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BACKGROUND: Corynebacterium spp. are widely disseminated in the environment, and they are part of the skin and mucosal microbiota of animals and humans. Reports of human infections by Corynebacterium spp. have increased considerably in recent years and the appearance of multidrug resistant isolates around the world has drawn attention. OBJECTIVES: To describe a new species of Corynebacterium from human tissue bone is described after being misidentified using available methods. METHODS: For taxonomic analyses, phylogenetic analysis of 16S rRNA and rpoB genes, in silico DNA-DNA hybridization, average nucleotide and amino acid identity, multilocus sequence analysis, and phylogenetic analysis based on the complete genome were used. FINDINGS: Genomic taxonomic analyzes revealed values of in silico DNA-DNA hybridization, average nucleotide and amino acids identity below the values necessary for species characterization between the analyzed isolates and the closest phylogenetic relative Corynebacterium aurimucosum DSM 44532T. MAIN CONCLUSIONS: Genomic taxonomic analyzes indicate that the isolates analyzed comprise a new species of the Corynebacterium genus, which we propose to name Corynebacterium hiratae sp. nov. with isolate 332T (= CBAS 826T = CCBH 35,014T) as the type strain.
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Infecções por Corynebacterium , Corynebacterium , DNA Bacteriano , Filogenia , RNA Ribossômico 16S , Corynebacterium/genética , Corynebacterium/classificação , Corynebacterium/isolamento & purificação , Humanos , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Infecções por Corynebacterium/microbiologia , Osso e Ossos/microbiologia , Tipagem de Sequências Multilocus , Genoma Bacteriano , Técnicas de Tipagem Bacteriana , Hibridização de Ácido NucleicoRESUMO
Equine strangles is a prevalent disease that affects the upper respiratory in horses and is caused by the Gram-positive bacterium Streptococcus equi. In addition to strangles, other clinical conditions are caused by the two S. equi subspecies, equi and zooepidemicus, which present relevant zoonotic potential. Treatment of infections caused by S. equi has become challenging due to the worldwide spreading of infected horses and the unavailability of effective therapeutics and vaccines. Penicillin treatment is often recommended, but multidrug resistance issues arised. We explored the whole genome sequence of 18 S. equi isolates to identify candidate proteins to be targeted by natural drug-like compounds or explored as immunogens. We considered only proteins shared among the sequenced strains of subspecies equi and zooepidemicus, absent in the equine host and predicted to be essential and involved in virulence. Of these, 4 proteins with cytoplasmic subcellular location were selected for molecular docking with a library of 5008 compounds, while 6 proteins were proposed as prominent immunogens against S. equi due to their probabilities of behaving as adhesins. The molecular docking analyses revealed the best ten ligands for each of the 4 drug target candidates, and they were ranked according to their binding affinities and the number of hydrogen bonds for complex stability. Finally, the natural 5-ring compound C25H20F3N5O3 excelled in molecular dynamics simulations for the increased stability in the interaction with UDP-N-acetylenolpyruvoylglucosamine reductase (MurB). This research paves the way to developing new therapeutics to minimize the impacts caused by S. equi infections.Communicated by Ramaswamy H. Sarma.
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C-di-GMP is a bacterial second messenger with central role in biofilm formation. Spirochete bacteria from Leptospira genus present a wide diversity, with species of medical importance and environmental species, named as saprophytic. Leptospira form biofilms in the rat's reservoir kidneys and in the environment. Here, we performed genomic analyses to identify enzymatic and effector c-di-GMP proteins in the saprophytic biofilm-forming species Leptospira biflexa serovar Patoc. We identified 40 proteins through local alignments. Amongst them, 16 proteins are potentially functional diguanylate cyclases, phosphodiesterases, or hybrid proteins. We also identified nine effectors, including PilZ proteins. Enrichment analyses suggested that c-di-GMP interacts with cAMP signaling system, CsrA system, and flagella assembly regulation during biofilm development of L. biflexa. Finally, we identified eight proteins in the pathogen Leptospira interrogans serovar Copenhageni that share high similarity with L. biflexa c-di-GMP-related proteins. This work revealed proteins related to c-di-GMP turnover and cellular response in Leptospira and their potential roles during biofilm development.
