RESUMO
In the present research work, we propose a new antimicrobial treatment for pyoderma via cutaneous permeation of bacteriophage particles conveyed in a hydroxyethylcellulose (HEC) gel integrating ionic liquid as a permeation enhancer. Ionic liquids are highly viscous fluids constituted exclusively by ions, that are usually hydrolytically stable and promote solubilization of amphipathic molecules such as proteins, hence serving as green solvents and promoting the transdermal permeation of biomolecules. In the research effort entertained herein, the synthesis and use of choline geranate for integrating a HEC gel aiming at the structural and functional stabilization of a cocktail of isolated lytic bacteriophage particles was sought, aiming at transdermal permeation in the antimicrobial treatment of animal pyoderma. The results obtained showed a high ability of the ionic liquid in enhancing transdermal permeation of the bacteriophage particles, with concomitant high potential of the HEC gel formulation in the antimicrobial treatment of animal skin infections.
Assuntos
Celulose/análogos & derivados , Colina/química , Terapia por Fagos/veterinária , Staphylococcus intermedius/virologia , Administração Cutânea , Animais , Bacteriófagos , Linhagem Celular , Sobrevivência Celular , Celulose/química , Cães/microbiologia , Cavalos/microbiologia , Humanos , Líquidos Iônicos/química , Líquidos Iônicos/metabolismo , Testes de Mutagenicidade , Permeabilidade , Pioderma/tratamento farmacológico , Pioderma/veterinária , Pele/metabolismo , SolventesRESUMO
Methicillin resistance in the Staphylococcus intermedius group (SIG) has emerged in small animal practice. Methicillin-resistant SIG (MRSIG) members have been implicated as causes of infections in both companion animals and humans. Staphylococcal cassette chromosome mec (SCCmec) elements carry the mecA/C genes, which encode for the transpeptidase PBP2a (PBP2) responsible for β-lactam antibiotic resistance in staphylococci. This study examined the SCCmec types of MRSIG isolates from different clinical specimens of dogs that exhibited methicillin MIC ≥ 0.5 μg/mL by an automated identification and susceptibility system in a Center for Veterinary Diagnostics in São Paulo, Brazil. Susceptibility to methicillin was determined by broth microdilution testing, and Oxoid® M.I.C.Evaluator® strips. PBP2a production was detected using a latex agglutination assay. SCCmec typing was performed according to the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) guidelines. SCCmec type II (2A), SCCmec type III (3A), composite SCC structures consisting of a class A mec gene complex in addition to multiple ccr gene complexes, and non-typable SCCmec elements were reported in these MRSIG isolates. SCCmec type variants differing from those so far acknowledged by IWG-SCC were found, indicating new rearrangements in the genetic context of mecA in these canine MRSIG isolates.(AU)
A resistência à meticilina no grupo Staphylococcus intermedius (GSI) tem aumentado na clínica de pequenos animais. Membros GSI resistentes à meticilina (GSIRM) têm sido causas de infecções tanto em animais de companhia e humanos. Cassetes cromossômicos estafilocócicos mec (SCCmec) carregam os genes mecA/C, que codificam a transpeptidase PBP2a (PBP2) responsável pela resistência aos antibióticos β-lactâmicos em estafilococos. Nosso objetivo foi investigar os elementos SCCmec de GSIRM isolados de diferentes amostras clínicas de cães que exibiram CIM de meticilina ≥ 0,5 μg/mL por meio de um sistema automatizado em um Centro Veterinário de Diagnósticos em São Paulo, Brasil. A sensibilidade à meticilina foi determinada por meio do teste de microdiluição em caldo e fitas Oxoid® M.I.C.Evaluator®. A produção de PBP2a foi detectada usando um ensaio de aglutinação de látex. A tipagem dos elementos SCCmec foi realizada de acordo com as diretrizes do International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). SCCmec tipo II (2A), SCCmec tipo III (3A), SCC compostos de um complexo mec de classe A com múltiplos complexos ccr, e elementos SCCmec não tipáveis foram encontrados nesses isolados GSIRM. Variantes que diferem dos elementos SCCmec reconhecidos até o momento pelo IWG-SCC foram encontradas, indicando novos rearranjos no contexto genético de mecA nesses isolados GSIRM caninos.(AU)
