Chemotherapy of surra in horses and mules with diminazene aceturate

During June–July 2000, an outbreak of surra occurred on an equine breeding farm in Khonkaen Province, Thailand. Forty-two percent of pregnant mares aborted or gave stillbirth and 40% (19/47) of horses and 10% (1/10) of mules died from surra. In August 2000 Trypanosoma evansi were detected in the remaining animals (28 horses and nine mules) on the farm by blood smear and/or the haematocrit centrifuge technique. All animals were treated with diminazene aceturate at 3.5 mg/kg body weight by intramuscular injection on days 0 and 41 of the study. Blood samples of eight randomly selected horses and mules were collected on days 0, 1 and once a week until day 56 and examined for T. evansi by various parasitological techniques. The sera were tested for antibodies against T. evansi using an indirect enzyme linked immunosorbent assay (ELISA).

The results revealed that diminazene aceturate at 3.5 mg/kg appeared to be effective in the first treatment of horses and mules infected with T. evansi. Parasites were cleared from the peripheral blood of horses on days 1 and 7 and mules on days 1 and 14. Thereafter the number of positive animals increased. After the second treatment, 50% of horses and 25% of mules were still positive to surra 24 h after treatment demonstrating that diminazene had no protective effect. Mild to severe toxicity of diminazene was seen in the horses and mules after injection.     

Comparative efficacy assessment of pentamidine isethionate and diminazene aceturate in the chemotherapy of Trypanosoma brucei brucei infection in dogs

The chemotherapeutic efficacy of diminazene aceturate (Berenil)--a standard veterinary trypanocide and pentamidine isethionate (PMI)--a human trypanocide was compared in dogs experimentally infected with Trypanosoma brucei brucei. Also, the activities of the drugs on some serum liver enzymes were evaluated before and after treatment to ascertain the relative safety of the drugs. Fifteen local dogs (mongrels) were used for the study. Three of the dogs were uninfected controls, and twelve were infected with a stock of T. brucei brucei. Three of the infected dogs were untreated controls, three were given diminazene aceturate (DA) at 7 mg/kg body weight intramuscularly (i/m), another three received pentamidine isethionate (PMI) at 4 mg/kg i/m on days 14, 17, 19, 27, 29, and 31 post infection (PI) and the remaining three dogs were also given same dose of PMI on days 14, 16, 18, 20, 22, 24 and 26 PI. Both trypanocides effectively cleared the parasites from the blood of the infected treated dogs. However, the infection subsequently relapsed at day 42 PI in one of the dogs in the DA treated group which later died at day 70 PI. Relapse infection was not recorded with the PMI treated groups although two dogs died in the PMI treated group II (treatment at days 14, 17, 19, 27, 29, and 31 PI) without showing relapsed parasitaemia. The packed cell volume (PCV), red blood cell (RBC) count, and haemoglobin (Hb) level which decreased significantly following infection, were reversed by the trypanocidal treatment. The reversal in the red cell values was faster in the PMI treated groups than in the DA treated group. The serum alkaline phosphate (SAP), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels increased following infection and drug administration. The increase in the enzyme levels was greater in the DA treated groups than PMI treated groups. It was thus concluded that PMI given at 4 mg/kg i/m at days 14, 16, 18, 20, 22, 24, and 26 PI constituted a safe and efficient trypanocide and exhibited a superior trypanocidal action than DA in T. brucei brucei infected dogs.     

Effects of the combination of DL-alpha-difluoromethylornithine and diminazene aceturate in Trypanosoma congolense infection of dogs

The therapeutic activity of a combination of difluoromethylornithine (DFMO) with diminazene aceturate was investigated in mongrel dogs experimentally infected with Trypanosoma congolense. The criteria used in the assessment of the trypanocidal effect of the therapy include the examination of the blood for parasites, as well as clinical and haematological changes at intervals following treatment. Diminazene aceturate and DFMO alone and in combination produced intermittent aparasitaemia in the dogs. Although relapse infection occurred with all three treatment regimes, the drug combination gave the best result. The packed red cell volume, haemoglobin concentrations and red blood cell values decreased significantly following parasite inoculation but increased after treatment. The total leucocyte counts decreased in all the infected dogs but improved with treatment, and the differential leucocyte counts indicated neutropenia in all the infected animals prior to treatment.     

DL-alpha-difluoromethylornithine (DFMO)-Berenil combination: therapeutic and prophylactic activity against Trypanosoma brucei brucei infection in mice.

The therapeutic and prophylactic activity of difluoromethylornithine (DFMO) (2% in drinking water for 4 days) and Berenil (7 mg/kg live weight intraperitoneally) combination was investigated in mice infected with Trypanosoma brucei brucei. Using a previously described mouse model of the African trypanosomosis of the central nervous system, it was demonstrated that the combination was curative and acted synergistically. However, if used prophylactically it had no advantage over Berenil alone.     

