Diminazene resistance in Trypanosoma congolense is not caused by reduced transport capacity but associated with reduced mitochondrial membrane potential.

Trypanosoma congolense is a principal agent causing livestock trypanosomiasis in Africa, costing developing economies billions of dollars and undermining food security. Only the diamidine diminazene and the phenanthridine isometamidium are regularly used, and resistance is widespread but poorly understood. We induced stable diminazene resistance in T. congolense strain IL3000 in vitro. There was no cross-resistance with the phenanthridine drugs, melaminophenyl arsenicals, oxaborole trypanocides, or with diamidine trypanocides, except the close analogs DB829 and DB75. Fluorescence microscopy showed that accumulation of DB75 was inhibited by folate. Uptake of [3 H]-diminazene was slow with low affinity and partly but reciprocally inhibited by folate and by competing diamidines. Expression of T. congolense folate transporters in diminazene-resistant Trypanosoma brucei brucei significantly sensitized the cells to diminazene and DB829, but not to oxaborole AN7973. However, [3 H]-diminazene transport studies, whole-genome sequencing, and RNA-seq found no major changes in diminazene uptake, folate transporter sequence, or expression. Instead, all resistant clones displayed a moderate reduction in the mitochondrial membrane potential Ψm. We conclude that diminazene uptake in T. congolense proceed via multiple low affinity mechanisms including folate transporters; while resistance is associated with a reduction in Ψm it is unclear whether this is the primary cause of the resistance.

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

African animal trypanosomiasis is a major cause of mortality and economic losses for the livestock industry in Nigeria. Chemotherapy has been the most reliable option for cattle herders, and the most commonly found drug on the market is diminazene aceturate. To ascertain the long-term efficacy of this compound, we sampled a cattle herd in Ogun State, Nigeria, 2 months after they were treated with diminazene aceturate. The ITS-PCR results revealed 19 positives for trypanosome DNA out of the 79 samples tested (24.1%, 95% CI 16.0-34.5). Seventeen out of the total 19 positives were Trypanosoma congolense (21.5%, 95% CI 13.9-31.8). Mixed infections were also observed. Therefore, the persistence of bovine trypanosomiasis at this Nigerian cattle farm despite treatment could be due to diminazene aceturate resistant trypanosomes being present in the herd.

Practices of cattle keepers of southwest Nigeria in relation to bovine trypanosomosis.

Significant increases in human and livestock populations coupled with agricultural practices have changed the socioeconomic perspectives of livestock diseases. Evaluating the socioeconomic impact of bovine trypanosomosis and its vectors (Glossina, Tabanus and Stomoxys) from the perspective of the livestock owners is of great significance. Participatory rural appraisal was conducted among 209 livestock owners (focus groups) to determine the behavioural practices of animal husbandary to bovine trypanosomosis. In Nigeria, common Trypanosoma species found in cattle are Trypanosoma vivax, Trypanosoma congolense and Trypanosoma brucei. Trypanosomosis peaks were reported by owners to be in the months of March-August. A total of 70.8% (95%CI 64.32-76.56%) cattle owners perceived trypanosomosis as a major disease in their herd, 13.4% (95%CI 9.43-18.68%) practiced transhumance in the wet season and 93.9% (95%CI 88.58-96.92%) make use of trypanocides, and approximately US$ 8.4 million is spent annually on trypanocides in southwest Nigeria livestock industry. About 60.5% (95%CI 51.84-68.48) make use of insecticides against transmitting vectors, and only 1.9% (95%CI 0.75-4.82%) have ever heard of any form of government intervention scheme. Estimated losses ≥ US$ 426 (80-100% loss) can be incurred on a single animal depending on the size and market value. There is significant increase (16.2%, 95%CI 11.15-23.00%, P < 0.05) in the mortality rate of bovine trypanosomosis when compared to other livestock diseases. It will therefore be useful to involve the livestock owners with devising new and integrated measures for reducing the impact of this trypanosomosis.

A 3D-QSAR Study on the Antitrypanosomal and Cytotoxic Activities of Steroid Alkaloids by Comparative Molecular Field Analysis.

