A study was conducted to determine the incidence of trypanosome infections in cattle in tsetse-free and tsetse-infested zones of the Amhara Region of northwest Ethiopia. A total of six sentinel herds were established and the cattle observed during a period of 8 consecutive months. The prevalence of seropositive cattle was high in both the tsetse-free and tsetse-infested zones. The average monthly incidence of trypanosome infection, determined using molecular diagnostic tools, was 20.9% and 25.7% in the tsetse-free and the tsetse-infested zones, respectively. In the tsetse-free, Trypanosoma vivax was responsible for 90.9% of the cattle trypanosome infections. In the tsetse-infested zone, Trypanosoma congolense and T. vivax contributed almost equally to the trypanosome infections in cattle. Trypanosome infection, regardless of species, resulted in anaemia as evidenced by a significant decrease in the packed cell volume of the infected animal. The outcome of this longitudinal study suggests that control of trypanosomiasis in the Amhara Region cannot be achieved by tsetse control alone. Supplemental measures to include drug therapy and biting fly control are discussed.
This is the first case report for the positive Trypanosoma evansi incident in Kelantan, Malaysia confirmed through protozoa detection in a Siam B mare. The horse was presented with complaints of lethargy and inappetence and it collapsed on the day of visit. Normal saline and dextrose solution were administered intravenously, while multivitamins and nerve supplements were given intramuscularly to stabilise the horse before further treatment. Haematological findings showed normocytic hypochromic anaemia and are suggestive of regenerative anaemia. Thin blood smear and examination revealed the presence of Trypanosoma sp., and it was confirmed as T. evansi through molecular identification. The horse was found dead 2 days after and post-mortem was conducted. Histopathology revealed that the horse had developed a neurological form of the disease, besides the detection of the protozoa in heart, spleen and kidney tissue. This first positive Surra case in Kelantan, Malaysia, that is bordering Thailand confirms the increasing concern of transboundary infections. In conclusion, Surra is a potential emerging disease and should be considered as differential diagnosis in horses with pale mucous membrane. This condition is particularly imperative in horses found in these regions as Surra is endemic.
A total of 719 wild rats were captured from four localities representing the west (Kuala Lumpur), east (Kuantan), north (Georgetown) and south (Malacca) to determine the diversity of blood protozoan from the urban wild rat population in peninsular Malaysia. Five rat species were recovered with Rattus rattus diardii being the most dominant species, followed by Rattus norvegicus, Rattus exulans, Rattus annandalei and Rattus argentiventer. Two blood protozoan species were found infecting the rodent population namely, Plasmodium sp. (42.1%) and Trypanosoma lewisi (25.0%). This study reports the presence of Plasmodium sp. for the first time in the rodent population in Malaysia. Two main intrinsic factors were identified affecting the parasitic infections. Trypanosoma lewisi infections were influenced by host age and sex with infections observed higher in male and juvenile rats meanwhile Plasmodium sp. infections were observed almost similar in both sexes. However, infections were higher in sub-adult rats.
Trypanosoma evansi, the causative agent of "surra", infects many species of wild and domestic animals worldwide. In the current study, the aqueous and ethanolic extracts of six medicinal plants, namely, Aquilaria malaccensis, Derris elliptica, Garcinia hombroniana, Goniothalamus umbrosus, Nigella sativa, and Strobilanthes crispus were screened in vitro for activity against T. evansi. The cytotoxic activity of the extracts was evaluated on green monkey kidney (Vero) cells using MTT-cell proliferation assay. The median inhibitory concentrations (IC50) of the extracts ranged between 2.30 and 800.97 μg/ml and the median cytotoxic concentrations (CC50) ranged between 29.10 μg/ml and 14.53 mg/ml. The aqueous extract of G. hombroniana exhibited the highest selectivity index (SI) value of 616.36, followed by A. malaccensis aqueous extract (47.38). Phytochemical screening of the G. hombroniana aqueous extract revealed the presence of flavonoids, phenols, tannins, and saponins. It is demonstrated here that the aqueous extract of G. hombroniana has potential antitrypanosomal activity with a high SI, and may be considered as a potential source for the development of new antitrypanosomal compounds.
The quantitative buffy coat (QBC) technique and conventional Giemsa thin blood smear was compared to determine the sensitivity and specificity of the technique in detecting blood parasitic infection of the rodent populations from four urban cities in Peninsular Malaysia. A total of 432 blood samples from four rat species (Rattus norvegicus, Rattus rattus diardii, Rattus exulans and Rattus argentiventer) were screened using both techniques and successfully detected two blood protozoan species (Trypanosoma lewisi and Plasmodium sp.) with Trypanosoma lewisi predominantly infecting the population. Results showed that Giemsa-stained thin film (GTF) was the better detection method on blood parasitemia (46.7%) compared to Quantitative Buffy Coat method (38.9%) with overall detection technique sensitivity and specificity at 83.2% and 74.8% respectively. The sensitivity in detection of Trypanosoma lewisi was 84.4% with value slightly lower for Plasmodium sp. infections at 76.6%. Statistical analysis proved that GTF technique was significantly more sensitive in the detection of blood protozoan infections in the rodent population compared to QBC (p<0.05).
