Trypanosoma brucei gambiense infection in vervet monkeys: a potential model for early-stage disease.

BACKGROUND: Non-human primates are important experimental models for human African trypanosomiasis. METHODS: Six monkeys were intravenously inoculated with 10(5) trypanosomes of Trypanosoma brucei gambiense IL 3253. They were monitored for 180 days for parasitemia, hematology, clinical and biochemical profiles. RESULTS: The pre-patent period was 2-3 days. From 33 to 123 dpi, the parasitemia was low and only detectable by the hematocrit centrifugation technique. Thereafter, to the end of the experimental period, the parasitemia was undetectable by parasitological methods. Clinical signs observed were lymphadenopathy and splenomegaly. Hematological changes included a decline in hemoglobin occurring between 14 and 56 dpi and a significant decline in platelet counts after infection. The levels of total protein, albumin and globulins increased from 26 dpi for the rest of the experimental period. No parasites were detected in cerebrospinal spinal fluid, and no brain pathology was observed. CONCLUSION: This vervet monkey model can only be used for early-stage disease Gambian sleeping sickness.

AN APPRAISAL OF RABIES OCCURRENCE AND CONTROL IN KISUMU MUNICIPALITY, KENYA.

BACKGROUND: Rabies is a serious neglected disease which occurs worldwide. In urban areas, the control of the diseaseis expected to be hampered by several unique constraints. OBJECTIVE: To appraise the occurrence and control of rabies. STUDY DESIGN: The tools used in the study included key informant interviews with senior veterinary and medical officers and a questionnaire survey on 15 veterinary officers and 9 human physicians. The occurrence of animal bites and costs involved in treatment was also investigated using retrospective hospital data for the year 2007. SETTING: Kisumu Municipality, Kenya. RESULTS: The control of rabies was under the Veterinary Department through dog vaccinations and baiting, and also under the Department of Public Health (DPH) through people vaccinations and post exposure treatments. The constraints observed were poor coordination in rabies control efforts, understaffing and limited resources. Animal bites were common with 1270 cases reported during 2007. The post-bite rabies vaccines were mainly purchased from local chemists at KES 7,500 per full dose. This amounted to an annual total cost of KES 9,525,000 (136,071 USD). The bites were mainly from domestic dogs (89%) and occurred commonly in highly populated areas. The mean age of people with bites was 21 (2.5 - 78) years, showing that young people had a higher risk. Males (59%) were at higher risk of bites than females (41%). CONCLUSIONS: It would be important for the relevant departments to harmonise their rabies control strategies and also address the prevalent constraints by using local networks. The incidence of rabies and the associated epidemiology needs proper updated documentation with a view of carrying out mass vaccination of dogs at times free of cost to the economically disadvantaged owners.

Lipid metabolism and other metabolic changes in vervet monkeys experimentally infected with Trypanosoma brucei rhodesiense.

BACKGROUND: Human African trypanosomiasis is associated with metabolic changes which have not been well characterized. METHODS: Chlorocebus aethiops were experimentally infected with Trypanosoma brucei rhodesiense and late-stage disease induced at 28 days post-infection. Ear prick blood for glucose determination and blood samples were obtained at weekly intervals for 56 days. Analysis was carried out using dry chemistry analysis. RESULTS: In early infection, there was a significant increase in creatine kinase, while during early and transitional stage of infection there was a significant decrease in glucose and high-density lipoprotein and an increase in triglyceride levels. In the late stage, there was a significant increase in both total cholesterol and LDL levels. CONCLUSIONS: Further investigations should focus on levels of total cholesterol during the follow-up period in curatively treated vervet monkeys. Apart from their importance in disease staging, the changes in lipids levels may also affect the pharmacokinetics of some trypanocides.

Prevalence and types of coinfections in sleeping sickness patients in kenya (2000/2009).

