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.     

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.     

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.     

Parasitaemia and clinical manifestations in Trypanosoma brucei infected dogs.

Four dogs were infected with Trypanosoma brucei (Mkar strain) while another four were used as uninfected controls. Two of the dogs showed acute disease and died in the first wave of parasitaemia on days 7 and 8 post infection (PI) while the other two died from the sub-acute disease on days 24 and 28 PI corresponding to the second wave of parasitaemia. In the first wave of parasitaemia there was a sharp decrease in the packed cell volume, red blood cell, haemoglobin, total leucocytes, eosinophil, neutrophil and lymphocyte values, but during the period of low parasitaemia there was a slight recovery of the values of total leucocytes and lymphocytes although these and the other values showed a continuous decrease during the second wave of parasitaemia. In contrast, there was a consistent monocytosis in both acute and sub-acute diseases. The general picture was that of loss of condition, anaemia, leucopenia, monocytosis, ocular impairment, elevated temperature, pulse and respiratory rates, the difference between the acute and sub-acute diseases being in the degree of intensity. The degree of anaemia noted and the circulatory disturbances associated with the infection could have caused the death of all the infected dogs.     

Relapse infection after chemotherapy in dogs experimentally infected with Trypanosoma brucei brucei

Studies on relapsing Trypanosoma brucei brucei infections in dogs after Berenil treatment revealed that the first relapse occurred 13 to 64 days after chemotherapy and 36 to 79 days after inoculation. A second relapse infection was observed in two dogs 43 and 60 days after a second Berenil treatment. During the aparasitaemic period following chemotherapy in four dogs, successful transmission (as evidenced by subsequent parasitaemia) following the intraperitoneal inoculation of homogenate of brain from two of the dogs into recipient rats was obtained. Transmission with blood collected just before the animals were sacrificed was, however, negative. Hornogenates of other organs (liver, spleen, eyes, testes, kidneys, heart and lymph node) were also non-infective. One dog inoculated with relapsed trypanosomes and treated with Berenil soon after showing parasitaemia was completely cured of the infection. It was considered that the brain is the source of relapse in T b brucei infection after Berenil therapy and that the relapse was not due to drug resistance.     

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.     

Erythrocyte response to Trypanosoma brucei in experimentally infected dogs.

Trypanosoma brucei infection produced an acute and fatal disease in Nigerian mongrel dogs due to a rapidly developing anaemia. Infected dogs responded with increased reticulocytosis, which was not sustained with chronicity. In comparison the response to artificially-induced haemolytic anaemia was progressive, marked and sustained. The anaemia of T. brucei infection of dogs was either normocytic normochromic in acute infection or microcytic normochromic in chronic infection. Artificially-induced haemolytic anaemia was either macrocytic normochromic or normocytic normochromic. The erythropoietic potential of plasma in vivo in mice increased in T. brucei-infected dogs except at the terminal parasitaemia. The anaemia in Trypanosoma brucei-infected dogs is therefore initially responsive but becomes poorly involved with chronicity.     

Trypanosoma congolense infection in two dogs

Trypanosomiasis, caused by Trypanosoma congolense, was diagnosed for the first time in Israel in two boxer dogs imported from Kenya. The dogs developed clinical signs two days after arrival and succumbed to the disease within four days. The major clinical and clinicopathological findings included anaemia, haemorrhages, lymphadenomegaly, hepatosplenomegaly and neurological signs. Histopathology showed lymphocytic-plasmacytic infiltration in the skin, brain, meninges, kidney and liver.     

The pathogenesis of anaemia in goats experimentally infected with Trypanosoma congolense or Trypanosoma brucei: use of the myeloid:erythroid ratio

The present study examined the development of anaemia in Small East African goats experimentally infected with Trypanosoma congolense or Trypanosoma brucei. Experimental goats received a primary trypanosome challenge on day 0, treated with diminazene aceturate on day 49 and received a secondary trypanosome challenge on day 77 of the 136-day experiment. Both primary and secondary challenges were characterised by reduced peripheral erythrocyte counts, fall in packed cell volume (PCV), hypohaemoglobinaemia and reductions in the myeloid:erythroid ratios (M:E) compared with the uninfected goats. The progressive reduction in the M:E ratios denoted increased erythrogenesis in response to increased destruction of erythrocytes in blood by infecting trypanosomes or their products. The more rapid fall in M:E ratio in T. congolense infections shows that this parasite causes more severe clinical pathological effects in goats than T. brucei.     

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.     

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.     

Pathogenicity of Trypanosoma brucei brucei in experimentally infected pigs.

