RESUMO
Endometritis is the leading cause of mare subfertility. Most mares respond to standard therapy, but alternative therapies have been developed for mares failing to respond. This study aimed to investigate a commercially available, yet unassessed, product labeled as a uterine sanitizer to determine the in vitro antimicrobial activity against microorganisms associated with endometritis and its in vitro stability to dilute antibiotics. In experiment 1, the microdilution broth technique and antimicrobial effects were assessed against Escherichia sp, Staphylococcus sp., Klebsiella sp., Pseudomonas sp., and Candida sp. Percentage inhibition was calculated by comparing the optical density. The minimum inhibitory concentration (MIC) 100% was determined using the resazurin dye technique. MIC 50% and 90% were determined using a dose-response non-linear regression. In experiment 2, the uterine sanitizer was used to dilute commonly used antibiotics achieving a final volume of 90 mL at 5°C, 21°C, and 37°C. The pH was measured at 0, 1, 3, 6, and 24 h after dilution. The uterine sanitizer had inhibitory properties against all microorganisms; Escherichia sp. being the most susceptible, and Pseudomonas sp. the most resistant. The uterine sanitizer had an acidic pH=4; however, when combined with the antibiotics, the pH of the antibiotic remained unchanged with the different temperatures and did not precipitate. In conclusion, the uterine sanitizer showed antimicrobial effects against endometritis-causing microorganisms. The dilution of antibiotics in the uterine sanitizer was stable and this association could potentiate the antimicrobial effects. Uterine sanitizer's safety and clinical efficacy in vivo remain to be tested.
Assuntos
Antibacterianos , Bactérias , Endometrite , Doenças dos Cavalos , Testes de Sensibilidade Microbiana , Feminino , Animais , Endometrite/tratamento farmacológico , Endometrite/microbiologia , Endometrite/veterinária , Cavalos , Antibacterianos/farmacologia , Antibacterianos/química , Doenças dos Cavalos/tratamento farmacológico , Doenças dos Cavalos/microbiologia , Bactérias/efeitos dos fármacos , Estabilidade de MedicamentosRESUMO
In this study, the main agents associated with endometritis in cows in the state of Santa Catarina, Brazil, were identified and the resistance profile and virulence mechanisms of the bacterial isolates were evaluated. Isolates of Escherichia coli and Trueperella pyogenes were tested for their biofilm forming ability and the antimicrobial action of bromhexine hydrochloride in combination with other antimicrobials. A total of 37 uterine lavage samples were collected from cows with endometritis. Of the 55 bacteria isolated, 25.4% were identified as T. pyogenes and 16.3% as E. coli. The bacterial isolates showed greater resistance to sulfamethoxazole + trimethoprim (58.2%) and tetracycline (56.3%). Among the species, E. coli showed the highest resistance rates, with 100% of isolates showing resistance to amoxicillin, streptomycin, and gentamicin. The results of the minimum inhibitory concentration for the T. pyogenes isolates showed that 91.6% of the isolates were resistant to enrofloxacin and tetracycline, and 75% were resistant to ceftiofur and sulfamethoxazole + trimethoprim. All E. coli and T. pyogenes isolates showed biofilm forming ability. The plo, fimA, and nanH genes were identified in 100% of T. pyogenes isolates. In parallel, 100% of E. coli isolates had the fimH gene, and 11.1% had the csgD gene. Bromhexine hydrochloride showed antimicrobial activity against 100% of E. coli isolates and 66.6% of T. pyogenes isolates. Furthermore, when associated with antimicrobials, bromhexine hydrochloride has a synergistic and additive effect, proving to be an option in the treatment of endometritis in cows and an alternative for reducing the use of antimicrobials.