The occurrence of coinfections in human African trypanosomiasis (HAT) patients was investigated using a retrospective data of hospital records at the National Sleeping Sickness Referral Hospital in Alupe, Kenya. A total of 31 patients, 19 males and 12 females, were diagnosed with HAT between the years 2000 and 2009. The observed co-infections included malaria (100%), helminthosis (64.5%), typhoid (22.5%), urinary tract infections (16.1%), HIV (12.9%), and tuberculosis (3.2%). The species of helminthes observed included Ancylostoma duodenale (38.7%), Ascaris lumbricoides (45.7%), Strongyloides stercoralis (9.7%), and Taenia spp. (3.2%). The patients were also infected with Entamoeba spp. (32.3%) and Trichomonas hominis (22.6%) protozoan parasites. The main clinical signs observed at the point of admission included headache (74.2%), fever (48.4%), sleep disorders (45.2%), and general body pain (41.9%). The HAT patients were treated with suramin (early stage, 9/31) and melarsoprol (late stage, 22/31). In conclusion, the study has shown that HAT patients have multiple co-infections which may influence the disease pathogenesis and complicate management of HAT.

Seroprevalence of Cysticercus cellulosae and associated risk factors in free-range pigs in Kenya.

Porcine cysticercosis is an emerging zoonosis with public health and economic importance. A cross-sectional study was undertaken to investigate the disease in free-range pigs on 182 smallholder farms in Busia District, Kenya. The survey households were selected using a snowballing technique. Serum samples were obtained from 284 pigs of all ages at farm level and 37 pigs from slaughter slabs in the study area. The samples were analysed for the presence of cysticercus antigen using an antigen enzyme-linked immunosorbent assay (ELISA). A structured questionnaire was administered to determine the risk factors for porcine cysticercosis on the study farms. At pig level, the total number of pigs testing positive were 11, resulting in a seroprevalence of 4% (95% confidence interval (CI): 1.9-6.2%), while the farms with a positive pig were 9% (95% CI: 3.9-14.1%). All pigs examined in the slaughter slab survey were seronegative. The distribution of possible risk factors for porcine cysticercosis that were observed at farm level was as follows: free-range pig keeping (100%), history of human taeniosis infection in a family (51%), slaughtering of pigs at home (20%), lack of meat inspection (15%) and absence of latrines (15%). The only significant (χ2 = 4.4, P = 0.034, odds ratio (OR) = 3.8) risk factor associated with the occurrence of cysticercosis was lack of latrines at household level. The study shows that porcine cysticercosis is prevalent in free-range pigs in Busia District, Kenya and thus control measures need to be instituted.

Questionnaire survey on urban and peri-urban livestock farming practices and disease control in Kisumu municipality, Kenya.

To characterise the urban livestock keeping practices and constraints in Kisumu municipality, Kenya, a questionnaire survey was carried out. Thirty-four contact farmers were interviewed on general farm characteristics and production constraints. The farming activities were categorised as either livestock only (41%), or mixed crops and livestock (59%). The surveyed farmers kept mainly cattle (100%), chickens (82%) and goats (74%). Most (94%) of the farmers had kept livestock for prolonged periods mainly for income generation (97%) and domestic consumption (59%). These data show that livestock keeping was popular and could be harnessed to increase food security, although the farmers kept mainly low-producing indigenous cattle (98%) which were grazed on unutilised land. The main production constraints mentioned by farmers included diseases (100%), poor fertility (68%) and lack of feed (56%). The diseases varied with species of ruminants and included lumpy skin disease (71%), diarrhoea (65%) and helminthosis (62%). The source of advice on management and treatment of the livestock was almost equally from private and government veterinary personnel. To improve livestock productivity, it is recommended that key stakeholders address the constraints mentioned in this study and in particular that the occurrence of diseases should be investigated with a view to developing sustainable control strategies.

Occurrence of risk factors for zoonoses in Kisumu City, Kenya: a questionnaire survey.

