Molecular characterization of Trypanosoma evansi isolates from water buffaloes (Bubalus bubalis) in the Philippines

Villareal, Marjo V.; Mingala, Claro N.; Rivera, Windell L.

doi:10.2478/s11686-013-0110-5

Cited by 19 publications

(15 citation statements)

References 57 publications

(46 reference statements)

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“…This was supported by study result of Masiga et al (2006) who found a genetic diversity of T. evansi from Kenya according to the amplified fragment length polymorphism/AFLP test (Masiga et al 2006). A genetic variable of T. evansi was also reported based on the expression-siteassociated gene e 6/7 (ESAG6/7) as a marker (Mekata et al 2009;Villareal et al 2013). Trypanosome is able to avoid the immune attack of the host by changing the expression of Variant Surface Glycoprotein (VSG) genes.…”

Section: Introductionsupporting

confidence: 50%

Genetic variability of ESAG6/7 gene Trypanosoma evansi

Sawitri¹,

Wardhana²

2018

JITV

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ABSTRAKSawitri DH, Wardhana AH. 2017. Variabilitas genetik dari gen ESAG 6/7 Isolat T. evansi. JITV 22(1): 38-50. DOI: http://dx.doi.org/10.14334/jitv.v22i1.1638Trypanosoma evansi (T. evansi) sebagai agen penyakit Sura merupakan salah satu penyakit parasitik penting untuk diperhatikan karena dapat menimbulkan kerugian ekonomi yang sangat besar di Indonesia. Parasit ini memerlukan zat besi untuk fase propagasi yang diperoleh dari pejamu melalui reseptor transferrin (Tf) yang dikode oleh Expression Site Associated Genees (ESAGs). ESAG6/7 dilaporkan mengkode reseptor transferrin pada afinitas yang berbeda pada pejamu yang berbeda.Adanya perbedaan patogeneitas T. evansi diduga menyebabkan variabilitas pada gene ESAG 6/7. Penelitian ini bertujuan untuk melihat variabilitas gene ESAG6 T. evansi dengan virulensi yang berbeda pada mencit. Penelitian ini dilakukan dengan 2 tahap yaitu uji patogeneitas T. evansi pada mencit dan analisis sekuen gene ESAG6/7. Uji patogeneitas dilakukan dengan melihat median lama hidup mencit setelah masing-masing kelompok diinfeksi dengan 25 T. evansi asal kerbau dari berbagai kondisi geografi. Hasil penelitian uji patogeneitas menunjukkan adanya perbedaan virulensi pada 25 isolat T.evansi pada mencit. Trypanosoma evansi as an agent of Surra is one of the crucial parasitic diseases that cause great economic losses in Indonesia. These parasites need iron for growth and propagation phase which is obtained by receptor-mediated uptake of host transferin. The transferrin receptors are encoded by Expression Site Associated Genees (ESAGs). ESAG6/7 encodes transferrin receptors which reported have different affinities of a different host. The distinction of T. evansi pathogenicity is supposed to cause variability in the ESAG6/7 gene. This research was aimed to investigate the variability of genes ESAG6/7 T. evansi with different virulence in mice. This research was conducted in two steps: bioassay pathogeneicity in mice and analysis of ESAG6/7 gene sequences. The median survival time of mice was investigated after each group of mice infected by 25 T. evansi isolates from buffaloes where its geographically differ. The test results showed a difference of pathogenic virulence on 25 T. evansi isolates in mice. Sequence analysis of the ESAG6/7 gene from 25 T. evansi isolates origin from Indonesia tended to be homogeneous on the transferrin binding site but there was variability in the hypervariable site. These changes are able to separate high and low virulence of the T. evansi isolates. Phylogenetic tree analysis was formed 11 clades of 25 T. evansi. High virulence T. evansi was included in clades 7 and 10, while low virulence T. evansi was included in clade 5 and 11 and the moderate virulence was divided into those four clades.

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Section: Introductionsupporting

confidence: 50%

Genetic variability of ESAG6/7 gene Trypanosoma evansi

Sawitri¹,

Wardhana²

2018

JITV

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“…A partial sequence of RoTat 1.2 VSG gene was identical to the T. evansi sequences reported from India and Kenya, but the varied similarity was seen when aligned with other Egyptian T. evansi sequences [22]. Whilst, analysis of transferrin receptor-encoding gene (ESAG6) showed variable repertoire in the studied isolates, which may indicate to a novel structure of T. evansi population from Egypt and/or a difference in host range [18]. In the present study, the results of the phylogenies obtained from the Maximum Parsimony (MP) method suggest that T. evansi isolates from HSA (Egypt), America and CIV (India) are more diverse than other isolates.…”

Section: Discussionmentioning

confidence: 82%

“…Thus, this study intended to apply TR3/TR4 as a molecular tool to characterize trypanosomes in Egypt. The current search was the third study on T. evansi genotypes in imported dromedary camels in Egypt after [18,22], but it was the first studied T. evansi genotypes in both imported and local camels. A partial sequence of RoTat 1.2 VSG gene was identical to the T. evansi sequences reported from India and Kenya, but the varied similarity was seen when aligned with other Egyptian T. evansi sequences [22].…”

