA, which was confirmed in 65 goats [84.41% (95% CI: 76.30-92.50)]. Sex (P = 0.044) and age (P = 0.002) were associated with infection. While the sex of the animals affected carcass and viscera weights and carcass yield, their age affected live weight, carcass, organ, and viscera weights; conversely, the incidence of CLA did not affect these variables. The average price in Real (R$) of a kilogram (kg) of goat meat cuts and “offal” was used to calculate the economic loss. Considering the average carcass weight (11,485 kg), organ weight (1,085 kg), and viscera weight (2,013 kg), 4,433.232 kg of meat was produced, resulting in R$ 72,945.43. The condemnations resulted in the loss of 269.894 kg, which is equivalent to R$ 4,540.33. CLA is responsible for a negative impact of 6.09% on production. Implementing control measures for this disease is important to increase the financial return in goat farming.(AU)

Linfadenite Caseosa (LC) causada pela bactéria patogênica Corynebacterium pseudotuberculosis resulta em perda econômica na caprinocultura. No rebanho brasileiro sua prevalência varia de 5 a 50%, gerando gastos com tratamento, queda de produção, condenação de carcaça e órgãos, e desvalorização da pele. O objetivo deste trabalho foi quantificar a perda associada à LC na caprinocultura de corte por condenação em abatedouro do semiárido, bem como caracterizar a distribuição das lesões. O trabalho foi realizado no Abatedouro Municipal da cidade de Patos, Estado da Paraíba, Nordeste do Brasil, onde ocorre abate de caprinos provenientes desta região geográfica intermediária. No ano de 2017 foram abatidos 3.662 animais, uma média de 305 por mês. Durante o período de estudo, março a agosto do mesmo ano, avaliou-se 304 caprinos sem padrão racial definido, ambos os sexos e idades diversas. No exame ante mortem foram submetidos à inspeção e palpação de linfonodos superficiais; post mortem, órgãos e vísceras. O material caseoso coletado foi submetido à análise microbiológica para diagnóstico. Dos 304 indivíduos, 227 [74,67% (IC 95%: 69,50-79,23)] não apresentaram qualquer lesão, enquanto 77 [25,33% (IC 95%: 20,77-30,50)] evidenciaram abscessos sugestivos de LC, com confirmação em 65 [84,41% (IC 95%: 76,30-92,50)]. Constatou-se associação de sexo (P = 0,044) e idade (P = 0,002) à infecção. Foi verificado efeito do sexo dos animais nas variáveis peso de carcaça, peso de vísceras e rendimento de carcaça, bem como houve efeito da idade nas variáveis peso vivo, peso de carcaça, peso de órgãos e peso de vísceras; no entanto, não houve efeito da ocorrência de LC nessas variáveis. Para cálculo de perda econômica adotou-se o preço médio em Real (R$) do quilograma (kg) dos cortes de carne caprina, e “miúdos”. Considerando o peso médio de carcaça (11,485 kg), órgãos (1,085 kg) e vísceras (2,013 kg), houve produção de 4.433,232 kg, resultando em R$ 72.945,43. As condenações subtraíram 269,894 kg, equivalente a R$ 4.540,33. Atribui-se à LC um impacto negativo de 6,09% na produção. Implementar medidas de controle para essa doença é importante para que se amplie o retorno financeiro à atividade.(AU)

rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay.

Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.Trial registration AEC No 4958051018. 12/18/2018, retrospectively registered.

In silico functional prediction of hypothetical proteins from the core genome of Corynebacterium pseudotuberculosis biovar ovis.

