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Trypanosome diversity in wildlife species from the serengeti and Luangwa Valley ecosystems.

Auty H, Anderson NE, Picozzi K, Lembo T, Mubanga J, Hoare R, Fyumagwa RD, Mable B, Hamill L, Cleaveland S, Welburn SC - PLoS Negl Trop Dis (2012)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Centre for Infectious Diseases, College of Medicine and Veterinary Medicine, The University of Edinburgh, Edinburgh, United Kingdom.

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.

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Related in: MedlinePlus

Unrooted neighbour-joining dendrogram of ITS1, 5.8S and ITS2 sequences for Trypanosoma vivax.Bootstrap values are shown at nodes with >70% support. Sequences generated in this study shown in black; other sequences retrieved from Genbank and listed in Table 3. Sequence locations are shown by colour: Tanzania (this study, black); Kenya (blue); Mozambique (green); Nigeria (purple); South America (red). Host species from which sequence was amplified are indicated. Accession numbers for reference sequences are: IL3905 cl8, Genbank ID:DQ316040; IL3905 cl4Ro, DQ316043; IL3905 cl4, DQ316039; IL3905 cl3Ro, DQ316042; IL3905 cl2Ro, DQ316041; IL3905 cl5Ro, DQ316044; IL3905 cl2 DQ316037; IL3905 cl3, DQ316038; TviBrMi cl4, DQ316048; TviBrPo cl13, DQ316049; TviBrCa cl2, DQ316045; Y485, U22316; TviBrMi cl2, DQ316047; TviVeMe cl1, DQ316051; TviVeMe cl12, DQ316052; TviBrCa cl13, DQ316046; TviBrPo cl6, DQ316050; TviMzNy cl5, EU482080; TviMzNy cl2, EU482079; TviMzNy cl8, EU482082; TviMzNy cl1, EU482078; TviMzNy cl6, EU482081.
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pntd-0001828-g002: Unrooted neighbour-joining dendrogram of ITS1, 5.8S and ITS2 sequences for Trypanosoma vivax.Bootstrap values are shown at nodes with >70% support. Sequences generated in this study shown in black; other sequences retrieved from Genbank and listed in Table 3. Sequence locations are shown by colour: Tanzania (this study, black); Kenya (blue); Mozambique (green); Nigeria (purple); South America (red). Host species from which sequence was amplified are indicated. Accession numbers for reference sequences are: IL3905 cl8, Genbank ID:DQ316040; IL3905 cl4Ro, DQ316043; IL3905 cl4, DQ316039; IL3905 cl3Ro, DQ316042; IL3905 cl2Ro, DQ316041; IL3905 cl5Ro, DQ316044; IL3905 cl2 DQ316037; IL3905 cl3, DQ316038; TviBrMi cl4, DQ316048; TviBrPo cl13, DQ316049; TviBrCa cl2, DQ316045; Y485, U22316; TviBrMi cl2, DQ316047; TviVeMe cl1, DQ316051; TviVeMe cl12, DQ316052; TviBrCa cl13, DQ316046; TviBrPo cl6, DQ316050; TviMzNy cl5, EU482080; TviMzNy cl2, EU482079; TviMzNy cl8, EU482082; TviMzNy cl1, EU482078; TviMzNy cl6, EU482081.

Mentions: Blast results suggested that T. vivax sequences generated in this study did not closely match existing sequences. To assess the phylogenetic relationships within the T. vivax clade, ITS1, 5.8S and ITS2 sequences were aligned for all sequences clustering in this group, together with all T. vivax sequences available in Genbank for this region, and an unrooted neighbour-joining tree constructed using a HKY model of substitution in Geneious, with bootstrap values calculated for 1000 replicates. Accession numbers for all T. vivax reference sequences are listed in Figure 2.


Trypanosome diversity in wildlife species from the serengeti and Luangwa Valley ecosystems.

Auty H, Anderson NE, Picozzi K, Lembo T, Mubanga J, Hoare R, Fyumagwa RD, Mable B, Hamill L, Cleaveland S, Welburn SC - PLoS Negl Trop Dis (2012)

Unrooted neighbour-joining dendrogram of ITS1, 5.8S and ITS2 sequences for Trypanosoma vivax.Bootstrap values are shown at nodes with >70% support. Sequences generated in this study shown in black; other sequences retrieved from Genbank and listed in Table 3. Sequence locations are shown by colour: Tanzania (this study, black); Kenya (blue); Mozambique (green); Nigeria (purple); South America (red). Host species from which sequence was amplified are indicated. Accession numbers for reference sequences are: IL3905 cl8, Genbank ID:DQ316040; IL3905 cl4Ro, DQ316043; IL3905 cl4, DQ316039; IL3905 cl3Ro, DQ316042; IL3905 cl2Ro, DQ316041; IL3905 cl5Ro, DQ316044; IL3905 cl2 DQ316037; IL3905 cl3, DQ316038; TviBrMi cl4, DQ316048; TviBrPo cl13, DQ316049; TviBrCa cl2, DQ316045; Y485, U22316; TviBrMi cl2, DQ316047; TviVeMe cl1, DQ316051; TviVeMe cl12, DQ316052; TviBrCa cl13, DQ316046; TviBrPo cl6, DQ316050; TviMzNy cl5, EU482080; TviMzNy cl2, EU482079; TviMzNy cl8, EU482082; TviMzNy cl1, EU482078; TviMzNy cl6, EU482081.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3475651&req=5

pntd-0001828-g002: Unrooted neighbour-joining dendrogram of ITS1, 5.8S and ITS2 sequences for Trypanosoma vivax.Bootstrap values are shown at nodes with >70% support. Sequences generated in this study shown in black; other sequences retrieved from Genbank and listed in Table 3. Sequence locations are shown by colour: Tanzania (this study, black); Kenya (blue); Mozambique (green); Nigeria (purple); South America (red). Host species from which sequence was amplified are indicated. Accession numbers for reference sequences are: IL3905 cl8, Genbank ID:DQ316040; IL3905 cl4Ro, DQ316043; IL3905 cl4, DQ316039; IL3905 cl3Ro, DQ316042; IL3905 cl2Ro, DQ316041; IL3905 cl5Ro, DQ316044; IL3905 cl2 DQ316037; IL3905 cl3, DQ316038; TviBrMi cl4, DQ316048; TviBrPo cl13, DQ316049; TviBrCa cl2, DQ316045; Y485, U22316; TviBrMi cl2, DQ316047; TviVeMe cl1, DQ316051; TviVeMe cl12, DQ316052; TviBrCa cl13, DQ316046; TviBrPo cl6, DQ316050; TviMzNy cl5, EU482080; TviMzNy cl2, EU482079; TviMzNy cl8, EU482082; TviMzNy cl1, EU482078; TviMzNy cl6, EU482081.
Mentions: Blast results suggested that T. vivax sequences generated in this study did not closely match existing sequences. To assess the phylogenetic relationships within the T. vivax clade, ITS1, 5.8S and ITS2 sequences were aligned for all sequences clustering in this group, together with all T. vivax sequences available in Genbank for this region, and an unrooted neighbour-joining tree constructed using a HKY model of substitution in Geneious, with bootstrap values calculated for 1000 replicates. Accession numbers for all T. vivax reference sequences are listed in Figure 2.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Centre for Infectious Diseases, College of Medicine and Veterinary Medicine, The University of Edinburgh, Edinburgh, United Kingdom.

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.

Show MeSH
Related in: MedlinePlus