American Journal of Epidemiology and Infectious Disease
ISSN (Print): 2333-116X ISSN (Online): 2333-1275 Website: Editor-in-chief: Apply for this position
Open Access
Journal Browser
American Journal of Epidemiology and Infectious Disease. 2014, 2(3), 74-82
DOI: 10.12691/ajeid-2-3-2
Open AccessArticle

Genotyping and Phylogenetic Analysis of Cystic Echinococcosis Isolated from Camels and Humans in Egypt

Nashwa O Khalifa1, Hanem F. Khater2, , Hanan A. Fahmy3, Mervat E.I.Radwan4 and Jehan S.A. Afify5

1Zoonoses Department, Faculty of Veterinary Medicine, Benha University, Toukh, Egypt

2Parasitology Department, Faculty of Veterinary Medicine, Benha University, Toukh, Egypt

3Biotechnology Department, Animal Health Research Institute (AHRI), Giza, Egypt

4Department of infectious Diseases, Faculty of Veterinary Medicine, Benha University, Toukh, Egypt

5Department of Food Hygiene, Faculty of Veterinary Medicine, Benha University, Toukh, Egypt

Pub. Date: May 13, 2014

Cite this paper:
Nashwa O Khalifa, Hanem F. Khater, Hanan A. Fahmy, Mervat E.I.Radwan and Jehan S.A. Afify. Genotyping and Phylogenetic Analysis of Cystic Echinococcosis Isolated from Camels and Humans in Egypt. American Journal of Epidemiology and Infectious Disease. 2014; 2(3):74-82. doi: 10.12691/ajeid-2-3-2


The objectives of the present study were to investigate strain identification of Echinococcus granulosus infecting camel and human in . Therefore partial sequences were generated after gel purification of nested PCR amplified products of mitochondrial NADH 1gene of Echinococcus granulosus complex. Sequences were further examined by sequence analysis and subsequent phylogeny to compare these sequences to those from known strains of E.granulosus circulating globally and retrieved from GenBank. All isolates are homologous to the camel strain, E. canadensis (G6) genotype. Nucleotide mutations generate polymorphism at position of 275 nucleotide, where a thymine replaced a cytosine and at the levels of 385 and 386 nucleotides, where two cytosine substituted a guanine and a thymine respectively. KF815488 Egypt showed typical identity (99.5%) with JN637176 Sudan, HM853659 Iran, AF386533 France and AJ237637 Poland with 0.5% diversion.. Phylogenetic analysis showed a robust tree clustering all isolates with sequences belonging to the camel genotype (G6) variant with strong bootstrap values at relevant nodes and the evolutionary distance between groups is very short. There are two mutations in the sequences of amino acids at the position of 92, where an Alanine is changed to a Valine and at the position of 129, where a Valine is transformed to a Proline. Our record of a single genotype determined a strain which could be incriminated for camel and human infectivity and responsible for its persistence in the endemic areas. Such epidemiological data could guide the application of efficient control strategies of hydatidosis in Egypt.