Relapses in dogs experimentally infected with Trypanosoma brucei and treated with diminazene aceturate or isometamidium chloride

Twenty dogs of mixed local East African breeds were used. Five of the dogs were uninfected controls and 15 were infected with T. brucei (ILRAD 273). Five of the infected dogs were untreated controls, five were treated with a high curative dose of diminazene aceturate, (7 mg kg-1 body weight (wt.), and five were given a subcurative dose of isometamidium chloride (1 mg kg-1 body wt.). The drugs, given at 8 days post infection (d.p.i..), led to apparent recovery. The antibody titres, however, remained high in both groups and at 42-49 d.p.i. there was at least one relapse in each treatment group. Parasite populations from relapsed animals were more resistant to the drugs than the original infecting populations. The implications of these findings are discussed.     

Comparative efficacy of diminazene diaceturate and diminazene aceturate for the treatment of babesiosis in horses

The present work was conducted to study the comparative efficacy of two anti-protozoan (babesicidal) drugs on the recovery and health of horses. A total of 80 horses, showing typical clinical symptoms of the disease, were selected for this study; the presence of babesiosis was confirmed through blood smear examination. These animals were divided into two groups i.e. A and B. Horses of Group A, were treated with diminazene diaceturate, while horses of Group B were treated with diminazene aceturate. Efficacy of the drugs was determined by the reversal of clinical signs and a negative blood smear examination. The efficacy of diminazene diaceturate was demonstrated to be 80% while diminazene aceturate was found to have 90% efficacy against babesiosis.     

The efficacy of berenil (diminazene aceturate) against Trypanosoma evansi infection in mice

The efficacy of berenil (diminazene aceturate) was studied in experimental Trypanosoma evansi infection in albino mice. The criteria used for the assessment of the anti-trypanosomal effect of berenil included the examination of the blood and tissues for T. evansi and the clinical, pathological and enzyme histochemical changes seen in the lungs, heart, liver, kidney and spleen at intervals after treatment. Single doses of 10 and 20 mg/kg of berenil given intraperitoneally or intramuscularly to infected mice produced a complete elimination of the protozoon and caused a slow tissue recovery mirrored in the persistence of lesions in different organs. Single doses of 3.5 mg/kg of berenil were less effective and none of the three dose levels of the drug used induced toxic effects in albino mice.     

Effect of tetracycline administration on the efficacy of diminazene aceturate therapy and prophylaxis in Trypanosoma brucei infections of mice

The simultaneous administration, to Trypanosoma brucei infected mice, of rolitetracycline or oxytetracycline and diminazene aceturate appeared to have no effect on the initial trypanocidal action of the diminazene aceturate in that trypanosomes were cleared from the circulation. It also had no effect on the duration of the aparasitaemic period which follows diminazene aceturate treatment and the mice remained free of circulating trypanosomes for some time. However, if used prophylactically, relatively large amounts of tetracyclines (4 x 10 mg kg-1) administered with 40 mg kg-1 diminazene aceturate caused a reduction of the prophylactic period compared with those mice given only diminazene aceturate. This reduction in the prophylactic period is unlikely to have any practical significance in the combination diminazene aceturate/tetracycline treatment of domestic animals as diminazene aceturate is used therapeutically and not prophylactically in the control of trypanosomiasis.     

Use of monoclonal antibodies for detecting T brucei brucei infection in splenectomised dogs

A monoclonal antibody against a plasma membrane antigen of Trypanosoma rhodesiense was used for the detection of T brucei group-specific circulating antigen in 24 adult local dogs experimentally infected with T brucei brucei strain 8/18. Ten of the dogs were splenectomised and the remainder non-splenectomised (intact). Five dogs each from the splenectomised and intact groups were inoculated intravenously with try-panosomes. The infected dogs developed trypanosomiasis between days 4 and 8 after infection. The circulating antigens were detected as early as six days after infection and remained high until two weeks after treatment, when the circulating antigen declined. The detection of the antigens showed the existence of infection unlike the antibody test. The treatment of the infected dogs with diminazene aceturate (Berenil; Hoechst) at a dose of 7-0 mg/kg on day 21 after infection cleared all the parasites but elevated the circulating antigen levels. The antigen capture enzyme-linked immunosorbent assay is a useful diagnostic tool for complementing parasitological diagnosis, for detecting infection in the field and for ascertaining the efficacy of trypanocidal drugs.     