Corynebacterium pseudotuberculosis is a pathogen of veterinary relevance diseases, being divided into two biovars: equi and ovis; causing ulcerative lymphangitis and caseous lymphadenitis, respectively. The isolation and sequencing of C. pseudotuberculosis biovar ovis strains in the Northern and Northeastern regions of Brazil exhibited the emergence of this pathogen, which causes economic losses to small ruminant producers, and condemnation of carcasses and skins of animals. Through the pan-genomic approach, it is possible to determine and analyze genes that are shared by all strains of a species-the core genome. However, many of these genes do not have any predicted function, being characterized as hypothetical proteins (HP). In this study, we considered 32 C. pseudotuberculosis biovar ovis genomes for the pan-genomic analysis, where were identified 172 HP present in a core genome composed by 1255 genes. We are able to functionally annotate 80 sequences previously characterized as HP through the identification of structural features as conserved domains and families. Furthermore, we analyzed the physicochemical properties, subcellular localization and molecular function. Additionally, through RNA-seq data, we investigated the differential gene expression of the annotated HP. Genes inserted in pathogenicity islands had their virulence potential evaluated. Also, we have analyzed the existence of functional associations for their products based on protein-protein interaction networks, and perform the structural prediction of three targets. Due to the integration of different strategies, this study can underlie deeper in vitro researches in the characterization of these HP and the search for new solutions for combat this pathogen.

Factors associated with the seroprevalence of caseous lymphadenitis in sheep from Northeastern Brazil.

Caseous lymphadenitis (CL) is one of the main infectious diseases in sheep, responsible for great economic losses, mainly in Northeast region of Brazil, which has the largest sheep flock in the country. Thus, the aim of this survey was to determine the factors associated with the seroprevalence of this disease in sheep from Northeastern Brazil using a planned sample of flocks and animals. Samples were collected from 2312 adult and young sheep of both sex from 196 farms, located in 51 municipalities in five Northeastern Brazilian states (Ceará, Paraíba, Piauí, Rio Grande do Norte, and Sergipe). Serological diagnosis was performed using the indirect ELISA technique. Factors associated with seropositivity were identified by variables extracted from epidemiological questionnaires administered to breeders, using univariable and multivariable analyses. Factors associated with prevalence were purebred sheep (Prevalence ratio - PR = 1.189; P = 0.017), ram acquisition from animal expositions (PR = 1.192; P = 0.020), offspring is separated from ewes (PR = 1.132; P = 0.048), water supplied to sheep from ponds (PR = 1.365; P = 0.002), and delayed disposal of infected animals (PR = 1.263; P = 0.027). Flock-level and animal-level prevalences were 93.88 % (95 % CI = 89.60-96.46%) and 37.46 % (95 % CI = 35.51-39.45%), respectively. In view of the high prevalence found here and based on the associated factors, the implementation of an efficient CL control programme in Northeastern Brazil is recommended.

Co-Expression Networks for Causal Gene Identification Based on RNA-Seq Data of Corynebacterium pseudotuberculosis.

Corynebacterium pseudotuberculosis is a Gram-positive bacterium that causes caseous lymphadenitis, a disease that predominantly affects sheep, goat, cattle, buffalo, and horses, but has also been recognized in other animals. This bacterium generates a severe economic impact on countries producing meat. Gene expression studies using RNA-Seq are one of the most commonly used techniques to perform transcriptional experiments. Computational analysis of such data through reverse-engineering algorithms leads to a better understanding of the genome-wide complexity of gene interactomes, enabling the identification of genes having the most significant functions inferred by the activated stress response pathways. In this study, we identified the influential or causal genes from four RNA-Seq datasets from different stress conditions (high iron, low iron, acid, osmosis, and PH) in C. pseudotuberculosis, using a consensus-based network inference algorithm called miRsigand next identified the causal genes in the network using the miRinfluence tool, which is based on the influence diffusion model. We found that over 50% of the genes identified as influential had some essential cellular functions in the genomes. In the strains analyzed, most of the causal genes had crucial roles or participated in processes associated with the response to extracellular stresses, pathogenicity, membrane components, and essential genes. This research brings new insight into the understanding of virulence and infection by C. pseudotuberculosis.

