RESUMO
Mastitis is a major disease affecting dairy sheep. It is caused by microorganisms that generate inflammation of the mammary gland in response to tissue invasion. This syndrome affects the welfare of ewes, as well as the production and quality of the milk, thereby reducing its productive efficiency. Because mastitis causes inflammation process, it also increases the production of free radicals that cause lesions via lipoperoxidation, causing damage to proteins, cells and tissues. One way to minimize the impact of the disease is antimicrobial treatment. Nevertheless, the continuous use of antimicrobials contributes to microbial resistance, in addition to producing residues in the milk and derivatives if not given during the grace period. Therefore, the objective of this study was to evaluate the consequences of subclinical mastitis on ewe health, milk production, milk composition and quality. We also evaluated the susceptibility of the bacteria in vitro using disk diffusion antibiograms. Finally, we performed two-way testing of efficacy of treatment in Lacaune ewes using the same agents. In the first stage of the study, 30 lactating ewes (±90 days) were used, 10 of which were negative on the CMT (California Mastitis Test) used as control group (CG) and 20 sheep with subclinical mastitis diagnosed by CMT (MG). Samples were collected and several analyses were performed on the milk and blood. We found that ewes in the MG had higher lipid peroxidation in serum and milk, as well as lower production, with reduction of the total dry extract in milk. There were 15 isolates of Staphylococcus hyicus, four isolates of each S. epidermidis and S. intermedius, and two isolates of Corynebacterium spp. The primary hematological result was leukocytosis in ewes with mastitis. Based on the antibiogram, we chose ceftiofur for in vivo tests. In this stage, we divided the sheep with subclinical mastitis into two subgroups of 10 ewes each, to receive drug by two routes: intramuscular (IM) and intramammary (IMM). In the IMM group, of the 10 CMT-positive ewes at the beginning of the experiment, seven were already negative by the racket test 120â¯h after the last application (70% efficacy). In the IM group, of the 10 positive ewes, only four were negative after 120â¯h of the final application, a low efficacy treatment (40%). We evaluated antimicrobial residues in the milk of treated animals. We found this material within 5 days after treatment in the two forms used; despite the fact that the product's stated withholding period is 3 days. We conclude that ewes with mastitis produce less milk of lower quality. We also conclude that, although ceftiofur is 100% effective in vitro, when used in ewes with mastitis, the efficacy did not exceed 70%, and was more efficient when administered via the intramammary route.
Assuntos
Cefalosporinas/farmacologia , Cefalosporinas/uso terapêutico , Mastite/tratamento farmacológico , Mastite/microbiologia , Leite/microbiologia , Estresse Oxidativo , Animais , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Corynebacterium/isolamento & purificação , Feminino , Qualidade dos Alimentos , Glândulas Mamárias Animais/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Ovinos , Doenças dos Ovinos/tratamento farmacológico , Doenças dos Ovinos/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus epidermidis/isolamento & purificação , Staphylococcus hyicus/isolamento & purificação , Staphylococcus intermedius/isolamento & purificação , Infecções Estreptocócicas/tratamento farmacológico , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/veterinária , Resultado do TratamentoRESUMO
Hyicin 3682, the first bacteriocin reported for Staphylococcus hyicus, is a BsaCOL variant produced by S. hyicus 3682, a strain isolated from bovine milk. Hyicin 3682 is found in the culture supernatant, is bactericidal and its producing strain exhibits a much broader spectrum of antimicrobial activity than the producing strain of BsaCOL against several Gram-positive bacteria, which include foodborne pathogens, food-spoilage microorganisms and bacterial species of medical and veterinary importance. Sequencing of the genome of S. hyicus 3682 provided the nucleotide sequence of the entire gene cluster involved in hyicin 3682 production, which seems to be located on pRJ109, the single plasmid carried by this strain. This gene cluster is expressed and consists of 8525bp and of eight genes (hyiA, hyiB, hyiC, hyiD, hyiP, hyiF, hyiE and hyiG) encoded on the same DNA strand. The mature lantibiotic exhibits 91% identity to BsaCOL and its molecular mass was found to be â¼26Da higher due to two amino acid substitutions. S. hyicus 3682 proved to be only partially immune to its cognate bacteriocin up to 1024 AU/ml. Therefore, hyicin 3682, the first Bsa variant reported in coagulase-negative staphylococci, does exhibit antimicrobial and siblicidal activities.