The influence of Trypanosoma congolense infection on the disposition kinetics of diminazene aceturate in the dog

Diminazene aceturate was administered intravenously at 3.5 mg/kg body weight to mongrel dogs before and after infection with Trypanosoma congolense. Plasma and urine were collected at varying intervals thereafter and analysed for the compound. The mean are under the concentration-time curve (AUC) of diminazene in healthy dogs was 25.8 h.g/ml but was significantly increased (p<0.05) to 35.7 h.g/ml after infection with T. congolense. The distribution half-life was significantly reduced (p<0.05) in dogs after infection, being 0.12 h compared to 0.17 h in the same dogs before infection. The mean proportion of the diminazene recovered in the urine of infected dogs (25.1%) was not significantly different from that recovered in the urine of healthy dogs (26.8%). These results indicate that infection with T. congolense increases the rate at which diminazene is distributed in the body but that the infection has no marked influence on the urinary excretion of the drug.     

The pharmacokinetics of diminazene aceturate after intramuscular administration in healthy dogs

The pharmacokinetics of diminazene aceturate following intramuscular (i.m.) administration at 4.2 mg/kg was evaluated in 8 healthy German Shepherd dogs. Blood samples were collected at 19 intervals over a period of 21 days. Diminazene plasma concentrations were measured using a validated HPLC method with UV detection and a sensitivity of 25 ng/ml. The in vitro and in vivo binding of diminazene to blood elements was additionally determined. Diminazene pharmacokinetics showed a large inter-individual variation after i.m. administration. It had a short absorption half-life (K01-HL of 0.11 +/- 0.18 h), resulting in a C(max) of 1849 +/- 268.7 ng/ml at T(max) of 0.37 h and a mean overall elimination half-life (T1/2beta) of 5.31 +/- 3.89 h. A terminal half-life of 27.5 +/- 25.0 h was measured. At 1 h after i.m. injection, 75% of the diminazene in whole blood was in the plasma fraction. The results of this study indicate that diminazene is rapidly distributed and sequestered into the liver, followed by a slower terminal phase during which diminazene is both redistributed to the peripheral tissues and/or renally excreted. It is recommended that diminazene administered i.m. at 4.2 mg/kg should not be repeated within a 21-day period.     

Diminazene aceturate residues in the tissues of healthy, Trypanosoma congolense and Trypanosoma brucei brucei infected dogs

The tissue distribution and residue profile of diminazene aceturate was investigated in healthy dogs and in dogs infected with Trypanosoma congolense and Trypanosoma brucei brucei. The drug was administered at 3.5 mg/kg i.m. and tissue samples were taken post mortem from the animals at 48, 72, 120, 168 and 240 h after injection. The drug was distributed to various organs and tissues of the body with the highest concentrations occurring in liver and kidney. Higher drug levels were obtained in the tissues of healthy dogs compared with trypanosome infected animals except in the brain. The levels of residues in the healthy animals were significantly different (P less than 0.05) from those of the infected dogs. The drug residues were still detectable in the tissues of the animals 10 days after drug administration.     

Protective efficacy of isometamidium chloride and diminazene aceturate against natural Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax infections in cattle under a suppressed tsetse population in Uganda

The protective efficacy of isometamidium chloride (ISMM) and diminazene aceturate (DIM) against Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax infections in cattle under a suppressed tsetse population was assessed in southeast Uganda. A total of 66 and 57 trypanosome-infected cattle were treated with ISMM and DIM, respectively together with 177 trypanosome-free animals not treated were followed for 12 months, checked every 4 weeks. There was no statistical difference in the mean time to infection with any trypanosome species in animals treated with ISMM or DIM. However, the mean time to trypanosome infection was significantly longer for treated animals than controls. The mean time to infection with each of the three trypanosome species differed significantly, with the average time to T. vivax infection the lowest, followed by T. congolense and then T. brucei. The protective efficacy of DIM was as good as that of ISMM; implying curative treatments against trypanosomosis are sufficient for combination with tsetse control. Isometamidium chloride or DIM had the highest impact on T. brucei and T. congolense infections in cattle.     

Response of Trypanosoma congolense in goats to single and double treatment with diminazene aceturate.

Diminazene aceturate is one of a limited number of compounds currently marketed for treatment of trypanosomiasis in cattle, sheep and goats. The pharmacokinetics of the compound in goats suggest that double treatment with diminazene aceturate might enhance the compound's therapeutic activity. A study was therefore conducted in goats using two clones of Trypanosoma congolense, IL 3274 and IL 1180, which were previously shown to be resistant and sensitive, respectively, to single treatment with diminazene aceturate. The results indicated that, as compared to single treatment, double treatment with diminazene aceturate at a dose of 7.2 mg kg-1 bodyweight, at either eight or 24 hour intervals, did not greatly enhance the therapeutic activity of the drug. Furthermore, treatment with the same drug dose eliminated infections with T congolense IL 3274 when treatment was administered 24 hours after infected Glossina morsitans centralis had fed, but failed to do so if treatment was delayed until after goats were detected to be parasitaemic. This suggests that failure of T congolense IL 3274 to respond to treatment with diminazene may not be due to drug resistance per se.     

