Abstract
The current study was contributed to the analysis of the nucleotide sequence pattern of the nucleotide sequence of the tissue DNA isolates based on the Internal Transcribed Spacer ribosomal DNA (ITS2rDNA) gene and Cytochrome c oxidase subunit 1 mitochondrial DNA (COX1 mtDNA) using the traditional polymerase chain reaction (150 samples of abomasum collected directly from camel carcasses after working in Al-Najaf slaughterhouse from 1/10/2019 to 1/2/2020). ITS2rDNA were well suited for the prepared primer with size783 bp and identical ratio ranged 81.17-99.73% of the same species, as indicated a high similarity of the isolates taken from two-humped camels in China or less related to Parabronema skrjabini in sheep and goat. In addition, the study identified the number of the mutations within the four COX1 gene and ITS-2 region, which were the most conservative region of the host's species, supporting the concept of host specificity with Parabronema skrjabini. The COX1 gene and ITS-2 region applied to confirm the diagnosis using a universal primer, as it included eight isolates with a size of 689 bp, identical values were ranged from 84.99-98.02% depending on the multiple sequence alignment and showed an increase in the substitution level among isolates at an upper taxonomic level. Studying of the COX1 gene and ITS-2 region in Parabronema skrjabini demonstrated a significant relation in the cluster and an early common ancestor with isolates of the two-humped camel (China). As for the COX1 gene and ITS-2 region, the phylogenetic relationship supported the ribosomal gene results, especially with Habronema muscae or related species such as Habronema majus, Dirofilaria repens, Dipetalonema evansi, Setaria tundra, Cercopithifilaria sp towards the root node. Therefore, considering COX1 gene and ITS-2 region as an ideal tool in determining the phylogenetic history of the sequence maps, but less conservative mode than the ITS2 ribosome gene based to a taxonomic species level.
Highlights
Parabronema skrjabini: one of the spirurida nematodes that infect abomasum of the domesticated ruminants such as camel, sheep and goats [1]
Considering COX1 gene and ITS-2 region as an ideal tool in determining the phylogenetic history of the sequence maps, but less conservative mode than the ITS2 ribosome gene based to a taxonomic species level
The PCR primers used in this analysis to the amplify ITS2rDNA gene of P. skrjabini, it is being designed according to [9], while COX1mtDNA gene was prepared as according to the molecular study of other Spirurida [19], these primers were produced from Bioneer company, Korea as following table 1, and reaction conditions of PCR for amplification showed as in the table 2
Summary
Parabronema skrjabini: one of the spirurida nematodes that infect abomasum of the domesticated ruminants such as camel, sheep and goats [1]. P. skrjabini is isolated from sheep and recorded high prevalence rate of the gastrointestinal nematodes during December (100%), less in March about 44.47% [6]. Different factors effect on the prevalence rate of abomasal worms (age, sex, body condition and immunity status) Borji [9], and he observed high prevalence of gastrointestinal nematode in sheep's females than males, adult sheep with high ability to resistance nematode's invasion compared to the young ages due to development of immunity response, in other hand the seasonal fluctuation associated with the activation of the intermediate host [10]. There are an increased the economic importance of camels has increased with utilization of their products such as milk, meat and skin [12], due to its geographical distribution, few studies were concentrated on gastrointestinal nematodes in camels and difficulties in taking samples due to the fact that the Bedouin majority is rising [13], Al-Taif [14] was the first study on one-humped camel's infection in Iraq
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