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Proteínas de Escherichia coli , Leptospira , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Spirochaetales/metabolismo , Proteínas de Escherichia coli/genética , Bactérias/metabolismo , Leptospira/genética , Leptospira/metabolismo , Genômica , Biofilmes , Regulação Bacteriana da Expressão GênicaRESUMO
Staphylococcus aureus is the main etiological agent of mastitis in small ruminants worldwide. This disease has a difficult cure and possible relapse, leading to significant economic losses in production, milk quality and livestock. This study performed comparative genomic analyses between 73 S. aureus genomes from different hosts (human, bovine, pig and others). This work isolated and sequenced 12 of these genomes from ovine. This study contributes to the knowledge of genomic specialization and the role of specific genes in establishing infection in ovine mastitis-associated S. aureus. The genomes of S. aureus isolated from sheep maintained a higher representation when grouped with clonal complexes 130 and 133. The genomes showed high genetic similarity, the species pan-genome consisting of 4200 genes (central = 2008, accessory = 1559 and unique = 634). Among these, 277 unique genes were related to the genomes isolated from sheep, with 39.6 % as hypothetical proteins, 6.4 % as phages, 6.4 % as toxins, 2.9 % as transporters, and 44.7 % as related to other proteins. Furthermore, at the pathogen level, they showed 80 genes associated with virulence factors and 19 with antibiotic resistance shared in almost all isolates. Although S. aureus isolated from ovine showed susceptibility to antimicrobials in vitro, ten genes were predicted to be associated with antibiotic inactivation and efflux pump, suggesting resistance to gentamicin and penicillin. This work may contribute to identifying genes acquired by horizontal transfer and their role in host adaptation, virulence, bacterial resistance, and characterization of strains affecting ovine.
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Mastite Bovina , Infecções Estafilocócicas , Feminino , Animais , Bovinos , Ovinos/genética , Humanos , Suínos , Fatores de Virulência/genética , Staphylococcus aureus/genética , Adaptação ao Hospedeiro , Infecções Estafilocócicas/genética , Infecções Estafilocócicas/veterinária , Infecções Estafilocócicas/microbiologia , Ruminantes/genética , Genômica , Sequências Repetitivas Dispersas , Mastite Bovina/genética , Mastite Bovina/microbiologiaRESUMO
Biochemical, serological, and molecular methods have been developed for the laboratory diagnosis of diseases caused by C. pseudotuberculosis (CP), but the identification of the pathogen and biovars differentiation may be time-consuming, expensive, and confusing compared with other bacteria. This study aimed to evaluate MALDI Biotyper and Overall Genome Relatedness Index (OGRI) analysis to optimize the identification and differentiation of biovars of C. pseudotuberculosis. Out of 230 strains isolated from several hosts and countries, 202 (87.8%) were precisely classified using MALDI Biotyper and the BioNumerics platform. The classification accuracies for the Ovis and Equi biovars were 80 (88.75%) and 82 (92.68%), respectively. When analyzing a sampling of these strains by Average Nucleotide Identity based on BLAST and TETRA analyses using genomic sequence data, it was possible to differentiate 100% of the strains in Equi and Ovis. Our data show that MALDI Biotyper and OGRI analysis help identify C. pseudotuberculosis at the species and biovar levels.