Apart from occasional reports of clinical disease affecting horses, there is no information about Trypanosoma evansi in horses in Peninsula Malaysia. Thus, a cross-sectional study was conducted in eight states in Peninsula Malaysia to determine the active presence of T. evansi in horses. A total of 527 blood samples were obtained and examined by haematocrit centrifugation technique (HCT), Giemsa-stained thin blood smear (GSS), morphometric measurements, polymerase chain reaction (PCR) and cloning of PCR products. The results showed an overall parasitological prevalence of 0.57% (3/527, CI: 1.6-0.19%) with both HCT and GSS. Morphometric study revealed the mean total length of the trypanosomes including the free flagellum was 27.94 ± 2.63 μm. PCR successfully amplified a trypanosome specific 257 bp in 1.14% of samples (6/527, CI: 2.4-0.52%) and was confirmed by nucleotide sequences. The mean packed cell volume (PCV) for the positive cases detected by HCT was lower (23% ± 7.00) compared to the positive cases detected by PCR alone in the state of Terengganu (35% ± 4.73). In conclusion, this study showed T. evansi infection occurred in low frequency in horses in Peninsula Malaysia, and anaemia coincided with parasitaemic animals. PCR is considered as a sensitive diagnostic tool when parasitaemia is undetectable. The slight lengthier mean of parasite and anaemia may indicate a virulent strain of T. evansi circulating throughout the country. Thus, it's highly recommended to shed light on host-parasite relationship for better epidemiological understanding.
Essential oil from Cymbopogon nardus was evaluated for activity against Trypanosoma brucei brucei BS221 (IC50 = 0.31 ± 0.03 μg/mL) and cytotoxic effect on normal kidney (Vero) cells (IC50 = >100 μg/mL). The crude essential oil was subjected to various chromatography techniques afforded active sub fractions with antitrypanosomal activity; F4 (IC50 = 0.61 ± 0.06 μg/mL), F6 (IC50= 0.73 ± 0.33 μg/mL), F7 (IC50 = 1.15 ± 0 μg/mL) and F8 (IC50 = 1.11 ± 0.01 μg/mL). These active fractions did not exhibit any toxic effects against Vero cell lines and the chemical profiles investigation indicated presence of α-and γ-eudesmol, elemol, α-cadinol and eugenol by GC/MS analysis.
A comparative seroprevalence study on bovine trypanosomiasis and anaplasmosis was conducted. Sera of adult cattle and buffaloes of different breeds from farms from five different states in Malaysia were collected and tested for the presence of Trypanosoma evansi antibodies by CATT and Anaplasma marginale antibodies by c-ELISA. Of the 116 samples, 14.7% tested positive for bovine trypanosomiasis and 77.6% for bovine anaplasmosis.
Trypanosomiasis is a disease caused by a flagellate protozoon called Trypanosoma and can be mechanically transmitted by vectors to humans and animals. Various species of Trypanosoma are found in livestock and poultry, which include Trypanosoma evansi, T. brucei, T. vivax and T. congolense. The camel is the most sensitive livestock for T. evansi, so the exact identification of infection is very important for epidemiological studies and the design of control programs. The present study was conducted with the aim of molecular detection of camel trypanosomiasis in the Sistan region in 2015. Previous studies have shown that internal transcribed spacer one (ITS1) of the ribosomal DNA is a reliable genetic marker for carrying out systematic molecular studies of trypanosomes. In order to investigate infections of camels with T. evansi, a total of 113 blood samples were collected randomly and the presence of parasites in each sample was evaluated using the microscopic method and polymerase chain reaction (PCR) test. Genomic DNA was extracted and the ITS-1 was amplified by PCR. In comparison to the nucleotide sequence obtained with the sequences recorded in GenBank, it was determined that there is a 99% homology with the recorded sequence of T. evansi. The obtained sequence was registered in Gen Bank with kx900449 code. The T. evansi sequences from different countries such as India, Taiwan, Thailand, the Philippines, China and Argentina and etc., were extracted from the Gene bank and aligned using the ClustalW2 sequence alignment tool and MEGA software. In this study the prevalence of T. evansi infection using the molecular method was 6.19% and no positive samples were found by microscopic observation.