Assuntos
Animais , Cães , Doenças do Cão , Staphylococcus intermedius/genética , Staphylococcus intermedius/isolamento & purificação , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/prevenção & controle , Infecções Estafilocócicas/veterinária , Resistência a MeticilinaRESUMO
Methicillin resistance in the Staphylococcus intermedius group (SIG) has emerged in small animal practice. Methicillin-resistant SIG (MRSIG) members have been implicated as causes of infections in both companion animals and humans. Staphylococcal cassette chromosome mec (SCCmec) elements carry the mecA/C genes, which encode for the transpeptidase PBP2a (PBP2) responsible for β-lactam antibiotic resistance in staphylococci. This study examined the SCCmec types of MRSIG isolates from different clinical specimens of dogs that exhibited methicillin MIC ≥ 0.5 μg/mL by an automated identification and susceptibility system in a Center for Veterinary Diagnostics in São Paulo, Brazil. Susceptibility to methicillin was determined by broth microdilution testing, and Oxoid® M.I.C.Evaluator® strips. PBP2a production was detected using a latex agglutination assay. SCCmec typing was performed according to the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) guidelines. SCCmec type II (2A), SCCmec type III (3A), composite SCC structures consisting of a class A mec gene complex in addition to multiple ccr gene complexes, and non-typable SCCmec elements were reported in these MRSIG isolates. SCCmec type variants differing from those so far acknowledged by IWG-SCC were found, indicating new rearrangements in the genetic context of mecA in these canine MRSIG isolates.
A resistência à meticilina no grupo Staphylococcus intermedius (GSI) tem aumentado na clínica de pequenos animais. Membros GSI resistentes à meticilina (GSIRM) têm sido causas de infecções tanto em animais de companhia e humanos. Cassetes cromossômicos estafilocócicos mec (SCCmec) carregam os genes mecA/C, que codificam a transpeptidase PBP2a (PBP2) responsável pela resistência aos antibióticos β-lactâmicos em estafilococos. Nosso objetivo foi investigar os elementos SCCmec de GSIRM isolados de diferentes amostras clínicas de cães que exibiram CIM de meticilina ≥ 0,5 μg/mL por meio de um sistema automatizado em um Centro Veterinário de Diagnósticos em São Paulo, Brasil. A sensibilidade à meticilina foi determinada por meio do teste de microdiluição em caldo e fitas Oxoid® M.I.C.Evaluator®. A produção de PBP2a foi detectada usando um ensaio de aglutinação de látex. A tipagem dos elementos SCCmec foi realizada de acordo com as diretrizes do International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). SCCmec tipo II (2A), SCCmec tipo III (3A), SCC compostos de um complexo mec de classe A com múltiplos complexos ccr, e elementos SCCmec não tipáveis foram encontrados nesses isolados GSIRM. Variantes que diferem dos elementos SCCmec reconhecidos até o momento pelo IWG-SCC foram encontradas, indicando novos rearranjos no contexto genético de mecA nesses isolados GSIRM caninos.
Assuntos
Animais , Cães , Doenças do Cão , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/prevenção & controle , Infecções Estafilocócicas/veterinária , Resistência a Meticilina , Staphylococcus intermedius/genética , Staphylococcus intermedius/isolamento & purificaçãoRESUMO
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
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
The study evaluated cefoxitin disk diffusion tests breakpoints and their correlation to mecA gene PCR results for detecting Methicillin-resistant Staphylococcus intermedius Group (MRSP) isolates from dogs in Brazil. Agreement using proposed breakpoint (resistant ≤ 30 mm) was encouraging. The current study reinforces that an epidemiological breakpoint can be established to predict presence of MRSP.