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Corynebacterium pseudotuberculosis , Ovinos , Animais , Corynebacterium pseudotuberculosis/genética , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis in small ruminants, a chronic disease characterized by the development of granulomas in superficial and visceral lymph nodes as well as in several organs. An important characteristic of the infection with this bacterium is the formation of a biofilm and the absence of effective antibiotic therapy against the disease. From this scenario, the objective of this study was to evaluate the susceptibility of C. pseudotuberculosis to conventional antibiotics and to red, green, and brown propolis extracts obtained by the supercritical and ethanolic extraction methods as well as its activity in the bacterial biofilm. The results of the sensitivity test using antibiotics indicated a sensitivity of C. pseudotuberculosis strains to the antimicrobial agents. The ethanolic extract of green propolis and the supercritical red propolis extract showed the best antibacterial activities against planktonic C. pseudotuberculosis. A lower antimicrobial activity of the brown propolis extract was identified. Propolis extracts were effective in interfering with the formation of the C. pseudotuberculosis biofilm but had little activity on the consolidated biofilm. In conclusion, propolis extracts are more effective against C. pseudotuberculosis in the planktonic stage, being able to interfere with the formation of bacterial biofilm. However, the action of propolis extracts in a sessile and structured microbial biofilm is reduced.
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Caseous lymphadenitis (CLA) is a chronic disease that affects small ruminants and causes economic losses in the associated breeding system. The causative agent of CLA is Corynebacterium pseudotuberculosis, a Gram-positive bacterium that exhibits tropism for external and internal lymph nodes and induces abscess formation in the host. Bacterial communities often produce a biofilm matrix that serves various functions, including protection against hostile environmental conditions, antibiotics, and the host immune response. Although biofilm formation has been reported for C. pseudotuberculosis, not all strains demonstrate this property in culture. In this work, we report the first comparative proteomic analysis of one biofilm-forming (CAPJ4) and one biofilm-non-forming strain (CAP3W) of C. pseudotuberculosis isolated from goats. Bacterial whole cell protein extracts were obtained for mass spectrometry analyses. Using LC-MS/MS, our studies reveal three and four proteins exclusively found in the CAPJ4 and CAP3W proteome, respectively. In addition, label-free quantitative analysis identified 40 proteins showing at-least 2-fold higher values in CAPJ4 compared CAP3W proteome Notably, CAPJ4 differentially synthesized the penicillin-binding protein, which participates in the formation of peptidoglycans. CAPJ4 also exhibited upregulation of N-acetylmuramoyl-L-alanine amidase and galactose-1-phosphate uridylyltransferase, which are involved in biofilm formation and exopolysaccharide biosynthesis. Here, we demonstrate that biofilm formation in C. pseudotuberculosis is likely associated with specific proteins, some of which were previously shown to be associated with virulence and biofilm formation in other organisms. Our findings may drive studies related to the bacterial mechanisms involved in the biofilm formation, in addition to providing targets for the treatment of CLA.
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The genus Klebsiella comprises species that cause nosocomial and community-acquired infections. A dataset was created to compile the sequence type (ST) and capsule type (K-locus) information predicted for 172 worldwide isolates of Klebsiella spp. whose complete genomes could be retrieved from the GenBank (NCBI) repository. The dataset also includes information related to one multidrug-resistant strain (B31) isolated from a patient who was admitted to an intensive care unit in the Northeast region of Brazil. This strain was phenotypically characterized and submitted to whole-genome sequencing and comparative genomics analysis as we recently reported [1]. The dataset also compiles information on Pathogenicity Islands (PIs), Resistance Islands (RIs) and Miscellaneous Islands (MIS) present in the genome of strain B31. The information provided here may support outbreak prevention policies and future epidemiological studies involving Klebsiella spp.
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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.
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The emergence of community acquired infections increases the public health concern on K. pneumoniae and closely related bacteria among which antimicrobial resistance spreads. We report a multidrug-resistant K. pneumoniae isolate, B31, of a patient infected in the community and admitted to an intensive care unit in Northeast Brazil. Antimicrobial susceptibility and genome information were thoroughly investigated to characterize B31 in front of 172 sequenced strains of different countries. Assigned to the Sequence Type 15, which is globally spread, B31 presented extended spectrum beta-lactamase, tigecycline and ciprofloxacin resistance. Genome sequencing revealed most resistance genes being carried by plasmids with high dissemination potential. The absence of main virulence factors, like yersiniabactin and colibactin, apparently suggests a mild pathogenic strain which, on the contrary, persisted and caused severe infection in a previously healthy patient. The present study contributes to unveil the unclear genomic scenario of virulent and multidrug-resistant K. pneumoniae in Brazil.