Seroprevalence of Corynebacterium pseudotuberculosis and Toxoplasma gondii in sheep in semi-arid region of Ceará state / Soroprevalência de Corynebacterium pseudotuberculosis de Toxoplasma gondii em ovinos de região semi-árida do estado do Ceará

The present study was conducted to investigate in 20 extensive sheep farms for the seroprevalence of Corynebacterium pseudotuberculosis (n=402) and Toxoplasma gondii (n=228). Enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies to C. pseudotuberculosis/T. gondii. It was observed that C. pseudotuberculosis showed the highest prevalence in the region (34.07%) with statistically significant presence (p 0.05) in ewes. Antibodies to T. gondii was reported in 14.91% of the animals studied. About C. pseudotuberculosis/T. gondii coinfection the categories of rams showed significant (p 0.05) differences, suggesting that this gender could perpetuate the diseases in the flocks. It was concluded that the knowledge about the diseases in the region under study would facilitate the execution of prophylactic measures, especially against the diseases that pose risks to the public health and cause damages to the producer.(AU)

O presente estudo foi conduzido para investigar a soroprevalência em 20 fazendas de criação extensiva de ovinos quanto à presença de anticorpos para Corynebacterium pseudotuberculosis (n=402) e Toxoplasma gondii (n=228). Ensaio de imunoabsorção enzimático (ELISA) foi utilizado para a detecção de anticorpos contra C. pseudotuberculosis/T. gondii. Observou-se que C. pseudotuberculosis apresentou a maior prevalência na região (34,07%), com presença estatisticamente significante (p 0,05) nas categorias de ovelha. Anticorpos contra T. gondii foram encontrados em 14,91% dos animais estudados. Sobre a coinfecção de C. pseudotuberculosis/T. gondii, as categorias carneiro apresentaram diferenças significativas (p 0,05), sugerindo que esse gênero poderia perpetuar as doenças nos rebanhos. Concluiu-se que o conhecimento sobre as doenças na região em estudo facilitaria a execução de medidas profiláticas, principalmente contra as doenças que apresentam riscos à saúde pública e causam danos ao produtor.(AU)

Clinical-epidemiological study of caseous lymphadenitis in goats in the state of Pernambuco / Estudo clínico-epidemiológico da linfadenite caseosa em caprinos no estado de Pernambuco

Caseous lymphadenitis (CL), a contagious infectious disease caused by Corynebacterium pseudotuberculosis, stands out with great economic relevance for goat breeding in the country because it is one of the main causes of carcass condemnation and intense depreciation of the leather of small ruminants. The objective was to carry out a descriptive study of the clinical and epidemiological aspects of CL in goats in the state of Pernambuco and also to evaluate the detection of C. pseudotuberculosis in milk, blood, and stool samples by real-time PCR. Six hundred and forty-three goats were evaluated, of which 76 (11.8%) presented clinical signs suggestive of CL. And 88.9% (24/27) of the breeding was considered focus. Among symptomatic animals, 25 were selected and submitted to the collection of abscess content for bacteriological isolation. Milk, blood, and stool samples were analyzed for PLD gene detection by real-time polymerase chain reaction (PCR). Out of the 25 samples of abscess content, 72% (18/25) were positive. The PLD gene was detected in the frequencies of 8% (2/25) and 4% (1/25) in milk and blood samples, respectively. Caseous lymphadenitis is possibly widespread in the dairy herds of goats in the state of Pernambuco, especially in adult animals, with a higher occurrence of submandibular lymph node abscesses. The preliminary detection of C. pseudotuberculosis by mammary and