Assuntos
Antibacterianos/farmacologia , Cefoxitina/farmacologia , Genes Bacterianos , Resistência a Meticilina , Reação em Cadeia da Polimerase/métodos , Staphylococcus intermedius/efeitos dos fármacos , Staphylococcus intermedius/genética , Animais , Brasil , Cães , Testes de Sensibilidade Microbiana/métodosRESUMO
The study evaluated cefoxitin disk diffusion tests breakpoints and their correlation to mecA gene PCR results for detecting Methicillin-resistant Staphylococcus intermedius Group (MRSP) isolates from dogs in Brazil. Agreement using proposed breakpoint (resistant < 30 mm) was encouraging. The current study reinforces that an epidemiological breakpoint can be established to predict presence of MRSP.(AU)
Assuntos
Animais , Cães , Cefoxitina , Staphylococcus intermedius , Farmacorresistência Bacteriana , Resistência a Meticilina , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão/veterinária , Reação em Cadeia da PolimeraseRESUMO
The study evaluated cefoxitin disk diffusion tests breakpoints and their correlation to mecA gene PCR results for detecting Methicillin-resistant Staphylococcus intermedius Group (MRSP) isolates from dogs in Brazil. Agreement using proposed breakpoint (resistant < 30 mm) was encouraging. The current study reinforces that an epidemiological breakpoint can be established to predict presence of MRSP.
Assuntos
Animais , Cães , Cefoxitina , Farmacorresistência Bacteriana , Resistência a Meticilina , Staphylococcus intermedius , Reação em Cadeia da Polimerase , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão/veterináriaRESUMO
Este trabalho objetivou determinar os microrganismos bacterianos no saco conjuntival de cães clinicamente normais e cães com ceratoconjuntivite seca (CCS), atendidos no Hospital de Medicina Veterinária da Universidade Federal da Bahia. Foram incluídos na pesquisa 30 animais sadios e 28 com CCS. Foi observado crescimento bacteriano em 60% das amostras oculares dos cães sadios e em 89,29% dos cães com CCS. Houve predomínio de espécies Gram positivas em ambos os grupos. Nos animais normais as bactérias mais prevalentes foram Staphylococcus intermedius (26,83%), seguido por Staphylococcus aureus (21,95%) e Staphylococcus epidermidis (19,51%). Houve identificação de leveduras em 38,34% das amostras. Dos 30 animais sadios estudados, 46,66% apresentaram cultura positiva bilateral, 26,67% cultura positiva unilateral e 26,67% cultura negativa. Nos animais com CCS, as bactérias de maior prevalência foram o Streptococcus b-hemolítico (16,13%), seguido por Staphylococcus intermedius (12,90%) e o Staphylococcus aureus (11,83%). Ocorreu identificação de leveduras em 19,64% das amostras. Dos 28 animais com CCS, observou-se que 82,14% apresentaram cultura positiva bilateral e 17,86% cultura positiva unilateral.(AU)
This study aimed to determine the bacterial microorganisms in the conjunctival sac of clinically normal dogs and of dogs with keratoconjunctivitis sicca (KCS) attended at the Veterinary Hospital of the Federal University of Bahia. Thirty health animals and 28 with KCS were included in the research. Bacterial growth was observed in 60% of the healthy dogs and in 89.29% of the dogs with KCS. There was a predominance of Gram-positive species in both groups. In normal animals, Staphylococcus intermedius (26.83%), followed by Staphylococcus aureus (21.95%) and Staphylococcus epidermidis (19.51%) were the most prevalent bacteria and the identification of yeasts in the samples of this group was 38.34%. Of the 30 healthy animals studied, 46.66% showed bilateral positive culture, 26.67% had unilateral positive culture and 26.67% had negative culture. In animals with KCS, Streptococcus b-hemolytic (16.13%), followed by Staphylococcus intermedius (12.90%) and Staphylococcus aureus (11.83%) were the most prevalent bacteria. Yeast identification in this group occurred in 19.64% of the samples. Of the 28 animals with KCS, 82.14% showed bilateral positive culture and 17.86% unilateral positive culture.(AU)
Assuntos
