Lenita M. Stefani

In vitro and in vivo action of terpinen-4-ol, γ-terpinene, and α-terpinene against Trypanosoma evansi.

This study aimed to evaluate the susceptibility in vitro and in vivo of Trypanosoma evansi to terpinen-4-ol, γ-terpinene and α-terpinene, the three main compounds of tea tree oil (Melaleuca alternifolia) with known efficacy in the treatment of trypanosomosis. In vitro, a trypanocidal effect of terpinen-4-ol, γ-terpinene, and α-terpinene was observed when used alone or associated at 0.5, 1 and 2% concentrations i.e., the α-terpinene showed a faster trypanocidal effect when compared to chemotherapy (diminazene aceturate - D.A.). In vivo studies were performed in two experiments: I and II where experiment I used T. evansi infected mice treated with terpinen-4-ol, γ-terpinene and α-terpinene alone (at a dose of 1.0 mL kg(-1)) or associated (two compounds, dose of 0.5 mL kg(-1) of each compound; tree compounds, dose of 0.335 mL kg(-1) of each compound); Treatment with α-terpinene was able to extend animal longevity, but showed no curative efficacy. In experiment II, T. evansi infected mice were treated with D.A. associate with α-terpinene, where a curative efficacy of 57.14% was found, a much better result when D.A. was used alone (14.28%). In summary, α-terpinene associated with D.A. can be used as an alternative treatment for T. evansi infection. The compound α-terpinene from M. alternifolia essential oil is the one responsible for the trypanocidal effect, a fact confirmed by in vitro results and the increased longevity observed on treated mice.

Insecticidal and repellent effects of tea tree and andiroba oils on flies associated with livestock

This study aimed to evaluate the insecticidal and repellent effects of tea tree, Melaleuca alternifolia (Myrtales: Myrtaceae), and andiroba, Carapa guianensis (Sapindales: Meliaceae), essential oils on two species of fly. For in vitro studies, free‐living adult flies were captured and reared in the laboratory. To evaluate the insecticidal effects of the oils, adult flies of Haematobia irritans (L.) and Musca domestica L. (both: Diptera: Muscidae) were separated by species in test cages (n = 10 per group), and subsequently tested with oils at concentrations of 1.0% and 5.0% using a negative control to validate the test. Both oils showed insecticidal activity. Tea tree oil at a concentration of 5.0% was able to kill M. domestica with 100.0% efficacy after 12 h of exposure. However, the effectiveness of andiroba oil at a concentration of 5.0% was only 67.0%. The insecticidal efficacy (100.0%) of both oils against H. irritans was observed at both concentrations for up to 4 h. The repellency effects of the oils at concentrations of 5.0% were tested in vivo on Holstein cows naturally infested by H. irritans. Both oils demonstrated repellency at 24 h, when the numbers of flies on cows treated with tea tree and andiroba oil were 61.6% and 57.7%, respectively, lower than the number of flies on control animals. It is possible to conclude that these essential oils have insecticidal and repellent effects against the species of fly used in this study.

Pseudomonas aeruginosa strain PA01 impairs enzymes of the phosphotransfer network in the gills of Rhamdia quelen

Integration of mitochondria with cytosolic ATP-consuming/ATP-sensing and substrate supply processes is critical for gills bioenergetics, since this tissue plays an important role in the respiratory energy metabolism. The effects of bacterial infection on gills remain poorly understood, limited only to histopathological analyses. Thus, the aim of this study was to investigate whether experimental infection by Pseudomonas aeruginosa strain PA01 alters the enzymes of the phosphoryltransfer network (adenylate kinase (AK), pyruvate kinase (PK) and cytosolic and mitochondrial creatine kinase (CK)) in gills of silver catfish (Rhamdia quelen). The animals were divided into two groups with six fish each: uninfected (negative control) and infected (positive control). On day 7 post-infection (PI), animals were euthanized and the gills collected. AK, PK, and cytosolic and mitochondrial CK activities in gills decreased in infected compared to uninfected animals. Also, severe gill damage and destruction in the primary and secondary lamellae was observed in the infected animals. Therefore, we have demonstrated, for the first time, that experimental infection by P. aeruginosa inhibits key enzymes linked to the production and utilization of metabolic energy in silver catfish, and consequently, impairs cellular energy homeostasis, which may contribute to disease pathogenesis.