An experimental infection of 4-to 5-month old pigs with a stock of Trypanosoma brucei brucei resulted in a high parasitaemia, anorexia, pyrexia and a decline in the packed cell volume by one third. Nervous sign of circling and wobbling of the hind legs occurred in one of the pigs which at necropsy revealed a very severe meningo-encephalitis and the presence of trypanosomes in the brain. These results confirm that T. b. brucei might cause a severe disease in pigs.     

伊氏锥虫、马媾疫锥虫、布氏锥虫、刚果锥虫的18S rDNA部分序列测定与系统发育关系

对伊氏锥虫（Trypanosoma evansi）：新疆株（XJCA）、湖北株（HBM）、云南株（YNB）、广东株（GDB2）；马媾疫锥虫（Trypanosoma equiperdum）、布氏锥虫（Trypanosoma brucei）、刚果锥虫（Trypanosoma congolense）提取基因组DNA，根据已报道的伊氏锥虫株18SrDNA基因序列设计合成引物，用PCR扩增了锥虫虫株基因组DNA,伊氏锥虫新疆株、湖北株、云南株、广东株、布氏锥虫、刚果锥虫均为373bp的片段；马媾疫锥虫为372bp的片段，PCR产物经电泳鉴定后用试剂盒回收纯化，纯化后PCR产物经连接、转化后测序，将测得的序列用DNAMAN软件分析并与国外已发表的相应序列进行了同源性比较，并绘制了系统发育进化树。结果与国外AJ009153、AJ223564、D89527株同源性达到99％～100％，与另外11株同源性75％。本研究为锥虫分子流行病学研究及分类研究打下基础。     

Recirculation of Trypanosoma brucei brucei in cattle after T. congolense challenge by tsetse flies

The effect of challenging cattle, chronically infected with Trypanosoma brucei brucei, with T. congolense on the development of the T. b. brucei infection was investigated. For this purpose, nine experimental animals were first infected with T. b. brucei through the bites of infected tsetse flies. Once the T. b. brucei had developed into a chronic infection, that was difficult to detect using routine parasitological diagnostic tools, seven of the experimental animals were challenged by tsetse flies infected with T. congolense. Two of the animals infected with T. b. brucei were kept as control. The infection with T. congolense resulted in a sudden increase in the parasitaemia of T. b. brucei. In the T. b. brucei control animals, on the other hand, the parasitaemia remained below the level of detection. The epidemiological repercussions of this increase in the parasitaemia of T. b. brucei after infection with T. congolense are discussed.     

Interference in the establishment of tsetse-transmitted Trypanosoma congolense, T. brucei or T. vivax superinfections in goats already infected with T. congolense or T. vivax

An interference phenomenon that delays superinfection with a trypanosome species different from that used for the initial infection has been found to occur in goats. Following tsetse transmission of Trypanosoma brucei to goats already infected with T. congolense, there was a delay in chancre development, as well as in the appearance of T. brucei and anti-T. brucei antibodies in the blood when compared to previously uninfected goats. However, there was no delay in the establishment of a tsetse-transmitted superinfection with T. vivax in goats already infected with either T. congolense or in animals already infected with a different serodeme of T. vivax.     

Pathological changes in male genitalia of cattle infected with Trypanosoma vivax and Trypanosoma congolense

Samples for histological studies were taken from the genitalia of 14 bulls (five infected with Trypanosoma vivax, five with T. congolense and four uninfected control animals), slaughtered 12, 22 or 30 weeks post-infection. Infection with Y58 strain of T. vivax and strain 2295 of T. congolense caused various grades of lesions in the male reproductive organs, especially the testes and epididymides. T. congolense produced more severe degenerative changes than T. vivax. It is concluded that in long-standing cases, the result of trypanosome infection is either serious infertility or even sterility.     

Depressed immunoconglutinin responses in calves experimentally infected with Trypanosoma congolense

Contrary to expectation, immunoconglutinin levels failed to rise significantly in calves infected with Trypanosoma congolense. In addition, it was shown that trypanosome infection appeared to inhibit the immunoconglutinin response to Brucella abortus strain 19. The possible reasons for these findings are discussed.     

Trypanosomes in the lymph nodes of cattle and sheep infected with Trypanosoma congolense

The prefemoral lymph nodes of two calves and a sheep infected with a stock of Trypanosoma congolense transmitted by Glossina morsitans were examined histologically for the presence of trypanosomes. Ten days after infection trypanosomes were found in the subcapsular sinuses of the nodes of a calf and the sheep but parasites were absent from the blood at this time. Trypanosomes were also detected in the prefemeral lymph node of the other calf on examination 30 days after infection, when parasites were also present in the blood. These observations provide further evidence that extravascular foci of trypanosomes develop in infections with T congolense and indicate that it should not be regarded as a strict plasma parasite.