Genetic diversity and phylogenetic positioning of Entamoeba histolytica isolates from stool samples of patients from few Hospitals at Diyala governorate
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Abstract
Entamoeba histolytica is a protozoan parasite that causes amoebiasis, an infection primarily affecting the intestines. The study aimed to determine the phylogenetic placement of six isolates of E. histolytica that were amplified from three genetic loci, rRNA, cysteine proteinase, and amoebapore C to evaluate the genetic variation pattern of each sequence. DNA extracted from cultured and frozen stool samples using PrestoTM Stool DNA Extraction Kit and Geneaid kits was analyzed for E. histolytica diversity in amoebiasis patients at Baquba General Hospital from March-June 2023. Polymerase chaing kreaction (PCR) and sequencing of virulence genes (rRNA, cysteine proteinase, amoebapore C) followed by phylogenetic analysis revealed genetic variation and potential implications for pathogenicity. The conducted sequencing reactions confirmed the precise identification of the examined samples, which were determined to be attributed to variable isolates of E. histolytica. Aligning the rRNA sequences of samples A1 and A2 with the most closely related rRNA sequences of E. histolytica (GenBank acc. no. OP925909.1) revealed the presence of two nucleic acid variants (212C>T in A1, and 427G>C in A1 and A2) not present in the reference sequences. Aligning the cysteine proteinase sequences of samples B1 and B2 with the most closely related genomic DNA sequences of E. histolytica (GenBank acc. no. X91642.1) revealed the presence of four nucleic acid variants (95A>T and 96A>T in B1 and B2, and 246A>T and 247A>T in B2) not present in the reference sequences. Translation of nucleic acid sequences to amino acid residues showed that both 95A>T and 96A>T variants caused a missense effect of p.46E>V, while both 246A>T and 247A>T variants showed another missense effect of p.97T>S. Aligning the amoebapore C sequences of samples C1 and C2 with the most closely related genomic sequences of E. histolytica (GenBank acc. no. X76903.1) revealed their entire similarity. It was inferred from the tree that the investigated protozoan samples occupied distinct phylogenetic positions and were suitable for the vicinity of various clinical isolates that were isolated from several locations worldwide. However, the rRNA-based tree provided ample genetic diversity with more effective discrimination compared with the other used loci.
Article Details
Article Details
Amoebapore C, Cysteine proteinase, Entamoeba histolytica, Genetic diversity, Phylogenetic positioning, rRNA
Al-Damerchi, A. T. N. & Al-Ebrahimi, H. N. (2016). Detection of major virulence factor of Entamoeba histolytica by using PCR technique. Al-Qadisiyah Medical Journal, 12(21), 36–45.
Al-Kaeebi, S. R. A. & Al-Difaie, R. S. S. (2016). PCR conventional for detecting AP and PLA virulence Use factors of Entamoeba histolytica in patients stool samples in Al-Qadisiyah Province. Journal of Wasit for Science and Medicine, 8(4), 101–102.
Alwan, H., Torabi, M., Nourani, H. & Al-Shuhaib, M. B. S. (2023). The emergence of novel Iranian variants in sheeppox and goatpox viral envelope proteins with remarkably altered putative binding affinities with the host receptor. Virus Genes, 59(3), 437–448.
Argüello-García, R., Carrero, J. C. & Ortega-Pierres, M. G. (2023). Extracellular Cysteine Proteases of Key Intestinal Protozoan Pathogens—Factors Linked to Virulence and Pathogenicity. International Journal of Molecular Sciences, 24(16), 12850.
Bakr, M. M., Taher, H. M. & Mohamed, A. H. (2022). The Effect of Entamoeba Histolytica Infection on Levels of Adiponectin and Histamine in Children. Bahrain Medical Bulletin, 44(2), 961–963.
Carrero, J. C., Reyes-López, M., Serrano-Luna, J., Shibayama, M., Unzueta, J., Leon-Sicairos, N. & de la Garza, M. (2020). Intestinal amoebiasis: 160 years of its first detection and still remains as a health problem in developing countries. International Journal of Medical Microbiology, 310(1), 151358.
Castellanos-Castro, S., Bolaños, J. & Orozco, E. (2020). Lipids in Entamoeba histolytica: Host-dependence and virulence factors. Frontiers in Cellular and Infection Microbiology, 10, 75.