Lice Outbreak in Buffaloes: Evidence of Anaplasma marginale Transmission by Sucking Lice Haematopinus tuberculatus

Abstract: Lice infestations are commonly seen in buffaloes, causing damage directly to the animal, i.e., itching, skin lesions, and anemia. In addition, these insects can also be vectors for infectious diseases. The present study describes an outbreak of lice in buffaloes as well as evidence for Haematopinus tuberculatus acting as a vector of anaplasmosis. Lice and blood were collected from 4 young buffaloes (2- to 4-mo-old) and a molecular analysis for the presence of Anaplasma marginale was conducted. DNA of A. marginale was detected in the blood of all 4 animals. Twelve lice were collected and separated in 4 groups, with 3 insects each, to comprise a pool of samples. After DNA extraction and molecular analysis, a positive PCR for A. marginale was found in all pooled samples. These results identify sucking lice as potential vectors of anaplasmosis. However, additional studies are necessary to fully evaluate the vector potential of H. tuberculatus for A. maginale transmission.

Melaleuca alternifolia essential oil enhances the non-specific immune system and prevents oxidative damage in Rhamdia quelen experimentally infected by Aeromonas hydrophila: Effects on cholinergic and purinergic systems in liver tissue

Abstract The aim of this study was to evaluate the effects of M. alternifolia essential oil used to treat silver catfish (Rhamdia quelen) experimentally infected by Aeromonas hydrophila on oxidative stress variables, and for the first time, on hepatic enzymes of the cholinergic and adenosinergic systems. For that, fish were divided into six groups (A‐F), each containing seven animals. Groups A, B and C were composed of uninfected animals, while animals in groups D, E and F were intramuscularly inoculated with A. hydrophila. Groups B and E received a prophylactic bath with M. alternifolia essential oil (50 &mgr;L/L, diluted in ethanol) for seven days, while groups C and F were exposed to ethanol. After the prophylactic baths, groups D, E and F were inoculated with 100 &mgr;L of A. hydrophila solution (2.1 × 109 colony‐forming unit). Two days after inoculation, the animals were euthanized and liver samples were collected. Infected animals (the group D) showed increased TBARS and protein carbonylation levels, while CAT, AChE and ADA activities decreased compared to uninfected animals (the group A). The prophylactic treatment with M. alternifolia essential oil (the group E) prevented the alterations caused by A. hydrophila, but it did not change AChE activity. Thus, the prophylactic treatment prevents damage caused by lipids and proteins, as well as alterations of the adenosinergic system, demonstrating that the anti‐inflammatory effect of TTO is mediated by the adenosinergic pathway. In addition, TTO prophylactic treatment might be considered an important approach to prevent the hepatic damage caused by A. hydrophila. HighlightsAeromonas hydrophila cause injury on antioxidant system in liver tissue.Hepatic AChE and ADA activities decrease in infected fish with A. hydrophila.Treatment with TTO prevents the injury on antioxidant system.The anti‐inflammatory effect of TTO is mediated by the adenosinergic pathway.TTO might be considered an important approach to prevent the damage caused by bacteria.

Fowl typhoid in laying hens cause hepatic oxidative stress

The aim of this study was to analyses nitric oxide, antioxidant status, and oxidative profile in the liver of laying hens naturally infected by Salmonella enterica subsp enterica serovar Gallinarum (S. Gallinarum). The nitrite/nitrate (NOx), reactive oxygen species (ROS), thiobarbituric acid-reactive substances (TBARS), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione S-transferase (GST) activities were measured in liver samples, as well the biomarkers of hepatic function (alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), total protein and albumin levels measured in serum. NOx levels and CAT activity were reduced in hepatic tissue of infected hens. On the other hand, TBARS and ROS levels, GR, GPx and GST activities were higher in infected animals. On biomarkers of tissue damage, ALT, AST, GGT and total protein levels were higher in serum of infected hens, and showed decreased albumin levels. In summary, ROS and TBARS production lead to damage on the membrane lipids that alter activities of antioxidant enzymes CAT, GR, GPx and GSH, an adaptive response against S. Gallinarum infection, contributing to the pathophysiology and clinical signs of the disease.

Butyrylcholinesterase as a marker of inflammation and liver injury in the acute and subclinical phases of canine ehrlichiosis.