As part of our research for new leads against human African trypanosomiasis (HAT), we report on a 3D-QSAR study for antitrypanosomal activity and cytotoxicity of aminosteroid-type alkaloids recently isolated from the African medicinal plant Holarrhena africana A. DC. (Apocynaceae), some of which are strong trypanocides against Trypanosoma brucei rhodesiense (Tbr), with low toxicity against mammalian cells. Fully optimized 3D molecular models of seventeen congeneric Holarrhena alkaloids were subjected to a comparative molecular field analysis (CoMFA). CoMFA models were obtained for both, the anti-Tbr and cytotoxic activity data. Model performance was assessed in terms of statistical characteristics (R², Q², and P² for partial least squares (PLS) regression, internal cross-validation (leave-one-out), and external predictions (test set), respectively, as well as the corresponding standard deviation error in prediction (SDEP) and F-values). With R² = 0.99, Q² = 0.83 and P² = 0.79 for anti-Tbr activity and R² = 0.94, Q² = 0.64, P² = 0.59 for cytotoxicity against L6 rat skeletal myoblasts, both models were of good internal and external predictive power. The regression coefficients of the models representing the most prominent steric and electrostatic effects on anti-Tbr and for L6 cytotoxic activity were translated into contour maps and analyzed visually, allowing suggestions for possible modification of the aminosteroids to further increase the antitrypanosomal potency and selectivity. Very interestingly, the 3D-QSAR model established with the Holarrhena alkaloids also applied to the antitrypanosomal activity of two aminocycloartane-type compounds recently isolated by our group from Buxus sempervirens L. (Buxaceae), which indicates that these structurally similar natural products share a common structure⁻activity relationship (SAR) and, possibly, mechanism of action with the Holarrhena steroids. This 3D-QSAR study has thus resulted in plausible structural explanations of the antitrypanosomal activity and selectivity of aminosteroid- and aminocycloartane-type alkaloids as an interesting new class of trypanocides and may represent a starting point for lead optimization.

Applicability of plant-based products in the treatment of Trypanosoma cruzi and Trypanosoma brucei infections: a systematic review of preclinical in vivo evidence.

Chagas disease and sleeping sickness are neglected tropical diseases closely related to poverty, for which the development of plant-derived treatments has not been a promising prospect. Thus, we systematicaly review the preclinical in vivo evidence on the applicability of plant-based products in the treatment of Trypanosoma cruzi and Trypanosoma brucei infections. Characteristics such as disease models, treatments, toxicological safety and methodological bias were analysed. We recovered 66 full text articles from 16 countries investigating 91 plant species. The disease models and treatments were highly variable. Most studies used native (n = 36, 54·54%) or exotic (n = 30, 45·46%) plants with ethnodirected indication (n = 45, 68·18%) for trypanosomiasis treatment. Complete phytochemical screening and toxicity assays were reported in only 15 (22·73%) and 32 (48·49%) studies, respectively. The currently available preclinical evidence is at high risk of bias. The absence of or incomplete characterization of animal models, treatment protocols, and phytochemical/toxicity analyses impaired the internal validity of the individual studies. Contradictory results of a same plant species compromise the external validity of the evidence, making it difficult determine the effectiveness, safety and biotechnological potential of plant-derived products in the development of new anti-infective agents to treat T. cruzi and T. brucei infections.

Trypanosomiasis challenge estimation using the diminazene aceturate (Berenil) index in Zebu in Gabon.

A longitudinal study was conducted within a cattle ranch in Gabon to determine the diminazene aceturate (Berenil) index (DAI) in a group of Zebu, raised under low tsetse density; this measure providing an assessment of trypanosomiasis risk. The objective was to evaluate the trypanosomiasis pressure thus informing trypanosomiasis control methods and cattle management. Twenty female adult Zebu were monitored for 24 weeks during the dry season. Blood samples were collected on a weekly basis and subjected to parasitological and haematological analysis (n = 480), using the buffy-coat method and the packed cell volume value (PCV), respectively, infected animals were treated with a single intramuscular injection of diminazene aceturate (8 mg/kg). Twenty-nine single infectious events were recorded and a DAI of 1.45 was calculated. Two trypanosome species were identified: Trypanosoma congolense (96.2%) and Trypanosoma vivax (3.8%). The mean PCV value of the infected animals was lower (26.6) compared to non-infected animals (32.0). This study shows that DAI may be a useful tool to assess trypanosomiasis. However, this is a time-consuming method that may be improved by using randomly selected sentinel animals to adapt the chemoprophylactic schemes, hence decreasing the costs and the drug resistance risk.

