Detection of Molecular Typing of the Intestinal Parasite E. histolytica in Humans

Maged Hameed Salman; Abdulkhalik .A Mohemeed; Anas Y. Al-Hayawi

Maged Hameed.Salman Tikrit University/ College of Pure Sciences/ Departement of Biology
Abdulkhalik .A Mohemeed College of Science, Tikrit University, Iraq
Anas Y. Al-Hayawi Tikrit University/ College of Pure Sciences/ Departement of Biology

Keywords: Molecular Profiling, E.histolytica, Entamoeba, Parasite

Abstract

This study was conducted on patients visiting Tikrit Teaching Hospital and some private laboratories in Tikrit from the beginning of January 2023 to the end of December 2023. A total of 220 stool samples were collected, with a percentage of 22.72% positive for Entamoeba histolytica, based on 50 positive microscopic tests. The results demonstrated the success of the Polymerase Chain Reaction (PCR) technique in amplifying the gene responsible for encoding the Galactose/N-acetyl-D-galactosamine lectin enzyme from E. histolytica samples indicating the method's effectiveness and efficiency in diagnosing the targeted genetic sequence.

Downloads

Download data is not yet available.

References

A., Qureshi, A. W., Khan, L., Mansoor, A., Ullah, S., & Irfan, M. (2018). Prevalence of Entamoeba histolytica in district Buner Khyber Pakhtunkhwa, Pakistan. Journal of Entomology and Zoology Studies, 6(1), 185- 188.FOOD-HANDLERS IN ERBIL CITY, IRAQ. Plant Archives, 21(1), 1367-1372.

Abhilash, M. (2010). Potential applications of Nanoparticles. Int. J. Pharm. Biol. Sci., 1(1):1-12.

Ahmed, S.; Annu; Chaudhry, S.A.; Ikram, S. (2017) A review on biogenic synthesis of ZnO nanoparticles using plant extracts and microbes: A prospect towards green chemistry. J. Photochem. Photobiol.

Al-Areeqi, M. A. ; Sady, H. ;Al-Mekhlafi, H. M. ;Anuar, T. S. ;Al-Adhroey, A. H. ; Atroosh, W. M. and Lau, Y.

L. (2017). First molecular epidemiology of Entamoeba histolytica, E. dispar and E. moshkovskii infections in Yemen: different species-specific associated risk factors. Tro Med & Inte H, 22(4), 493-504.

Aldaba-Muruato, L. R., Muñoz-Ortega, M. H., Macías-Pérez, J. R., Pulido-Ortega, J., Martínez-Hernández, S. L., and Ventura-Juárez, J. (2017). Adrenergic regulation during acute hepatic infection with Entamoeba histolytica in the hamster: involvement of oxidative stress, Nrf2 and NF-KappaB. Parasite, 24

Al-Damerchi, A.T.N. , and Al-Ebrahimi , H.N. (2016). Detection of major virulence factor of Entamoeba histolytica by using PCR technique, AL-Qadisiya Medical Journal Vol.12 No.21.PP: 36-45.

Al-Delaimy, A. K., Al-Mekhlafi, H. M., Nasr, N. A., Sady, H., Atroosh, W. M., Nashiry, M.,... & Mahmud, R. (2014). Epidemiology of intestinal polyparasitism among Orang Asli school children in rural Malaysia. PLoS neglected tropical diseases, 8(8), e3074.

Al-Kaeebi, S.R.A and Al-Difaie ,R.S.S. (2016). Use PCR conventional for detecting AP and PLA virulence factors of Entamoeba histolytica in patients stool samples in Al-Qadisiyah Province. Wasit Journal for Science & Medicine , 8(4): (102- 110).

Alshaibani, H. A. A. (2024). A review of entamoebahistolytica genome, virulence factors, phylogenetic tree and treatment. World Journal of Current Medical and Pharmaceutical Research, 26-35.

Al-Zahrani, S. S. (2019). Silver nanoparticles (AgNPs) from plant extracts.Journal of Natural Sciences, Life and Applied Sciences, 3(2) , 95-70

Al-Zubadi, W. F., Al-Masoudi, H. K., & Abdul-Lateef, L. A. (2021). Detection and Sequencing of Iron Superoxide Dismutase Gene in Entamoeba histolytica Isolated from Patients with Diarrhea in Iraq. Archives of Razi Institute, 76(5), 1165-1171.

