Ozone therapy by rectal insufflation in dogs: safety and oxidative stress - a randomized cross-over study.

Ozone therapy acts in the body inducing controlled oxidative stress, thereby improving the antioxidant, immune and circulatory responses. However, very little is known about how this therapy affects oxidative stress indicators in dogs. We aimed to assess the clinical, hematological, biochemical and oxidative stress parameters of healthy dogs subjected to ozone therapy and oxygen therapy by rectal insufflation. Ten healthy dogs were allocated into three experimental groups in a cross-over design: control, without intervention; ozone, which received 100 µg of O3/kg through rectal insufflation; and oxygen, which received an ozone-equivalent volume of medicinal O2 through rectal insufflation. Dogs received four applications weekly and were followed up until the seventh week. Ozone therapy significantly increased the weight, mean corpuscular volume and mean platelet volume and decreased total cholesterol of treated dogs. Regarding oxidative stress, ozone therapy reduced total antioxidant capacity by ferric reduction (TAC-FRAP) in D7 compared with baseline and the control, significantly increased total antioxidant capacity by cupric reduction (TAC-CUPRAC) in D42 and D49 compared with the control group, caused an increase in uric acid compared with the oxygen group and decreased lipid peroxidation on D21 compared with the control group. In conclusion, ozone therapy through rectal insufflation causes transient oxidative stress followed by an antioxidant response and discreetly interferes with a few clinical, hematological and biochemical variables in healthy dogs, although variables still remained within the reference ranges for the species, thus proving the safety of the therapy. Furthermore, oxygen therapy causes oxidative stress without inducing a subsequent antioxidant response.

Amphotericin B-loaded natural latex dressing for treating Candida albicans wound infections using Galleria mellonella model.

Amphotericin B (AmB) is the gold standard for antifungal drugs. However, AmB systemic administration is restricted because of its side effects. Here, we report AmB loaded in natural rubber latex (NRL), a sustained delivery system with low toxicity, which stimulates angiogenesis, cell adhesion and accelerates wound healing. Physicochemical characterizations showed that AmB did not bind chemically to the polymeric matrix. Electronic and topographical images showed small crystalline aggregates from AmB crystals on the polymer surface. About 56.6% of AmB was released by the NRL in 120 h. However, 33.6% of this antifungal was delivered in the first 24 h due to the presence of AmB on the polymer surface. The biomaterial's excellent hemo- and cytocompatibility with erythrocytes and human dermal fibroblasts (HDF) confirmed its safety for dermal wound application. Antifungal assay against Candida albicans showed that AmB-NRL presented a dose-dependent behavior with an inhibition halo of 30.0 ± 1.0 mm. Galleria mellonella was employed as an in vivo model for C. albicans infection. Survival rates of 60% were observed following the injection of AmB (0.5 mg.mL-1) in G. mellonella larvae infected by C. albicans. Likewise, AmB-NRL (0.5 mg.mL-1) presented survival rates of 40%, inferring antifungal activity against fungus. Thus, NRL adequately acts as an AmB-sustained release matrix, which is an exciting approach, since this antifungal is toxic at high concentrations. Our findings suggest that AmB-NRL is an efficient, safe, and reasonably priced ($0.15) dressing for the treatment of cutaneous fungal infections.

Feline obesity causes hematological and biochemical changes and oxidative stress - a pilot study.

Obesity, an extremely important factor in feline clinical practice, is estimated to affect up to one third of the feline population. Moreover, it can trigger chronic inflammation, which could predispose to oxidative stress by increasing reactive oxygen species, thereby generating potentially irreversible cellular damage. This study analyzed hematological, biochemical and oxidative stress profiles at various degrees of feline obesity. Forty-five cats were selected and divided into three groups: control (n = 17), overweight (n = 13) and obese (n = 15), after clinical and laboratory evaluation and body condition score. Biochemical and oxidative stress analyses were performed using a photocolorimeter and hematological analyses were performed in a veterinary cell counter. Obese cats showed increased mean corpuscular volume (MCV), red cell distribution width (RDW), HDL cholesterol and triglycerides and decreased activity of gamma-glutamyl transferase (GGT) than control cats, although within the reference ranges for the species. As for oxidative stress, obese cats showed higher total antioxidant capacity (TAC), by the inhibition of 2,2'-Azino-Bis-3-Ethylbenzthiazoline-6-Sulfonic Acid (ABTS), inhibition of ABTS associated with horseradish peroxidase (ABTS + HRP), cupric ion reducing antioxidant capacity (CUPRAC) and ferric reducing antioxidant power (FRAP) methods, while overweight cats had a higher TAC-ABTS + HRP and TAC-FRAP than control cats. We conclude that the conditions of natural obesity and overweight in the feline species alter its hematological, biochemical and oxidative stress parameters.

Epidemiology of canine visceral leishmaniasis in a vulnerable region in Brazil.

Visceral leishmaniasis (VL) is a neglected and endemic zoonosis that occurs throughout Brazil; nevertheless, few studies have focused on the early detection of the disease. The municipality of Ourinhos is a non-receptive, silent and vulnerable area for VL, where the seroprevalence of this disease has so far not been investigated. The present study aimed to determine the seroprevalence of canine VL in Ourinhos-SP, and to identify the presence of risk factors. Blood samples were obtained from 604 dogs during a rabies vaccination campaign together with application of a socioeconomic questionnaire, environmental and animal characteristics and tutor's knowledge about the disease. The samples were subjected to indirect ELISA and new samples were collected from reactive and suspect animals, including whole blood and lymph node aspiration evaluated by parasitological method, complete blood count and PCR. No animal was diagnosed as positive based on the combination of direct and indirect tests and the tutors' answers indicated little knowledge about leishmaniasis, being often confused with other diseases transmitted by arthropods; hence, according to the proposed methods, the presence of canine leishmaniasis in the city of Ourinhos was not confirmed and health education campaigns about the disease should be carried out.