A linfadenite caseosa (LC), doença infectocontagiosa ocasionada pelo Corynebacterium pseudotuberculosis, se destaca com grande relevância econômica para a caprinocultura no país por ser umas das principais causas de condenação de carcaças e intensa depreciação do couro de pequenos ruminantes. Objetivou-se realizar um estudo descritivo dos aspectos clínico-epidemiológicos da LC em caprinos no estado de Pernambuco e, adicionalmente, avaliar a detecção do C. pseudotuberculosis em amostras de leite, sangue e fezes por PCR em tempo real. Foram avaliados 643 caprinos, dos quais 76 (11,8%) apresentaram sinais clínicos sugestivos de LC. E 88,9% (24/27) das propriedades foram consideradas foco. Dentre os animais sintomáticos, 25 foram selecionados e submetidos à colheita de conteúdo de abscesso para isolamento bacteriológico. Amostras de leite, sangue e fezes foram analisadas para detecção do gene PLD pela reação em cadeia da polimerase (PCR) em tempo real. Das 25 amostras de conteúdo de abscesso, 72% (18/25) resultaram positivas. O gene PLD foi detectado nas frequências de 8% (2/25) e 4% (1/25) em amostras de leite e sangue, respectivamente. A linfadenite caseosa possivelmente encontra-se disseminada nos rebanhos leiteiros de caprinos do estado de Pernambuco, sobretudo em animais adultos, com maior ocorrência de abscessos em linfonodos submandibulares. A detecção preliminar do C. pseud

Assessment of caseous lymphadenitis in goats in a slaughterhouse in the Brazilian semi-arid region and estimates of economic losses due to carcass condemnation / Avaliação da linfadenite caseosa em caprinos em um abatedouro na região do semiárido brasileiro e estimativas de perdas econômicas por condenação de carcaças

Caseous lymphadenitis (CLA) caused by the pathogenic bacterium, Corynebacterium pseudotuberculosis, results in economic loss in goat farming. Its prevalence in Brazilian herds varies from 5 to 50%, generating expenses with treatment, loss in production, carcass and organ condemnation, and reduced price of the hide. This study aimed to describe the distribution of lesions and quantify the loss associated with CLA in goat breeding due to condemnation in a slaughterhouse located in a semiarid region. The study was conducted at the municipal slaughterhouse of the city of Patos, State of Paraíba, Northeastern Brazil, where goats from this intermediate geographical region are slaughtered. In 2017, 3,662 animals were slaughtered, an average of 305 per month. During the study period, from March to August 2017, 304 goats of both sexes and different ages of undefined breed were assessed. In the ante-mortem examination, inspection and palpation of the superficial lymph nodes was done; in the post-mortem examination, organ and viscera were assessed. The caseous material collected was sent for microbiological analysis. Of the 304 individuals, 227 [74.67% (95% CI: 69.50-79.23)] did not have any lesions, while 77 [25.33% (95% CI: 20.77-30.50)] showed abscesses suggestive of CLA, which was confirmed in 65 goats [84.41% (95% CI: 76.30-92.50)]. Sex (P = 0.044) and age (P = 0.002) were associated with infection. While the sex of the animals affected carcass and viscera weights and carcass yield, their age affected live weight, carcass, organ, and viscera weights; conversely, the incidence of CLA did not affect these variables. The average price in Real (R$) of a kilogram (kg) of goat meat cuts and “offal” was used to calculate the economic loss. Considering the average carcass weight (11,485 kg), organ weight (1,085 kg), and viscera weight (2,013 kg), 4,433.232 kg of meat was produced, resulting in R$ 72,945.43. The condemnations resulted in the loss of 269.894 kg, which is equivalent to R$ 4,540.33. CLA is responsible for a negative impact of 6.09% on production. Implementing control measures for this disease is important to increase the financial return in goat farming.

