Lucas T. Gressler

Trypanosoma evansi: hematologic changes in experimentally infected cats.

This study aimed at evaluating hemogram and erythropoietic changes in cats experimentally infected with Trypanosoma evansi. Thirteen adult female non-breeding Felix catus were separated into two groups: seven animals were infected with 10(8) trypomastigotes each, and six animals were used as negative controls. Animals were kept in air-conditioned rooms and blood smears were performed daily for 49 days. Blood samples were collected from the jugular vein at days 0, 7, 21, 35 and 49 and stored in blood-collecting tubes containing anticoagulant. Bone marrow was collected from the proximal epiphysis of the right femur at days 14 and 42 post-inoculation (PI). Total erythrocyte count, hematocrit and hemoglobin showed statistical differences among groups from the seventh day PI onwards (P<0.05). The mean corpuscular volume and the mean corpuscular hemoglobin concentration remained normal, characterizing a normocytic-normochromic anemia. Reticulocyte count increased in the infected group from the 21st day onwards, but remained near normal values suggesting a mild regenerative anemia. Moreover, the myeloid:erythroid ratio significantly reduced at day 42 PI, evidencing a bone marrow hematopoietic response. Based on these results we conclude that cats infected with T. evansi have normocytic, normochromic, regenerative anemia.

Susceptibility of Trypanosoma evansi to propolis extract in vitro and in experimentally infected rats

Current therapy of Trypanosoma evansi infections is not effective for the vast majority of animals with relapsing parasitemia and clinical signs. Recently, attention is being focused on the antiparasitic activity of propolis. This study evaluated the susceptibility of T. evansi to propolis extract in vitro and in vivo. A dose-dependent trypanocidal activity of propolis extract was observed in vitro. All trypomastigotes were killed 1 h after incubation with 10 μg mL(-1) of the extract. In vivo, the concentrations of 100, 200, 300 and 400 mg kg(-1) administered orally for 10 consecutive days showed no curative effect, and the rats died from the disease. However, rats treated with the two highest concentrations of propolis extract showed higher longevity than the other groups. Based on these data, we concluded that T. evansi is susceptible to propolis in vitro. Despite the lack of curative efficacy observed in vivo at the concentrations tested, the propolis extract can prolong life in rats infected with the protozoan.

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.

Serum levels of LH, FSH, estradiol and progesterone in female rats experimentally infected by Trypanosoma evansi.

The goal of this study was to evaluate reproductive hormones in sera samples of female rats experimentally infected by Trypanosoma evansi during different phases of the estrous cycle. For that, 64 animals were divided into two groups: 24 rats for the control group (uninfected), and 40 animals were infected by T. evansi. These groups were divided into subgroups according to the time of infection (days 5 and 15 post-infection; PI) and the phase of the estrous cycle (proestrus, estrus, metestrus and diestrus). Serum was collected at days 5 and 15 PI and the levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone and estradiol were assessed by enzyme immunoassay technique. The concentration of nitrite/nitrate (NOx), advanced oxidation protein products (AOPP), and thiobarbituric acid reactive substances (TBARS) were measured in ovaries and uteruses in these same periods. Infected females showed significant decrease (P<0.05) of LH, FSH, estradiol and progesterone in different periods and phases of the estrous cycle when compared to uninfected rats. In addition, it was observed an increase in the concentration of NOx, AOPP, and TBARS in the ovaries, which is indicative of cell damage. Therefore, our experimental study showed that T. evansi infection in female rats may cause changes in LH, FSH, estradiol, and progesterone levels regardless of the time of infection or phase of the estrous cycle.

Biochemical changes in cats infected with Trypanosoma evansi

This study aimed at evaluating biochemical changes of cats (Felis catus) experimentally infected with Trypanosoma evansi. Seven animals were infected with 10(8) blood trypomastigotes per animal and six were used as controls. Blood smears were performed daily for 56 days and the hepatic, renal and muscular parameters in blood serum were evaluated at days 0, 7, 21, 35 and 49. The protozoan was found in the bloodstream 24-48 h post-inoculation (PI) and irregular peaks of parasitemia were observed throughout the experiment. Muscular enzymatic activities (aspartate aminotransferase and creatine kinase) were increased in infected cats compared to controls. Increased concentrations of total proteins and globulins and decreased levels of albumin and albumin/globulin ratio were observed in infected group versus the controls values (P<0.05). No alteration in serum activity of alanine aminotransferase, gamma-glutamyltransferase, creatinine and urea was observed in both groups.

Susceptibility of Trypanosoma evansi to human blood and plasma in infected mice.

Around 1900 Laveran and Mesnil discovered that African trypanosomes do not survive in the blood of some primates and humans. The nature of the trypanolytic factor present in these sera has been the focus of a long-standing debate between different groups. The aim of this study was to investigate the susceptibility of T. evansi isolates to therapy using human blood and plasma in experimentally infected mice. Forty-eight 2-month-old female mice (Mus musculus) were divided into six groups of eight animals per group (A, B, C, D, E and F). Plasma was obtained after blood collection in order to perform therapy. Animals from group A (positive control) were inoculated with T. evansi and treated with 0.2mL of saline solution. Animals from groups B and C were infected with the flagellate and received a curative treatment with 0.2mL of human blood (group B) and 0.2mL of human plasma (group C), 24h after infection. Animals from groups D and E received a prophylactic treatment with 0.2mL of human blood and 0.2mL of human plasma, respectively, 24h prior to the infection. Animals from group F (negative control) were not infected and received 0.2mL of saline solution. The four treatments (B, C, D and E) increased animals longevity when compared to group A. Prepatency period was longer in groups D (15 days) and E (37.7 days) under prophylactic immunotherapy. Moreover, no parasites were found in most of the animals 60 days post-inoculation (PI). Besides the longer longevity, treatments were capable of curing 50% of mice of group B, 37.5% of group C, 37.5% of group D and 25% of the animals from group E.