Section: Discussionmentioning

confidence: 99%

“…During their journey in Egypt, they mix with local animals at borders leading to mixed genotyping and strain difference might occur between Egyptian sub-populations and imported strains of T. evansi [10,24]. A few studies on genetic variability and characterization of T. evansi isolate in Sudanese originating camels and in other domestic animals were conducted [10,18,22], but the genetic information is lacking in local dromedary camels. The objective of this study was to ascertain the usefulness of sequence analysis of the region targeted by TR3/TR4 in assessing the genetic variability and characterization of Trypanosoma circulating in imported and local camels in Egypt, and as a tool to study phylogenetic and potential relationships of various T. evansi isolates.…”

Section: Introductionmentioning

confidence: 99%

“…Few studies have been carried out to investigate whether T. evansi populations are genetically sub-structured using the internal transcribed spacer (ITS) region including (ITS1 and ITS2), that are located between the repeating array of nuclear 18S, 5.8S, and 28S ribosomal RNA genes [13][14][15][16][17]. In addition, diversity of the transferrin receptor-encoding gene located at expression-site-associated (ESAGs 6 and 7) genes [18][19][20][21] are utilized in molecular classification of trypanosomes. Those studies demonstrated that the first and second internal transcribed spacers (ITS-1 and ITS-2, respectively) of the rDNA are useful targets for species delineation and for inferring phylogenetic relationships of Trypanosoma spp.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Characterization and Phylogenetic Analysis of Trypanosoma evansi from Local and Imported Camels in Egypt

Barghash¹,

Darwish²

2016

J Phylogen Evolution Biol

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Trypanosoma evansi, the agent of trypanosomiasis commonly known as Surra or Guffar, is regarded as one of the most economically important animal parasitic pathogen affecting livestock in Egypt. The current study aims to discuss genetic characterization and phylogenetic analysis of Trypanosoma isolates from local and imported naturally infected camels in Egypt. The study was initially started with parasitological and molecular surveillance on 411 native and 117 imported camels by using PCR-RoTat 1.2 VSG gene targeting 205 bp. Further, the molecular characterization and sequencing were achieved on four positive samples using PCR-TR3/TR4 primers that derived from a trypanosomespecific repetitive nucleotide sequence fragment. Product sequences were aligned against the corresponding GenBank sequences of known isolates of T. evansi and subjected to phylogenetic analysis. Results revealed that T. evansi was present in 66.67% and 74.36% of the local and imported camels respectively, regardless of age and sex factors. Basic Local Alignment Search Tool (BLAST) data of the obtained PCR TR3/TR4 gene sequences revealed that they corresponded to those of T. evansi, with the homology of 93% to 99%. Phylogenetic and molecular analyses of this gene showed that three genotypes of T. evansi in Egypt are present showed two common SNPs (G136A and G189T) in all samples, two SNPs in ISM and ISD (C3T and A207G) and six SNPs in HSA (T12C, C14T, T15C, G19C, C21G, and G22C). We conclude that T. evansi is described as presenting genotype variability among its isolates according to geographical distribution in Egypt.

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Molecular study of Trypanosoma caninum isolates based on different genetic markers

Barros¹,

Toma

Madeira

2015

Parasitol Res

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Trypanosoma caninum is a parasite recently described in dogs, whose life cycle is rather unknown. Here, we performed a genetic study with T. caninum samples obtained in different Brazilian regions. The study was based on PCR assays target to small and large subunit ribosomal DNA (rDNA) (18S rDNA and 24Sα rDNA), cytochrome B (Cyt b), and internal transcribed spacer 1 rDNA (ITS1 rDNA) following by the sequence analysis. Additionally, we used primers for the variable regions of kinetoplast DNA (kDNA) minicircles and endonucleases restriction in the ITS1 rDNA amplification product. T. caninum samples displayed the same patterns. Tree construction confirmed the close relationship between T. caninum samples, regardless of the molecular target used and endonuclease restriction digestion revealed that all samples have the same restriction profile. Therefore, T. caninum seems to be a genetically homogeneous specie. In the kDNA assay, T. caninum possessed a different molecular size profile with respect to others trypanosomes, 330 and 350 bp. This study provides nucleotide sequences from different regions of the genome of T. caninum that certainly facilitate future studies.

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Molecular characterization of Trypanosoma evansi isolates from water buffaloes (Bubalus bubalis) in the Philippines

Cited by 19 publications

References 57 publications

Genetic variability of ESAG6/7 gene Trypanosoma evansi

Genetic variability of ESAG6/7 gene Trypanosoma evansi

Molecular Characterization and Phylogenetic Analysis of Trypanosoma evansi from Local and Imported Camels in Egypt

Molecular study of Trypanosoma caninum isolates based on different genetic markers

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