Multiplex PCR-based amplification of the ITS2 region of rDNA and

phylogenetic characterization of zoonotic Trichostrongylus species

isolated from goats for the first time in Bangladesh

Nurnabi Ahmed, Babul Chandra Roy, Mohammad Manjurul Hasan, Hiranmoy Biswas, Md.
Makshuder Rahman Zim and Md. Hasanuzzaman Talukder*.
Department of Parasitology, Bangladesh Agricultural University, Mymensingh-2202,
Bangladesh.
*Corresponding author: E-mail: talukdermhasan@bau.edu.bd

Abstract:

Trichostrongylus is one of the most important trichostrongylid nematodes, infecting livestock and
humans, which causes a zoonotic disease known as trichostrongyliasis. The study was conducted
to identify Trichostrongylus species from goats by morphology-morphometry and multiplex PCR
followed by sequence analysis. A total of 124 goat’s viscera were collected, processed and
trichostrongylid parasites were isolated and identified. A total of 39 out of the 124 viscera of goats
(31.45%) were infected. Morphometry revealed two Trichostrongylus species. Based on multiplex
PCR, T. vitrinus (150 bp) and T. colubriformis (232 bp) were identified. Sequence analysis also
revealed these two species. ITS2 partial nucleotide sections of T. vitrinus and T. colubriformis
isolates displayed 3 and 4 haplotypes, respectively. Seven single nucleotide polymorphisms were
detected (3 transitions and 4 transversions) in these risen haplotypes. A phylogenetic tree was
constructed with 36 sequences (eight studied genotypes and 28 reference sequences from the
GenBank database). The neighbor-joining tree represented that the studied T. colubriformis and T.
vitrinus isolates were grouped with reference T. colubriformis and T. vitrinus isolates with strong
nodal support in clade A and B without any geographical boundaries. This is the first study on
the genetic analysis of Trichostrongylus species in Bangladesh. These findings will provide
baseline data on the species confirmation and genetic variations of Trichostrongylus species.
Further, extensive investigation is imperative to reveal the population genetic structure of this
parasite in livestock and humans in Bangladesh.
Keywords: T. colubriformis, T. vitrinus, Morphology, Morphometry, Multiplex PCR, ITS2 gene
sequencing, Goat, Bangladesh.