Trypanosome Diversity in Wildlife Species from the Serengeti and Luangwa Valley Ecosystems

Abstract

Background

The importance of wildlife as reservoirs of African trypanosomes pathogenic to man and livestock is well recognised. While new species of trypanosomes and their variants have been identified in tsetse populations, our knowledge of trypanosome species that are circulating in wildlife populations and their genetic diversity is limited.

Methodology/Principal Findings

Molecular phylogenetic methods were used to examine the genetic diversity and species composition of trypanosomes circulating in wildlife from two ecosystems that exhibit high host species diversity: the Serengeti in Tanzania and the Luangwa Valley in Zambia. Phylogenetic relationships were assessed by alignment of partial 18S, 5.8S and 28S trypanosomal nuclear ribosomal DNA array sequences within the Trypanosomatidae and using ITS1, 5.8S and ITS2 for more detailed analysis of the T. vivax clade. In addition to Trypanosoma brucei, T. congolense, T. simiae, T. simiae (Tsavo), T. godfreyi and T. theileri, three variants of T. vivax were identified from three different wildlife species within one ecosystem, including sequences from trypanosomes from a giraffe and a waterbuck that differed from all published sequences and from each other, and did not amplify with conventional primers for T. vivax.

Conclusions/Significance

Wildlife carries a wide range of trypanosome species. The failure of the diverse T. vivax in this study to amplify with conventional primers suggests that T. vivax may have been under-diagnosed in Tanzania. Since conventional species-specific primers may not amplify all trypanosomes of interest, the use of ITS PCR primers followed by sequencing is a valuable approach to investigate diversity of trypanosome infections in wildlife; amplification of sequences outside the T. brucei clade raises concerns regarding ITS primer specificity for wildlife samples if sequence confirmation is not also undertaken.

Author Summary

The trypanosomes include a number of species that cause disease in livestock. In recent years, several trypanosomes have been identified which do not fit into the classic trypanosome classification system. However, previous work has focused on trypanosomes identified in the tsetse vector, with little information available on trypanosomes found in their natural hosts, wildlife. We studied trypanosome sequences from wildlife in Serengeti National Park in Tanzania and the Luangwa Valley in Zambia and found a number of trypanosome species pathogenic to livestock were circulating in these areas. For Trypanosoma vivax, one of the causes of trypanosomiasis in cattle, variants were identified in giraffe and waterbuck that were different from all published sequences and from each other. These variants did not test positive with the molecular tests usually used to identify T. vivax suggesting that T. vivax may often be under-diagnosed in Tanzania. The trypanosome classification system is facing challenges as molecular data are incorporated into a system that historically was based on factors such as morphology, host range and geographical distribution.