Assuntos
Antibacterianos/metabolismo , Bacteriocinas/genética , Bacteriocinas/metabolismo , Staphylococcus hyicus/genética , Staphylococcus hyicus/metabolismo , Sequência de Aminoácidos , Animais , Antibacterianos/química , Bacteriocinas/química , Vias Biossintéticas/genética , Bovinos , Ordem dos Genes , Genes Bacterianos , Genoma Bacteriano , Bactérias Gram-Positivas/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Leite/microbiologia , Dados de Sequência Molecular , Peso Molecular , Família Multigênica , Análise de Sequência de DNA , Staphylococcus hyicus/isolamento & purificaçãoRESUMO
This research aimed to detect coagulase-positive Staphylococcus (CPS) directly in samples of artificially contaminated milk, using multiplex PCR (mPCR). Standard and isolated bacterial strains of S. aureus, S. epidermidis, S. hyicus, S. intermedius, Listeria monocytogenes and Escherichia coli species were used, evaluating the specificity and detection limit of mPCR, for artificially contaminated UHT milk. Primers specific for the nuc gene (NUC1-NUC2 were used for S. aureus, NUC3-NUC4 for S. hyicus and NUC5-NUC6 for S. intermedius). It was possible to detect the three target species by mPCR, directly from bovine whole milk, with adequate specificity and acceptable detention limit for identification of coagulase-positive Staphylococcus (CPS) in foods. The specificity was determined by the amplification of species-specific fragments, and the detection limit was assessed by the detection thresholds obtained for the three species (103 CFU mL-1). From these results, the mPCR described, with the proposed set of primers, has the potential for use in precise identification and differentiation between CPSs in milk samples.(AU)
Esta pesquisa teve como objetivo detectar diretamente em amostras de leite contaminado artificialmente Staphylococcus coagulase positiva (ECP) por multiplex PCR (mPCR). Cepas padrão e isolados de S. aureus, S. epidermidis, S. hyicus, S. intermedius, Listeria monocytogenes e Escherichia coli foram utilizados no estudo. Foram utilizados primers específicos para o gene nuc (NUC1-NUC2 para o S. aureus, NUC3-NUC4 para o S. hyicus e NUC5-NUC6 para o S. intermedius ). Foi possível detectar as três espécies-alvo por mPCR, formar diretamente nas amostras de leite integral bovino, com especificidade adequada e limite de detecção aceitável para identificação de espécies de Staphylococcus coagulase positiva (ECP) em alimentos. A especificidade foi determinada por meio da amplificação de fragmentos específicos das espécies e o limite de detecção foi avaliado pelos limiares de detecção obtidos para as três espécies (103 UFC mL-1 para as espécies presentes nas amostras de leite contaminadas artificialmente).(AU)
Assuntos
Leite/microbiologia , Staphylococcus aureus/patogenicidade , Staphylococcus epidermidis/patogenicidade , Staphylococcus hyicus/patogenicidade , Staphylococcus intermedius/patogenicidade , Listeria monocytogenes/patogenicidade , Escherichia coli/patogenicidade , Reação em Cadeia da Polimerase Multiplex/métodos , Contaminação de Alimentos/análise , Contaminação de Alimentos/prevenção & controleRESUMO
Staphylococcal infection is a severe bacterial infection. Finding satisfactory predictable biological markers is essential for the treatment of this condition. In this study, we applied a 32-marker sandwich ELISA-based antibody array to evaluate cytokine changes in Staphylococcus hyicus-inoculated BALB/c mice at 24 and 48 h post infection. Among the cytokines detected, the expression levels of granulocyte colony stimulating factor (G-CSF), interleukin 6 (IL-6), macrophage inflammatory protein 2, and keratinocyte chemoattractant (KC) were increased to levels more than twice higher than those in the control group 24 h after infection, while the expression of interleukin 12p40p70 (IL-12p40p70) was decreased to less than half the level measured in the controls. The expression of G-CSF, IL-6, monocyte chemoattractant protei-5, chemokine ligand 11, and KC was upregulated 48 h post infection, whereas IL-12p40p70 expression was still significantly lower (P < 0.05). Among the detected cytokines, the expression levels of G-CSF, IL-6, and KC were constantly upregulated, while IL-12p40p70 was downregulated. This result was then validated by an ELISA assay analysis, which confirmed that G-CSF, IL-6, KC, and IL- 12p40p70 expression levels were specifically modulated after an S. hyicus bacterial infection, while granulocyte monocyte colony stimulation factor, IL-12, and IFNγ levels were significantly increased after a viral infection. Our study indicated the potential of cytokines G-CSF, IL-6, KC, and IL- 12p40p70 as markers for detecting S. hyicus infection. The results of this study may provide useful data for the appropriate use of medication following S. hyicus infection.