Assuntos
Actinomycetaceae , Infecções por Actinomycetales , Antibacterianos , Doenças dos Bovinos , Endometrite , Infecções por Escherichia coli , Escherichia coli , Testes de Sensibilidade Microbiana , Bovinos , Animais , Endometrite/microbiologia , Endometrite/veterinária , Endometrite/tratamento farmacológico , Feminino , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Antibacterianos/farmacologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/tratamento farmacológico , Actinomycetaceae/efeitos dos fármacos , Actinomycetaceae/genética , Actinomycetaceae/isolamento & purificação , Actinomycetaceae/classificação , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Actinomycetales/microbiologia , Infecções por Actinomycetales/veterinária , Infecções por Actinomycetales/tratamento farmacológico , Biofilmes/efeitos dos fármacos , Brasil , Farmacorresistência BacterianaRESUMO
Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a mucosal commensal of the lower genital tract in horses and is the most isolated bacterium causing endometritis in mares. The aim of this study was to determine the molecular diversity of S. zooepidemicus obtained from endometritis in mares in Buenos Aires province, Argentina. Thirty isolates obtained from the uterus of mares in 2005 and 2017 were studied. The MLST scheme was applied to identify the Argentinian genotypes and the clonal relationships and patterns of evolutionary descent were identified using the eBURST algorithm - goeBURST. Twenty six different Sequence types (STs) were identified, being only 11 of them previously reported in horses and also, from several host species and tissues. The other 15 STs were reported in Argentinian reproductive strains of mares in our study for the first time. The genotypes obtained from uterus in Argentina were not evenly distributed when all the published S. zooepidemicus STs were analysed, thus, it was not possible to establish that the same lineage circulates in our equine population. The fact that the identified genotypes were also reported in other countries, diverse samples and host species suggest that there is not a host, and an anatomical niche adaptation. Finally, the isolation of the same genotype in the vagina/clitoris and the uterus of the same mare highlights the versatility of S. zooepidemicus and its role as an opportunistic pathogen.
Assuntos
Endometrite , Genótipo , Doenças dos Cavalos , Infecções Estreptocócicas , Animais , Cavalos/microbiologia , Doenças dos Cavalos/microbiologia , Feminino , Argentina , Endometrite/veterinária , Endometrite/microbiologia , Infecções Estreptocócicas/veterinária , Infecções Estreptocócicas/microbiologia , Variação Genética , Tipagem de Sequências Multilocus/veterinária , Útero/microbiologia , Streptococcus/genética , Streptococcus/isolamento & purificação , Streptococcus/classificação , Streptococcus equi/genética , Streptococcus equi/isolamento & purificação , Streptococcus equi/classificaçãoRESUMO
Endometritis, the inflammation of the endometrium, is the leading cause of subfertility in mares, and therefore responsible for major economic losses in the horse industry worldwide. It is generally treated with uterine lavages combined with ecbolic agents and local or systemic antibiotics. However, since antibiotic overuse has been associated with antimicrobial resistance in mares with persistent endometritis, new prevention and treatment alternatives are needed. One such alternative could be the use of probiotic lactic acid bacteria (LAB) isolated from the host. Thanks to their species specificity, resident microbiota may restore ecological equilibrium within the host, and therefore, help prevent infections and improve physiological functions. In the present study, 257 bacterial strains were isolated from 77 healthy mares, and 88.76% (n = 228) of them were phenotypically classified as LAB. Within this group, 65.79% were able to inhibit at least one strain from each of the genera that most commonly cause equine endometritis (Streptococcus equi subsp. zooepidemicus, Escherichia coli, and Staphylococcus spp.). Five strains (RCE11, RCE20, RCE91, RCE99, and RCE167) were selected on the basis of their beneficial properties: ability to autoaggregate and adhere to equine epithelial cells, high inhibition of and co-aggregation with all the bacteria isolated from clinical cases of endometritis evaluated, and negative co-inhibition between one another. All five were finally identified as Enterococcus spp., namely E. faecium (two strains), E. hirae (two strains), and E. gallinarum (one strain). Further studies will assess their safety and biotechnological potential for the design of a multi-strain probiotic formula to prevent equine endometritis.