OBJECTIVE: To determine the occurrence of risk factors amongst farmers keeping livestock in Kisumu municipality, Kenya. METHODS: Thirty four contact house-holds involved in livestock keeping constituted the study group. The head of the household was interviewed using a structured questionnaire relating to public health issues affecting water, food and sanitation RESULTS: The water consumed at the household level originated from the taps (56%), bore holes (18%), local rivers (15%) and wells (15%). The wells were shallow and open to contamination. Fourty one percent (41%) of the households consumed water without boiling or any other form of treatment. Farmers had a history of slaughtering cattle (79%) and pigs (66%) at their homes and these animals were not inspected by legalized government officers. During the last five years, porcine cysticercosis and taeniosis (9%) were reported to occur amongst the homesteads. Although toilets were found in most homes, some were overflowing (27%), while children below 4 years (mean) of age defaecated in the bushes/grass within the neighbourhood. Most farmers (62%) disposed the manure next to the animal boma resulting in creation of small manure 'hills' that were a nuisance and breeding ground for vector flies. Other risk factors for disease transmission included consumption of unwashed raw food (48%), dried raw fish (21%) and failure to wash cows' udder before milking (29%). CONCLUSION: The study shows that risk factors for transmission of zoonoses are common in house holds keeping livestock in the study area and highlights the need for public health education to raise awareness of these factors. Further studies should be conducted to determine the possibilities of zoonoses occurrences among the human population.

Immunospecific immunoglobulins and IL-10 as markers for Trypanosoma brucei rhodesiense late stage disease in experimentally infected vervet monkeys.

OBJECTIVE: To determine the usefulness of IL-10 and immunoglobulin M (IgM) as biomarkers for staging HAT in vervet monkeys, a useful pathogenesis model for humans. METHODS: Vervet monkeys were infected with Trypanosoma brucei rhodesiense and subsequently given sub-curative and curative treatment 28 and 140 days post-infection (dpi) respectively. Matched serum and CSF samples were obtained at regular intervals and immunospecific IgM, immunoglobulin G (IgG) and IL-10 were quantified by ELISA. RESULTS: There was no detectable immunospecific IgM and IgG in the CSF before 49 dpi. CSF IgM and IgG and serum IgM were significantly elevated with peak levels coinciding with meningoencephalitis 98 dpi. The serum IL-10 was upregulated in both early and late disease stage, coinciding with primary and relapse parasitaemia respectively. CSF white cell counts (CSF WCC) were elevated progressively till curative treatment was given. After curative treatment, there was rapid and significant drop in serum IgM and IL-10 concentration as well as CSF WCC. However, the CSF IgM and IgG remained detectable to the end of the study. CONCLUSIONS: Serum and CSF concentrations of immunospecific IgM and CSF IgG changes followed a pattern that mimics the progression of the disease and may present reliable and useful biomarkers of the disease stage. Due to rapid decline, serum IgM and IL-10 are, additionally, potential biomarkers of the success of chemotherapy.

Aggravation of pathogenesis mediated by ochratoxin A in mice infected with Trypanosoma brucei rhodesiense.

Mice fed 1.5 mg ochratoxin A (OTA) per kg body weight and infected with Trypanosoma brucei rhodesiense were compared with trypanosome-infected placebo-fed and uninfected OTA-fed controls. Uninfected OTA-fed mice showed fever, lethargy, facial and eyelid oedemas, mild hepatitis and nephritis, and high survival. Infected placebo-fed controls had mean pre-patent period (PPP) of 3.26 days, lethargy, dyspnoea, fever, facial and scrotal oedema, survival of 33-65 days, reduced red cell counts (RCC: 10.96-6.87x106 cells/microl of blood), packed cell volume (PCV: 43.19-26.36%), haemoglobin levels (Hb: 13.37-7.92 g/dL) and mean corpuscular volume (MCV) of 37.96-41.31 fL, hepatosplenomegaly, generalized oedemas, heart congestion, hepatitis and nephritis. Compared to infected placebo-fed controls, infected OTA-fed mice had significantly (P<0.05) shorter mean PPP (2.58 days), reduced survival (6-47 days), more pronounced fever and dyspnoea. The latter had significantly (P<0.05) reduced RCC (10.74-4.56x106 cells/microl of blood), PCV (43.90-20.78%), Hb (13.06-5.74 g/dL), increased MCV (39.10-43.97 fL), severe generalized oedemas, haemorrhages, congestion, hepatic haemosiderosis, hepatitis, nephritis, endocarditis, pericarditis and exclusively, splenic macrophage and giant cell hyperplasia, expanded red pulp and splenic erythrophagocytosis. It was concluded that OTA aggravated the pathogenesis of T. b. rhodesiense infection in mice, and should therefore be taken into consideration during trypanosomosis control programmes.