Amer, O. S., Al-Malki, E. S., Waly, M. I., AlAgeel, A., and Lubbad, M. Y. (2018). Prevalence of Intestinal Parasitic Infections among Patients of King Fahd Medical City in Riyadh Region, Saudi Arabia: A 5-Year Retrospective Study. J parres.

Apel ,K and Hirt, H. (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399.

Armeni, T., and Principato, G. (2020). Glutathione, an over one billion years ancient molecule, is still actively involved in cell regulatory pathways. In The First Outstanding 50 Years of “Università Politecnica delle Marche” (pp. 417-429). Springer, Cham.

Arredondo, J. L., González, M. B., Coria, A. L., Ortega, J. E., Vargas, J., Villarreal, J. L., & Vallarta, M. M. (2014). Entamoeba histolytica: trophozoite, precyst and cyst studied by atomic force microscopy.

Banisch, D. M., El-Badry, A., Klinnert, J. V., Ignatius, R., & El-Dib, N. (2015). Simultaneous detection of Entamoeba histolytica/dispar, Giardia duodenalis and cryptosporidia by immunochromatographic assay in stool samples from patients living in the Greater Cairo Region, Egypt. World Journal of Microbiology and Biotechnology, 31(8), 1251-1258.

Bansal, D.; Malla, N. and Mahajan, R.C. (2006). Drug resistance in amoebiasis. Indian J. Med. Res., 23: 115-118.

Bazzaz, A. A., Shakir, O. M., & Alabbasy, R. H. (2017). Prevalence of two gastrointestinal parasites Entamoeba histolytica and Giardia lamblia within Samarra city, Iraq. Advances in Bioscience and Biotechnology, 8(11), 399- 410.

Begum, S.; Gorman, H.; Chadha, A. and Chadee, K. (2021). Entamoeba histolytica. Trends in Parasitology, 37(7), 676-677.

Ben Ayed, L., & Sabbahi, S. (2017). Entamoeba histolytica global water pathogens project. www. waterpathogens. org/book/entamoeba-histolytica, Michigan State University, E. Lansing, MI, UNESCO.

Bharadwaj, R., Kushwaha, T., Ahmad, A., Inampudi, K. K., & Nozaki, T. (2021). An atypical EhGEF regulates phagocytosis in Entamoeba histolytica through EhRho1. PLoS pathogens, 17(11), e1010030.

Bilici, M. (2014). The Importance of Oxidative stress in Early Week Pregnancy Losses. Crescent Journal of Medical and Biological seiences. 1(4) : 151 – 153 .

Bogitsh, B. J., Carter, C. E., and Oeltmann, T. N. (2018). Human parasitology. Academic Press.

Brooks, G.F. ;Butel, J.S. and Morse, S.A .(2004). Medical microbiology, 23th ed. McGraw-Hill companies, United State of America, New York, 671-674.

Buzea, C., Pacheco, I. I., & Robbie, K. (2007). Nanomaterials and nanoparticles: sources and toxicity. Biointerphases, 2(4), MR17-MR71.

Chachain, N. F.; Hussein, A. J. and Shaima, D. S. (2019). Investigation the level of IL-10 and IFN-γ in mice

infected with Entamoeba histolytica and treated with earthworm powder. Al-Mustansiriyah J. Sci. 30(1): 52-57.

Chauhan, R., Reddy, A., and Abraham, J. (2015). Biosynthesis of silver and zinc oxide nanoparticles using Pichia fermentans JA2 and their antimicrobial property. Journal of Applied nanoscience, 5(1): 63-71.Sastry, A. S., and Bhat, S. (2018). Essentials of medical parasitology. Jaypee Brothers, Second Edition, Medical Publishers Pvt. Limited.

Chin, Y.T, ; Lim, Y.A. and Chong, C.W, et al.(2016). Prevalence and risk factors of intestinal parasitism among two indigenous sub-ethnic groups in peninsular Malaysia. Infect Dis Poverty; 5:77.

Choi, M. H. ; Sajed , D. ; Poole , L. (2005). An usual surface peroxiredoxin protects invasive Entamoeba histolytica

from oxidant attak. Mol. Bio. 10;9689-9699.

clinical importance of amoebiasis in Malaysia: a review. Trop. Biomed, 28,194-222.