Patogenicidade de um isolado de Trypanosoma evansi em ratos inoculados com o parasito em sangue in natura e criopreservado

This study aimed to evaluate the Trypanosoma evansi strain pathogenicity (LPV-2005) in rats under passive immunity influence, of different concentrations and media preservation. Thirty six adult female Rattus norvergicus were separated in six equal groups. Groups A and B were inoculated with 105 T. evansi and groups C and D with 106 blood trypomastigotes per animal. Groups E and F were used as negative control in which the animals were inoculated with fresh and cryopreserved blood, without the parasite. Group A were composed of T. evansi infected born rats and cured females. Groups B, C and D were composed with animals never exposed to the LPV-2005 strain. All groups B and C animals received different doses of blood trypomastigotes kept in Wistar rats, while animals from group D were infected with cryopreserved blood kept in liquid nitrogen. The the strain pathogenicity was estimated by prepatency evaluation period, levels of parasitemia and animals longevity. Group D showed a longer prepatency period in comparison with other groups. The longevity of group D (27.8 days) was significantly different (P<0.05) from group C (4.8 days). Rats of the control group were euthanized 50 days postinfection. In conclusion, the tested inoculum-preservation methods and the infective dose of T. evansi influenced the pathogenicity of the LPV-2005 strain in rats. The presence of maternal antibodies did not prevent the infection and mortality of the rats by T. evansi.

Relationship between behavioral alterations and activities of adenylate kinase and creatine kinase in brain of rats infected by Trypanosoma evansi

The aim of this study was to investigate the behavioral assessment and activities of important enzymes involved in the phosphoryl transfer network in rat brains that were experimentally infected with Trypanosoma evansi. Behavioral assessment (cognitive performance), pro-inflammatory cytokines in serum and activities of adenylate kinase (AK), pyruvate kinase (PK), and creatine kinase (CK) in brain were evaluated at 5 and 15 days post-infection (PI). Here we demonstrate a cognitive impairment in the rats infected with T. evansi. At 5 and 15 days PI, a memory deficit and a depressant activity were demonstrated by an inhibition avoidance test and increase in the immobility time in a tail suspension test, respectively. On day 5 PI, a decrease in the CK activity and an increase in the AK activity were observed. On day 15 PI, an increase in the CK activity and a decrease in the AK activity were observed. Considering the importance of energy metabolism for brain functioning, it is possible that the changes in the activity of enzymes involved in the cerebral phosphotransfer network and an increase in the proinflammatory cytokines (TNF and IFN) may be involved at least in part in the cognitive impairment in infected rats with T. evansi.

Experimental infection by Haemonchus contortus in lambs: influence of disease on purine levels in serum.

The aim of this study was to evaluate the purine levels of lambs experimentally infected with Haemonchus contortus. A total of 12 healthy lambs were divided into two groups, composed of 6 animals each: Group A represented the healthy animals (uninfected), while in Group B the animals were infected with 15 000 larvae of H. contortus. Blood was drawn on days 15, 45 and 75 post-infection (PI) in order to perform the purine analysis (ATP, ADP, AMP, adenosine, inosine, hypoxanthine, xanthine and uric acid) by high pressure liquid chromatography (HPLC) in serum. On day 15 PI a significant (P<0·05) increase in the levels of ATP and inosine was observed in the infected animals, unlike the levels of ADP, adenosine, xanthine and uric acid which were reduced. On day 45 PI a significant (P<0·05) increase in the ATP and xanthine levels in infected animals was observed, contrasting with reduced levels of ADP and uric acid. Finally, on day 75 PI an increase occurred in the levels of ATP, adenosine and hypoxanthine in infected lambs, concomitant with a reduction in the levels of ADP and uric acid (P<0·05). These changes in purine levels may influence the inflammatory process and the pathological events.

Gastrointestinal parasites of cavy (Cavia aperea aperea) in southern Brazil

The aim of this study was to evaluate the gastrointestinal parasitism in Cavia aperea aperea (cavy), captured in the central region of Rio Grande do Sul State. Fecal samples from five free-living cavies were collected for research of parasites. Samples were analyzed by the centrifugal-flotation method with zinc sulfate and parasites were identified microscopically based on (oo)cyst and egg size and morphology. Cysts of Giardia sp. and (oo)cysts of Cryptosporidium sp. and Cystoisospora sp. were observed in one or more cavies. Eggs of Paraspidodera uncinata were observed in three of the five rodents. All infected animals showed mild infection by parasite. This is the first report of Giardia sp., Cryptosporidium sp. and Cystoisospora sp. in Cavia a. aperea.