Echinococcus granulosus sequences phylogeny nucleotide mutation

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Omer, R.A., Dinkel, A., Romig, T., Mackenstedt, U., Elnahas, A.A., Aradaib, I.E., Ahmed, M.E., Elmalik, K.H., Adam, A. (2010). A molecular survey of cystic echinococcosis in Sudan. Veterinary Parasitology 169, 340-346.
[2]  Ibrahim, K., Thomas, R., Peter, K. and Omer, R.A. (2011). A molecular survey on cystic echinococcosis in Sinnar area,Blue Nile state (Sudan). Chinese Medical Journal (Engl.) 124, 2829-2833.
[3]  Salih, M., Degefu, H. and Yohannes, M. (2011). Infection rates, cyst fertility and larval viability of hydatid dsease in camels (Camelus dromedarius) from Borena, Kereyu and Harar aeas of Ethiopia. Global Veterinaria 7, 518-522.
[4]  Sadjjadi, M.S. (2006). Present situation of echinococcosis in the Middle East and Arabic North Africa. Parasitology international 55 (S3), 197-202.
[5]  Njoroge, E.M., Mbithi, P.M., Gathuma, J.M., Wachira, T.M., Gathura, P.B., Magambo, J.K. and Zeyhle, E. (2002). A study of cystic echinococcosis in slaughter animals in three selected areas of northern Turkana, Kenya. Veterinary parasitology 104, 85-91.
[6]  Ahmadi, N.A. (2005). Hydatidosis in camels (Camelus dromedarius) and their potential role in the epidemiology of Echinococcus granulosus in Iran. Journal of Helminthology 79, 119-125.
[7]  Torgerson, P.R., Oguljahan, B., Muminov, A.E., Karaeva, R.R., Kuttubaev, O.T., Aminjanov, M. and Shaikenov, B. (2006). Present situation of cystic echinococcosis in Central Asia. Parasitology International 55 (Suppl), S207-S212.
[8]  Ibrahim, M.M. (2010). Study of cystic echinococcosis in slaughtered animals in Al Baha region, Saudi Arabia: interaction between some biotic and abiotic factors. Acta Tropica 113, 26-33.
[9]  Epsinosa, S., Salas, A.M., Vargas, A. Freire, V., Diaz, E., Sánchez, G. and Venegas, J. (2014). Detection of G3 genotype of Echinococcus granulosus from hydatid cysts of Chilean cattle using COX1 and ND1 mitochondrial markers. Parasitology Research 113 (1), 139-147.
[10]  Jenkins, D.J. (2006). Echinococcus granulosus in Australia, widespread and doing well! Parasitology International 55 (Suppl), S203-S206.
[11]  Torgerson, P.R. (2003). Economic effects of echinococcosis. Acta Tropica 85 (2), 113-118(6).
[12]  Naseri– Moghddam, S., Abrishami, A., Taefi, A. and Malekzadeh, R. (2011). Percutaneous needle aspiration injection, and re-aspiration with or without benzimidazole coverage for uncomplicated hepatic hydatid cysts. Cochrane Database. Syst. Rev., pp: CD003623.
[13]  Jenkins, D.J., Romig, T. and Thompson, R.C. (2005). Emergence/re-emergence of Echinococcus spp. – a global update. International Journal for Parasitology 35, 1205-1219.
[14]  Budke, C.M., Deplazes, P. and Torgerson, P.R. (2006). Global socioeconomic impact of cystic echinococcosis. Emerging Infectious Diseases 12, 296–303.
[15]  Sako, Y., Tappe, D., Fukuda, K., Kobayashi, Y. and Ito A (2011). Immunochromatographic test with recombinant Em18 antigen for the follow- up study of alveolar Echinococcosis. Clinical and Vaccine Immunology 18 (8), 1302-1305.
[16]  Sarkari, B., Sadjjadi, S.M., Abidi, H. and Rafati, A. (2007). Application of western blotting using native antigen B for serodiagnosis of human cystic echinococcosis. Iranian Journal of Parasitology 2(3), 7-12.
[17]  Sako,Y., Nakao, M., Nakaya, K., Yamasaki, H., Gottstein, B. and Ito, A. (2002). Alveolar echinococcosis: characterization of diagnostic antigen Em18 and serological evaluation of recombinant Em18. Journal of Clinical Microbiology 40, 2760-2765.
[18]  Moro, P.L. and Schantz, P.M. (2009). Echinococcosis: a review. International Journal of Infectious Diseases 13, 125-133.
[19]  Cardona, G.A. and Carmena, D. (2013). A review of the global prevalence, molecular epidemiology and economics of cystic echinococcosis in production animals. Veterinary parasitology 192, 10-32.
[20]  Huttner, M., Nakao, M., Wassermann, T., Siefert, L., Boomker, J.D., Dinkel, A., Sako, Y., Mackenstedt, U., Romig, T. and Ito, A. (2008). Genetic characterization and phylogenetic position of Echinococcus felidis (Cestoda:Taeniidae) from the African lion. International Journal for Parasitology 38, 861-868.
[21]  Thompson, R.C., McManus, D.P., 2002. Towards a taxonomic revision of the genus Echinococcus. Trends in Parasitology 18 (10), 452-457.