Linfadenite Caseosa (LC) causada pela bactéria patogênica Corynebacterium pseudotuberculosis resulta em perda econômica na caprinocultura. No rebanho brasileiro sua prevalência varia de 5 a 50%, gerando gastos com tratamento, queda de produção, condenação de carcaça e órgãos, e desvalorização da pele. O objetivo deste trabalho foi quantificar a perda associada à LC na caprinocultura de corte por condenação em abatedouro do semiárido, bem como caracterizar a distribuição das lesões. O trabalho foi realizado no Abatedouro Municipal da cidade de Patos, Estado da Paraíba, Nordeste do Brasil, onde ocorre abate de caprinos provenientes desta região geográfica intermediária. No ano de 2017 foram abatidos 3.662 animais, uma média de 305 por mês. Durante o período de estudo, março a agosto do mesmo ano, avaliou-se 304 caprinos sem padrão racial definido, ambos os sexos e idades diversas. No exame ante mortem foram submetidos à inspeção e palpação de linfonodos superficiais; post mortem, órgãos e vísceras. O material caseoso coletado foi submetido à análise microbiológica para diagnóstico. Dos 304 indivíduos, 227 [74,67% (IC 95%: 69,50-79,23)] não apresentaram qualquer lesão, enquanto 77 [25,33% (IC 95%: 20,77-30,50)] evidenciaram abscessos sugestivos de LC, com confirmação em 65 [84,41% (IC 95%: 76,30-92,50)]. Constatou-se associação de sexo (P = 0,044) e idade (P = 0,002) à infecção. Foi verificado efeito do sexo dos animais nas variáveis peso de carcaça, peso de vísceras e rendimento de carcaça, bem como houve efeito da idade nas variáveis peso vivo, peso de carcaça, peso de órgãos e peso de vísceras; no entanto, não houve efeito da ocorrência de LC nessas variáveis. Para cálculo de perda econômica adotou-se o preço médio em Real (R$) do quilograma (kg) dos cortes de carne caprina, e “miúdos”. Considerando o peso médio de carcaça (11,485 kg), órgãos (1,085 kg) e vísceras (2,013 kg), houve produção de 4.433,232 kg, resultando em R$ 72.945,43. As condenações subtraíram 269,894 kg, equivalente a R$ 4.540,33. Atribui-se à LC um impacto negativo de 6,09% na produção. Implementar medidas de controle para essa doença é importante para que se amplie o retorno financeiro à atividade.

Clinical-epidemiological study of caseous lymphadenitis in goats in the state of Pernambuco / Estudo clínico-epidemiológico da linfadenite caseosa em caprinos no estado de Pernambuco

Caseous lymphadenitis (CL), a contagious infectious disease caused by Corynebacterium pseudotuberculosis, stands out with great economic relevance for goat breeding in the country because it is one of the main causes of carcass condemnation and intense depreciation of the leather of small ruminants. The objective was to carry out a descriptive study of the clinical and epidemiological aspects of CL in goats in the state of Pernambuco and also to evaluate the detection of C. pseudotuberculosis in milk, blood, and stool samples by real-time PCR. Six hundred and forty-three goats were evaluated, of which 76 (11.8%) presented clinical signs suggestive of CL. And 88.9% (24/27) of the breeding was considered focus. Among symptomatic animals, 25 were selected and submitted to the collection of abscess content for bacteriological isolation. Milk, blood, and stool samples were analyzed for PLD gene detection by real-time polymerase chain reaction (PCR). Out of the 25 samples of abscess content, 72% (18/25) were positive. The PLD gene was detected in the frequencies of 8% (2/25) and 4% (1/25) in milk and blood samples, respectively. Caseous lymphadenitis is possibly widespread in the dairy herds of goats in the state of Pernambuco, especially in adult animals, with a higher occurrence of submandibular lymph node abscesses. The preliminary detection of C. pseudotuberculosis by mammary and(AU)

A linfadenite caseosa (LC), doença infectocontagiosa ocasionada pelo Corynebacterium pseudotuberculosis, se destaca com grande relevância econômica para a caprinocultura no país por ser umas das principais causas de condenação de carcaças e intensa depreciação do couro de pequenos ruminantes. Objetivou-se realizar um estudo descritivo dos aspectos clínico-epidemiológicos da LC em caprinos no estado de Pernambuco e, adicionalmente, avaliar a detecção do C. pseudotuberculosis em amostras de leite, sangue e fezes por PCR em tempo real. Foram avaliados 643 caprinos, dos quais 76 (11,8%) apresentaram sinais clínicos sugestivos de LC. E 88,9% (24/27) das propriedades foram consideradas foco. Dentre os animais sintomáticos, 25 foram selecionados e submetidos à colheita de conteúdo de abscesso para isolamento bacteriológico. Amostras de leite, sangue e fezes foram analisadas para detecção do gene PLD pela reação em cadeia da polimerase (PCR) em tempo real. Das 25 amostras de conteúdo de abscesso, 72% (18/25) resultaram positivas. O gene PLD foi detectado nas frequências de 8% (2/25) e 4% (1/25) em amostras de leite e sangue, respectivamente. A linfadenite caseosa possivelmente encontra-se disseminada nos rebanhos leiteiros de caprinos do estado de Pernambuco, sobretudo em animais adultos, com maior ocorrência de abscessos em linfonodos submandibulares. A detecção preliminar do C. pseud(AU)