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Infecções Comunitárias Adquiridas/microbiologia , Farmacorresistência Bacteriana Múltipla , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/classificação , Sequenciamento Completo do Genoma/métodos , Adulto , Ciprofloxacina/farmacologia , Feminino , Genoma Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Klebsiella pneumoniae/genética , Tipagem de Sequências Multilocus , Plasmídeos/genética , Tigeciclina/farmacologiaRESUMO
Caseous lymphadenitis (CLA) is an infectious disease caused by Corynebacterium pseudotuberculosis in small ruminants and is characterized by the development of granulomas in the lymph nodes, spleen, liver, and lungs. Although little is known about the host-pathogen relationship of this bacterium, it was previously reported that the pathogen's lipids are important for its taxonomic classification and survival inside macrophages. However, there are no studies regarding the composition of these molecules. In this study, cell wall glycolipids from two C. pseudotuberculosis strains presenting different virulence profiles were purified and its composition was characterized. A difference was observed between the electrophoretic and chromatogram profiles for cell wall components from the two strains, mainly among molecules with low molecular weights. IgM from sheep with acute CLA recognized antigens with an estimated molecular weight of 11 kDa of the low-pathogenicity strain, while low-molecular weight antigens from the high-pathogenicity strain presented a lower recognition by these antibodies. Mass spectrometry analysis showed that the cell wall of the high-pathogenicity strain contained glycolipids with high amounts of unsaturated fatty acids and glycerophosphoinositols, which may contribute to the capacity of this strain to cause severe disease. In conclusion, it is indicated that cell wall non-protein antigens can play a key role in C. pseudotuberculosis virulence.
Assuntos
Anticorpos Antibacterianos/imunologia , Antígenos de Bactérias/imunologia , Parede Celular/química , Corynebacterium pseudotuberculosis/química , Glicolipídeos/imunologia , Linfadenite/veterinária , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/química , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/imunologia , Corynebacterium pseudotuberculosis/patogenicidade , Glicolipídeos/química , Doenças das Cabras/imunologia , Doenças das Cabras/microbiologia , Cabras/microbiologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Linfonodos/imunologia , Linfonodos/microbiologia , Linfadenite/imunologia , Linfadenite/microbiologia , Ovinos , Doenças dos Ovinos/microbiologia , VirulênciaRESUMO
Caseous lymphadenitis (CL) is an infectious and zoonotic disease characterized by the development of granulomas in the lymph nodes and internal organs of small ruminants. The etiological agent of this disease is Corynebacterium pseudotuberculosis, a Gram-positive and facultative intracellular bacterium. The conventional treatment for CL consists of drainage and chemical cauterization of the lesions using a 10% iodine solution. However, this type of treatment is not effective, due to iodine's cytotoxic profile and low antibacterial activity. Currently, silver nanoparticles (AgNPs) can be seen as an alternative treatment for CL due to their antimicrobial activity and wound healing effects. Therefore, the present study aimed to evaluate AgNPs as a post-surgical treatment for CL. Twenty-nine goats and sheep with clinical signs of CL were selected. Surgical intervention was performed to excise the caseous lesions. To treat the lesions, an ointment formulation based on AgNP mixed with natural waxes and oils was used in the experimental group, and the conventional treatment with 10% iodine was used in the control group. Bacteria were isolated from the excised caseous material. The animals were observed for 8 weeks after the surgical treatment, and blood samples were taken weekly. The surgical wounds of sheep treated with AgNP healed faster, and the surgical wound area was smaller during the observation period; the latter effect was also observed in goats. AgNP-treated animals also had less purulent discharge and less moisture in the surgical wounds. The AgNP-treated animals had lower leukocyte counts and lower titers of anti-C. pseudotuberculosis antibodies. There was no statistically significant difference between the groups with regard to the hemogram results. The results of the susceptibility testing of C. pseudotuberculosis strains (T1, 1002, FRC41, and VD57 strains) and clinical isolates to AgNPs showed growth inhibition, even at low concentrations. It can be concluded that post-surgical treatment of CL using the AgNP-based ointment may be a promising tool in the control of CL, through faster healing, decreased wound contamination, and no apparent toxic effects.
