Régis Adriel Zanette

Aceturato de diminazeno e dipropionato de imidocarb no controle de infecção por Trypanosoma evansi em Rattus norvegicus infectados experimentalmente

The aim of this study was to evaluate the efficacy of diminazene aceturate and imidocarb dipropionate in the control of Trypanosoma evansi infection in rats (Rattus norvegicus) experimentally infected. Fifty-four male rats were inoculated through intraperitoneal route with 104 T. evansi trypomastigotes. The rats were evaluated daily by periferic blood smears examination and treated when eight flagellated parasites were observed in 1000x microscopic field. Two therapeutics protocols were used. The first one included Groups A, B, C, D in which the rats were submitted to a single dose of the testing drugs administered by intramuscular route at the day 0 and again when T. evansi was observed in the blood smears. The rats of the second protocol (Groups E, F, G, H) were submitted to the same treatment by five consecutive days. Four rats (Group I) were used as control and were not submitted to any treatment. Tested drugs did not show any curative effect when used in the first protocol, since parasitaemia was evident few days after treatment. The use of diminazene aceturate in the second protocol resulted in elimination of the trypomastigotes from circulation. In this case the rats were euthanized at the day 90. The infection recurred 30 days after the administration of imidocarb dipropionate. Histologically, no lesions were found in the liver or kidney. Diminazene aceturate is effective in treating trypanosomosis in rats when used five days consecutively.

In vitro susceptibility of fluconazole-susceptible and -resistant isolates of Malassezia pachydermatis against azoles

OBJECTIVES The first aim of this study was to evaluate the in vitro efficacies of fluconazole, ketoconazole, itraconazole and voriconazole on M. pachydermatis growth inhibition. This study also evaluated M. pachydermatis azole cross-resistance, comparing wild clinical isolates and the same isolates with in vitro-induced fluconazole resistance. METHODS Two techniques were used: (1) a broth microdilution method based on protocol M27-A3 from the Clinical and Laboratory Standards Institute to determine the minimum inhibitory concentration (MIC) and (2) the Fekete-Forgács method to induce fluconazole resistance in vitro. The isolates were divided into two groups: group 1 included fluconazole-susceptible clinical isolates (n=30) and group 2 contained the same isolates with in vitro-induced fluconazole resistance (n=30). RESULTS The two groups exhibited differences in susceptibility (p<0.001). Group 1 isolates were susceptible to azoles: ketoconazole (MIC 0.01-1.0 μg/mL), itraconazole (MIC 0.01-1.0 μg/mL), voriconazole (MIC 0.01-4.0 μg/mL), and fluconazole (MIC 0.01-4.0 μg/mL). Group 2 isolates demonstrated a wider range of MICs to azoles: ITZ (MIC 0.06-64.0 μg/mL), KTZ (MIC 0.25-32.0 μg/mL), VRZ (MIC 2.0-128.0 μg/mL), and FLZ (MIC 64.0-128.0 μg/mL). CONCLUSIONS It was shown that FLZ-resistant M. pachydermatis isolates exhibit cross-resistance to other azoles, reinforcing the importance of susceptibility tests as a guide for the therapeutic prescription of antifungals in medical and veterinary mycology.

Influence of Trypanosoma evansi in blood, plasma, and brain cholinesterase of experimentally infected cats.

Changes in blood, plasma and brain cholinesterase activities in Trypanosoma evansi-infected cats were investigated. Seven animals were infected with 10(8) trypomastigote forms each and six were used as control. Animals were monitored for 56 days by examining daily blood smears. Blood samples were collected at days 28 and 56 post-inoculation to determine the activity of acetylcholinesterase (AChE) in blood and the activity of butyrylcholinesterase (BChE) in plasma. AChE was also evaluated in total brain. The activity of AChE in blood and brain, and the activity of BChE in plasma significantly reduced in the infected cats. Therefore, the infection by T. evansi influenced cholinesterases of felines indicating changes in the responses of the cholinergic system.

In vitro activity of terbinafine associated to amphotericin B, fluvastatin, rifampicin, metronidazole and ibuprofen against Pythium insidiosum.

We evaluated the in vitro activities of terbinafine alone and in combination with amphotericin B, fluvastatin, rifampicin, metronidazole or ibuprofen against 17 clinical isolates of Pythium insidiosum. The assays were based on technique M38-A2, as well as the checkerboard microdilution method. The main synergism observed was by combination of terbinafine plus amphotericin B (41.18%). Antagonisms were observed in combinations of terbinafine with fluvastatin (35.30%) or rifampicin (5.88%).

