To determine and compare the prevalence of trypanosome infections in different livestock species (cattle, pigs and goats) in areas where game animals are scarce and livestock constitute the main food source of tsetse, a survey was conducted on the plateau of the Eastern Province of Zambia in Katete and Petauke districts where Glossina morsitans morsitans is the only tsetse species present. Blood was collected from a total of 734 cattle, 333 goats and 324 pigs originating from 59 villages in both districts and was examined using the buffy coat method and the PCR-RFLP as diagnostic tools.The prevalence of trypanosome infections differed substantially between livestock species. Using microscopic diagnostic methods, trypanosome infections were detected in 13.5% of the cattle and 0.9% of the pigs. All goats were parasitologically negative. The PCR-RFLP analyses increased the trypanosomiasis prevalence to 33.5, 6.5 and 3.3% in cattle, pigs and goats respectively. The majority of the infections (91.2%) were due to Trypanosoma congolense. The presence of a trypanosome infection in cattle and pigs resulted in a significant decline in the packed cell volume. The outcome of the study clearly shows that despite the availability of goats and pigs, cattle seem to be the major livestock species affected by the disease in trypanosomiasis endemic areas. The high proportion of infections in cattle could be partly attributed to their higher availability and attractiveness to tsetse.
In this study, packed cell volume-values (PCV) are evaluated as indicator of trypanosomiasis infections in cattle. A total of 734 blood samples were collected in 11 different sampling sites in eastern Zambia: 84 calves (<1 year), 52 young females and 40 young males (between 1 and 3 years), 228 cows, 317 oxen and 13 bulls (>3 years). All samples were subjected to three diagnostic tests: parasitological examination using the buffy coat method, PCR/RFLP and PCV determination. The results were compared and analysed in a Bayesian model, which allowed the estimation of the infection prevalence and the respective test sensitivities and specificities. The presence of a trypanosomal infection significantly reduced the PCV, independently of the age and sex of the infected animal. The estimated prevalence of trypanosomal infections in the study area was 34% (95% credibility interval: 30-38%). While the specificity of both the parasitological and the PCR/RFLP tests were set to 1, the parasitological diagnosis had a low sensitivity (37%) compared to the PCR/RFLP (96%). When using a cut-off value of 24, the PCV had a high specificity (98%) but a rather low sensitivity (53%) for identifying trypanosomiasis infections. Using 26% as a cut-off increased the sensitivity to 76% without much affecting the specificity (94%). A parallel combination of the parasitological diagnosis and the PCVimproved the diagnostic sensitivity (74% and 89% for PCV cut-off values of 24% or 26%, respectively) while specificity remained high (98% and 94% for PCV cut-off values of 24% or 26%, respectively). These results suggest that such a combination could advantageously be used for the diagnosis of cattle trypanosomiasis in the field: it is much www.elsevier.com/locate/prevetmed more sensitive than parasitological examination alone and it is much cheaper than molecular tests. However, the value of this approach depends largely on the determination of an appropriate cut-off value to consider a sample positive, depending on the required test sensitivities and specificities. #
BackgroundLoop-mediated isothermal amplification (LAMP) is a novel strategy which amplifies DNA with high sensitivity and rapidity under isothermal conditions. In the present study, the performance of the repetitive insertion mobile element (RIME)-LAMP and human serum resistance-associated gene (SRA)-LAMP assays were evaluated using clinical specimens obtained from four male patients from Luangwa and Zambezi valleys in Zambia and Zimbabwe, respectively.FindingsThe cases reported in this preliminary communication were all first diagnosed by microscopy, through passive surveillance, and confirmed by both RIME-LAMP and SRA-LAMP. A good correlation between microscopy and LAMP was observed and contributed to staging and successful treatment of patient. RIME-LAMP and SRA-LAMP complimented each other well in all the cases.ConclusionsBoth RIME-LAMP and SRA-LAMP were able to detect Trypanosoma brucei rhodesiense DNA in patient blood and CSF and hence confirmed HAT in the parasitaemic patients. Our study indicates that the LAMP technique is a potential tool for HAT diagnosis, staging and may be useful for making therapeutic decisions. However, no statistically significant conclusion may be drawn due to the limited sample size used in the present study. It is thus imperative to conduct a detailed study to further evaluate the potential of LAMP as a bedside diagnostic test for HAT.
BackgroundBecause of the low sensitivity of conventional rapid diagnostic tests (RDTs) for malaria infections, the actual prevalence of the diseases, especially those caused by non-Plasmodium falciparum (non-Pf) species, in asymptomatic populations remain less defined in countries lacking in well-equipped facilities for accurate diagnoses. Our direct blood dry LAMP system (CZC-LAMP) was applied to the diagnosis of malaria as simple, rapid and highly sensitive method as an alternative for conventional RDTs in malaria endemic areas where laboratory resources are limited.ResultsLAMP primer sets for mitochondria DNAs of Plasmodium falciparum (Pf) and human-infective species other than Pf (non-Pf; P. vivax, P. ovale, P. malariae) were designed and tested by using human blood DNA samples from 74 residents from a malaria endemic area in eastern Zambia. These malaria dry-LAMPs were optimized for field or point-of-care operations, and evaluated in the field at a malaria endemic area in Zambia with 96 human blood samples. To determine the sensitivities and specificities, results obtained by the on-site LAMP diagnosis were compared with those by the nested PCR and nucleotide sequencing of its product. The dry LAMPs showed the sensitivities of 89.7% for Pf and 85.7% for non-Pf, and the specificities of 97.2% for Pf and 100% for non-Pf, with purified blood DNA samples. The direct blood LAMP diagnostic methods, in which 1 μl of anticoagulated blood were used as the template, showed the sensitivities of 98.1% for Pf, 92.1% for non-Pf, and the specificities of 98.1% for Pf, 100% for non-Pf. The prevalences of P. falciparum, P. malariae and P. ovale in the surveyed area were 52.4, 25.3 and 10.6%, respectively, indicating high prevalence of asymptomatic carriers in endemic areas in Zambia.ConclusionsWe have developed new field-applicable malaria diagnostic tests. The malaria CZC-LAMPs showed high sensitivity and specificity to both P. falciparum and non-P. falciparum. These malaria CZC-LAMPs provide new means for rapid, sensitive and reliable point-of-care diagnosis for low-density malaria infections, and are expected to help update current knowledge of malaria epidemiology, and can contribute to the elimination of malaria from endemic areas.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1949-8) contains supplementary material, which is available to authorized users.
a b s t r a c tThe control of bovine trypanosomiasis could be improved by using the available control tools during periods when the incidence of the disease is highest. The present study assessed the monthly risk of bovine trypanosomiasis in 85 sentinel cattle kept on the tsetse-infested eastern plateau of Zambia during a period of 19 consecutive months. To avoid problems associated with persistence of infections because of trypanocidal drug resistance and/or the time lag between sampling and molecular analysis, a survival analysis and the subsequent calculation of risk was used as an indicator of challenge. Results showed that the average monthly risk of infection (92.3% due to Trypanosoma congolense) was 6%. It was significantly higher (7.7%) during the beginning of the rainy season (December-February). According to the outcome of the study, bovine trypanosomiasis control in the study area can be improved through increasing control efforts during this period of highest challenge.
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