Assuntos
Citocinas/sangue , Infecções Estafilocócicas/sangue , Animais , Camundongos , Camundongos Endogâmicos BALB C , Staphylococcus hyicus , Regulação para CimaRESUMO
Staphylococcus hyicus is an important microorganism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the species characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.
Staphylococcus hyicus é um micro-organismo de importância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O presente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de patogenicidade e sua ocorrência em animais, alimentos e seres humanos.
Assuntos
Humanos , Animais , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/virologia , Infecções , Microbiologia de Alimentos , Staphylococcus hyicus/patogenicidade , VirulênciaRESUMO
Staphylococcus hyicus is an important microorganism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the species characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.(AU)
Staphylococcus hyicus é um micro-organismo de importância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O presente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de patogenicidade e sua ocorrência em animais, alimentos e seres humanos.(AU)
Assuntos
Humanos , Animais , Virulência , Infecções , Staphylococcus hyicus/patogenicidade , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/virologiaRESUMO
Staphylococcus hyicus é um micro-organismo de importância em Medicina Veterinária e saúde pública, tendo em vista sua capacidade de causar doenças em animais e seres humanos. O presente trabalho teve como objetivo realizar uma revisão de literatura a fim de reunir as informações mais atuais sobre o S. hyicus. Para tanto, foram abordados: características da espécie, fatores de patogenicidade e sua ocorrência em animais, alimentos e seres humanos.(AU)
Staphylococcus hyicus is an important micro organism Veterinary Medicine and public health, because of its ability to cause disease in animals and humans. The present study aims to conduct a literature review to bring together the most current information about S. hyicus. Thus, the specie's characteristics, the pathogenicity factors and their occurrence in animals, food and humans were approached.(AU)
Assuntos
Humanos , Animais , Virulência , Staphylococcus hyicus/patogenicidade , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/virologia , InfecçõesRESUMO
Bovine mastitis is an important disease in dairy farming, not only by promoting direct economic losses, but also for indirect losses and the potential risk to public health. The main causes of intramammary infections include the bacteria of the genus Staphylococcus spp., Staphylococcus aureus being the predominant etiologic agent in subclinical mastitis. The aim of this study was to determine the frequency of subclinical mastitis in eight herds from southern Rio Grande do Sul (Brazil) and the relationship of the disease with the presence of S. aureus. In addition, we checked for the presence of S. intermedius and S. hyicus in the milk samples obtained. For identification of the disease, we used the California Mastitis Test (CMT). Identification of Staphylococcus spp. species was made in Barid-Parker agar culture medium, with subsequent confirmation of suspected colonies by way of Gram stain and catalase test along with free-coagulase and thermonuclease research. Subclinical mastitis was identified in 53.6% of animals tested. The presence of coagulase-positive Staphylococcus was identified in 12.6% of animals with subclinical mastitis. In these same animals, bacteria identified as S. aureus were the etiologic agent present in 17.6% of cases. Additionally, it was revealed that among the group identified as coagulase positive, 85.7% corresponded to S. aureus, while 8.5% had biochemical characteristics consistent with S. intermedius and 5.8% were considered S. hyicus.(AU)