The animal trypanosomiases and their chemotherapy: a review.

Pathogenic animal trypanosomes affecting livestock have represented a major constraint to agricultural development in Africa for centuries, and their negative economic impact is increasing in South America and Asia. Chemotherapy and chemoprophylaxis represent the main means of control. However, research into new trypanocides has remained inadequate for decades, leading to a situation where the few compounds available are losing efficacy due to the emergence of drug-resistant parasites. In this review, we provide a comprehensive overview of the current options available for the treatment and prophylaxis of the animal trypanosomiases, with a special focus on the problem of resistance. The key issues surrounding the main economically important animal trypanosome species and the diseases they cause are also presented. As new investment becomes available to develop improved tools to control the animal trypanosomiases, we stress that efforts should be directed towards a better understanding of the biology of the relevant parasite species and strains, to identify new drug targets and interrogate resistance mechanisms.

Economic burden of bovine trypanosomosis in three villages of Metekel zone, northwest Ethiopia.

The study was carried out to assess the economic burden of trypanosomosis in three villages of the Metekel zone in 2009. The disease was found to cause substantial economic losses through cattle mortality, drug purchase, and draft power loss of infected oxen. The farmers in the area were spending a significantly (p < 0.05) higher amount of money for the treatment of trypanosomosis than all other diseases combined. The overall mortality rate of cattle due to trypanosomosis was 4.4%. The mortality was significantly higher (p < 0.05) in an area where trypanosomosis prevalence was also higher. Many of the farmers prioritized losses of draft power as the most important impact of the disease. The overall prevalence of the disease was 12.1%. The disease burden was significantly (p < 0.05) higher in the rainy season than at other times of the year. In general, farmers had good knowledge on the signs and seasonality of trypanosomosis. Thus, tsetse suppression activities that involve the local community can be an important tool towards minimizing the economic burden of the disease in the area.

An evaluation of melarsomine hydrochloride efficacy for parasitological cure in experimental infection of dairy cattle with Trypanosoma evansi in Thailand.

Melarsomine hydrochloride can cure Trypanosoma evansi infection in camels at a dose of 0·25 mg/kg, but at that dose relapses occur in cattle. In our study, the efficacy of an intramuscular injection of melarsomine hydrochloride at 0·5 mg/kg was assessed in 3 normal and 3 splenectomized dairy cattle experimentally infected with a stock of T. evansi from Thailand. The animals were monitored for 5 months by haematocrit centrifugation, blood- or cerebrospinal fluid-mouse inoculation, polymerase chain reaction, the card agglutination test (CATT) for T. evansi, and the enzyme-linked immunosorbent assay­T. evansi. Parasitological and DNA tests became and remained negative just after treatment. By the end of the experiment, CATT was negative and ELISA scores were below or very close to the cut-off value. One of the splenectomized cattle died from anaplasmosis during the experiment, but tested negative for surra. It was concluded that the parasites had been cleared from the cattle, and melarsomine hydrochloride at 0·5 mg/kg can be recommended for treatment against T. evansi infection in dairy cattle in Thailand. Further work is necessary to validate the efficacy of the treatment in the event of confirmed CSF-infection.

Composition, and antimicrobial and remarkable antiprotozoal activities of the essential oil of rhizomes of Aframomum sceptrum K. Schum. (Zingiberaceae).

