Screening the Medicines for Malaria Venture Pathogen Box against piroplasm parasites.

Diminazene aceturate (DA) and imidocarb dipropionate are commonly used in livestock as antipiroplasm agents. However, toxic side effects are common in animals treated with these two drugs. Therefore, evaluations of novel therapeutic agents with high efficacy against piroplasm parasites and low toxicity to host animals are of paramount importance. In this study, the 400 compounds in the Pathogen Box provided by the Medicines for Malaria Venture foundation were screened against Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi. A fluorescence-based method using SYBR Green 1 stain was used for initial in vitro screening and determination of the half maximal inhibitory concentration (IC50). The initial in vitro screening performed using a 1 µM concentration as baseline revealed nine effective compounds against four tested parasites. Two "hit" compounds, namely MMV021057 and MMV675968, that showed IC50 < 0.3 µM and a selectivity index (SI)> 100 were selected. The IC50s of MMV021057 and MMV675968 against B. bovis, B. bigemina, T. equi and B. caballi were 23, 39, 229, and 146 nM, and 2.9, 3, 25.7, and 2.9 nM, respectively. In addition, a combination of MMV021057 and DA showed additive or synergistic effects against four tested parasites, while combinations of MMV021057 with MMV675968 and of MMV675968 with DA showed antagonistic effects. In mice, treated with 50 mg/kg MMV021057 and 25 mg/kg MMV675968 inhibited the growth of Babesia microti by 54 and 64%, respectively, as compared to the untreated group on day 8. Interestingly, a combination treatment with 6.25 mg/kg DA and 25 mg/kg MMV021057 inhibited B. microti by 91.6%, which was a stronger inhibition than that by single treatments with 50 mg/kg MMV021057 and 25 mg/kg DA, which showed 54 and 83% inhibition, respectively. Our findings indicated that MMV021057, MMV675968, and the combination treatment with MMV021057 and DA are prospects for further development of antipiroplasm drugs.

Clinical, haematological and therapeutic studies on tropical theileriosis in water buffaloes (Bubalus bubalis) in Egypt.

Thirty buffaloes naturally infected with Theileria annulata and 10 parasitologically free controls were used to determine the potential clinical, haematological and therapeutic impact of tropical theileriosis in Egypt. The clinical signs in the infected buffaloes were pyrexia (40.5-41.5 degrees C), enlargement of superficial lymph nodes, slight nasal and ocular discharges, salivation, anaemia and respiratory distress. Eye lesions also were recorded. There was a significant decrease in erythrocyte counts and haemoglobin content and a significant decrease in total leucocyte counts in infected buffaloes compared to controls. Early treatment with buparvaquone was 100% effective in eliminating the protozoan parasites from the blood and lymph nodes and led to an improvement in the clinical state whereas treatment in the later stages of the disease whilst eliminating the parasites failed to improve the clinical condition of the animal.

Biochemical profiles in Friesian cattle naturally infected with Theileria annulata in Saudi Arabia.

The biochemical parameters were studied in adult and young Friesian cattle naturally infected with Theileria annulatta in the Qassim Region, Saudi Arabia. Forty-three clinical cases of tropical theileriosis were studied, together with 40 clinically healthy Friesian cattle. Cattle clinically infected with T. annulata had significantly lower serum total protein, albumin, globulin, creatinine, calcium, phosphorus, magnesium, potassium, iron and copper concentrations and significantly higher AST activity and bilirubin concentration than the healthy cattle.

Immunisation against Theileria parva in eastern Zambia: influence of maternal antibodies and demonstration of the carrier status.

Immunisation of calves by the infection and treatment method (I & T) has been extensively used in the eastern province of Zambia to control East Coast fever (ECF), a protozoan tick-borne disease. This paper presents the results of a field longitudinal study, which included a total of 148 Angoni calves. After immunisation against ECF, they were monitored for a full rainy season, coinciding with the main peak of activity of the vector of Theileria parva, the tick Rhipicephalus appendiculatus. Dysimmunisation (acute reaction generated by I & T immunisation), seroconversion and mortality are among the parameters recorded. The effect of maternal antibodies on these parameters was analysed and also studied in experimental conditions on two calves. Before immunisation, young calves had a higher seroprevalence than older animals (maternal antibodies) but their post-immunisation seroprevalence was lower. There was no evidence that their immunoprotection was weaker but this indicates that the post-immunisation seroconversion is probably not a reliable tool to monitor the efficacy of calf immunisation. The carrier state of cattle after immunisation was investigated in experimental conditions on three bovines whereas in the field, the infection prevalence in the ticks was estimated using the relation between the tick burden and the T. parva contacts with the calves. The ability of larval and nymphal R. appendiculatus ticks to pick-up T. parva from carriers and to transmit it to naïve animals after moulting was assessed. It was found that both instars are able to transmit clinical and lethal ECF but that the prevalence of T. parva infection in nymphs is much lower than in adults, confirming the primary role of adults in the transmission of ECF in endemic conditions. Similar results were obtained from the field whereby the ECF peak corresponds with the peak of adult R. appendiculatus activity. The infection prevalence in the ticks was however much lower in the field than in experimental conditions indicating that an important proportion of them feed on alternative hosts. Old ticks seemed to have lost part of their infectivity.

Changes in blood chemical components during experimentally induced Theileria annulata infections in cattle.

During the course of infection of calves with Theileria annulata levels of glucose, calcium, proteins, phosphorus, magnesium and potassium decreased and those of bilirubin, cholesterol and alkaline phosphatase increased with no changes in sodium and acid phosphatase levels. Several of these biochemical alterations have been explained in relation to the injury to the liver and other organs.

A comparative study of the disease in cattle caused by Theileria parva or T. lawrencei: II. Hematology, clinical chemistry, coagulation studies and complement.

Theileria lawrencei tick-derived stabilate infection of 8 cattle resulted in the development of panleukopenia and hypoproteinemia. In addition to these changes, T. parva infection caused mild normocytic, normochromic, non-responsive anemia at either of two dose rates. Disseminated intravascular coagulation, as indicate by positive protamine paracoagulation tests, prolonged prothrombin and partial thromboplastin times, and thrombocytopenia, developed in cattle infected with either of the Theileria spp., and was probably an important intermediary mechanism leading to death. Infection of calves with a high dose of T. parva stabilate resulted in more rapid onset of clinico-pathologic changes than did the low dose infection. Theileria lawrencei infection produced a severe, acute syndrome, the clinico-pathologic alterations of which varied in time of onset and severity between those of the T. parva high dose and low dose groups.