Seroprevalence of Corynebacterium pseudotuberculosis and Toxoplasma gondii in sheep in semi-arid region of Ceará state / Soroprevalência de Corynebacterium pseudotuberculosis de Toxoplasma gondii em ovinos de região semi-árida do estado do Ceará

ABSTRACT: The present study was conducted to investigate in 20 extensive sheep farms for the seroprevalence of Corynebacterium pseudotuberculosis (n=402) and Toxoplasma gondii (n=228). Enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies to C. pseudotuberculosis/T. gondii. It was observed that C. pseudotuberculosis showed the highest prevalence in the region (34.07%) with statistically significant presence (p<0.05) in ewes. Antibodies to T. gondii was reported in 14.91% of the animals studied. About C. pseudotuberculosis/T. gondii coinfection the categories of rams showed significant (p<0.05) differences, suggesting that this gender could perpetuate the diseases in the flocks. It was concluded that the knowledge about the diseases in the region under study would facilitate the execution of prophylactic measures, especially against the diseases that pose risks to the public health and cause damages to the producer.

RESUMO: O presente estudo foi conduzido para investigar a soroprevalência em 20 fazendas de criação extensiva de ovinos quanto à presença de anticorpos para Corynebacterium pseudotuberculosis (n=402) e Toxoplasma gondii (n=228). Ensaio de imunoabsorção enzimático (ELISA) foi utilizado para a detecção de anticorpos contra C. pseudotuberculosis/T. gondii. Observou-se que C. pseudotuberculosis apresentou a maior prevalência na região (34,07%), com presença estatisticamente significante (p <0,05) nas categorias de ovelha. Anticorpos contra T. gondii foram encontrados em 14,91% dos animais estudados. Sobre a coinfecção de C. pseudotuberculosis/T. gondii, as categorias carneiro apresentaram diferenças significativas (p <0,05), sugerindo que esse gênero poderia perpetuar as doenças nos rebanhos. Concluiu-se que o conhecimento sobre as doenças na região em estudo facilitaria a execução de medidas profiláticas, principalmente contra as doenças que apresentam riscos à saúde pública e causam danos ao produtor.

Brazilian Green Propolis as a Therapeutic Agent for the Post-surgical Treatment of Caseous Lymphadenitis in Sheep.

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.

Acute death of kangaroos in a zoo due to highly pathogenic Corynebacterium pseudotuberculosis and Yersinia pseudotuberculosis