Assuntos
Endometrite , Doenças dos Cavalos , Probióticos , Animais , Cavalos , Feminino , Endometrite/veterinária , Endometrite/prevenção & controle , Endometrite/microbiologia , Doenças dos Cavalos/prevenção & controle , Doenças dos Cavalos/microbiologia , Probióticos/farmacologia , Probióticos/administração & dosagem , Lactobacillales/isolamento & purificação , Lactobacillales/fisiologia , Genitália Feminina/microbiologiaRESUMO
The aim of this study was to identify the impact of Trueperella pyogenes in cows with clinical endometritis (CE) on reproductive performance and milk production in affected cows. In total, 230 lactating Holstein dairy cows from six commercial dairy herds were sampled once between 28 and 33 days post-partum. Cows included in the present study did not receive antibiotic or anti-inflammatory treatments prior to the experimental period. Clinical endometritis (CE) was characterized as cow with vaginal mucus score = 3 (>50% of purulent vaginal discharge) and >18% polymorphonuclear neutrophilic leukocyte (PMNL). The body condition scores (BCS) and milk production were evaluated at the time of enrolment. The identification of isolated bacteria was carried out through the analysis of MALDI-TOF MS (matrix-assisted laser desorption ionization time-of-flight mass spectrometry). According to uterine health, three groups of dairy cows were formed: healthy control cows without T. pyogenes (n = 147), CE cows with T. pyogenes (n = 22) and CE cows without T. pyogenes (n = 61). CE cows with T. pyogenes had lower BCS, milk production and conception at first AI (p < .01) than CE cows without T. pyogenes and control cows. Furthermore, CE cows with T. pyogenes had higher (p < .01) service per pregnancy and had greater (p < .01) days to get pregnant than CE cows without T. pyogenes and control cows. This study demonstrates that CE cows with T. pyogenes had impaired reproductive performance and milk production when compared to cows without CE and CE cows without T. pyogenes. This information can contribute to a strategic treatment in cows affected by clinical endometritis, favouring the rational use of antibiotics on dairy farms.
Assuntos
Actinomycetaceae/patogenicidade , Infecções Bacterianas/veterinária , Doenças dos Bovinos , Endometrite , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Endometrite/microbiologia , Endometrite/veterinária , Feminino , Lactação , Leite , Gravidez , ReproduçãoRESUMO
A endometrite é a principal responsável pela diminuição nas taxas de fertilidade e recuperação embrionária em éguas e em parte, pode ser associada a contaminações por falhas na antissepsia das biotécnicas reprodutivas aplicadas à espécie. O objetivo deste trabalho foi avaliar os níveis de contaminação decorrentes de diferentes protocolos de antissepsia da genitália externa de éguas doadoras de embrião. Éguas sadias tiveram a vulva higienizada conforme um dos três grupos experimentais: (G1) sabão de côco; (G2) detergente neutro e (G3) clorexidina degermante 2%. Foi realizada a coleta de material da região da vulva, vestíbulo e útero com auxílio de swab estéril. As amostras foram processadas com objetivo de se determinar os principais microgranismos presentes, além da avaliação de Unidades Formadoras de Colônia (UFC). O G1 mostrou-se mais efetivo quanto à redução nas contagens de UFC em todas amostras. Por outro lado, G2 apresentou eficiência de 62%, sendo o tratamento menos eficiente na redução da contaminação das diferentes regiões avaliadas. A escolha adequada do agente sanitizante utilizado na higienização de períneo previamente a intervenções ginecológicas são de suma importância afim de se evitar contaminações ascendentes que levem a endometrites.
Endometritis is mainly responsible for the decrease in fertility and embryonic recovery rates in mares and, in part, it can be associated with contaminations due to failures in the antisepsis of reproductive biotechniques applied to the specie. The objective of this study was to evaluate the levels of contamination resulting from different protocols for antisepsis of the external genitalia of embryo donor mares. Healthy mares had their vulva sanitized according to one of the three experimental groups: (G1) coconut soap; (G2) neutral detergent and (G3) 2% chlorhexidine degerming. It was made the collection of material from the vulva, vestibule and uterus region with the aid of a sterile swab. The samples were processed in order to determine the main microorganisms present, in addition to the evaluation of Colony Forming Units (CFU). G1 was more effective in reducing CFU counts in all samples. On the other hand, G2 showed an efficiency of 62%, being the treatment less efficient in reducing contamination in the different regions evaluated. The proper choice of the sanitizing agent used to clean the perineum prior to gynecological interventions is of paramount importance in order to avoid ascending contamination that lead to endometritis.
