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The use of molecular technology to investigate trypanosome infections in tsetse flies at Liwonde Wild Life Reserve.
Malawi Med J. 2019 12; 31(4):233-237.MM

Abstract

Background

Trypanosomes are protozoan flagellates that cause human African trypanosomiasis (HAT) and African animal trypanosomiasis (AAT). HAT is caused by Trypanosoma brucei rhodesiense in East and Central Africa and T.b. gambiense in West Africa, whereas AAT is caused by a number of trypanosome species, including T. brucei brucei, T. evansi, T. vivax, T. congolense, T. godfreyi and T. simiae. The aim of this study was to establish if tsetse flies at Liwonde Wild Life Reserve (LWLR) are infected with these trypanosomes and thus pose a risk to both humans and animals within and surrounding the LWLR.

Methods

A total of 150 tsetse flies were caught. Of these, 82 remained alive after capture and were dissected such that the mid-gut could be examined microscopically for trypanosomes. DNA extractions were performed from both mid-guts and the 68 dead flies using a Qiagen Kit. Amplification techniques involved the Internal Transcriber Spacer 1 (ITS 1) conventional polymerase chain reaction (PCR) with primers designed to identify trypanosome species, and Repetitive Insertion Mobile Element - Loop Mediated Isothermal Amplification (RIME LAMP), a sequence specific to T. brucei.

Results

Analysis showed that 79/82 (96.3%) of the mid-guts examined microscopically were positive for trypanosomes and that 75/150 (50%) of the DNA extracts (from the mid-gut, and tsetse fly carcasses) were positive for T. brucei, as determined by the RIME LAMP method. ITS1 PCR further showed that 87/150 (58.0%) flies were positive for trypanosomes, of which 56/87 (64.4%) were T. brucei, 9/87 (10.3%) were T. vivax; 7/87 (8.1%) were T. simiae; 6/87 (6.9%) were T. congolense, and 6/87 (6.9%) were T. godfreyi. Ten samples had a mixture of infections.

Conclusion

Our analysis demonstrated a mixture of infections from trypanosome species in tsetse flies at LWLR, and that T. brucei, the species that causes HAT, was the most common. Our study successfully used molecular techniques to demonstrate the presence of T. b. rhodesiense at LWLR, a species that causes HAT in both East and Central Africa.

Authors+Show Affiliations

College of Medicine, Blantyre, Malawi.College of Medicine, Blantyre, Malawi.College of Medicine, Blantyre, Malawi. Malawi-Liverpool Wellcome Trust (MLW), Blantyre, Malawi.College of Medicine, Blantyre, Malawi.College of Medicine, Blantyre, Malawi.College of Medicine, Blantyre, Malawi. Malawi-Liverpool Wellcome Trust (MLW), Blantyre, Malawi.Mikolongwe Veterinary College of Agriculture and Food Security, Limbe, Malawi.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32133052

Citation

Nayupe, Symon F., et al. "The Use of Molecular Technology to Investigate Trypanosome Infections in Tsetse Flies at Liwonde Wild Life Reserve." Malawi Medical Journal : the Journal of Medical Association of Malawi, vol. 31, no. 4, 2019, pp. 233-237.
Nayupe SF, Simwela NV, Kamanga PM, et al. The use of molecular technology to investigate trypanosome infections in tsetse flies at Liwonde Wild Life Reserve. Malawi Med J. 2019;31(4):233-237.
Nayupe, S. F., Simwela, N. V., Kamanga, P. M., Chisi, J. E., Senga, E., Musaya, J., & Maganga, E. (2019). The use of molecular technology to investigate trypanosome infections in tsetse flies at Liwonde Wild Life Reserve. Malawi Medical Journal : the Journal of Medical Association of Malawi, 31(4), 233-237. https://doi.org/10.4314/mmj.v31i4.3
Nayupe SF, et al. The Use of Molecular Technology to Investigate Trypanosome Infections in Tsetse Flies at Liwonde Wild Life Reserve. Malawi Med J. 2019;31(4):233-237. PubMed PMID: 32133052.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The use of molecular technology to investigate trypanosome infections in tsetse flies at Liwonde Wild Life Reserve. AU - Nayupe,Symon F, AU - Simwela,Nelson V, AU - Kamanga,Peace M, AU - Chisi,John E, AU - Senga,Edward, AU - Musaya,Janelisa, AU - Maganga,Emmanuel, PY - 2020/3/6/entrez PY - 2020/3/7/pubmed PY - 2020/5/27/medline KW - African Trypanosomiasis KW - DNA KW - Malawi KW - Polymerase Chain Reaction KW - Southern Africa KW - Tsetse Flies SP - 233 EP - 237 JF - Malawi medical journal : the journal of Medical Association of Malawi JO - Malawi Med J VL - 31 IS - 4 N2 - Background: Trypanosomes are protozoan flagellates that cause human African trypanosomiasis (HAT) and African animal trypanosomiasis (AAT). HAT is caused by Trypanosoma brucei rhodesiense in East and Central Africa and T.b. gambiense in West Africa, whereas AAT is caused by a number of trypanosome species, including T. brucei brucei, T. evansi, T. vivax, T. congolense, T. godfreyi and T. simiae. The aim of this study was to establish if tsetse flies at Liwonde Wild Life Reserve (LWLR) are infected with these trypanosomes and thus pose a risk to both humans and animals within and surrounding the LWLR. Methods: A total of 150 tsetse flies were caught. Of these, 82 remained alive after capture and were dissected such that the mid-gut could be examined microscopically for trypanosomes. DNA extractions were performed from both mid-guts and the 68 dead flies using a Qiagen Kit. Amplification techniques involved the Internal Transcriber Spacer 1 (ITS 1) conventional polymerase chain reaction (PCR) with primers designed to identify trypanosome species, and Repetitive Insertion Mobile Element - Loop Mediated Isothermal Amplification (RIME LAMP), a sequence specific to T. brucei. Results: Analysis showed that 79/82 (96.3%) of the mid-guts examined microscopically were positive for trypanosomes and that 75/150 (50%) of the DNA extracts (from the mid-gut, and tsetse fly carcasses) were positive for T. brucei, as determined by the RIME LAMP method. ITS1 PCR further showed that 87/150 (58.0%) flies were positive for trypanosomes, of which 56/87 (64.4%) were T. brucei, 9/87 (10.3%) were T. vivax; 7/87 (8.1%) were T. simiae; 6/87 (6.9%) were T. congolense, and 6/87 (6.9%) were T. godfreyi. Ten samples had a mixture of infections. Conclusion: Our analysis demonstrated a mixture of infections from trypanosome species in tsetse flies at LWLR, and that T. brucei, the species that causes HAT, was the most common. Our study successfully used molecular techniques to demonstrate the presence of T. b. rhodesiense at LWLR, a species that causes HAT in both East and Central Africa. SN - 1995-7270 UR - https://www.unboundmedicine.com/medline/citation/32133052/The_use_of_molecular_technology_to_investigate_trypanosome_infections_in_tsetse_flies_at_Liwonde_Wild_Life_Reserve_ DB - PRIME DP - Unbound Medicine ER -