The aim of this study was to evaluate the role of butyrylcholinesterase (BChE) as a marker of inflammation and liver injury in the acute and subclinical phases of canine ehrlichiosis. Forty-two serum samples of dogs naturally infected with Ehrlichia canis were used, of which 24 were from animals with the acute phase of the disease and 18 with subclinical disease. In addition, sera from 17 healthy dogs were used as negative controls. The hematocrit, BChE activity, hepatic injury (alanine aminotransferase (ALT) and aspartate aminotransferase (AST)), nitric oxide, and cytokines levels were evaluated. The BChE activity was significantly elevated (P<0.05) in dogs with the acute phase of the disease when compared to healthy animals. However, there was a reduction on BChE activity on dogs with subclinical disease compared to the other two groups. AST and ALT levels were significantly higher (P<0.05) in the acute phase, as well as the inflammatory mediators (NOx, TNF-α, INF-γ, IL-4, IL-6) when compared to the control group. On the other hand, IL-10 levels were lower in the acute phase. Based on these results, we are able to conclude that the acute infection caused by E. canis in dogs leads to an increase on seric BChE activity and some inflammatory mediators. Therefore, this enzyme might be used as a marker of acute inflammatory response in dogs naturally infected by this bacterium.

Effect of tea tree oil (Melaleuca alternifolia) on the longevity and immune response of rats infected by Trypanosoma evansi

This study aimed to evaluate the effect of tea tree oil (TTO - Melaleuca alternifolia) on hepatic and renal functions, and the immune response of rats infected by Trypanosoma evansi. A pilot study has shown that rats treated with TTO orally (1 ml kg(-1)) had increased survival rate without curative effect. In order to verify if increased longevity was related to a better immune response against T. evansi when using tea tree oil, a second experiment was conducted. Thus, twenty-four rats were divided into four groups. The groups A and B were composed of uninfected animals, and the groups C and D had rats experimentally infected by T. evansi. Animals from the groups B and D were treated orally with TTO (1 ml kg(-1)) for three days. Blood samples were collected to verify humoral response analysis for immunoglobulins (IgA, IgM, IgE, and IgG) and cytokines (TNF-α, INF-γ, IL-1, IL-6, IL-4, and IL-10) at days 0, 3, 5 and 15 post-infection (PI). TTO treatment caused changes in the immunoglobulins and cytokines profile, as well as the course of T. evansi infection in rats. It was found that the TTO was not toxic, i.e., hepatic and renal functions were not affected. Therefore, it is possible to conclude that TTO influences the levels of inflammatory mediators and has trypanocidal effect, increasing life expectancy of rats infected by T. evansi.

Increase nitric oxide and oxidative stress in dogs experimentally infected by Ehrlichia canis: Effect on the pathogenesis of the disease

The aim of this study was to evaluate nitric oxide levels, lipid peroxidation, protein oxidation and glutathione reductase activity in serum of dogs experimentally infected by Ehrlichia canis. Banked serum samples of dogs divided into two groups were used: negative control (n=5) and infected by E. canis (n=5). The concentration of nitrite/nitrate (NOx), lipid peroxidation (TBARS), advanced oxidation protein products (AOPP), and glutathione reductase (GR) activity in sera were evaluated. Samples were collected on days 0, 3, 6, 18 and 30 post-infection (PI). NOx and TBARS levels were significantly (P<0.05) higher in the infected group at 18 and 30 days PI, as well as AOPP levels at 30 days PI when compared to samples from control group. The GR activity was significant (P<0.05) increased in serum of dogs infected by E. canis on days 18 and 30 PI. Based on the increased levels of NOx, TBARS, AOPP and GR activity we concluded that dogs experimentally infected by E. canis develop a state of redox imbalance and that these changes might be involved in the pathophysiology of the disease.

Trypanocidal activity of free and nanoencapsulated curcumin on Trypanosoma evansi.

This study aimed to evaluate in vitro and in vivo trypanocidal activity of free and nanoencapsulated curcumin against Trypanosoma evansi. In vitro efficacy of free curcumin (CURC) and curcumin-loaded in lipid-core nanocapsules (C-LNCs) was evaluated to verify their lethal effect on T. evansi. To perform the in vivo tests, T. evansi-infected animals were treated with CURC (10 and 100 mg kg(-1), intraperitoneally [i.p.]) and C-LNCs (10 mg kg(-1), i.p.) during 6 days, with the results showing that these treatments significantly attenuated the parasitaemia. Infected untreated rats showed protein peroxidation and an increase of nitrites/nitrates, whereas animals treated with curcumin showed a reduction on these variables. As a result, the activity of antioxidant enzymes (superoxide dismutase and catalase) differs between groups (P<0.05). Infected animals and treated with CURC exhibited a reduction in the levels of alanine aminotransferase and creatinine, when compared with the positive control group. The use of curcumin in vitro resulted in a better parasitaemia control, an antioxidant activity and a protective effect on liver and kidney functions of T. evansi-infected adult male Wistar rats.