Delialioglu , N..; Alsan , G. and Sozen , M , et al. (2004). Detection of Entamoeba histolytica/Entamoeba dispar in stools specimen by using enzyme - linked immunosorbent assay Mem. Inst. Oswaldo. Cruz , 99 (7) 769-772.

Delialioglu, N. ; Aslan, G. ; Sozen, M. ; Babur, C. ; Kanik, A. and Emekdas, G. (2004). Detection of Entamoeba histolytica/Entamoeba dispar in stools specimen by using enzyme-linked immunosorbent assay. Mem. Inst. Oswaldo. Cruz., 99(7): 769-772.

Dhubyan Mohammed Zaki, Z. (2021). The Prevalence of Entamoeba histolytica and Giardia Lamblia Associated with Diarrhea among Children in lbn Al-Atheer Hospital in Mosul City-Iraq. Archives of Razi Institute.

Diamond L.S. and Clark C.G. (2007). A Redescription of Entamoeba histolytica Schaudinn, (Emended Walker, 1911) Separating It From Entamoeba dispar Brumpt, J. of Euk

Dincer, M., Kahveci, K., Doger, C., & Yarici, A. K. (2017). Long-term care hospitals in Turkey: a review. Eastern Mediterranean Health Journal, 23(8), 564-570.

Dos Santos ,R. M.A.; Da Silva ,P.B.; Spósito, L. De ;Toledo, L.G.; Bonifácio ,B.V.; Rodero ,C.F.; Dos Santos, K.C.

;Chorilli ,M. and Bauab ,T.M. (2018) “Nanotechnology-based drug delivery systems for control of microbial

biofilms: a review”. Int J Nanomedicine.13,1179-1213

El-Dib, N. A. (2017). Entamoeba histolytica: an overview. Current Tropical Medicine Reports, 4(1), 11-20.

Fallahizadeh, S., Feiz-Haddad, M. H., Kazemi, F., and Afrisham, R. (2017). Prevalence of Intestinal Parasitic Infections in Shush County, Southwest of Iran during 2014-2016. Int J Inf, 4(3).

Ferror-Miliani L., Nielsen O. H., Andersen P. S. and Girardin S. E. (2007). Chronic inflammation: importance of NOD2 and NALP3 in interleukin – 1 beta generation. Clin. Exp. Immunol. 147 (2)

Forrester, S.J.; Kikuchi, D.S.; Hernandes, M.S.; Xu, Q. and Griendling, K.K. (2018). Reactive oxygen species in metabolic and inflammatory signaling. Circulation Research; 122(6): 877-902.

Fotedar , R .; Stark , D .; Beebe , N .; Marriott , D .; Ellis , J. and Harkness , J. (2007b) . PCR detection of Entamoeba histolytica , Entamoeba dispar , and Entamoeba moshkovskii in stool samples from Sydney , Australia . J. Clin Microbiol . , 45 ( 3 ) : 1035 1037 .

Fotedar, R.; Stark, D.; Beebe, N.; Marriott, D.; Ellis, J. and Harkness, J. (2007). Laboratory diagnosis techniques for Entamoeba histolytica. J. Clin. Microbiol. Rev., 20(3): 511-532.

Garcia, L. S., & Procop, G. W. (2016). Diagnostic medical parasitology. Manual of Commercial Methods in Clinical Microbiology: International Edition, 284-308.

Garvey, J. S., Cremer, N. E., & Sussdorf, D. H. (1977). Methods in immunology: a laboratory text for instruction and research. Addison-wesley.

Ghosh, S., Padalia, J., and Moonah, S. (2019). Tissue destruction caused by Entamoeba histolytica parasite: cell death, inflammation, invasion, and the gut microbiome. Current clinical microbiology reports, 6(1), 51-57.

Gillespie, S. & Richard, D. P. (2001). Principles and Practice of Clinical

Gliszczynska-Swiglo, A. (2006). Antioxidant activity of water soluble vitamins in the TEAC (trolox equivalent antioxidant capacity) and the FRAP (ferric reducing antioxidant power) assays. Food Chemistry, 96, 131–136.

Gopal, N.; Selvam, A.; Srinivasan, A. R.; Saha S.; and Muddegowda, P. H. (2012). Serum Gamma Glutamyl Transferase levels in Obese South Indian adults with reference to atherogenic lipid risk factors and lipid peroxides. Inter. J. Med. Health Sci. 1(1):2- 14.

Halliwell, B.,and Gutteridge, J.M.C. (2015). Free radicals in biology and medicine, 5th edn. Oxford University Press, Oxford.