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BACKGROUND: Corynebacterium pseudotuberculosis biovar ovis, a facultative intracellular pathogen, is the etiologic agent of caseous lymphadenitis in small ruminants. During the infection process, C. pseudotuberculosis changes its gene expression to resist different types of stresses and to evade the immune system of the host. However, factors contributing to the infectious process of this pathogen are still poorly documented. To better understand the C. pseudotuberculosis infection process and to identify potential factors which could be involved in its virulence, experimental infection was carried out in a murine model using the strain 1002_ovis and followed by a comparative proteomic analysis of the strain before and after passage. RESULTS: The experimental infection assays revealed that strain 1002_ovis exhibits low virulence potential. However, the strain recovered from the spleen of infected mice and used in a new infection challenge showed a dramatic change in its virulence potential. Label-free proteomic analysis of the culture supernatants of strain 1002_ovis before and after passage in mice revealed that 118 proteins were differentially expressed. The proteome exclusive to the recovered strain contained important virulence factors such as CP40 proteinase and phospholipase D exotoxin, the major virulence factor of C. pseudotuberculosis. Also, the proteome from recovered condition revealed different classes of proteins involved in detoxification processes, pathogenesis and export pathways, indicating the presence of distinct mechanisms that could contribute in the infectious process of this pathogen. CONCLUSIONS: This study shows that C. pseudotuberculosis modifies its proteomic profile in the laboratory versus infection conditions and adapts to the host context during the infection process. The screening proteomic performed us enable identify known virulence factors, as well as potential proteins that could be related to virulence this pathogen. These results enhance our understanding of the factors that might influence in the virulence of C. pseudotuberculosis.
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Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/metabolismo , Corynebacterium pseudotuberculosis/patogenicidade , Proteômica/métodos , Virulência , Animais , Proteínas de Bactérias/análise , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Proteoma/genética , Proteoma/metabolismo , Baço/microbiologia , Fatores de Virulência/genética , Fatores de Virulência/metabolismoRESUMO
Exfoliative toxins are serine proteases secreted by Staphylococcus aureus that are associated with toxin-mediated staphylococcal syndromes. To date, four different serotypes of exfoliative toxins have been identified and 3 of them (ETA, ETB, and ETD) are linked to human infection. Among these toxins, only the ETD structure remained unknown, limiting our understanding of the structural determinants for the functional differentiation between these toxins. We recently identified an ETD-like protein associated to S. aureus strains involved in mild mastitis in sheep. The crystal structure of this ETD-like protein was determined at 1.95 Å resolution and the structural analysis provide insights into the oligomerization, stability and specificity and enabled a comprehensive structural comparison with ETA and ETB. Despite the highly conserved molecular architecture, significant differences in the composition of the loops and in both the N- and C-terminal α-helices seem to define ETD-like specificity. Molecular dynamics simulations indicate that these regions defining ET specificity present different degrees of flexibility and may undergo conformational changes upon substrate recognition and binding. DLS and AUC experiments indicated that the ETD-like is monomeric in solution whereas it is present as a dimer in the asymmetric unit indicating that oligomerization is not related to functional differentiation among these toxins. Differential scanning calorimetry and circular dichroism assays demonstrated an endothermic transition centered at 52 °C, and an exothermic aggregation in temperatures up to 64 °C. All these together provide insights about the mode of action of a toxin often secreted in syndromes that are not associated with either ETA or ETB.