[22]  Abdel Aaty, H.E., Abdel-Hameed, D.M., Alam-Eldin, Y.H. and El- Shenawy, S.F. Aminou H.A., Makled S.S., Darweesh,S.K. (2012). Molecular genotyping of E. granulosus in animal and human isolates from Egypt. Acta Tropica 121(2), 125-128.
[23]  Osman, A.M., Aradaib, I.E., Ashmaig, A.K. and Gameel, A.A. (2009). Detection and differentiation of Echinococcus granulosus- complex using a simple PCR –based assay. International Journal of Tropical Medicine 4 (1), 21-26.
[24]  Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. and Kumar, S. (2011). MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution 28, 2731-2739
[25]  Bowles. J. and McManus, D.P. (1993). NADH dehydrogenase 1 gene sequences compared for species and strains of the genus Echinococcus. International Journal for Parasitology 23, 969-972.
[26]  Abushhewa, M.H., Abushhiwa, M.H., Nolan, M.J., Jex, A.R., Campbell, B.E., Jabbar, A. and Gasser, R.B. (2010). Genetic classification of Echinococcus granulosus cysts from humans, cattle and camels in Libya using mutation scanning-based analysis of mitochondrial loci. Molecular and Cellular Probes 24, 346-351.
[27]  Bardonnet, K., Benchikh-Elfegoun, M.C., Bart, J.M., Harraga, S., Hannache, N., Haddad, S., Dumon, H., Vuitton, D.A. and Piarroux, R. (2003). Cystic echinococcosis in Algeria: cattle act as reservoirs of a sheep strain and may contribute to human contamination. Veterinary parasitology 116, 35-44.
[28]  Maillard, S., Benchikh-Elfegoun, M.C., Knapp, J., Bart, J.M., Koskei, P., Gottstein, B. and Piarroux, R. (2007). Taxonomic position and geographical distribution of the common sheep G1 and camel G6 strains of Echinococcus granulosus in three African countries. Parasitology Research 100, 495–503.
[29]  M’Rad, S., Filisetti, D., Oudni, M., Mekki, M., Belguith, M., Nouri, A., Sayadi, T., Lahmar, S., Candolfi, E., Azaiez, R, Mezhoud, H. and Babba, H. (2005). Molecular evidence of ovine (G1) and camel (G6) strains of Echinococcus granulosus in Tunisia and putative role of cattle in human contamination. Veterinary parasitology 129, 267-272.
[30]  Lahmar, S., Debbek, H., Zhang, L.H., McManus, D.P., Souissi, A., Chelly, S. and Torgerson, P.R. (2004). Transmission dynamics of the Echinococcus granulosus sheep–dog strain (G1 genotype) in camels in Tunisia. Veterinary parasitology 121, 151-156.
[31]  Dinkel, A., Njoroge, E.M., Zimmermann, A., Walz, M., Zeyhle, E., Elmahdi, I.E., Mackenstedt, U. and Romig, T. (2004). A PCR system for detection of species and genotypes of the Echinococcus granulosus-complex, with reference to the epidemiological situation in eastern Africa. International Journal for Parasitology 34, 645-653.
[32]  Bardonnet, K., Piarroux, R., Dia, L., Schneegans, F., Beurdeley, A., Godot, V. and Vuitton, D.A. (2002). Combined eco-epidemiological and molecular biology approaches to assess Echinococcus granulosus transmission to humans in Mauritania: occurrence of the ‘camel’ strain and human cystic echinococcosis.
[33]  Farjallah, S., Busi, M., Mahjoub, M.O., Slimane, B.B., Said, K. and D’Amelio, S. (2007). Molecular characterization of Echinococcus granulosus in Tunisia and Mauritania by mitochondrial rrnS gene sequencing. Parassitologia 49 (4), 239-246.
[34]  Addy, F., Alakonya, A., Wamae, N., Magambo, J., Mbae, C., Mulinge, E., Zeyhle, E., Wassermann, M., Kern, P. and Romig, T. (2012). Prevalence and diversity of cystic echinococcosis in livestock in Maasailand, Kenya. Parasitology Research 111(6), 2289-94
[35]  Harandi, M.F., Hobbs, R.P., Adams, P.J., Mobedi, I., Morgan-Ryan, U.M. and Thompson, R.C. (2002). Molecular and morphological characterization of Echinococcus granulosus of human and animal origin in Iran. Parasitology 125, 367-373.
[36]  Ahmadi, N. and Dalimi A (2006). Characterization of Echinococcus granulosus isolates from human, sheep and camel in Iran. Infection Genetics and Evolution 6, 85-90.
[37]  Sharbatkhori, M., Mirhendi, H., Jex, A.R., Pangasa, A., Campbell, B.E., Kia, E.B., Eshraghian, M.R., Harandi, M.F. and Gasser, R.B. (2009). Genetic categorization of Echinococcus granulosus from humans and herbivorous hosts in Iran using an integrated mutation scanning-phylogenetic approach. Electrophoresis 30, 2648-2655.
[38]  Sharbatkhori, M., Mirhendi, H., Harandi, M.F., Rezaeian, M., Mohebali, M., Eshraghian, M., Rahimi, H. and Kia, E.B. (2010). Echinococcus granulosus genotypes in livestock of Iran indicating high frequency of G1 genotype in camels. Experimental Parasitology 124, 373-379.