Hamad, N. R., & Ramzy, I. A. (2012). Epidemiology of Entamoeba histolytica among children in Erbil province, Kurdistan Region-Iraq. Journal of research in Biology, 1, 057-062.

hamady obeid Al-Taei, A. (2019, September). The prevalence of intestinal parasite among the attending peoples to Al-Hashimyah hospitals for seven years, Babylon province, Iraq. In Journal of Physics: Conference Series (Vol. 1294, No. 6, p. 062022). IOP Publishing.

Hamzah, Z.; Petmitr, S.; Mungthin, M.; Leelayoova, S. and Chavalitshewinkoon-Petmitr, P. (2010). Development of multiplex real-time polymerase chain reaction for detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in clinical specimens. The American journal of tropical medicine and hygiene.83(4): 909-913.

Haque, R.; Mollah, N. U.; Ali, I. K. M.; Alam, K.; Eubanks, A.; Lyerly, D. and Petri, W. A. (2000). Diagnosis of amebic liver abscess and intestinal infection with the TechLab Entamoeba histolytica II antigen detection and antibody tests. J Cli Mic, 38(9): 3235-3239.

Hemphill, A., Müller, N., & Müller, J. (2019). Comparative pathobiology of the intestinal protozoan parasites Giardia lamblia, Entamoeba histolytica, and Cryptosporidium parvum. Pathogens , 8 (3), 116.

Hofman, P. (Ed.). (2016). Infectious Disease and Parasites. Springer Reference.

Jaeffer, H.S. (2011). Prevalence of Gairdia lamblia and Entamoeba histolytic/ Entamoeba dispar infections among Children in AL- Shulaa and AL-khadimya –Baghdad-Iraq, J. of Al-Anbar University. for pure science: 5(2): 6-10.

Johan Wiley and Sons. Ltd. England. Parasitology; Amoebiasis: 179-214.

John, D.T. and Petri W.A.(2006).Medical Parasitology. 9th edn. Elsevier, Amstdan, Thewatherlands: 461.

John, D.T. and Petri, W.A. (2006). Markell and Voge Medical Parasitology. 9th edn. Saunders Elsevier, St. Louis.

Johnson L. V., Anderson D. H., Mullins R. F. and Hageman G. S. (2002). A role for local inflammation in the formation of drusen in the aging eye. Am. J. Ophthalmol. 134 (3): 411 – 31.

Justin G . Wikes ; Matthew S.; Alexander and Joseph J.Cullen.(2017). Superoxide Dismutase in pancreatic Cancer. J. Antioxidants ;6(66).

Kadir, M. A. and Ali S. M.(2011). Nutritional status of children infected with Giardia lamblia and Entamoeba histolytica infections in Kalar town, Iraq. Tikri. J. Pharm. Scie. ;7(2): 162- 170.

Kadir, M. A., El-Yassin, S. T., and Ali, A. M. (2018). Detection of Entamoeba histolytica and Giardia lamblia in children with diarrhea in Tikrit city. Tikrit J Pur Sci, 23(6): 57-64.

Kadri, K. (2019). Polymerase chain reaction (PCR): principle and applications. In Synthetic Biology-New Interdisciplinary Science. IntechOpen.

Kamaly, N.; He, J.C.; Ausiello, D. A. and Farokhzad, O. C. (2016). Nanomedicines for renal disease: Current status and future applications. Nat. Rev. Nephrol., 12(12): 738-753.

Kang T, Guan R, Chen X, Song Y, Jiang H, Zhao J.(2013) In vitro toxicity of different-sized ZnO nanoparticles in Caco-2 cells. Nanoscale Res Lett. 8(1):1–8.

Kantor M., Abrantes A., Estevez A., Schiller A., Torrent J., Gascon J., Hernandez R., Ochner Ch. (2018). Entamoeba Histolytica: Updates in Clinical Manifestation, Pathogenesis, and Vaccine Development. Canadian Journal of Gastroenterology and Hepatology, vol. Article ID 4601420, 6.

Kaya S.; Sutcu R.; Cetin E S.; Arıdogan B C.;Delibas N.; and Demirci M.(2007). Lipid peroxidation level and antioxidant enzyme activities in the blood of patients with acute and chronic fascioliasis. International Journal of Infectious Diseases.; 11, 251—255.

Khairnar, K. and 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 Microbiol. 7: 47.