The essential oil from the rhizomes of Aframomum sceptrum (Zingiberaceae) was analyzed by GC/MS, and its major constituents were found to be ß-pinene (12.7%), caryophyllene oxide (10.0%), and cyperene (6.0%). The oil was also evaluated for antimicrobial activities, in comparison with ß-pinene, caryophyllene oxide, and the leaf essential oil of Melaleuca alternifolia (Myrtaceae). The A. sceptrum essential oil exhibited bacteriostatic activity against the Gram-positive bacteria Bacillus subtilis, Staphylococcus epidermidis, and S. aureus, but not against Gram-negative bacteria. Moreover, it showed mild fungicidal activity against Candida albicans and Aspergillus fumigates, and remarkable antiprotozoal activity against Trypanosoma brucei brucei (MLC of 1.51 µl/ml) and Trichomonas vaginalis (IC(50) of 0.12±0.02 and MLC of 1.72 µl/ml).

William Horner Andrews (1887-1953)- first professor of physiology at Onderstepoort.

W H Andrews qualified as a veterinarian in London in 1908 and was recruited soon after, in 1909, by Sir Arnold Theiler to join the staff of the newly established veterinary laboratory at Onderstepoort. After initial studies on the treatment of trypanosomosis and on snake venoms he was deployed by Theiler in 1911 to start research on lamsiekte (botulism)at a field station on the farm Kaffraria near Christiana, where he met and married his wife Doris. After a stint as Captain in the SA Veterinary Corps during World War I he succeeded D T Mitchell as head of the Allerton Laboratory in 1918, where he excelled in research on toxic plants, inter alia identifying Matricaria nigellaefolia as the cause of staggers in cattle. When the Faculty of Veterinary Science was established in 1920 he was appointed as the first Professor of Physiology. After the graduation of the first class in 1924, and due to health problems, he returned to the UK, first to the Royal Veterinary College and then to the Weybridge Veterinary Laboratories of which he became Director in 1927. After his retirement in 1947 he returned to South Africa as a guest worker at Onderstepoort where he again became involved in teaching physiology when Prof. Quin unexpectedly died in 1950. Andrews died in Pretoria in 1953 and was buried in the Rebecca Street Cemetery.

Follow-up of dairy cattle naturally infected by Trypanosoma vivax after treatment with isometamidium chloride.

Trypanosoma vivax infections cause nonspecific clinical signs in cattle associated with aparasitemic intervals, making disease diagnosis a challenge. In Brazil, diminazene aceturate and isometamidium chloride (ISM) are available to treat bovine trypanosomosis. The objective of this study was to follow-up, by molecular and serological techniques, dairy cattle naturally infected by T. vivax after ISM treatment. Thirty cattle naturally infected with T. vivax received two applications of ISM, at a dosage of 1.0 mg/kg intramuscularly, on days 0 and 150. For T. vivax diagnosis, EDTA-blood and serum samples were evaluated on 0, 7, 15, 30, 60, 90, 120, 150, 180, 210, and 240 days after treatment PCR, Loop-mediated isothermal amplification (LAMP) and ELISA. Animals with persistent detection of T. vivax DNA by both PCR and LAMP were found and continuous detection of anti-T. vivax IgG antibodies by ELISA, suggesting the presence of T. vivax resistance to ISM. The combination of LAMP and ELISA tests can prevent misdiagnosis of the parasite clearance in treated cattle, contributing to better disease control. This is the first experiment that demonstrates the persistence infection of T. vivax under ISM treatment in a natural infected herd and evidence of ISM chemotherapy-resistant T. vivax in Brazil.

Trypanosomiasis: an approach to chemotherapy by the inhibition of carbohydrate catabolism.

When the infected mammalian host of Trypanosoma brucei brucei is injected with a solution of the iron chelator salicyl hydroxamic acid and glycerol, the aerobic and anaerobic glucose catabolism of the parasite is blocked and the parasite is rapidly destroyed.

Economic assessment of the performance of trypanotolerant cattle breeds in a pastoral production system in Kenya.