Background: “Lumpy jaw” is disease effecting wallabies and kangaroos, particularly in Macropus rufus and Macropusgiganteus. In the most serious situations, additional tooth loss and fistulas follow, accompanied by a stench, weight loss,and eventually death due to sepsis or blood poisoning. “Lumpy jaw” disease has seriously affected the normal display andhealth of kangaroos, and cause a huge economic loss. There was an outbreak of jaw infection in kangaroos at the HongshanForest Zoo. Two Macropus giganteus and two Macropus rufus died of “lumpy jaw”. The main objective of the describingcase was to isolate pathogens, provide a basis for follow-up treatment, and serve to establish a disease prevention protocol.Case: Four grown-up kangaroos (two Macropus giganteus and two Macropus rufus) were raised in Hongshan Forest Zoo,which had obviously clinical symptoms, such as oral lesions of pus, necrotic tissue, rotting teeth, then died of “lumpyjaw”. Oral swab samples were collected from the lesion sites of the dying kangaroos. Mice experiments were conducted toexamine the pathogenicity of the strains. Tests of antimicrobial susceptibity were performed to prescribe with better drugtreatments for kangaroos. Corynebacterium pseudotuberculosis and Yersinia pseudotuberculosis were identified based onmorphology, culture characteristics and biochemical tests. Corynebacterium pseudotuberculosis (G+) in Sucrose, Mannitol,Lactose, Maltose, Glucose tubes were positive, that acids and gases both production, in Gelatin liquefaction, Indol test,MR were positive, that only acids production, others were negative; Yersinia pseudotuberculosis (G-) in Urea, MR werepositive, that only acids production, others were negative.The infected mice presented with gum erosion or ulcers whenthe two pathogens were injected subcutaneous at the oral regional by 2-3 point at 0.2 mL of individual strains 1.0×109CFU/mouse...(AU)

Characterization of caseous lymphadenitis in caprine animals slaughtered in a semi-arid region of Brazil / Caracterização da linfadenite caseosa em caprinos abatidos no semiárido brasileiro

In the Northeast of Brazil, goat breeding plays an important socioeconomic and strategic role in coexistence within the semi-arid region. The rural nature of the species and adaptations that are appropriate for the climate are some of the characteristics that favor the Northeast as the largest national producer of goats (93% of the national herd). In spite of this, sanitary deficiencies facilitate the appearance and diffusion of pathogenic agents that compromise the productivity, e.g., Corynebacterium pseudotuberculosis, which are widespread in the national herd and cause damages in the industry. The objective of this study was to characterize caseous lymphadenitis (CL) in lamb cuttings at semi-arid abattoirs. The research was carried out between March and August 2017 at the Municipal Abattoir of Patos, State of Paraíba, Brazil, where the caprines from this region are slaughtered. We studied 304 animals with no defined racial pattern, of both the sexes, characterized according to their age group as evidenced by their teeth, submitted to ante- and post-mortem examinations with microbiological confirmation. Cheek material was processed for diagnostic confirmation, using the techniques of isolation and microbiological identification (gold standard). The bacteriological examination confirmed the presence of the agent in 21.38% of the animals, in which 13.16% were females and 8.22% were males. Of those affected, 11.51% had the clinical form of the disease, while 9.87% had the subclinical form and 1.31% had both the clinical form as well as the internal impairment. The precrural (25.71%), mammary (11.43%), and prescapular (11.43%) lymph nodes were the most affected. Internally, the most affected were the liver (63.33%), lung (13.33%), and medium mediastinal lymph nodes (10%).[...]

No Nordeste do Brasil a caprinocultura desempenha importante função socioeconômica e estratégica na convivência em região semiárida. A boa adaptação ao clima somado a rusticidade da espécie são algumas das caraterísticas que favorecem o Nordeste como maior produtor nacional (93% do efetivo). Apesar dessa condição, deficiências sanitárias proporcionam surgimento e difusão de agentes patogênicos que comprometem a produtividade, destacando-se o Corynebacterium pseudotuberculosis, difundido no rebanho nacional e que gera prejuízos à atividade. O objetivo deste estudo foi caracterizar a Linfadenite Caseosa (LC) na caprinocultura de corte em abatedouro do semiárido. Desenvolveu-se a pesquisa entre março e agosto 2017 no Abatedouro Municipal de Patos, Estado da Paraíba, Brasil, onde ocorre abate de caprídeos provenientes desta região geográfica intermediária. Foram estudados 304 animais sem padrão racial definido, ambos os sexos, caracterizados conforme faixa etária evidenciada pelos dentes, submetidos a exames ante e post mortem com confirmação microbiológica. Processou-se material caseoso dos doentes; para confirmação de diagnóstico se empregou a técnica de isolamento e identificação microbiológica (padrão ouro). O exame bacteriológico confirmou presença do agente em 21.38% dos animais. As fêmeas representaram 13.16% dos doentes, enquanto os machos, 8.22%. Dos acometidos, 11.51% apresentaram a forma clínica, enquanto 9.87% a subclínica; 1.31% tiveram simultaneamente forma clínica e comprometimento interno. Os linfonodos pré-crurais 25.71%, mamários 11.43% e pré-escapulares 11.43% foram mais afetados; internamente, fígado 63.33%, pulmão 13.33% e linfonodo do mediastino médio 10%.[...]