Khan, U., Mirdha, B. R., Samantaray, J. C., & Sharma, M. P. (2006). Detection of Entamoeba histolytica using polymerase chain reaction in pus samples from amebic liver abscess. Indian Journal of Gastroenterology, 25(2), 55.

Kolls, J.K. Lindén, A. (2004) "Interleukin-17 family members and inflammation". Immunity;21(4):467–76

Krishnan, D. and S.K. Ghosh, (2018) Cellular events of multinucleated giant cells formation during the encystation of Entamoeba invadens. Frontiers in cellular and infection microbiology,. 8: p. 262.

Kumar , S. and Singh , V. A. (2016). Prevalence of Entamoeba histolytica and Giardia lamblia infection in arural area of Haryana , India. International Journal of Current Microbiology and Applied Sciences , 5 (6): 204-209.

Kumar, V. ; Abbas, A. K. ; Fausto N. and Mitchell, R. N.( 2007) Inflammation induces by high fat diet concomitant with mild fatty lippincott wilkin's PP:178-180.

Kumar, V. ; Abbas, A. K. ; Fausto N. and Mitchell, R. N.(2007). Inflammation induces by high fat diet concomitant with mild fatty lippincott wilkin's PP:178-180.

Latifah , I. ; Teoh, K.Y. ; Wan, K.L. ; Rahmah, M. ; Normaznah, Y. and Rohani, A.(2005). Detection of giardine gene in local isolates of Giardia duodenalis by polymerase chain reaction (PCR). Malaysian J. Pathol., 27(2) : 83 – 89.

Lee, C. Y., Horng, J. L., Chen, P. Y., & Lin, L. Y. (2019). Silver nanoparticle exposure impairs ion regulation in zebrafish embryos. Aquatic Toxicology, 214, 105263

Lee, J.; Koo, N. and Min, D. B. (2004). Reactive Oxygen Species, Aging, and Antioxidative Nutraceuticals. Compr. Rev. Food. Sci. F. 3: 21-33.

Lin ,T.C. ;Hung ,K.H. and Peng ,CH, (2015) “Nanotechnology-based drug delivery treatments and specific targeting therapy for age-related macular degeneration”. J Chin Med Assoc.78(11),635–641

Lohia, A.; Foster, PG.; Sicheritz-Ponten, T.; Weber, C.; Singh, U.; Mukherjee, C.; El-Sayed, NM.; Petri, WA Jr.; Clark, CG.; Embley, TM.; Barrell, B.; Fraser, CM.; and Hall, N. (2005). The genome of the protist parasite Entamoeba histolytica. Nature 433(7028):865–868.

Loker, E., and Hofkin, B. (2015). Parasitology: a conceptual approach. Garland Science.

Luna, L. G.( 1968). Manual of Histological Staining Methods, 3rd ed. McGraw – Hill Book Company, New York

: 258 .

Mahmmud, R., Lim, Y. A. L., and Amir, A. (2017). Medical Parasitology: A Textbook. Springer.

Mahmud, R., Lim, Y. A. L., and Amir, A.(2017). Medical Parasitology: A Textbook. Springer.

McPherson, R. A., Msc, M. D., & Pincus, M. R. (2021). Henry's clinical diagnosis and management by laboratory methods E-book. Elsevier Health Sciences.

Mehlhorn, H. (2016). Human Parasites. Springer International Publishing.

Mishra, A .C.; K. S. Negi.; H. Y. Shukla and A. K. Sharma, (2009). Effect of spacing on the performance of Rosemary ( Rosmarinus officinalis Linn.) blue flowered genotype (NIC-23416) in mid hills of Uttarakhand under rainfed conditions , Natural Produc Radiance, 8 (5) : 528-531.

Mitra, B.N., et al., (2010) Compounds of the upper gastrointestinal tract induce rapid and efficient excystation of Entamoeba invadens. International Journal for Parasitology, 2010. 40: p. 9.

Mohammadzadeh, A., Spotin, A., Mahami-Oskouei, M., Haghighi, A., Zebardast, N., & Kohansal, K. (2017). Gene migration for re-emerging amebiasis in Iran’s northwest–Iraq borders: a microevolutionary scale for reflecting epidemiological drift of Entamoeba histolytica metapopulations. Parasitology research, 116(1), 217- 224.