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.

Granulomatous rhinitis associated with Pythium insidiosum infection in sheep.

Pythium insidiosum is an aquatic organism classified in the kingdom Stramenopila, class Oomycetes ([De Cock and others 1987][1]). It is the aetiological agent of pythiosis in mammals; the occurrence of this disease is associated with contact between traumatic lesions in animals or human beings and

Diminazene aceturate in the control of Trypanosoma evansi infection in cats.

The aim of this study was to investigate the efficacy of diminazene aceturate in the control of the infection by Trypanosoma evansi in cats. Fourteen animals were infected with 10(8) trypomastigote forms each and six were used as negative control (group A). Seven of the infected cats were used as positive control (group B) and seven were treated with diminazene aceturate (3.5 mg kg(-1)) for 5 consecutive days (group C). Biochemical and hematological parameters were evaluated during the experiment. Blood with anticoagulant was collected at day 49 post-inoculation and preserved in ethanol for DNA extraction. Samples were analyzed using PCR T. evansi-specific to assess the effectiveness of treatment. The treatment with diminazene aceturate had an efficacy of 85.7%. Alanine aminotransferase, gamma-glutamyltransferase, urea, and creatinine values remained within the normal physiological range in the treated cats. Hemogram was normalized in all the cured animals. Therefore, the therapy used is effective in controlling T. evansi in cats.

In vitro and in vivo susceptibility of two-drug and three-drug combinations of terbinafine, itraconazole, caspofungin, ibuprofen and fluvastatin against Pythium insidiosum.

The present study investigated the in vitro inhibitory activity of terbinafine, itraconazole, caspofungin, fluvastatin and ibuprofen against 15 isolates of Pythium insidiosum in double and triple combinations and determined in vivo correlations using rabbits with experimental pythiosis. The minimal inhibitory concentration (MIC) was determined in accordance with the Clinical and Laboratory Standards Institute M 38-A2 protocol (2008), and the in vitro interactions were evaluated using a checkerboard microdilution method. For the in vivo study, 20 rabbits inoculated with P. insidiosum zoospores were divided into four groups: group 1 was treated with terbinafine and itraconazole; group 2 was treated with terbinafine, itraconazole and fluvastatin; group 3 was treated with terbinafine and caspofungin; and group 4 was the control group. Combinations of terbinafine with caspofungin or ibuprofen were synergistic for 47% of the isolates, and antagonism was not observed in any of the double combinations. The triple combinations were mostly indifferent, but synergism and antagonism were also observed. In the in vivo study, the histological aspect of the lesions was similar among the groups, but group 2 showed the lowest amount of hyphae and differed significantly from the other groups.

Horses naturally infected by Trypanosoma vivax in southern Brazil

In this study, we reported the first outbreak of the infection by Trypanosoma vivax in horses in southern Brazil, a non-endemic region where bovines have only recently been found infected by this trypanosome species. We evaluated 12 horses from a farm in southern Brazil, where four horses displayed pale mucous membranes, fever, weight loss, and swelling of abdomen, prepuce, or vulva. The diagnosis of T. vivax was confirmed in four horses by morphological parameters of trypomastigotes in blood smears and species-specific PCR. All T. vivax-infected animals showed anemia, and most showed increased levels of beta-1, beta-2, and gamma globulins. Horses were treated with diminazene aceturate, but cure was not achieved, and the disease relapsed after therapy. These findings demonstrated that Brazilian T. vivax isolates, which were already reported infecting cattle, buffaloes, goats, and sheep, can be highly pathogenic for horses, causing severe disease and even death of the animals due to the recurrence of the infection.

Trypanosoma evansi: levels of copper, iron and zinc in the bloodstream of infected cats.

The aim of this study was to evaluate the concentrations of copper, iron and zinc in blood serum of cats experimentally infected with Trypanosoma evansi. Animals were divided into two groups: control and infected with T. evansi. The animals were infected with 10(8) trypomastigotes each and parasitemia was estimated daily for 56 days by microscopic examination of smears. Hematological and biochemical parameters were evaluated for monitoring of the disease. Serum metal levels were determined in blood samples collected at days 7, 28 and 56 of the experiment. Inductively coupled plasma optical emission spectrometry was used to measure the levels of copper, iron and zinc. Significant differences were observed among groups (P<0.05). Increased levels of copper and decreased iron and zinc levels were observed. A decrease in the number of red blood cells was also observed 7 days after inoculation. Biochemical parameters were not altered. Therefore, the infection by T. evansi might alter the serum metal levels, causing metabolic disturbances in cats.