Assuntos
Feminino , Animais , Antissepsia , Cavalos/microbiologia , Endometrite/microbiologia , Genitália Feminina/microbiologia , Técnicas MicrobiológicasRESUMO
A endometrite é a principal causa de infertilidade em éguas, além de gerar grandes transtornos econômicos e produtivos. Objetivou-se com esse trabalho, realizar uma revisão sistemática sobre endometrite equina, sua etiologia, os mecanismos próprios de defesa uterina mecânicos e quimiotáticos. Avaliando ainda os principais microrganismos envolvidos no processo bem como sua virulência e resistência antimicrobiana. Apresentamos ainda uma proposta de abordagem diagnóstica contemplando a identificação dos agentes, susceptibilidade farmacológica associadas a comprovação da produção de biofilme, assim como sua graduação, possibilitando assim em caso de comprovação de produção de biofilme uma terapia alternativa eficaz para a quebra desse biofilme associado a um tratamento antibacteriano específico. A correlação desses testes possivelmente fornecerá subsídios ainda mais importantes para melhorar a eficiência reprodutiva dos rebanhos equinos.
Endometritis is the main cause of infertility in mares, in addition to promote major economic and productive disorders. The objective of this work was to carry out a systematic review of equine endometritis, its etiology, mechanical and chemotactic uterine defense mechanisms. Also evaluating the main microorganisms involved in the process as well as their virulence and antimicrobial resistance. We also present a proposal for a diagnostic approach contemplating the identification of agents, pharmacological susceptibility associated with presence of biofilm production, as well as its graduation, thus enabling, in case of biofilm present, an effective alternative therapy to break this biofilm associated with an specific antibacterial treatment. The correlation of these tests will possibly provide even more important subsidies to improve reproductive efficiency of equine herds.
Assuntos
Animais , Anti-Infecciosos/análise , Biofilmes , Endometrite/microbiologia , Endometrite/veterináriaRESUMO
A endometrite é a principal responsável pela diminuição nas taxas de fertilidade e recuperação embrionária em éguas e em parte, pode ser associada a contaminações por falhas na antissepsia das biotécnicas reprodutivas aplicadas à espécie. O objetivo deste trabalho foi avaliar os níveis de contaminação decorrentes de diferentes protocolos de antissepsia da genitália externa de éguas doadoras de embrião. Éguas sadias tiveram a vulva higienizada conforme um dos três grupos experimentais: (G1) sabão de côco; (G2) detergente neutro e (G3) clorexidina degermante 2%. Foi realizada a coleta de material da região da vulva, vestíbulo e útero com auxílio de swab estéril. As amostras foram processadas com objetivo de se determinar os principais microgranismos presentes, além da avaliação de Unidades Formadoras de Colônia (UFC). O G1 mostrou-se mais efetivo quanto à redução nas contagens de UFC em todas amostras. Por outro lado, G2 apresentou eficiência de 62%, sendo o tratamento menos eficiente na redução da contaminação das diferentes regiões avaliadas. A escolha adequada do agente sanitizante utilizado na higienização de períneo previamente a intervenções ginecológicas são de suma importância afim de se evitar contaminações ascendentes que levem a endometrites.(AU)
Endometritis is mainly responsible for the decrease in fertility and embryonic recovery rates in mares and, in part, it can be associated with contaminations due to failures in the antisepsis of reproductive biotechniques applied to the specie. The objective of this study was to evaluate the levels of contamination resulting from different protocols for antisepsis of the external genitalia of embryo donor mares. Healthy mares had their vulva sanitized according to one of the three experimental groups: (G1) coconut soap; (G2) neutral detergent and (G3) 2% chlorhexidine degerming. It was made the collection of material from the vulva, vestibule and uterus region with the aid of a sterile swab. The samples were processed in order to determine the main microorganisms present, in addition to the evaluation of Colony Forming Units (CFU). G1 was more effective in reducing CFU counts in all samples. On the other hand, G2 showed an efficiency of 62%, being the treatment less efficient in reducing contamination in the different regions evaluated. The proper choice of the sanitizing agent used to clean the perineum prior to gynecological interventions is of paramount importance in order to avoid ascending contamination that lead to endometritis.(AU)