Mojarad,E.N.;Nochi,Z.;Sahebekhtiari,N.;Nejad,M.R.;Dabiri,H .;Zali,M.R.; Kazemi,B.and Haghighi,A. (2009).Discrimination of Entamoeba moshkovskii in patients with gastrointestinal disorder by Single - PCR. Jpn. J. Infect. Dis.,63: 136-138.

Moncada,D.,Keller,K.,and chadee,K. (2003). Entamoeba histolytica cystiene proteinase disrupt the polymorphic structure of colonic mucin and alter its protective function.Infect.Immune.17(2),838-844.

Morga, S.; Ali, A. N., and Nineer, A. (2004). Primary pulmonaryamoebiasis, unusual cause of pulmonary consolidation, J. K. Mad. Assoc. ,42: 245 -246.

Muhajir, A. E. M., Hajissa, K., Mohamed, Z., and Aal, A. A. A. (2017). Prevalence of Intestinal Parasitic Infection among Children in Al-kalakla, Khartoum, Sudan. World Applied Sciences Journal, 35(2), 219-222.

Nagpal, M. L., Boldura, O. M., Balta, C., & Enany, S. (Eds.). (2020). Synthetic Biology: New Interdisciplinary Science. BoD–Books on Demand.

Nandi, A.; Yan, L. J.; Jana, C. K. and Das, N. (2019). Role of catalase in oxidative stress-and age-associated degenerative diseases. Oxidative medicine and cellular longevity, 2019.of Disease. Infect. Immun.,71(9):5355- 5359.

Nath, J., Ghosh, S. K., Singha, B., & Paul, J. (2015). Molecular epidemiology of amoebiasis: a cross-sectional study among North East Indian population. PLoS neglected tropical diseases, 9(12), e0004225.

Neves,S. F.L.; Gonçalves, M.D.S. and Soares, N.M.(2011).Validation and utilization of PCR for differential diagnosis and prevalence determination of Entamoeba histolytica/Entamoeba dispar in Salvador City, Brazil. Braz J Infect Dis.15(2).

Newton ,C.R. and Graham ,G.A. (1997). PCR.2nd Edition Introduction to biotechniques .Information Press

,Eynsham ,Oxon ,Uk.Stacey N., Haschek W. and Winder C. (1993). Systemic toxicology. In: Occupational Toxicology. Statacey, N. and London: Taylor and Francis Ltd, 2nd ed., Pp 37 – 76.

Ojo, O. and Ladeji O (2005). Hepatoprotective and antioxidant effect of Camellia sinensis (Black tea) in rats. Af J Biotechnology 4(11): 1432-1438.

Oliveira, FJA and Cechini, R. (2002).Oxidative stress of liver in hamsters infected with Leishmania (L.) chagasi; J Parasitol.;86: 1067–1072.

Olivos, A., Ramos, E., Nequiz, M., Barba, C., Tello, E., Castañón, G., and Pérez-Tamayo, R. (2005). Entamoeba histolytica: mechanism of decrease of virulence of axenic cultures maintained for prolonged periods. Exp.par., 110(3), 309-312.

Paiva, C.N. and Bozza, M.T.; (2014). Are reactive oxygen species always detrimental to pathogens? Antioxid Redox Signal 20(6):1000–1037.

Paniker, C. J. (2007). Textbook of medical parasitology (No. Ed. 6). Jaypee Brothers Medical Publishers (P) Ltd.

Paniker, C. J., and Ghosh, S. (2018). Paniker's Textbook of Medical Parasitology. Eighth Edition. JP Medical Ltd.

Paolella, P. (1998). Introducation to molecular biology. MCGrow. Hill Companies.

Pappu,R. Ramirez-Carrozzi, V. Sambandam, A. (2011). "The interleukin-17 cytokine family: critical players in host defence and inflammatory diseases".Immunology;134 (1): 8–16.

Plaa G. and Hewitt W. (1982). Detection and evaluation of chemically induced liver injury. In: Principles and methods of Toxicology. Raven Press, Pp 407 - 445.

Pokropivny, V.; Lohmus, R.; Hussainova, I.; Pokropivny, A. and Vlassov, S. (2007). Introduction in nanomaterials and nanotechnology. University of Tartu. P:225.

Proctor, E . M . (1991) . Laboratory diagnosis of amoebiasis. Clin Lab. Med .,11:829-859.