Effect of uterine ozone therapy and anticoagulant sampling on oxidative stress parameters in mares.

With the purpose of assessing the effects of uterine ozone therapy and anticoagulant sampling on oxidative stress (OS) parameters in mares, ten mares underwent three consecutive days of uterine ozone therapy by flushing the uterus with ozonated lactated Ringer's solution followed by insufflation with ozone­oxygen gas. Serum samples were obtained at baseline and days 3, 6, 10 and 17 to determine the effect of ozone therapy on OS markers. Plasma obtained with anticoagulants citrate, ethylenediaminetetraacetic acid (EDTA) and heparin were at baseline and 6 days following therapy to determine the effect of anticoagulant on OS parameters. Antioxidants albumin and uric acid, total antioxidant capacity (TAC) using four different methods, total oxidant capacity (TOC) and lipid peroxidation were determined through photocolorimetry. Statistical analyses comprised repeated measures ANOVA followed by Dunnett's test or Friedman followed by Dunn's post-hoc test. Differences were considered significant when p < 0.05. Uterine ozone therapy significantly decreased uric acid, TAC in all four different methods, concomitantly with an increase on TOC at days 3 and 6 following therapy. No changes were observed on albumin and lipid peroxidation levels. Anticoagulants prevented the detection of oxidative stress induced by uterine ozone therapy depending on the method of analysis. In conclusion, uterine ozone therapy causes systemic oxidative stress in mares and the choice of anticoagulant sampling interferes with laboratory tests.

Epidemiology of canine visceral leishmaniasis in a vulnerable region in Brazil / Soroepidemiologia da leishmaniose visceral canina em uma região vulnerável do Brasil

Abstract Visceral leishmaniasis (VL) is a neglected and endemic zoonosis that occurs throughout Brazil; nevertheless, few studies have focused on the early detection of the disease. The municipality of Ourinhos is a non-receptive, silent and vulnerable area for VL, where the seroprevalence of this disease has so far not been investigated. The present study aimed to determine the seroprevalence of canine VL in Ourinhos-SP, and to identify the presence of risk factors. Blood samples were obtained from 604 dogs during a rabies vaccination campaign together with application of a socioeconomic questionnaire, environmental and animal characteristics and tutor's knowledge about the disease. The samples were subjected to indirect ELISA and new samples were collected from reactive and suspect animals, including whole blood and lymph node aspiration evaluated by parasitological method, complete blood count and PCR. No animal was diagnosed as positive based on the combination of direct and indirect tests and the tutors' answers indicated little knowledge about leishmaniasis, being often confused with other diseases transmitted by arthropods; hence, according to the proposed methods, the presence of canine leishmaniasis in the city of Ourinhos was not confirmed and health education campaigns about the disease should be carried out.

Resumo A leishmaniose visceral (LV) é uma zoonose negligenciada e endêmica presente em todas as regiões do Brasil, mas mesmo assim poucos estudos têm objetivado a detecção inicial da doença. O município de Ourinhos - SP é uma área não receptiva, silenciosa e vulnerável à LV, não havendo até o momento estudos que tenham investigado a soroprevalência no município. Nesse sentido, o presente estudo objetivou determinar a soroprevalência da LV canina em Ourinhos-SP, bem como associar a presença de fatores de risco. Amostras sanguíneas de 604 cães foram obtidas juntamente com a aplicação de questionário socioeconômico, características ambientais e dos animais e conhecimento sobre a doença. As amostras foram submetidas à sorologia por ELISA e novas amostras coletadas de cães reagentes ou suspeitos foram analisadas por método parasitológico direto, hemograma e PCR. Nenhum animal foi considerado positivo na combinação de testes direto e indireto, e as respostas dos tutores indicaram pouco conhecimento sobre a leishmaniose, sendo muitas vezes confundida com outras doenças transmitidas por artrópodes. Dessa forma, de acordo com os métodos propostos, a presença de leishmaniose canina, na cidade de Ourinhos, não foi confirmada. Por isso campanhas de educação em saúde sobre a doença deveriam ser realizadas.

Vertical transmission of Neospora caninum in bovine fetuses from a slaughterhouse in Brazil.

Neospora caninum is considered as one of the main causes of reproductive failure in cattle. Vertical transmission is the main route of infection in the bovine host and plays an important role in maintaining the parasite in the herd. Molecular detection of N. caninum is important to determine the occurrence of the disease and to evaluate the genetic diversity of the parasite. The present study aimed at assessing the vertical transmission of N. caninum using molecular techniques to detect the parasite in tissue samples from bovine fetuses collected in a slaughterhouse in the state of São Paulo, Brazil. Seventy fetuses and 70 blood samples from pregnant cows were collected in a slaughtering line. Fresh samples of heart and brain tissue from fetuses were analyzed using molecular assays. Serum samples from fetuses and cows were subjected to an indirect fluorescent antibody test (IFAT) to detect antibodies against N. caninum. Nested PCR targeting the internal transcriber 1 (ITS1) region of the protozoan organism was used in the molecular testing. From the total of fetuses examined, 71.42% were positive for N. caninum by PCR. A higher number of heart samples (47.1%) were positive for the parasite using this technique. Antibodies against the protozoa were detected in 12.9% of serum samples of cows; 2.8% of fetuses were seropositive for this pathogen. Our results show that vertical transmission of N. caninum occurs in cattle from this region of Brazil, and that the use of different diagnostic techniques contributes to successful diagnosis of congenital transmission of the parasite in cattle.