Molecular characterization of pathogenic African trypanosomes in biting flies and camels in surra-endemic areas outside the tsetse fly belt in Kenya

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Authors

Getahun, Merid N.
Villinger, Jandouwe
Bargul, Joel L.
Muema, Jackson M.
Orone, Abel
Ngiela, John
Ahuya, Peter O.
Saini, Rajinder K.
Torto, Baldwyn
Masiga, Daniel K.

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Publisher

Springer

Abstract

African animal trypanosomosis (nagana) is becoming prevalent beyond its traditionally defined geographical boundaries in African tsetse belts. However, knowledge of clinically important trypanosomes and infection rate in non-tsetse hematophagous flies and domestic animals are limited. This study characterized the potential mechanical vectors, their host feeding patterns, and trypanosome infection in them and domestic animals outside the tsetse belt in northern Kenya. Field-trapped flies and blood from camels, cattle, donkeys, goats, and sheep were screened for trypanosome infection by microscopy and polymerase chain reaction (PCR) of the internal transcribed spacer 1 region. Blood-fed specimens were analysed using PCR-HRM and/or sequencing of 16S rRNA gene to identify vertebrate blood-meal host sources. Hippobosca camelina, Stomoxys calcitrans, Tabanus spp., and Pangonia rueppellii were identified as potential vectors of trypanosomes outside the tsetse belt in Marsabit County. The trypanosome species, Trypanosoma vivax, T. evansi, T. brucei, and T. congolense were recovered in biting flies as well as in camels (Camelus dromedarius). The diversity of parasites in the biting flies was similar to that detected in the tsetse fly Glossina pallidipes collected from the tsetse-infested Shimba Hills, in coastal Kenya, suggesting a wide geographic distribution of the trypanosomes in Kenya. The biting flies fed on camels, cattle, goats, and sheep. Furthermore, we identified diverse clinical outcomes based on PCV (anemia), heamorrhagia) associated with infection with disparate Trypanosoma species. Thus, infection of flies and camels by diverse Trypanosoma species could contribute to the complex epidemiology of observed trypanosomosis in camels.

Description

SUPPLEMENTARY FILE 1: FIGURE S1. PCR products were resolved 1% ethidium-bromide stained agarose gel (8V for 1.5 hrs) to check for any contamination. The DNA isolated from whole fly was amplified targeting trypanosomal ITS1 gene. Lane: M 10- bp marker, Bf (reaction buffer), wt (PCR water), TB (T. brucei ILTat 1.4) TV (T. vivax IL 2136), TC (T. congolense savannah (IL3000)), and TE (T. evansi KETRI 2479), F1- F10 DNA sample from H. camelina flies. The absence of PCR product under Bf, and wt show no contamination from extraction buffer.
SUPPLEMENTARY FILE 2: FIGURE S2. Number of H. camelina recaptured at the specified distance from pint of release. Number in parenthesis shows percentage of flies recaptured.
SUPPLEMENTARY FILE 3: FIGURE S3. (A) PCR products were resolved 1% ethidium-bromide stained agarose gel (8V for 1.5 hrs) to check for trypanosomes in blood and lymph node aspirate. The DNA isolated from blood and lymph node aspirate was amplified targeting trypanosomal ITS1 gene. Lane: M 10- bp marker, -Ve (reaction buffer), TE (T. evansi KETRI 2479) TV (T. vivax IL 2136), TC (T. congolense savannah (IL3000)), and LN_C1, LN_C2, DNA sample from two camel lymph node aspirate, B_C1 and B_C2 DNA from corresponding blood samples from the same camel. The result shows both samples of the lymph node aspirate were positive, while blood samples were negative from the same camel. (B) Five camels blood and lymph node aspirate were analysed, only camel five lymph node aspirate was positive for T.vivax but blood sample from the same camel was negative.
SUPPLEMENTARY FILE 4. TABLE S1. Trypanosomes identified based on ITS1 gene sequence from different host included in Fig.6.
DATA AVAILABILITY: All data generated or analysed in this study are included in the article and as additional files. The newly generated sequences were deposited in the NCBI Nucleotide database under the accession numbers listed in Supplementary table.

Keywords

Surra, Camel, Non-tsetse transmitted trypanosomes, Biting flies, Tsetse belt, Kenya

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Citation

Getahun, M.N., Villinger, J., Bargul, J.L. et al. Molecular characterization of pathogenic African trypanosomes in biting flies and camels in surra-endemic areas outside the tsetse fly belt in Kenya. International Journal of Tropical Insect Science 42, 3729–3745 (2022). https://doi.org/10.1007/s42690-022-00896-2.