Animais , Cães , Doenças do Cão/diagnóstico , Ceratoconjuntivite/epidemiologia , Ceratoconjuntivite/veterinária , Olho/microbiologia , Infecções Estreptocócicas/veterinária , Staphylococcus aureus , Staphylococcus epidermidis , Staphylococcus intermediusRESUMO
Este trabalho objetivou determinar os microrganismos bacterianos no saco conjuntival de cães clinicamente normais e cães com ceratoconjuntivite seca (CCS), atendidos no Hospital de Medicina Veterinária da Universidade Federal da Bahia. Foram incluídos na pesquisa 30 animais sadios e 28 com CCS. Foi observado crescimento bacteriano em 60% das amostras oculares dos cães sadios e em 89,29% dos cães com CCS. Houve predomínio de espécies Gram positivas em ambos os grupos. Nos animais normais as bactérias mais prevalentes foram Staphylococcus intermedius (26,83%), seguido por Staphylococcus aureus (21,95%) e Staphylococcus epidermidis (19,51%). Houve identificação de leveduras em 38,34% das amostras. Dos 30 animais sadios estudados, 46,66% apresentaram cultura positiva bilateral, 26,67% cultura positiva unilateral e 26,67% cultura negativa. Nos animais com CCS, as bactérias de maior prevalência foram o Streptococcus b-hemolítico (16,13%), seguido por Staphylococcus intermedius (12,90%) e o Staphylococcus aureus (11,83%). Ocorreu identificação de leveduras em 19,64% das amostras. Dos 28 animais com CCS, observou-se que 82,14% apresentaram cultura positiva bilateral e 17,86% cultura positiva unilateral.
This study aimed to determine the bacterial microorganisms in the conjunctival sac of clinically normal dogs and of dogs with keratoconjunctivitis sicca (KCS) attended at the Veterinary Hospital of the Federal University of Bahia. Thirty health animals and 28 with KCS were included in the research. Bacterial growth was observed in 60% of the healthy dogs and in 89.29% of the dogs with KCS. There was a predominance of Gram-positive species in both groups. In normal animals, Staphylococcus intermedius (26.83%), followed by Staphylococcus aureus (21.95%) and Staphylococcus epidermidis (19.51%) were the most prevalent bacteria and the identification of yeasts in the samples of this group was 38.34%. Of the 30 healthy animals studied, 46.66% showed bilateral positive culture, 26.67% had unilateral positive culture and 26.67% had negative culture. In animals with KCS, Streptococcus b-hemolytic (16.13%), followed by Staphylococcus intermedius (12.90%) and Staphylococcus aureus (11.83%) were the most prevalent bacteria. Yeast identification in this group occurred in 19.64% of the samples. Of the 28 animals with KCS, 82.14% showed bilateral positive culture and 17.86% unilateral positive culture.
Assuntos
Animais , Cães , Ceratoconjuntivite/epidemiologia , Ceratoconjuntivite/veterinária , Doenças do Cão/diagnóstico , Infecções Estreptocócicas/veterinária , Olho/microbiologia , Staphylococcus aureus , Staphylococcus epidermidis , Staphylococcus intermediusRESUMO
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
BACKGROUND: The aim of this study was to identify the aerobic bacteria of the preputial and vaginal microbiota in owl monkeys that have been raised in captivity and to evaluate the antimicrobial susceptibility of these bacteria by gender and social organization. METHODS: Thirty clinically healthy Aotus azarai infulatus were used. A total of 134 samples were collected, 60 from the preputial mucosa and 74 from the vaginal mucosa. An automated system of bacterial identification was used. RESULTS AND CONCLUSIONS: Staphylococcus intermedius and Proteus mirabilis were the microorganisms that were most frequently identified according to gender and social organization. The antimicrobial susceptibility of the isolated gram-positive bacteria was similar in both sexes. However, the gram-negative strains had some differences. The aerobic bacterial population of the vaginal and preputial microbiota is similar in owl monkeys, and there are no differences in the number and bacterial species according to sex and social organization.