Cattle are the major source of food security and income for pastoral farmers in sub-Saharan Africa. However, infectious and parasitic diseases remain a major constraint to improved cattle productivity in the region. The use of animal health economics to support decision-making on cost-effective disease control options is increasingly becoming important in the developing world. Trypanotolerant indigenous Orma/zebu cattle in a trypanosomosis-endemic area of Kenya were evaluated for economic performance using gross-margin analysis and partial-farm budgeting. Orma/zebu and Sahiwal/zebu cross-bred cattle were exposed to similar husbandry practices and monitored for growth rate, incidence of common infections (trypanosomosis, anaplasmosis, babesiosis, East Coast Fever and helminthosis) and the cost of treatment assessed. Interview questionnaires were also used to assess the preference rating of the 2 breeds. Results indicated that incidence of infection was trypanosomosis 3%, anaplasmosis 58%, babesiosis 11%, East Coast Fever 22% and helminthosis 28%, with no significant difference between breeds. The Orma/zebu and Sahiwal/zebu breeds had comparable economic benefits, hence a pastoralist in Magadi division is likely to get similar returns from both breeds. This study therefore recommends adoption of not only the Sahiwal/zebu but also the Orma/zebu breed for cattle improvement in trypanosomosis endemic areas and conservation of indigenous genetic resources.

How rational drug use reduces trypanosome infections in cattle in chemo-resistance hot-spot villages of northern Togo.

The study assessed an integrated trypanosomosis control strategy in drug-resistant hotspot villages of northern Togo. This strategy comprised (i) rational trypanocidal drug use in symptomatic cattle, (ii) vectors and ticks control by targeted bi-monthly insecticidal spraying of the lower body parts of cattle and (iii) strategic deworming with Albendazole in the beginning and the end of the rainy season. The program was implemented between June 2014 and October 2015 in four villages in northern Togo, which had been previously identified as drug resistant hotspots for diminazene diaceturate (DA) and isometamidium chloride (ISM). The integrated control strategy was implemented in eight cattle herds at risk of the disease from two villages. Twelve herds from two other villages served as controls where trypanosomosis management and deworming remained under control of the farmers. Trypanocidal drug use during the study period was recorded by the intervention team based on the farmers' reports and own observations. Cattle herds were followed-up for trypanosomosis symptoms which were recorded at 3 to 4-month intervals, while extensive trypanosome diagnostics and recording of the packed cell volume were done before and after the intervention. Intervention herds had a significantly lower risk of trypanosome infection with a risk ratio of 0.18 (95% CI: 0.04, 0.91; p = 0.03), but no significant effect on mean packed cell volume was observed. However, trypanocidal treatments per animal per year were lower in intervention herds compared to control herds (0.3 vs 5 for DA and 0.8 vs 2 for ISM). This study demonstrates that the implementation of an integrated best-bet strategy leads to a reduced trypanosome prevalence under lowered trypanocidal use.

African animal trypanosomosis (nagana) in northern KwaZulu-Natal, South Africa: Strategic treatment of cattle on a farm in endemic area.

African animal trypanosomosis (AAT) is caused by several species of the genus Trypanosoma, a parasitic protozoan infecting domestic and wild animals. One of the major effects of infection with pathogenic trypanosome is anaemia. Currently, the control policies for tsetse and trypanosomosis are less effective in South Africa. The only response was to block treat all infected herds and change the dip chemical to one which controls tsetse flies during severe outbreaks. This policy proved to be less effective as demonstrated by the current high level of trypanosome infections in cattle. Our objective was to study the impacts of AAT (nagana) on animal productivity by monitoring the health of cattle herds kept in tsetse and trypanosomosis endemic areas before and after an intervention that reduces the incidence of the disease. The study was conducted on a farm in northern KwaZulu-Natal which kept a commercial cattle herd. There was no history of any cattle treatment for trypanosome. All cattle were generally in poor health condition at the start of the study though the herd received regular anthelminthic treatment. A treatment strategy using two drugs, homidium bromide (ethidium) and homidium chloride (novidium), was implemented. Cattle were monitored regularly for 13 months for herd trypanosomosis prevalence (HP), herd average packed cell volume (H-PCV) and the percentage of the herd that was anaemic (HA). A total of six odour-baited H-traps were deployed where cattle grazed from January 2006 to August 2007 to monitor the tsetse population. Glossina brevipalpis Newstead and Glossina austeni Newstead were collected continuously for the entire study period. High trypanosomes HP (44%), low average H-PCV (29.5) and HA (24%) were rerecorded in the baseline survey. All cattle in the herd received their first treatment with ethidium bromide. Regular monthly sampling of cattle for the next 142 days showed a decline in HP of 2.2% - 2.8%. However, an HP of 20% was recorded by day 220 and the herd received the second treatment using novidium chloride. The HP dropped to 0.0% and HA to 0.0% by day 116 after the second treatment. The cow group was treated again by day 160 when the HP and HA were 27.3% and 11%, respectively. The same strategy was applied to the other two groups of weaners and the calves at the time when their HP reached 20%. Ethidium and novidium treatment protected cattle, that were under continuous tsetse and trypanosomosis challenge, for up to 6 months. Two to three treatments per year may be sufficient for extended protection. However, this strategy would need to be included into an integrated pest management approach combining vector control for it to be sustainable.