Trypanothione reductase is a key enzyme that upholds the redox balance in hemoflagellate protozoan parasites such as T. congolense. This study aims at unraveling the potency of Kolaviron against trypanothione reductase in T. congolense infection using Chrysin as standard. The experiment was performed using three different approaches; in silico, in vitro and in vivo. Kolaviron and Chrysin were docked against trypanothione reductase, revealing binding energies (-9.3 and -9.0 kcal/mol) and Ki of 0.211μM and 0.151μM at the active site of trypanothione reductase as evident from the observed strong hydrophobic/hydrogen bond interactions. Parasitized blood was used for parasite isolation and trypanothione reductase activity assay using standard protocol. Real-time PCR (qPCR) assay was implored to monitor expression of trypanothione reductase using primers targeting the 177-bp repeat satellite DNA in T. congolense with SYBR Green to monitor product accumulation. Kolaviron showed IC50 values of 2.64μg/ml with % inhibition of 66.78 compared with Chrysin with IC50 values of 1.86μg/ml and % inhibition of 53.80. In vivo studies following the administration of these compounds orally after 7 days post inoculation resulted in % inhibition of Chrysin (57.67) and Kolaviron (46.90). Equally, Kolaviron relative to Chrysin down regulated the expression trypanothione reductase gene by 1.352 as compared to 3.530 of the infected group, in clear agreement with the earlier inhibition observed at the fine type level. Overall, the findings may have unraveled the Kolaviron potency against Trypanosoma congolense infection in rats.
An investigation into the epidemiology of Trypansoma evansi infection in crossbred dairy cattle was conducted for a period of 12 months on a dairy cattle farm in Penninsular Malaysia. The prevalence of parasitaemia was highest in lactating animals (13.4%), followed by those in the dry herd (8.8%), late pregnant animals (8.1%), early pregnant animals (4.7%), calves (0.3%) and heifers (0.2%). The prevalence of antigenaemia was highest in the lactating animals (54.7%), followed by that in dry animals (53.7%), heifers (51.1%), late pregnant animals (47.7%), early pregnant animals (46.5%) and calves (24.2%).
During a period of four consecutive years, trypanosomosis surveys were conducted in a tsetse-infested and tsetse-free area of the Amhara Region of north-west Ethiopia. In each study area randomly selected communal cattle were sampled and their blood was investigated using parasitological diagnostic methods. At the same time the population of biting flies was sampled. The monthly average prevalence of trypanosome infections in cattle did not differ significantly between study areas. In both study areas, the prevalence of trypanosome infections was highest during the long rainy season. Trypanosome infections were mainly due to Trypanosoma vivax and they significantly reduced the average packed cell volume and the body condition of the animals. The monthly prevalence of infection was correlated with the density of biting flies, such as Tabanidae and Stomoxys spp., in the preceding month suggesting an important role of mechanical transmission in the epidemiology of trypanosomosis in both areas.
Systematic surveys of the wild macaques of South Asia by blood culture resulted in the discovery that trypanosomiasis is enzootic in the simians of Indonesia, Malaysia, India, and Thailand. The isolates obtained differ in morphology, metabolism, and ability to multiply in arthropods. Following this discovery, interest focused on possible transmissions of these trypanosomiases. Laboratory-reared and wild-caught insects were studied to determine which are satisfactory intermediate hosts and potential natural vectors. Successful results were obtained with insectary-reared reduviids and Indonesian isolates. In Rhodnius prolixus and Triatoma rubrofasciata the Indonesian trypanosomes multiply for periods which can exceed 40 days. The flagellate infections are in the digestive tract, whereas trypanosomes have never been seen in the salivary glands or in the hemolymph. The feces of trypanosome-carrying reduviids are infective, suggesting a stercoreal method of infection of mammals, and infection was produced in experiments in which feeding by the insects was not possible. The relevance of these findings to natural transmission in Indonesia is not known. Experiments with insects and all other trypanosomal isolates have been negative. The natural transmission mechanism(s) of the simian trypanosomiases in South Asia remains an unsolved problem.
A laboratory prototype system that correlates murine blood absorbance with degree of infection for Plasmodium berghei and Trypanosoma avensi has been designed, constructed and tested. A population (n = 6) of control uninfected, Plasmodium infected and Trypanosoma infected BALB/c mice were developed and spectral absorption measurements pre and post infection were made every 3 days. A fibre optic spectrometer set-up was used as the basis of a laboratory prototype biosensor that uses the Beer Lambert Law to relate Ultraviolet-Visible-Near-infrared absorbance data to changes in murine blood chemistry post infection. Spectral absorption results indicate a statistically relevant correlation at a 650 nm with infection for Plasmodium from between 4 and 7 sampling days' post infection, in spite of significant standard deviations among the sample populations for control and infected mice. No significant spectral absorption change for Trypanosoma infection was been detected from the current data. Corresponding stained slides of control and infected blood at each sampling date were taken with related infected cell counts determined and these correlate well for Plasmodium absorbance at 650 nm.