Qureshi H, Ali A, Baqai R, Ahmed W.(1997). Efficacy of a combined diloxanidefuroate-metronidazole preparation in the treatment of amoebiasis and giardiasis. J Int Med Res. 1997;25:167.

Raghunath ,A. and Perumal, E. (2017) “Metal oxide nanoparticles as antimicrobial agents: a promise for the

future”. Int. J. Antimicrob. Agents. 49(2),137-152.

Reed, S. L., and Gigli, I. (1990). Lysis of complement-sensitive Entamoeba histolytica by activated terminal complement components. Initiation of complement activation by an extracellular neutral cysteine proteinase. J. Clin. Invest., 86(6): 1815-22.

Ridley, J.W. (2012). Parasitology for Medical and Clinical Laboratory Professionals. Dickinson S, Bellegarde M. Delmar, Cengage Learning. (US) New York.

Roberts L.S. and Janovy J. (2005). Foundation of parasitology. 7thed. McGraw-Hill companies, London.:108- 111.

Roberts, L. S., Janovy, J., and Schmidt, G. D, Nadler, S. (2013). Foundation of parasitology. 9thed. McGraw-Hill companies, London.

Roshdy, M. H., El-Kader, N. M. A., Ali-Tammam, M., Fuentes, I., Mohamed, M. M., El-Sheikh, N. A., & Rubio,

J. M. (2017). Molecular diagnosis of Entamoeba spp. versus microscopy in the Great Cairo. Act par, 62(1), 188- 191.

Rostami, N. M. ; Dabiri, H. ;Zali, M.R. ; Kazemi, B. and ;Haghighi, A.(2010). Discrimination of Entamoeba moshkovskii in patients with gastrointestinal disorders by single-round PCR. Jpn. J. Infect. Dis., 63(2): 136-138.

Saad, A, H.A. ; Soliman, M.I . ;Azzam, A. M and ; MO, A.B. (2015). Antiparasitic activity of silver and copper oxide nanoparticles against Entamoeba histolytica and Cryptosporidium parvum cyst. J. Egypt. Soc. Parasitol. (JESP), 45(3) : 593-602

Saiki, R.K .; Gelfand, D.H. and Stoffel, S. (1988). Primer directed enzymatic amplification of DNA with hremostable DNA Polymerase. Science, 239 (4839): 487 – 491.

Sambrook ,J.;Fritsch , E. and Maniatis ,T. (1989).Molecular cloning and laboratory manual 2nd ed .Cold Spring Harbor Laboratory ,New York.

Samie, A., ElBakri, A., and AbuOdeh, R. E. (2012). Amoebiasis in the tropics: epidemiology and pathogenesis. Current topics in tropical medicine, 201-226.

Shakir, M.J. and Hussein, A.A. (2014). Frequency of intestinal parasitic infection among children under 5 years of age in Baghdad province. International Journal of Advanced Research. 29( 8): 332-337.

Shakir, O. M., & Abdulwahhab, I. G. (2019). Study of the Effect of Klebsiella pneumoniae Antigens on the Histological Structure of the Kidney in Laboratory Rabbits Infected with Entamoeba histolytica. Tikrit Journal for Agricultural Sciences, 18(4), 139-145.

Sharma, V.; Singh, P.; Pandey, A.K.and Dhawan, A. (2012). Induction of oxidative stress, DNA damage and apoptosis in mouse liver after sub-acute oral exposure to zinc oxide nanoparticles. Mutat. Res.,745: 84–91.

Shinjiro, H. ;Amon, A. ;Suzanne, E. ;Thomas,A. ; Edward, H. and Eric, H. (2006). Resistance of C57BL/6 mice to Amoebiasis is Mediated by non-hemopoietic cell but requires hemopoietic IL-10 production.American Association of Immunologists, 177: 1208-1213.

Shirley, DT.; Farr, L.; Watanabe, K. and Moonah, S. (2018). "A Review of the Global Burden, New Diagnostics, and Current Therapeutics for Amebiasis".

Siderovski. Dr. DP. G. (2013). Protein signaling in the parasite Entamoeba histolytica. West Virginia University School of Medicine, Morgantown, WV 26506-9229, USA.

Silva, G. A. (2003). Nanotechnology approaches to crossing the blood-brain barrier and drug delivery to CNS.BMC Neuro. Sc.,9(3): S4.