[39]  Shahnazi, M., Hejazi, H., Salehi, M., Andalib, A.R. (2011). Molecular characterization of human and animal Echinococcus granulosus isolates in Isfahan, Iran. Acta Tropica 117, 47-50.
[40]  Hajialilo, E., Harandi, M.F., Sharbatkhori, M., Mirhendi, H. and Rostami, S. (2012). Genetic characterization of Echinococcus granulosus in camels, cattle and sheep from the south-east of Iran indicates the presence of the G3 genotype. Journal of Helminthology 86, 263-270.
[41]  Rostami Nejad, M., Taghipour, N., Nochi, Z., Nazemalhosseini Mojarad, E., Mohebbi, S.R., Fasihi Harandi, M. and Zali, M.R. (2012). Molecular identification of animal isolates of Echinococcus granulosus from Iran using four mitochondrial genes. Journal of Helminthology 86(4), 485-492.
[42]  Simsek, S., Balkaya, I., Ciftci, A.T. and Utuk, A.E. (2011). Molecular discrimination of sheep and cattle isolates of Echinococcus granulosus by SSCP and conventional PCR in Turkey. Veterinary parasitology 178, 367-369.
[43]  Latif, A.A., Tanveer, A., Maqbool, A., Siddiqi, N., Kyaw-Tanner, M., Traub, R.J. (2010). Morphological and molecular characterisation of Echinococcus granulosus in livestock and humans in Punjab, Pakistan. Veterinary parasitology 70, 44-49.
[44]  Cruz-Reyes, A., Constantine, C.C., Boxell, A.C., Hobbs, R.P. and Thompson, R.C. (2007). Echinococcus granulosus from Mexican pigs is the same strain as that in Polish pigs. Journal of Helminthology 81, 287-292.
[45]  Kamenetzky, L., Gutierrez, A.M., Canova, S.G., Haag, K.L., Guarnera, E.A., Parra, A., Garcia, G.E. and Rosenzvit, M.C. (2002). Several strains of Echinococcus granulosus infect livestock and humans in Argentina. Infection, Genetics and Evolution 2, 129-136.
[46]  Bruzinskaite, R., Sarkunas, M., Torgerson, P.R., Mathis, A. and Deplazes, P. (2009). Echinococcosis in pigs and intestinal infection with Echinococcus spp. in dogs in southwestern Lithuania. Veterinary parasitology 160, 237-241
[47]  Azab, M.E., Bishara, S.A., Helmy, H., Oteifa, N.M., El-Hoseiny, L.M., Ramzy, R.M. and Ahmed, M.A. (2004). Molecular characterization of Egyptian human and animal Echinococcus granulosus isolates by RAPD-PCR technique. Journal of the Egyptian Society of Parasitology 34(1), 83–96.
[48]  Santivañez, S.J., Gutierrez, A.M., Rosenzvit, M.C., Muzulin, P.M., Rodriguez, M.L., Vasquez, J.C., Rodriguez, S., Gonzalez, A.E., Gilman, R.H., Garcia, H.H. and The Cysticercosis Working Group in Peru (2008). Human hydatid disease in Peru is basically restricted to Echinococcus granulosus Genotype G1. American Journal of Tropical Medicine and Hygiene, 89-92.
[49]  Casulli, A., Zeyhle, E., Brunetti, E., Pozio, E., Meroni, V., Genco, F. and Filice, C. (2010). Molecular evidence of the camel strain (G6 genotype) of Echinococcus granulosus in humans from Turkana, Kenya. Transactions of the Royal Society of Tropical Medicine and Hygiene 104 (1), 29-32.
[50]  Grosso, G., Gruttadauria, S., Biondi, A., Marventano, S. and Mistretta, A. (2012). Worldwide epidemiology of liver hydatidosis including the Mediterranean area. World Journal of Gastroenterology 18(13), 1425-1437.
[51]  Mogoye, B.K., Menezes, C.N., Wong, M.L., Stacey, S., von Delft, D., Wahlers, K., Wassermann, M., Romig, T., Kern, P., Grobusch, M.P. and Frean, J. (2013). First insights into species and genotypes of Echinococcus in South Africa. Veterinary Parasitology, in press.
[52]  Dousti, M., Abdi, J., Bakhtiyari, S., Mohebali, M., Mirhendi,S.H. and Rokni,M.B. (2013). Genotyping of Hydatid Cyst Isolated from Human and Domestic Animals in Ilam Province, Western Iran Using PCR-RFLP. Iranian Journal of Parasitology 8 (1), 47-52.
[53]  Dybicz, M., Gierczak, A., Dąbrowska, J., Rdzanek, L. and Michałowicz, B. (2013). Molecular diagnosis of cystic echinococcosis in humans from central Poland. Parasitology International 62, 364-367.
[54]  Dyab, K.A., Hassanein, R., Hussein, A.A., Metwally, S.E. and Gaad, H.M. (2005). Hydatidosis among man and animals in Assiut and Aswan Governorates. Journal of Egyptian society of parasitology 35, 157–166.
[55]  Rosenzvit, M.C., Zhang, L.H., Kamenetzky, L., Canova, S.G., Guarnera, E.A. and McManus, D.P. (1999). Genetic variation and epidemiology of Echinococcus granulosus in Argentina. Parasitology 118, 523-530
[56]  Romig, T. (2003). Epidemiology of echinococcosis. Langenbeck's Archives of Surgery 388 (4), 209-217.