A mastite bovina é uma doença importante na exploração leiteira, não apenas pelas perdas econômicas diretas que promove, mas também pelas perdas indiretas e o potencial risco à saúde pública. Dentre as principais causas de infecções intramamárias, destacam-se as bactérias do gênero Staphylococcus spp., sendo que Staphylococcus aureus é o agente etiológico predominante em mastite subclínica. O objetivo desse trabalho foi verificar a frequência de mastite subclínica em oito rebanhos localizados na região Sul do Rio Grande do Sul (Brasil) e a relação da enfermidade com a presença de S. aureus. Adicionalmente, pesquisou-se a presença de S. intermedius e S. hyicus nas amostras de leite obtidas. Para identificação da doença, utilizou-se o California Mastitis Test (CMT). A identificação da espécie de Staphylococcus spp. foi feita em meio de cultura ágar Baird-Parker, com posterior confirmação das colônias suspeitas em coloração de gram, prova de catalase, pesquisa de coagulase livre e pesquisa de termonuclease. A mastite subclínica foi constatada em 53,6% dos animais testados. A presença de Staphylococcus coagulase positiva foi identificada em 12,6% dos animais com mastite subclínica. Nesses mesmos animais, a bactéria identificada como S. aureus foi o agente etiológico presente em 17,6% dos casos. Adicionalmente, pode-se perceber que, dentre o grupo identificado como coagulase positiva, 85,7% corresponderam a S. aureus, enquanto 8,5% mostraram características bioquímicas compatíveis com S. intermedius e 5,8% foram consideradas S. hyicus.(AU)
Assuntos
Animais , Mastite Bovina , Coagulase , Infecções Estafilocócicas , Staphylococcus aureus , Staphylococcus hyicus , Staphylococcus intermediusRESUMO
Bacteriocins are peptides produced by bacteria and having inhibitory activity against other bacteria. Many of these substances may be useful as antibacterial agents for practical applications. In this study, 21 Staphylococcus spp. isolated from pigs, dogs and bovine milk in different states of Brazil were investigated for staphylococcin production. Hyicin 3682, a bacteriocin produced by one such strain, inhibited almost all strains tested, including Bacillus cereus, Listeria monocytogenes and Staphylococcus aureus. PCR experiments showed that hyicin 3682 is lantibiotic-related, but not identical, to both epidermin and Bsa. The maximum production of hyicin 3682 (6,400 AU/ml) was observed after 24 h of growth in BHI medium at 37 °C. Hyicin 3682 proved to be a cationic, small antimicrobial peptide with a molecular mass of 2,139 Da. It exhibited resistance to low pH and to heating at 65 °C, and partial sensitivity to proteolytic enzymes. Taken together, these results suggest that hyicin 3682, the first bacteriocin characterized in Staphylococcus hyicus, has potential biotechnological applications as a food preservative. Moreover, hyicin 3682 was able to inhibit its producer strain, suggesting that an effective immune system for specific protection against hyicin 3682 is not found in its producer strain, a characteristic not described thus far for other staphylococcins.
Assuntos
Antibacterianos/biossíntese , Bactérias/efeitos dos fármacos , Bacteriocinas/biossíntese , Conservação de Alimentos/métodos , Staphylococcus hyicus/metabolismo , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Bacillus cereus/efeitos dos fármacos , Bacteriocinas/isolamento & purificação , Bacteriocinas/farmacologia , Brasil , Bovinos , Meios de Cultura/química , Cães , Listeria monocytogenes/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus hyicus/crescimento & desenvolvimento , Staphylococcus hyicus/isolamento & purificaçãoRESUMO
Canine mastitis is an infrequent condition that occurs most commonly in the postpartum period due to ascending bacterial infection. This report aims to describe clinical, pathological and bacteriological aspects of mastitis caused by Staphylococcus hyicus accompanied by disseminated superficial lymphadenitis in a dog. In the postpartum period the animal showed clinical signs of prostration, dyspneia, and unresponsivess to environment. At physical examination, hypothermia, dehydration and subcutaneous edema of the limbs were detected. Gross and histologic features were consistent with marked purulent and necro-hemorrhagic mastitis accompanied by lymphangitis, lymphadenitis and thromboembolic pneumonia. Isolation and biochemical identification confirmed the infection by Staphylococcus hyicus. This study shows that one must be attentive not only of the risk of uterine infection postpartum, but also of the possibility of mammary gland infection.(AU)