Efficacy of the novel diamidine compound 2,5-Bis(4-amidinophenyl)- furan-bis-O-Methlylamidoxime (Pafuramidine, DB289) against Trypanosoma brucei rhodesiense infection in vervet monkeys after oral administration.

Owing to the lack of oral drugs for human African trypanosomiasis, patients have to be hospitalized for 10 to 30 days to facilitate treatment with parenterally administered medicines. The efficacy of a novel orally administered prodrug, 2,5-bis(4-amidinophenyl)-furan-bis-O-methlylamidoxime (pafuramidine, DB289), was tested in the vervet monkey (Chlorocebus [Cercopithecus] aethiops) model of sleeping sickness. Five groups of three animals each were infected intravenously with 10(4) Trypanosoma brucei rhodesiense KETRI 2537 cells. On the seventh day postinfection (p.i.) in an early-stage infection, animals in groups 1, 2, and 3 were treated orally with pafuramidine at dose rates of 1, 3, or 10 mg/kg of body weight, respectively, for five consecutive days. The animals in groups 4 and 5 were treated with 10 mg/kg for 10 consecutive days starting on the 14th day p.i. (group 4) or on the 28th day p.i. (group 5), when these animals were in the late stage of the disease. In the groups treated in the early stage, 10 mg/kg of pafuramidine completely cured all three monkeys, whereas lower doses of 3 mg/kg and 1 mg/kg cured only one of three and zero of three monkeys, respectively. Treatment of late-stage infections resulted in cure rates of one of three (group 4) and zero of three (group 5) monkeys. These studies demonstrated that pafuramidine was orally active in monkeys with early-stage T. brucei rhodesiense infections at dose rates above 3 mg/kg for 5 days. It was also evident that the drug attained only minimal efficacy against late-stage infections, indicating the limited ability of the molecule to cross the blood-brain barrier. This study has shown that oral diamidines have potential for the treatment of early-stage sleeping sickness.

Efficacy of the diamidine DB75 and its prodrug DB289, against murine models of human African trypanosomiasis.

The choice of drugs for the treatment of sleeping sickness is extremely limited. To redress this situation, the recently synthesised diamidine, 2,5-bis(4-amidinophenyl)-furan (DB75, furamidine) and its methamidoxime prodrug, 2,5-bis(4-amidinophenyl)-furan-bis-O-methylamidoxime (DB289, pafuramidine) were, together with pentamidine, evaluated for efficacy in acute rodent models. The activity was compared in three common mouse models that mimic the first stage of human African trypanosomiasis. The mice were infected with the pleomorphic T .b. rhodesiense strains KETRI2537 and STIB900 or with the monomorphic T. b. brucei strain STIB795. Importantly, DB75 showed activity superior to that of pentamidine at comparable doses in all three mouse models. Complete cures were achieved with oral dosing of the prodrug DB289 in all three models without any overt toxicity. This shows that the prodrug strategy was successful in terms of reducing toxicity and increasing efficacy and oral bioavailability.

Occurrence of multiple drug resistance in Trypanosoma brucei rhodesiense isolated from sleeping sickness patients.