Parasite specific 7SL-derived small RNA is an effective target for diagnosis of active trypanosomiasis infection.

Human and animal African trypanosomiasis (HAT & AAT, respectively) remain a significant health and economic issue across much of sub-Saharan Africa. Effective control of AAT and potential eradication of HAT requires affordable, sensitive and specific diagnostic tests that can be used in the field. Small RNAs in the blood or serum are attractive disease biomarkers due to their stability, accessibility and available technologies for detection. Using RNAseq, we have identified a trypanosome specific small RNA to be present at high levels in the serum of infected cattle. The small RNA is derived from the non-coding 7SL RNA of the peptide signal recognition particle and is detected in the serum of infected cattle at significantly higher levels than in the parasite, suggesting active processing and secretion. We show effective detection of the small RNA in the serum of infected cattle using a custom RT-qPCR assay. Strikingly, the RNA can be detected before microscopy detection of parasitaemia in the blood, and it can also be detected during remission periods of infection when no parasitaemia is detectable by microscopy. However, RNA levels drop following treatment with trypanocides, demonstrating accurate prediction of active infection. While the small RNA sequence is conserved between different species of trypanosome, nucleotide differences within the sequence allow generation of highly specific assays that can distinguish between infections with Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax. Finally, we demonstrate effective detection of the small RNA directly from serum, without the need for pre-processing, with a single step RT-qPCR assay. Our findings identify a species-specific trypanosome small RNA that can be detected at high levels in the serum of cattle with active parasite infections. This provides the basis for the development of a cheap, non-invasive and highly effective diagnostic test for trypanosomiasis.

Prevalence of bovine trypanosomosis and trypanocidal drug sensitivity studies on Trypanosoma congolense in Wolyta and Dawero zones of southern Ethiopia.

Cross-sectional studies were conducted in tsetse and non-tsetse-controlled areas of the Southern Nation Nationalities and Peoples Regional State (SNNPRS) of Ethiopia to determine the prevalence of bovine trypanosomosis as well as drug sensitivity tests on Trypanosoma congolense in both naturally and experimentally infected cattle and mice, respectively. A total trypanosome prevalence of 4.8% (95% CI: 1.8-7.5) and 20.4% (95% CI: 14-26.8) were recorded in the tsetse-controlled study area of Humbo district and the non-tsetse-controlled area of Mareka district, respectively, indicated statistically significant difference between the two areas (P<0.001). The mean PCV value for Humbo and Mareka was 26.2 (95%: 25.7-26.7) and 22.7 (95% CI: 22.1-23.3), respectively, which were also statistically significant (P<0.001). The prophylactic activity of isometamidium chloride (ISMM) was observed in Humbo on nine naturally positive zebu cattle. Breakthrough infections were recorded in (6/9) 66.7% of the cases in less than 5 weeks. A qualitative assay on mice was conducted on two T. congolense isolates obtained from the breakthrough cases with ranges of doses of ISMM and diminazene diaceturate (DA). Thereafter the mice were followed for relapse infection. ISMM at doses 0.5-4 mg/kg body weight (bw) and DA at doses of 3.5-28 mg/kg bw failed completely to cure T. congolense infections in any of the mice. A quantitative assay on mice was conducted on four T. congolense isolates obtained from Mareka. The four isolates were pooled into two pools (Pool-1 and Pool-2) for the quantitative assay on mice. The pooled isolates were tested with the same trypanocidal drugs and ranges of doses as it was used for the qualitative assay on mice. The minimum curative dose (MCD) of ISMM that cleared T. congolense infected mice was 4 and 2mg/kg bw for Pool-1 and Pool-2, respectively, whereas MCD of DA was 28 and 14 mg/kg bw, in Pool-1 and Pool-2, respectively. Although cloned populations were not used to prove whether the observed resistance was at the individual level or not, the results show that there is resistance to both ISMM and DA; failure of the "sanative pair".