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Exfoliatinas/química , Exfoliatinas/toxicidade , Staphylococcus aureus/química , Staphylococcus aureus/patogenicidade , Animais , Cristalografia por Raios X , Exfoliatinas/classificação , Feminino , Humanos , Modelos Moleculares , Simulação de Dinâmica Molecular , Conformação Proteica , Ovinos , Infecções Estafilocócicas/etiologia , Infecções Estafilocócicas/microbiologia , Eletricidade Estática , Homologia Estrutural de Proteína , SíndromeRESUMO
Epidemiological and health aspects of sheep husbandry were assessed on 213 sheep flocks in 142 municipalities from the state of Minas Gerais, southeastern Brazil. An updated questionnaire was filled out for each flock, requesting data on the farm, the flock and the farmer by the veterinarians of the State Government Agency for Animal Health (Instituto Mineiro de Agropecuária). Thirteen important variables were selected and scored to determine the technological level of the 117 farms; 0.9% of them was classified as high technological level, 45.3% as medium technological level and 53.0% as low technological level. Lamb production was the main objective of the farms and the main features were low-frequencies of individual identification of animals (16.9%), technical assistance (31.9%), use of quarantine for newly acquired animals (0.9%) the separation of animals by age group (3.7%) and requeste the sanitary certificate at purchasing of animals (11.7%). The main health problems reported were abortion (23.9%), keratoconjunctivitis (17.9%), contagious ecthyma (13.6%), pneumonia (10.3%), diarrhea (9.3%) and caseous lymphadenitis (6.1%). Information of the epidemiological situation and the mainly health measures used in the sheep farms are important to improve the productivity and quality of the lamb...
Os aspectos epidemiológicos e sanitários da ovinocultura foram levantados em 213 rebanhos ovinos em 142 municípios do estado de Minas Gerais, sudeste do Brasil. Um questionário atualizado foi preenchido para cada rebanho, com informações sobre a fazenda, o rebanho e do fazendeiro, por veterinários do Instituto Mineiro de Agropecuária. Treze variáveis importantes foram selecionadas e pontuadas para determinar o nível tecnológico em 117 fazendas; 0,9% foram classificadas como de alto nível tecnológico, 45,3% como de médio nível tecnológico e 53,0% como de baixo nível tecnológico. A produção de carne de cordeiro foi o principal objetivo das fazendas amostradas e as principais características foram baixa frequência de identificação individual dos animais (16,9%), assistência técnica (31,9%), uso de quarentena para os animais recém-adquiridos (0,9%), separação de animais por faixa etária (3.7%) e solicitação de certificados sanitários na compra de animais (11,7%). Os principais problemas sanitários relatados foram o aborto (23,9%), ceratoconjuntivite (17,9%), ectima contagioso (13,6%), pneumonia (10,3%), diarreia (9,3%) e linfadenite caseosa (6,1%). Informações sobre a situação epidemiológica e das principais medidas sanitárias utilizadas nas propriedades com ovinos são importantes para melhorar produtividade e a qualidade da carne ovina...