The occurrence of cross-resistance among melarsoprol-resistant Trypanosoma brucei rhodesiense isolates was investigated in this study. The isolates, T. b. rhodesiense KETRI 237, 2538, 1992, 2709, 2694 and 3530, had been obtained from sleeping sickness patients in Kenya and Uganda between 1960 and 1985. Five groups consisting of six mice each were inoculated intraperitoneally with 10(5) parasites of each isolate, and 24 h later treated with either melarsoprol, homidium chloride, diminazene aceturate or isometamidium chloride. The control group comprised infected but untreated mice. The mice were monitored for cure for a period of 60 days post-treatment. The mean prepatent period in the control mice was 5 days while the mean survival period was 22 days. Five of the stabilates, KETRI 237, 2538, 2709, 2694, and 3530, were confirmed to be melarsoprol resistant. Cross-resistance was observed, with the majority of the isolates being resistant to homidium chloride (5/6) and diminazene aceturate (5/6), but all were sensitive to isometamidium chloride (6/6). However T. b. rhodesiense KETRI 1992, which was previously considered as melarsoprol resistant, was sensitive to all the drugs tested. In conclusion, our study has revealed the existence of cross-resistance among the melarsoprol resistant isolates which could only be cured by isometamidium.

Hematological changes in vervet monkeys (Chlorocebus aethiops) during eight months' adaptation to captivity.

This study investigated fluctuations in hematological values of 50 wild-caught vervet monkeys (African green monkeys, grivets, Chlorocebus aethiops) during habituation to captivity. The monkeys were categorized into four groups according to age and sex viz adult males, adult females, juvenile males, and juvenile females. The erythrocyte values were significantly higher (P<0.05) in the adult males than in the other animals. There was an increase in most of the erythrocyte parameters studied during the monitoring period with the most significant being hemoglobin, hematocrit, and mean corpuscular volume. However, the red cell distribution widths, which were higher in adult females, declined. The total white blood cell (WBC) counts, which were higher in adult females than in the other animals, were closely correlated with granulocytes counts. The WBC levels decreased in all the animals throughout the 8 months study, indicating gradually decreasing stress, but they were relatively stable in males. The platelet counts declined significantly (P<0.05) and at 8 months post capture the counts were higher in females than in males. The juvenile female platelet counts were relatively stable during the monitoring period. The maintenance of the monkeys on an improved stable diet and in environment-controlled housing combined with progressing psycho-physiological adaptation may be important factors for the gradual improvements of the hematological values recorded. There were wide variations in these between individual animals emphasizing the need for long adaptation combined with establishment of individual baseline values before experimental studies.

The point prevalence of gastro-intestinal parasites in calves, sheep and goats in Magadi division, south-western Kenya.

Helminths cause great economic loss in livestock in Africa, and can be categorized as either direct or indirect losses. Arid and semi-arid lands (ASAL) in Kenya comprise 71% of total land area and harbour the largest population of cattle, sheep and goats. However, little information on the distribution and impact of gastro-intestinal (GIT) parasitism in these animals is available. This survey was conducted to establish the prevalence of GIT parasites infecting calves, sheep and goats and their relative importance in Magadi division, which is semi-arid. Faecal samples were obtained directly from the rectum of 109 calves, 133 goats and 20 sheep and submitted to the laboratory for faecal worm egg counts, and coccidial oocysts examination using a modified McMaster method. The significance of differences in mean egg count per gram (epg) between animal species and herds (farms) were assessed using analysis of variance. The overall prevalence of nematodes in the calves, sheep and goats was 69.2%, 80% and 82%, respectively. About 10% of sheep and goats had epgs higher than 1 000, the remainder having light to moderate infections. The overall prevalence of coccidial oocysts in calves, sheep and goats was 30%, 44% and 45%, respectively. Poor productivity in ASAL areas, where nutrition is often poor, is likely to be pronounced in the presence of parasite infections. These findings indicate that viable internal parasite control should be implemented in the study area in order to increase the productivity of the livestock there.