Assuntos
Animais , Feminino , Antissepsia , Genitália Feminina/microbiologia , Técnicas Microbiológicas , Endometrite/microbiologia , Cavalos/microbiologiaRESUMO
INTRODUCTION: Endometritis is a condition marked by inflammation of the endometrium that affects dairy cows from 21 days after parturition, causing damage to herd fertility and economic losses on farms. The use of active compounds obtained from plant sources has gained importance as disease treatment agents in farm animals due to the high resistance rates currently observed against traditional antibiotics commonly used. The study was carried out to examine the chemical composition and to investigate the antibacterial activity of rosemary, cinnamon, cloves, eucalyptus, lemon, oregano and thyme essential oils against the reference strain of Escherichia coli (ATCC 25922), Fusobacterium necrophorum (ATCC 25286), Trueperella pyogenes (ATCC 19411) and Staphylococcus aureus (ATCC 29213), considered as typical bacteria causing endometritis. METHODOLOGY: The chemical composition of the seven essential oils were analyzed by GC-MS and their antibacterial activity was evaluated by the disc diffusion method. RESULTS: Thirty-six components were identified in total using GC-MS analyzes. The main compounds were cinnamaldehyde (86.5% for cinnamon essential oil), eugenol (85.7% for clove essential oil), 1,8-cineol (80% for eucalyptus and 47.8% rosemary essential oils), limonene (65.5% for lemon essential oil), carvacrol (72.1% for oregano essential oil) and thymol (48.8% for thyme essential oil). The disc diffusion assay revealed that cinnamon, clove, oregano, and thyme essential oils showed the best results compared to the other three essential oils, showing the largest zone of inhibition against all bacteria evaluated. CONCLUSIONS: These findings indicated that essential oils are a potential agent to be used as an alternative for bovine endometritis treatment.
Assuntos
Infecções Bacterianas/veterinária , Doenças dos Bovinos/tratamento farmacológico , Endometrite/veterinária , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Actinomycetaceae/efeitos dos fármacos , Animais , Anti-Infecciosos/farmacologia , Infecções Bacterianas/tratamento farmacológico , Bovinos , Doenças dos Bovinos/microbiologia , Endometrite/tratamento farmacológico , Endometrite/microbiologia , Escherichia coli/efeitos dos fármacos , Feminino , Fusobacterium necrophorum/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Staphylococcus aureus/efeitos dos fármacosRESUMO
Endometritis is an inflammation of the endometrium associated with bacterial infection. The pathogenesis of endometritis in cows is still not completely understood. The combined analysis of the markers of inflammation and oxidative stress has contributed to a better understanding of disease mechanisms, but is still unexplored in uterine disorders. Moreover, research provides evidence about an important role of the vagus nerve in regulating the innate immune function through the cholinergic anti-inflammatory pathway in response to bacterial infections. This new pathway has demonstrated a critical role in controlling the inflammatory system. The aim of this study was to evaluate the activity of cholinesterase in total blood, lymphocytes, and serum of dairy cows with clinical and subclinical endometritis. Sixty-one Holstein cows, between 30 and 45 days in milk, were classified into 3 groups of animals: presenting clinical endometritis (n = 22), subclinical endometritis (n = 17), and healthy (n = 22). Mean leukocyte counts did not differ among groups, but the neutrophil number was significantly higher in cows with clinical endometritis than those in healthy animals. Also, serum concentration of interleukin-1beta (pg/mL) was significantly higher in cows with endometritis. The activity of acetylcholinesterase in blood and lymphocytes increased in both groups with endometritis. Animals with endometritis presented an increase in lipid peroxidation, but the antioxidant enzyme activity (catalase levels) was higher in endometritis groups than in normal cows. In conclusion, the inflammatory process of clinical and subclinical endometritis leads to systemic lipid peroxidation despite the compensatory increase of the antioxidant enzyme. These data also provide evidence of an important role of the cholinergic pathway in regulating dairy cows with clinical and subclinical endometritis.