Suspected resistance of Trypanosoma species to diminazene aceturate on a cattle farm in Nigeria

Tropical Animal Health and Production - African animal trypanosomiasis is a major cause of mortality and economic losses for the livestock industry in Nigeria. Chemotherapy has been the most...     

The effects of diminazene aceturate on systemic blood pressure in clinically healthy adult dogs

Diminazene aceturate is a commonly used antibabesial agent. It has been postulated that diminazine may induce a decrease in blood pressure and exacerbate the hypotension presented in dogs with babesiosis. This study was undertaken to assess the effect of diminazine aceturate on the blood pressure of healthy dogs. Six healthy German shepherd dogs between 18 and 24 months of age with a mean weight of 30.4 +/- 2.75 kg were used. Blood pressure was directly measured at the following time intervals: -5, 0, 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 90 and 120 minutes after treatment with diminazine aceturate (4.2 mg/kg) intramuscularly. No statistical difference (P > 0.05) was found in blood pressure between any of the time intervals. An increase in heart rate was seen 5 minutes after the administration of diminazine aceturate but no change in blood pressure was evident. This study concluded that diminazene aceturate in its current formulation with antipyrine does not alter blood pressure in healthy adult dogs.     

Validation of a polymerase chain reaction assay for monitoring the therapeutic efficacy of diminazene aceturate in trypanosome-infected sheep

The diagnostic performance of a polymerase chain reaction assay (PCR) for monitoring the effectiveness of aceturate diminazene treatment was compared with those of an antibody-detection ELISA test and the buffy-coat technique using sheep experimentally infected with either savannah-type or forest-type Trypanosoma congolense or T. vivax. Within the period of infection, the PCR using specific savannah-type T. congolense primers showed a significant higher diagnostic sensitivity (p<0.05) than the buffy-coat technique. Both techniques gave closed results for detecting forest-type T. congolense or T. vivax infections. Following trypanocidal treatment, the PCR showed that specific product disappeared definitively 1 or 2 days later in animals in which a decrease of the antibody level and a significant improvement of the red packed cell volume were observed. The occurrence of relapse infection was detected by the PCR in one animal infected by T. vivax on day 19 post-treatment and confirmed by the persistence and increasing antibody level whereas the buffy-coat technique detected parasites 42 days later. Then, the PCR signals remained positive on several occasions while parasitaemia was detected only two times.The application of PCR combined with the antibody detection appeared to provide a useful tool as compared to the buffy-coat technique for monitoring the effectiveness of trypanocidal treatment.     

Ranitidine bismuth citrate and clarithromycin, alone or in combination, for eradication of Helicobacter mustelae infection in ferrets

OBJECTIVE: To determine whether ranitidine bismuth citrate, clarithromycin, or a combination of ranitidine bismuth citrate and clarithromycin would be efficacious in eradication of Helicobacter mustelae infection in ferrets. ANIMALS: 60 seven-month-old ferrets. PROCEDURE: To determine dosages of clarithromycin and ranitidine bismuth citrate that would suppress growth of, but not eradicate infection with, H mustelae, ferrets (n = 6/group) were treated p.o. with clarithromycin or ranitidine bismuth citrate at various dosages. Efficacy of treatment was then determined by treating ferrets with clarithromycin alone, ranitidine bismuth citrate alone, or clarithromycin and ranitidine bismuth citrate. Gastric biopsy specimens were obtained before, during, and at various times after treatment and submitted for quantitative bacterial culture and histologic evaluation. Minimum concentrations of clarithromycin that inhibited 90% of the growth of isolates obtained before and after treatment were determined. RESULTS: Dosages of clarithromycin and ranitidine bismuth citrate that suppressed growth of H mustelae were 12.5 and 24 mg/kg of body weight, p.o., every 8 hours, respectively. Infection was not eradicated in ferrets treated with ranitidine bismuth citrate alone but was eradicated in all 6 ferrets treated with clarithromycin and ranitidine bismuth citrate and in 4 of 6 treated with clarithromycin alone. A decrease in susceptibility to clarithromycin was detected for H mustelae isolates obtained after treatment. Mild or moderate antral gastritis was observed even in ferrets from which infection was eradicated. CONCLUSIONS AND CLINICAL RELEVANCE: A combination of ranitidine bismuth citrate and clarithromycin was efficacious in eradicating H mustelae infection from ferrets.