Studies on effects of lactose on experimental Trypanosoma vivax infection in Zebu cattle. 2. Packed cell volume.

The ability of intravenously administered lactose in normal saline to prevent a decline in packed cell volume (PCV) during experimental trypanosomosis was studied in Zebu cattle. During the lactose infusion period, the PCV was stable up to Day 5 post-infection (p.i.) in a lactose-infused group, compared to that in an uninfused group in which the PCV dropped significantly (P < 0.05) as shown by the values of cumulative percentage change. Furthermore the mean rate of change in PCV was significantly (P < 0.05) higher in the uninfused group relative to the lactose-infused group during the same period. While the PCV fell markedly in the lactose-infused group a day after lactose infusion was stopped (Day 13 p.i.), subsequent PCV values were significantly (P < 0.05) higher compared to those in the uninfused group, up to the end of experiment on Day 17 p.i. However the mean rates of change in PCV did not vary significantly (P > 0.05) between the groups during the period in which lactose infusion was stopped. The mean levels of parasitaemic waves and parasitaemia were higher, more prolonged and more frequent in the lactose-infused group. It was inferred that the lactose was able to prevent an early onset of anaemia in the Trypanosoma vivax-infected Zebu cattle.

Studies on effects of lactose on experimental Trypanosoma vivax infection in Zebu cattle. 1. Plasma kinetics of intravenously administered lactose at onset of infection and pathology.

Lactose in normal saline was administered intravenously to a group of Zebu cattle infected with Trypanosoma vivax to determine the blood plasma kinetics at onset of an experimental infection and its ability to protect tissues against damage as part of preliminary studies to determine its suitability for use in the treatment of trypanosomosis. Significantly (P < 0.01) higher lactose concentrations were observed in the T. vivax-infected bulls at 30 min and 1 h (P < 0.05) post-infection (p.i.) and by 4 h p.i. the plasma lactose remained above the level prior to infusion, after which it fell slightly below the pre-infusion level in the uninfected group. Calculated pharmacokinetic parameters revealed delayed excretion of lactose in the T. vivax-infected group soon after infection. The total body clearance (Cl(B)) was significantly (P < 0.05) reduced. The biological half-life (t1/2), elimination rate constant (k(el)) and apparent volume of distribution (V(d)) were relatively decreased (P > 0.05) as a result of the T. vivax infection. Retention of lactose in the plasma was attributed to decreased plasma clearance. It is suggested that the presence of trypanosomes in circulation rather than organic lesions could have been responsible for the delay observed in the excretion of lactose. At 12 weeks p.i., when the experiment was terminated, the group infected and given lactose infusion (despite higher parasitaemia) had no gross or histopathological lesions in the brain, spleen, lymph nodes, heart, kidneys, liver and testes. However, the group infected but not infused with lactose were emaciated, had pale mucosae, watery blood, general muscular atrophy, serous atrophy of coronary fat and other adipose tissue, hepatomegaly, splenomegaly, swollen and oedematous lymph nodes, all of which are suggestive of trypanosomosis. Histopathological lesions included narrowing of Bowman's space and hypercellularity of glomerular tufts in the kidneys with the mean glomerular tuft nuclear indices (GTNs) in the group significantly higher (P < 0.01) than the mean GTNs of the lactose-infused and control bulls. Degenerative changes occurred in the myocardium, spleen, testes and epididymides. The tesicular and epididymal lesions are indicative of male reproductive dysfunction.