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Animais , Masculino , Feminino , Ligação do Par , Doenças Virais Sexualmente Transmissíveis/epidemiologia , Doenças Virais Sexualmente Transmissíveis/prevenção & controle , Ovinos , Quarentena/veterinária , Perfis SanitáriosRESUMO
Epidemiological and health aspects of sheep husbandry were assessed on 213 sheep flocks in 142 municipalities from the state of Minas Gerais, southeastern Brazil. An updated questionnaire was filled out for each flock, requesting data on the farm, the flock and the farmer by the veterinarians of the State Government Agency for Animal Health (Instituto Mineiro de Agropecuária). Thirteen important variables were selected and scored to determine the technological level of the 117 farms; 0.9% of them was classified as high technological level, 45.3% as medium technological level and 53.0% as low technological level. Lamb production was the main objective of the farms and the main features were low-frequencies of individual identification of animals (16.9%), technical assistance (31.9%), use of quarantine for newly acquired animals (0.9%) the separation of animals by age group (3.7%) and requeste the sanitary certificate at purchasing of animals (11.7%). The main health problems reported were abortion (23.9%), keratoconjunctivitis (17.9%), contagious ecthyma (13.6%), pneumonia (10.3%), diarrhea (9.3%) and caseous lymphadenitis (6.1%). Information of the epidemiological situation and the mainly health measures used in the sheep farms are important to improve the productivity and quality of the lamb.(AU)
Os aspectos epidemiológicos e sanitários da ovinocultura foram levantados em 213 rebanhos ovinos em 142 municípios do estado de Minas Gerais, sudeste do Brasil. Um questionário atualizado foi preenchido para cada rebanho, com informações sobre a fazenda, o rebanho e do fazendeiro, por veterinários do Instituto Mineiro de Agropecuária. Treze variáveis importantes foram selecionadas e pontuadas para determinar o nível tecnológico em 117 fazendas; 0,9% foram classificadas como de alto nível tecnológico, 45,3% como de médio nível tecnológico e 53,0% como de baixo nível tecnológico. A produção de carne de cordeiro foi o principal objetivo das fazendas amostradas e as principais características foram baixa frequência de identificação individual dos animais (16,9%), assistência técnica (31,9%), uso de quarentena para os animais recém-adquiridos (0,9%), separação de animais por faixa etária (3.7%) e solicitação de certificados sanitários na compra de animais (11,7%). Os principais problemas sanitários relatados foram o aborto (23,9%), ceratoconjuntivite (17,9%), ectima contagioso (13,6%), pneumonia (10,3%), diarreia (9,3%) e linfadenite caseosa (6,1%). Informações sobre a situação epidemiológica e das principais medidas sanitárias utilizadas nas propriedades com ovinos são importantes para melhorar produtividade e a qualidade da carne ovina.(AU)
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Animais , Masculino , Feminino , Ovinos , Ligação do Par , Doenças Virais Sexualmente Transmissíveis/epidemiologia , Doenças Virais Sexualmente Transmissíveis/prevenção & controle , Quarentena/veterinária , Perfis SanitáriosRESUMO
Despite the economic importance of caseous lymphadenitis (CLA), a chronic disease caused by Corynebacterium pseudotuberculosis, few genes related to the virulence of its etiologic agent have been characterized. The oligopeptide permease (Opp) transporters are located in the plasma membrane and have functions generally related to the uptake of peptides from the extracellular environment. These peptide transporters, in addition to having an important role in cell nutrition, also participate in the regulation of various processes involving intercellular signaling, including the control of the expression of virulence genes in pathogenic bacteria. To study the role of Opp in C. pseudotuberculosis, an OppD deficient strain was constructed via simple crossover with a nonreplicative plasmid carrying part of the oppD gene sequence. As occurred to the wild-type, the ΔoppD strain showed impaired growth when exposed to the toxic glutathione peptide (GSH), indicating two possible scenarios: (i) that this component can be internalized by the bacterium through an Opp-independent pathway or (ii) that there is toxicity while the peptide is extracellular. Additionally, the ΔoppD mutant presented a reduced ability to adhere to and infect macrophages compared to the wild-type, although both strains exhibit the same potential to colonize spleens and cause injury and death to infected mice.