Characterization of caseous lymphadenitis in caprine animals slaughtered in a semi-arid region of Brazil / Caracterização da linfadenite caseosa em caprinos abatidos no semiárido brasileiro

In the Northeast of Brazil, goat breeding plays an important socioeconomic and strategic role in coexistence within the semi-arid region. The rural nature of the species and adaptations that are appropriate for the climate are some of the characteristics that favor the Northeast as the largest national producer of goats (93% of the national herd). In spite of this, sanitary deficiencies facilitate the appearance and diffusion of pathogenic agents that compromise the productivity, e.g., Corynebacterium pseudotuberculosis, which are widespread in the national herd and cause damages in the industry. The objective of this study was to characterize caseous lymphadenitis (CL) in lamb cuttings at semi-arid abattoirs. The research was carried out between March and August 2017 at the Municipal Abattoir of Patos, State of Paraíba, Brazil, where the caprines from this region are slaughtered. We studied 304 animals with no defined racial pattern, of both the sexes, characterized according to their age group as evidenced by their teeth, submitted to ante- and post-mortem examinations with microbiological confirmation. Cheek material was processed for diagnostic confirmation, using the techniques of isolation and microbiological identification (gold standard). The bacteriological examination confirmed the presence of the agent in 21.38% of the animals, in which 13.16% were females and 8.22% were males. Of those affected, 11.51% had the clinical form of the disease, while 9.87% had the subclinical form and 1.31% had both the clinical form as well as the internal impairment. The precrural (25.71%), mammary (11.43%), and prescapular (11.43%) lymph nodes were the most affected. Internally, the most affected were the liver (63.33%), lung (13.33%), and medium mediastinal lymph nodes (10%).[...](AU)

No Nordeste do Brasil a caprinocultura desempenha importante função socioeconômica e estratégica na convivência em região semiárida. A boa adaptação ao clima somado a rusticidade da espécie são algumas das caraterísticas que favorecem o Nordeste como maior produtor nacional (93% do efetivo). Apesar dessa condição, deficiências sanitárias proporcionam surgimento e difusão de agentes patogênicos que comprometem a produtividade, destacando-se o Corynebacterium pseudotuberculosis, difundido no rebanho nacional e que gera prejuízos à atividade. O objetivo deste estudo foi caracterizar a Linfadenite Caseosa (LC) na caprinocultura de corte em abatedouro do semiárido. Desenvolveu-se a pesquisa entre março e agosto 2017 no Abatedouro Municipal de Patos, Estado da Paraíba, Brasil, onde ocorre abate de caprídeos provenientes desta região geográfica intermediária. Foram estudados 304 animais sem padrão racial definido, ambos os sexos, caracterizados conforme faixa etária evidenciada pelos dentes, submetidos a exames ante e post mortem com confirmação microbiológica. Processou-se material caseoso dos doentes; para confirmação de diagnóstico se empregou a técnica de isolamento e identificação microbiológica (padrão ouro). O exame bacteriológico confirmou presença do agente em 21.38% dos animais. As fêmeas representaram 13.16% dos doentes, enquanto os machos, 8.22%. Dos acometidos, 11.51% apresentaram a forma clínica, enquanto 9.87% a subclínica; 1.31% tiveram simultaneamente forma clínica e comprometimento interno. Os linfonodos pré-crurais 25.71%, mamários 11.43% e pré-escapulares 11.43% foram mais afetados; internamente, fígado 63.33%, pulmão 13.33% e linfonodo do mediastino médio 10%.[...](AU)