Detection of trypanosomes that cause disease in human beings and livestock within their tsetse fly hosts is an essential component of vector and disease control programmes. Several molecular-based diagnostic tests have been developed for this purpose. Many of these tests, while sensitive, require analysis of trypanosome DNA extracted from single flies, or from pooled tsetse fly heads and amplified trypanosome DNA. In this study, we evaluated the relative analytical and diagnostic sensitivities of two PCR-based tests (ITS and TBR) and a Trypanozoon specific LAMP assay using pooled whole tsetse flies and midguts spiked with serially diluted procyclics of a laboratory strain of Trypanosoma brucei brucei (KETRI 3386). Test sensitivity was also evaluated using experimentally infected tsetse flies. The aim was to determine the most appropriate pooling strategy for whole tsetse and midguts. RIME-LAMP had the highest diagnostic sensitivity (100%) followed by TBR-PCR (95%) and ITS-PCR (50%) in detecting trypanosome DNA from pooled tsetse midguts. RIME-LAMP also had the best diagnostic specificity (75%) followed by ITS-PCR (68%) and TBR-PCR (50%). The relative detection limit determined by serial dilution of procyclics was below 10(-6) (equivalent to 1parasite/ml). Using TBR-PCR, ITS-PCR and RIME-LAMP, it was possible to detect trypanosome DNA in single flies or in pools of 2, 3, 4, 5, 10, or 15 flies/midguts. The proportion of positive pools declined by up to 60% when testing pools of 15 whole flies as opposed to testing pools of 5-10 flies. Additionally, it was possible to detect DNA in a single infected tsetse fly in the background of 4, 9, or 14 uninfected tsetse flies. Averaged across pool sizes and tsetse species, RIME-LAMP detected the highest proportion of positive pools in spiked whole tsetse and midguts (86.6% and 87.2%) followed by TBR-PCR (78. 6% and 79.2%) and ITS-PCR (34.3% and 40.2%). There were no significant differences between the proportions of positive pools detected in whole flies and midguts. We conclude that pooling of whole tsetse/midguts is an effective strategy to reduce hands-on-time and hence has potential application in large scale xenomonitoring to generate epidemiological data for decision making. RIME-LAMP offers the best diagnostic sensitivity and specificity on pooled tsetse midguts, thus demonstrating its superior diagnostic performance when compared with TBR-PCR and ITS-PCR. Using pools of whole tsetse or midguts as source of DNA does not have any significant effect on test results and is more representative of the field conditions where the proportion of flies with infected midguts tends to be higher than flies with infected salivary glands. Therefore to save time and minimize costs, pooling of whole tsetse flies is recommended.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.vetpar.2013.05.022DOI Listing

Publication Analysis

Top Keywords

pooled tsetse
20
tsetse flies
20
tsetse midguts
20
trypanosome dna
16
tsetse
14
flies
12
flies midguts
12
tsetse fly
12
positive pools
12
midguts
9

Similar Publications

Article Synopsis
  • Bovine trypanosomosis significantly hampers livestock productivity in Ethiopia, affecting around 70 million cattle, prompting a review of research from the last decade to analyze its prevalence, risk factors, and vector density.
  • A meta-analysis of 26 studies found a 9% overall prevalence of bovine trypanosomosis, with the highest rates in Amhara and Oromia regions, indicating substantial variability among studies.
  • Important risk factors identified include poor body condition and coat color, with black-coated animals being much more vulnerable to the disease compared to other colors, highlighting the need for targeted management strategies.
View Article and Find Full Text PDF

A cross-sectional study aimed to elucidate the prevalence of bovine trypanosomosis and its potential risk factors was conducted in tsetse suppression and non-suppression areas of South Omo Zone, Southern Ethiopia from November 2018- May 2019. A total of 1284 blood samples from local zebu cattle (642 each in dry and wet season) were examined by using buffy coat technique and thin blood smear method. The overall prevalence was 11.

View Article and Find Full Text PDF

Background: Trypanosomosis caused by Trypanosoma vivax is one of the diseases threatening the health and productivity of livestock in Africa and Latin America. Trypanosoma vivax is mainly transmitted by tsetse flies; however, the parasite has also acquired the ability to be transmitted mechanically by hematophagous dipterans. Understanding its distribution, host range and prevalence is a key step in local and global efforts to control the disease.

View Article and Find Full Text PDF

Several types of odours are involved in the location of host animals by tsetse (Diptera: Glossinidae), a vector of animal African trypanosomiasis. Host animals' ageing urine has been shown to be the source of a phenolic blend attractive to the tsetse. Nevertheless, limited research has been performed on the microbial communities' role in the production of phenols.

View Article and Find Full Text PDF

Background: Surra is an animal trypanosomosis, caused by infection with Trypanosoma evansi and leading to severe economic loss due to mortality and morbidity. Compared to tsetse-transmitted animal trypanosomoses, little attention is given to the epidemiology and control of surra. Understanding its epidemiology is a first step in local and global efforts to control the disease.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!