Ozonated oil is effective at killing Candida species and Streptococcus mutans biofilm-derived cells under aerobic and microaerobic conditions.

This study explores the growth of bacterial, fungal, and interkingdom biofilms under aerobiosis or microaerobic conditions and the effect of ozonated sunflower oil on these biofilms. Candida species and Streptococcus mutans were used to study this interaction due to their importance in oral health and disease as these microorganisms display a synergistic relationship that manifests in the onset of caries and tooth decay. Biofilms were developed in a 96-well microtiter plate at 37ºC for 24 h, under aerobiosis or microaerobic conditions, and treated with ozonated oil for 5 to 120 min. All the microorganisms formed biofilms in both oxygenation conditions. Scanning electron microscopy was used to visualize biofilm morphology. Rodent experiments were performed to verify the oil-related toxicity and its efficacy in oral candidiasis. The growth of all Candida species was increased when co-cultured with S. mutans, whilst the growth of bacterium was greater only when co-cultured with C. krusei and C. orthopsilosis under aerobiosis and microaerobic conditions, respectively. Regardless of the oxygenation condition, ozonated oil significantly reduced the viability of all the tested biofilms and infected mice, showing remarkable microbicidal activity as corroborated with confocal microscopy and minimal toxicity. Thus, ozonated oil therapy can be explored as a strategy to control diseases associated with these biofilms especially in the oral cavity. LAY SUMMARY: We demonstrated that ozonated sunflower oil is effective at killing the biofilms formed by Candida species, by the bacterium Streptococcus mutans, or by both micoorganisms that can interact in the oral cavity, making it a potential therapeutic option for the treatment of these infections.

The In Vitro Effect of Ozone Therapy Against Equine Pythium insidiosum.

The goal of the present study was to characterize the antimicrobial action of different ozone (O3) presentations against Pythium insidiosum isolated from horses. In experiment 1, P. insidiosum was treated with ozonated distilled water, ozonated sunflower oils with distinct peroxide indexes or O3 gas (72 µg O3 mL-1). In experiment 2, samples were exposed one or three times to oxygen (O2) or O3 gas (72 µg O3 mL-1; 30 min/day). In experiment 3, P. insidiosum was treated with different concentrations of O3 gas (Ø, 32, 52, or 72 µg O3 mL-1) for three days (30 min/day). In experiment 4, samples were exposed to O2 or O3 gas (72 µg O3 mL-1) for 05, 15, or 30 minutes during three days. Posteriorly, all samples were cultured for two weeks, and the pathogen growth area was measured until D14. Samples with absence of growth on D14 were recultured to assess the germicidal or germistatic action of the treatment. In experiment 1, only ozonized sunflower oil with a high peroxide index had germicidal action against the pathogen. In experiment 2, samples exposed three times to O3 gas were inactivated. In experiment 3, the O3 therapy had germicidal action against P. insidiosum independently of the gas concentration (P > .1). In experiment 4, O3 treatments ≥15 minutes suppressed the pathogen development, whereas samples exposed to O3 gas for 5 minutes had progressive growth (P < .01). In conclusion, ozonated sunflower oil with a high peroxide index and multiple exposures to O3 gas mixtures were able to inactivate P. insidiosum isolated from horses.

Luteal changes after treatment with sub-luteolytic doses of prostaglandin (cloprostenol sodium) in cattle.