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Proteínas de Bactérias/genética , Transporte Biológico/genética , Corynebacterium pseudotuberculosis/genética , Linfadenite/genética , Proteínas de Membrana Transportadoras/genética , Animais , Proteínas de Bactérias/metabolismo , Corynebacterium pseudotuberculosis/enzimologia , Corynebacterium pseudotuberculosis/patogenicidade , Humanos , Linfadenite/microbiologia , Linfadenite/patologia , Proteínas de Membrana Transportadoras/metabolismo , Camundongos , Mutação , Óperon/genéticaRESUMO
This study involves the comparison between the exoproteomes of two different strains of Corynebacterium pseudotuberculosis, the etiologic agent of caseous lymphadenitis in small ruminants. In a previous study, based on a gel-free system (TPP-LC/MS(E)), 70 exoproteins for the strain 1002 and 67 for the strain C231, totaling 93 different extracellular proteins for C. pseudotuberculosis, were identified. In the present work, we have used 2D gel electrophoresis to resolve the extracellular proteins of both strains, which were then digested with trypsin, analyzed by MALDI-TOF/TOF and identified with the software MASCOT(®). A total of 45 extracellular proteins of C. pseudotuberculosis were identified by this approach. The comparative analysis between the strains 1002 and C231 identified 13 and 3 strain-specific proteins, respectively, 11 of which are novel. These newly identified proteins may play an important role in the physiology and virulence of C. pseudotuberculosis.
Assuntos
Proteínas de Bactérias/análise , Corynebacterium pseudotuberculosis/química , Corynebacterium pseudotuberculosis/classificação , Proteoma , Animais , Proteínas de Bactérias/química , Infecções por Corynebacterium/microbiologia , Infecções por Corynebacterium/veterinária , Eletroforese em Gel Bidimensional/métodos , Linfadenite/microbiologia , Linfadenite/veterinária , Proteômica/métodos , Especificidade da Espécie , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodosRESUMO
Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis a chronic infectious disease affecting small ruminants. The 2D-DIGE technique was used to compare the exoproteomes of two C. pseudotuberculosis biovar ovis strains isolated from goat (strain 1002) and sheep (strain C231). Seventeen proteins differentially produced were identified here. Nine proteins appeared over-produced in the exoproteome of 1002 goat strain and 8 in that of C231 sheep strain. These proteins were related to various biological functions, such as the cell envelope, respiratory metabolism and proteolysis. This proteomic analysis revealed strain-specific exoproteins although each of the corresponding genes was found in both strain genomes. Such differential expression pattern may reflect inter-strain differences in adaptation to a specific host, in pathogenicity and or in antigenicity of this pathogenic bacterium.
Assuntos
Proteínas de Bactérias/química , Infecções por Corynebacterium/veterinária , Corynebacterium pseudotuberculosis/isolamento & purificação , Corynebacterium pseudotuberculosis/metabolismo , Doenças das Cabras/microbiologia , Proteômica , Doenças dos Ovinos/microbiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Infecções por Corynebacterium/microbiologia , Corynebacterium pseudotuberculosis/química , Corynebacterium pseudotuberculosis/genética , Cabras , OvinosRESUMO
Corynebacterium pseudotuberculosis equi is a Gram-positive pathogenic bacterium which affects a variety of hosts. Besides the great economic losses it causes to horse-breeders, this organism is also known to be an important infectious agent to cattle and buffaloes. As an outcome of the efforts in characterizing the molecular basis of its virulence, several complete genome sequences were made available in recent years, enabling the large-scale assessment of genes throughout distinct isolates. Meanwhile, the RNA-seq stood out as the technology of choice for comprehensive transcriptome studies, which may bring valuable information regarding active genomic regions, despite of the still impeditive associated costs. In an attempt to increase the use of generated reads per instrument run, by effectively eliminating unwanted rRNAs from total RNA samples without relying on any commercially available kits, we applied denaturing high-performance liquid chromatography (DHPLC) as an alternative method to assess the transcriptional profile of C. pseudotuberculosis. We have found that the DHPLC depletion method, allied to Ion Torrent sequencing, allows mapping of transcripts in a comprehensive way and identifying novel transcripts when a de novo approach is used. These data encourage us to use DHPLC in future transcriptional evaluations in C. pseudotuberculosis.