Singh GR, Manjappara UV.(2020). Impromptu Effects of Nt8U with Soyasaponins on Obesity-related Lipid Parameters in High Fat Fed C57BL/6 Mice. International Journal of Peptide Research and Therapeutics. 26(1):405-11. 88 .

Souza ,D.S.M.; Barreiros ,J. T. ; Papp, K.M. ;Smario, S .Barardi,C.M. and Barardi , R.C. (2003). Comparison between lmmumomagnetic separstion coupled with immunflourescenc and the techniques of foust and tutz for the diagnosis of G.lamblia cyst in haman feces. Rev. Inst. Med. trop. S. Paulo 45 (6).

Steve ,C. S. and Chadee, K.(2017). Entamoeba histolytica: Host parasite interactions at the colonic epithelium.Tissue Barriers.5(1):1-4.

Tanyuksel, M. and W.A. Petri. (2003). Laboratory diagnosis of amebiasis. Clinical microbiology reviews, 16(4):

p. 713-729.

Tavares,P. ; Rigothier, M. ;Khun, H. ;Roux, P.; Huerre،M. and Guillen, N. (2005).Roles of cell adhesion and cytoskeleton activity in Entamoeba histolytica pathogenesis: a delicate balance. Infect.Immun. 73:1771-1778.

Temple, M.D.; Perrone, G.G.; and Dawes, I.W. (2005) Complex cellular responses to reactive oxygen species. Trends Cell Biol 15(6):319–326.

Tengku, S., & Norhayati, M. (2011). Review Paper Public health and clinical importance of amoebiasis in Malaysia: a review. Trop. Biomed, 28, 194-222.

Thibeaux, R.;Weber, C.; Hon, C.C.; Dillies, M.A.; Avé, P.; Coppée, J.Y.(2013). Labruyère, E.; Guillén, N. Identification of the Virulence Landscape Essential for Entamoeba histolytica Invasion of the Human Colon. PLoS Pathog. 9, e1003824. doi: 10.1371/journal.ppat.1003824.

Upcroft, P. and Upcroft, J.A. (2001). Drug targets and mechanisms of resistance in the anaerobic protozoa. Clin. Microbiol. Rev., 14:150-164.

Valko, M.; C. J. Rhodes.; J. Moncol.; M. Izokovic. And M. Mazur, (2006). Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chemico- Biological Interactions,160: 1-40.

Walochnik, J., & Duchêne, M. (2016). Molecular Parasitology. Springer.

Wesel, J.; Shuman, J.; Bastuzel, I.; Dickerson, J. and Ingram-Smith, C. (2021). Encystation of Entamoeba histolytica in Axenic Culture. Microorganisms, 9(4), 873.

WHO, (1987). Manual for Laboratory investigation of Acute Enteric / infections . W. H. O. Geneva.

WHO. (1991). Basic Laboratory Methods in Medical Parasitology : microscopic examination of wet mounts Geneva:10-4P.

Wilson ,K.and Walker ,J. (2004). Principles and techniques of practical biochemistry .Cambridge university Press.

Wu,D.Y.;Vgozzli,L.;Pal,B.K.,Qian,J.andWallace ,R.B.(1991). The effects of temperature and oligonucleotide primer length on the specificity and efficiency of the amplification by polymerase chain reaction . DNA and cell Boil .10:233-238.

Ximénez, C. ;Morn ,P. ;Rojas, L. ;Valadez ,A. ;Gomez ,A. ; Ramiro, M.; ;Cerritos, R. ; Gonzez, E. ;Hernndez, E. and Oswaldo ,P. (2011). Novelties on Amoebiasis: A Neglected Tropical Disease. J. Glob Infect Dis., 3(2): 166– 174.

Yazar, S. ; Kilic, E. ; Saraymen, R. and Sahin, I. (2003) Serum malodialdehyde levels in Toxoplasma Seropositive patients.Annals of Saudi Medicine.; 23 :413-415.

Yimer, M. ;Zenebe, Y. ;Mulu, W. ;Abera, B. and Saugar, J. M. (2017). Molecular prevalence of Entamoeba histolytica/dispar infection among patients attending four health centres in north-west Ethiopia. Tropical doctor, 47(1), 11-15.

Zeb, A., Qureshi, A. W., Khan, L., Mansoor, A., Ullah, S., & Irfan, M. (2018). Prevalence of Entamoeba histolytica in district Buner Khyber Pakhtunkhwa, Pakistan. Journal of Entomology and Zoology Studies, 6(1), 185-188.