This study characterizes the physiological and morphological changes related to partial luteolysis in bovine corpus luteum (CL) after challenges with sub-doses of cloprostenol sodium on Day 6 (D6) of the estrous cycle. Cows (n = 12/treatment) were treated as follows: Control (2 mL, saline, i.m.); 2XPGF (two treatments i.m. 500 µg of cloprostenol sodium 2 h apart) and 1/6PGF (83.3 µg of cloprostenol sodium, i.m., once). Plasma progesterone (P4) concentration, CL volume and blood flow were measured immediately before the treatments, then every 8 h (h) for 48 h. In the Control, P4 concentrations were higher at 48 h than at 0 h. P4 decreased 8h after 2XPGF treatment (P < 0.05), and remained low until the end of the trial. P4 decreased in 1/6PGF between 8 and 16 h (P < 0.05), then began to rebound at 24 h. Luteal volume was higher in Controls at 48 h than at 0 h. Under 1/6PGF, luteal volume decreased at 24 h (P < 0.05) and began to rebound at 32 h. Luteal volume and blood flow were reduced starting at 24 and 32 h, respectively, after 2XPGF treatment (P < 0.05). In this study, we were able to describe the partial luteolysis phenomenon, induced by a treatment of a D6CL with cloprostenol sub-dose.

Aplicação da ultrassonografia colorida doppler em programas de transferência de embriões equinos / Potential use of doppler ultrasound in equine embryo transfer programs

Os programas comerciais de transferência de embriões em éguas existem por mais de três décadas e são hoje uma das biotécnias mais utilizadas na reprodução assistida de equinos. O exame ultrassonográfico nos períodos pré e pós-cobertura de doadoras de embriões, assim como a avaliação de receptoras no momento da inovulação é de vital importância para o êxito de um programa de transferência de embriões. A ultrassonografia Doppler é uma técnica não-invasiva que permite a avaliação em tempo real da hemodinâmica do trato reprodutivo de animais de grande porte. Por fornecer detalhes anatômicos e informações imediatas sobre a fisiologia do fluxo sanguíneo de tecidos e órgãos, o exame doppler permite a avaliação do potencial ovulatório de folículos e do status funcional de corpo lúteo e útero, além de ser uma técnica auxiliar para o diagnóstico de distúrbios na hemodinâmica do sistema reprodutivo. Alterações na perfusão sanguínea dos futuros folículos dominantes são detectáveis em diferentes fases de seu desenvolvimento, como na divergência folicular e no período pré-ovulatório. A partir dessas informações, é possível determinar o princípio da atividade sexual e o momento ideal para o início de tratamentos superovulatórios e indutores de ovulação, assim como o momento mais apropriado para a realização de coberturas de éguas doadoras de embriões. A avaliação através do modo-Doppler do corpo lúteo e útero de éguas receptoras é também um instrumento auxiliar para a seleção de animais com perfil sérico de progesterona e ambiente uterino adequados para a sobrevivência do embrião e manutenção da gestação. Essa técnica pode ser útil ainda ao se avaliar a interação concepto-maternal. Apesar da aplicabilidade da tecnologia doppler dentro de programas de transferência de embriões, novos estudos visando a determinar padrões de normalidade e posterior caracterização de distúrbios de fluxo sanguíneo de trato reprodutivo ainda se fazem necessários.

Embryo transfer in mares has been commercially used for more than three decades and it is one of most frequently applied biotechnologies on equine reproduction. Ultrasonic doppler exam of donors during the pre and post-breeding and recipients evaluation at the embryo transfer moment is essential for embryo transfer program success. Doppler ultrasonography is a non-invasive technic that allows real-time evaluation of the reproductive system hemodynamics in large animal. For providing anatomical details and blood-flow physiologic information of vessels and tissues, doppler exam can be used for ovulation prognostication, and to evaluate uterine and luteal functional status. Additionally, it can serve as a diagnostic aid of hemodynamic disturbance in reproductive system. Vascularity changes of future ovulatory follicles precede the diameter deviation. Based on this, it is possible to estimate the beginning of the breeding season and the best moment to initiate superovulatory treatments. Moreover, follicular blood flow can be used to decide the most appropriated moment for ovulation induction and breeding of donors mares. Uterine and luteal evaluation using Doppler-mode in mares can be useful to select embryo recipients with satisfactory progesterone production and adequate uterine vascularity for embryo development and maintenance of pregnancy. The maternal recognition of pregnancy also can be evaluated. Although the applicability of the Doppler technology in embryo transfer programs, additional studies are necessary to determine the standards of normality and to characterize different blood-flow disturbances of the reproductive system.