Christofyllakis, K. (2021). Influence of Vitamin D on Genome-Wide Expression in Natural Killer Cells.
Dobell, C. (1919). The amoebae living in man. J. Bale & Danielsson.
Espinosa-Cantellano, M., Chávez-Munguía, B., Talamás-Lara, D., Acosta-Vírgen, K. B. & Martínez-Palomo, A. (2022). Entamoeba histolytica and Entamoeba dispar. In Lifecycles of Pathogenic Protists in Humans (pp. 443–464). Springer.
Khairnar, K., & Parija, S. C. (2007). A novel nested multiplex polymerase chain reaction (PCR) assay for differential detection of Entamoeba histolytica, E. moshkovskii and E. dispar DNA in stool samples. BMC Microbiology, 7, 1–9. https://doi.org/10.1186/1471-2180-7-47
Khalaf, M. S. & Rashid, S. A. (2018). Molecular study of Enamoeba dispar and Entamoeba moshkovskii isolated from amoeboid dysentery in comparison with Entamoeba histolytica infections. Journal of Pharmaceutical Sciences and Research, 10(9), 2129–2133.
Lazar, L. T. Y., Al-Juboury, S. A. & Maaroof, M. N. (2023). Molecular detection of the ability of Biosynthesized Titanium dioxide nanoparticles to curing some genes of virulence factors of Entamoeba histolytica. Baghdad Science Journal. https://doi.org/10.21123/bsj.2023.7205
Lozano-Mendoza, J., Ramírez-Montiel, F., Rangel-Serrano, Á., Páramo-Pérez, I., Mendoza-Macías, C. L., Saavedra-Salazar, F., Franco, B., Vargas-Maya, N., Jeelani, G. & Saito-Nakano, Y. (2023). Attenuation of In Vitro and In Vivo Virulence Is Associated with Repression of Gene Expression of AIG1 Gene in Entamoeba histolytica. Pathogens, 12(3), 489.
Lübbert, C., Wiegand, J. & Karlas, T. (2014). Therapy of Liver Abscesses. Viszeralmedizin, 30(5), 334–341. https://doi.org/10.1159/000366579
Mahmud, R., Ibrahim, J., Moktar, N. & Anuar, T.-S. (2013). Entamoeba histolytica in Southeast Asia. Parasites and Their Vectors: A Special Focus on Southeast Asia, 103–129.
Misra, G. & Srivastava, V. K. (2020). Molecular Advancements in Tropical Diseases Drug Discovery. Academic Press.
Nakada-Tsukui, K., Watanabe, N., Maehama, T. & Nozaki, T. (2019). Phosphatidylinositol kinases and phosphatases in Entamoeba histolytica. Frontiers in Cellular and Infection Microbiology, 9, 150.
Ngui, R., Ishak, S., Chuen, C. S., Mahmud, R. & Lim, Y. A. L. (2011). Prevalence and risk factors of intestinal parasitism in rural and remote West Malaysia. PLoS Neglected Tropical Diseases, 5(3), e974.
Obaid, H. M., Sale, S. S. & Boundenga, L. (2023). Pharmaceutical Activity of a Synthetic Heterocyclic (C15H12N5Ocl) Compound on Entamoeba histolytica and Giardia lamblia. Russian Journal of Infection and Immunity, 13(1), 119–126. https://doi.org/10.15789/2220-7619-PAO-2024
Oliveira, F. M. S., Neumann, E., Gomes, M. A. & Caliari, M. V. (2015). Entamoeba dispar: could it be pathogenic. Tropical Parasitology, 5(1), 9.
Pal, S., Ghosh, R. & Dam, S. (2022). Understanding Entamoeba histolytica Pathogenesis: In Light of Recent.
Rostami, S., Rezaeian, M., Jamali, R., Rezaie, S., Babaei, Z. & Hooshyar, H. (2017). Differences in Entamoeba histolytica cysteine proteinase 5 gene isolated from Bandar Abbas and Tabriz, Iran. Int J Enteric Pathog, 5(2), 49–53.
Roy, M., Rawat, A., Kaushik, S., Jyoti, A. & Srivastava, V. K. (2022). Endogenous cysteine protease inhibitors in upmost pathogenic parasitic protozoa. Microbiological Research, 261, 127061.

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