Abstract：
Infectious endometritis is considered one of the major causes of infertility and it can affect up to 60% of barren mares. It is characterized by the presence of one or more microorganisms in the reproductive tract and it is treated with the administration of antibiotics, ecbolic agents and uterine lavages. Ozone, thanks to its antimicrobial properties that are based on its high oxidative potential, could represent an effective alternative treatment for endometritis. The aim of this study was to test in vitro the bactericidal and fungicidal properties of different ozone formulations, either as gas (experiment 1) or dissolved in two liquid matrices (experiment 2), specifically distilled water or oil (Neozone 4000, Cosmoproject, Parma, Italy), onto 6 different species of microorganisms isolated from mares with clinical endometritis, namely Escherichia coli, Staphylococcus aureus, Streptococcus equi subsp. Zooepidemicus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Candida albicans. In the first experiment, 3 clinical antibiotic-resistant strains per each species were exposed to different conditions: to O2O3 gas mixtures (15 and 40 μg/ml for 1, 3 and 5 min), to 100 % O2 or left untreated. The results showed a reduction of the microbial count of over 99,9% for every pathogen, time and concentration of O2O3 gas mixtures tested. Furthermore, gaseous ozone showed both a time-dependant effect (5 vs 3 vs 1 min of exposure) and a concentration-dependant effect (40 vs 15 μg/ml) at 1 and 3 min, while after 5 min no differences were observed. In the second experiment, minimum inhibitory concentration (MIC), and minimum bactericidal/fungicidal concentration (MBC, MFC) of ozonated distilled water and ozonated oil were evaluated. Ozonated oil showed a bactericidal/fungicidal activity against all the strains tested (MIC range 12.5-25 % v/v, MBC/MFC range 12.5-50 % v/v) while ozonated distilled water didn\'t show an observable antimicrobial effect, discouraging its use as an antimicrobial agent for the treatment of endometritis. The results of this in vitro study indicate that both gaseous ozone and ozonated oil exerted remarkable antimicrobial activities and are promising alternative treatments for infectious endometritis, even when caused by antibiotic-resistant bacteria, and encourage further experiments in an effort to scale down or even prevent the use of antibiotics in equine reproduction.
摘要：
感染性子宫内膜炎被认为是不孕的主要原因之一，它可以影响高达60％的贫瘠的母马。它的特征是在生殖道中存在一种或多种微生物，并通过施用抗生素进行治疗，栓塞剂和子宫灌洗。臭氧,由于其高氧化潜能的抗菌特性，可能是子宫内膜炎的有效替代疗法。这项研究的目的是在体外测试不同臭氧制剂的杀菌和杀菌性能，作为气体(实验1)或溶解在两种液体基质中(实验2)，特别是蒸馏水或油(Neozone4000，Cosmoproject，帕尔马,意大利),从临床子宫内膜炎的母马中分离出6种不同的微生物，即大肠杆菌，金黄色葡萄球菌,马链球菌亚种。动物流行病,铜绿假单胞菌,肺炎克雷伯菌和白色念珠菌。在第一个实验中,每个物种的3种临床抗生素抗性菌株暴露于不同的条件：O2O3气体混合物（15和40μg/ml，持续1、3和5分钟），至100%O2或不处理。结果表明，每种病原体的微生物数量减少了超过99,9％，O2O3气体混合物的时间和浓度测试。此外，气态臭氧在1和3分钟时显示出时间依赖性效应（5vs3vs1分钟暴露）和浓度依赖性效应（40vs15μg/ml），而5分钟后没有观察到差异。在第二个实验中,最小抑制浓度（MIC），和最低杀菌/杀真菌浓度(MBC，MFC）的臭氧化蒸馏水和臭氧化油进行了评估。臭氧化油对所有测试菌株均具有杀菌/杀真菌活性（MIC范围12.5-25％v/v，MBC/MFC范围12.5-50％v/v），而臭氧蒸馏水未显示可观察到的抗菌作用，不鼓励将其用作治疗子宫内膜炎的抗菌剂。这项体外研究的结果表明，气态臭氧和臭氧油都具有显着的抗菌活性，并且是感染性子宫内膜炎的有希望的替代疗法。即使是由抗生素抗性细菌引起的，并鼓励进一步的实验，以减少甚至防止在马繁殖中使用抗生素。
