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Molecular study on parasitic nematodes infection in the abomasum of sheep in Ilam, Iran.

ABSTRACT: Parasitic nematodes of ovine abomasum are of economic and hygienic importance throughout the world and Iran. This study was aimed to evaluate molecular identity and species diversity of parasitic nematodes in the abomasum of slaughtered sheep in Ilam, Iran. In this study, a total number of 240 of abomasa were randomly collected from the slaughtered sheep at industrial slaughterhouses in Ilam in all seasons between 2017 and 2018. The abomasum content and abomasal mucosa were removed and washed. The collected nematodes were morphologically identified. The genomic DNA was extracted and a 300 bp-fragment-length from internal transcribed spacer 2 ribosomal ribonucleic acid (ITS2-rRNA) gene was amplified. Overall prevalence was 66.70% (160/240). Five species of four genera of nematodes including Marshallagia marshalli (43.70%), Ostertagia circumcincta (15.50%), Parabronema skrjabini (5.00%), M. occidentalis (2.50%), and Haemonchus contortus (0.04%) were identified. Ostertagia circumcincta and H. contortus were found to be different in two nucleotides. There was no nucleotide difference between M. marshalli and M. occidentalis. This study revealed a significant prevalence of parasitic nematodes in sheep abomasum and species diversity of Trichostrongylid nematodes in the region.

SUBMITTER: Nazarbeigy M

PROVIDER: S-EPMC8328256 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundThis study was carried out to investigate the prevalence and analyze the molecular characteristics based on the internal transcribed spacer (ITS) 2 region of the ribosomal RNA (RNA) gene of trichostrongylid nematodes in different ruminants from Guilan province, northern of Iran.MethodsThe gastrointestinal tracts of 144 ruminants including 72 cattle, 59 sheep, and 13 goats were collected from an abattoir in Guilan province during July to September 2018. After isolation the helminths, male specimens were identified based on morphological parameters. PCR and partial sequencing of the ITS2 fragment were conducted. After phylogenetic analysis, the intraspecific and interspecific differences were calculated.ResultsThe prevalence of total infections with the nematodes was 38.9, 74.6 and 84.6% among cattle, sheep and goats, respectively. Eleven species of trichostrongylid nematodes including Haemonchus contortus, Marshallagia marshalli, Trichostrongylus axei, T. colubriformis, T. vitrinus, Ostertagia trifurcata, Teladorsagia circumcincta, Marshallagia occidentalis, O. lyrata, O. ostertagi, and Cooperia punctate were recovered from the ruminants. The most prevalent trichostrongyloid nematodes in cattle, sheep and goats were O. ostertagi (26.4%), M. marshalli (64.4%) and T. circumcincta (69.2%), respectively. Phylogenetic tree was discriminative for Trichostrongylidae family, while phylogenetic analysis of the ITS2 gene represented low variations and no species identification of Haemonchidae and Cooperiidae families.ConclusionsThis study suggests the high prevalence and species diversity of trichostrongyloid nematodes in different ruminants, indicating the importance of implement antiparasitic strategies in north regions of Iran. As well, this study showed that the ITS2 fragment is not a discriminative marker for Haemonchidae and Cooperiidae families, and investigation of other genetic markers such as mitochondrial genes would be more valuable for better understanding of their phylogenetic relationships.

Project description:BackgroundMolecular advances have accelerated our understanding of nematode systematics and taxonomy. However, comparative analyzes between various genetic markers have led to discrepancies in nematode phylogenies. This study aimed to evaluate the suitability of using mitochondrial 12S and 16S ribosomal RNA genes for nematode molecular systematics.MethodsTo study the suitability of mitochondrial 12S and 16S ribosomal RNA genes as genetic markers for nematode molecular systematics, we compared them with the other commonly used genetic markers, nuclear internal transcribed spacer 1 and 2 regions, nuclear 18S and 28S ribosomal RNA genes, and mitochondrial cytochrome c oxidase subunit 1 gene. After that, phylum-wide primers for mitochondrial 12S and 16S ribosomal RNA genes were designed, and parasitic nematodes of humans and animals from 75 taxa with 21 representative species were inferred through phylogenetic analyzes. Phylogenetic analyzes were carried out using maximum likelihood and Bayesian inference algorithms.ResultsThe phylogenetic relationships of nematodes based on the mitochondrial 12S rRNA gene supported the monophyly of nematodes in clades I, IV, and V, reinforcing the potential of this gene as a genetic marker for nematode systematics. In contrast, the mitochondrial 16S rRNA gene only supported the monophyly of clades I and V, providing evidence that the 12S rRNA gene is more suitable for nematode molecular systematics. In this study, subclades of clade III containing various nematode families were not monophyletic when the 16S or 12S rRNA gene was used as the genetic marker. This is similar to the phylogenetic relationship revealed by previous studies using whole mitochondrial genomes as genetic markers.ConclusionsThis study supports the use of the 12S rRNA gene as a genetic marker for studying the molecular systematics of nematodes to understand intra-phyla relationships. Phylum-wide primers for nematodes using mitochondrial ribosomal genes were prepared, which may enhance future studies. Furthermore, sufficient genetic variation in the mitochondrial 12S and 16S rRNA genes between species also allowed for accurate taxonomy to species level, revealing the potential of these two genes as genetic markers for DNA barcoding.

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Molecular study on parasitic nematodes infection in the abomasum of sheep in Ilam, Iran.

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