Sequencing and phylogenic variation of ITS-2 region and rrnL gene in Toxocara canis of Iraqi isolation

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Research Paper 01/07/2018
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Sequencing and phylogenic variation of ITS-2 region and rrnL gene in Toxocara canis of Iraqi isolation

Balkes Fadel Hade, Shurook M. K. Saadedin, Amer Murhum Abd Al-Amery, Halah Khalid Ibrahim
J. Bio. Env. Sci.13( 1), 72-83, July 2018.
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Abstract

Toxocara canis nematode roundworm, the main causative agent of zoonotic disease of canids and Toxocariasis in humans.This study designed to detect the ITS-2 region and rrnL gene Iraqi isolate and to characterize and analyse the genetic variation inT. canis  adult nematode which collected from puppies in different geographical areas of Baghdad city. DNA extracted from worms then detected by PCR followed by sequencing and phylogenetic tree analysis using BLASTed sequences (Neighbor-Joining method) and Phylogeny.fr program(Advance method). Two aligned sequences namely Balkes I (Ba I) for (ITS-2) region and Balkes II (Ba II) for (rrnL) gene were deposited in NCBI with accession numbers LC328970, LC328971, respectively and at DDBJ and ENA database for the first time in Iraq. BLASTed sequences results indicate clustered the aligned sequences of Ba I and Ba II with T. canis specie other than Toxocara species with low genetic changes (0.08) and (0.04) respectively, while phylogenetic analysis results indicated that Iraqi Ba I and Ba II aligned sequences are clustered with other sequences of T. canis isolates retrieved from databases and recorded low genetic changes of (0.05) and (0.007).The current study concluded that the ITS-2 region and rrnL gene can be used as genetic markers for studying specific genetic identification and diagnosis and study genetic variation of ascaridoid nematodes.

VIEWS 22

Awad A, Suzan A, Al-Azizz A. 2017. ‘Immunological study of Toxocara canis in dogs and mice at Basrah city’, AL-qadisyah medical journal,3, 236-352. www.qu.edu.iq/journalmed/index.php/QMJ/article/view/474

Bajalan MM. 2010. ‘Prevalence of intestinal helminths in stray dogs of Kalar city/Sulaimani province’,Iraqi Journal Veterinary Medicine34, 151-57.

Betson M, Peter N, Julia LH, Claire G, Aaron A, Moses A, Moses A, Andrew R, Grace T, Narcis BK. 2012. ‘Genetic diversity of Ascaris in southwestern Uganda’, Transactions of the Royal Society of Tropical Medicine and Hygiene,106, 75-83. https://doi.org/10.1016/j.trstmh.2011.10.011

Carvalho  EAA, Rocha L. 2011. ‘Toxocariasis: visceral larva migrans in children’, Jornal de pediatria, 87: 100-10. http://dx.doi.org/10.1590/S002175572011000200004

Cassenote AJF, Alba RAL, José MPN, Rubinsky-Elefant G. 2014. ‘Seroprevalence and modifiable risk factors for Toxocara spp. in Brazilian schoolchildren’, PLoS neglected tropical diseases, 8, e2830. https://doi.org/10.1371/journal.pntd.0002830.

Chang QC, Jun Feng G, Zhong Hua S, Yan L, Xu Z, Chun-Ren W. 2015. ‘Sequence variability in three mitochondrial genes among four roundworm species from wild animals in China’, Mitochondrial DNA, 26, 75-78. https://doi.org/10.3109/19401736.2013.823171.

Chen J, Zhou DH, Alasdair JN, Xu MJ, Huang SY, Li MW, Wang CR, Zhu XQ. 2012. ‘Advances in molecular identification, taxonomy, genetic variation and diagnosis of Toxocara spp’, Infection, Genetics and Evolution,12, 1344-48. https://doi.org/10.1016/j.meegid.2012.04.019.

Despommier D. 2003. ‘Toxocariasis: clinical aspects, epidemiology, medical ecology, and molecular aspects’, Clinical microbiology reviews,16, 265-72. http://cmr.asm.org/content/16/2/265.short.

Ellis, RE, Sulston JE, Coulson AR. 1986. ‘The rDNA of C. elegans: sequence and structure’, Nucleic acids research, 14, 2345-64. https://doi.org/10.1093/nar/14.5.2345.

Fogt-Wyrwas R, Mizgajska-Wiktor H, Pacoń J, Jarosz W. 2013. ‘Intraspecific variation between the ITS sequences of Toxocara canis, Toxocara cati and Toxascaris leonina from different host species in south-western Poland’, Journal of helminthology, 87, 432-42. https://doi.org/10.1017/S0022149X1200048X.

Hadi AM. 2011. ‘Isolation and identification of intestinal parasites from vegetables from different markets of Iraq’, Bulletin of the Iraq Natural History Museum, 11, 17-25.

Hadi AM. 2017. An ELISA Serodiagnostic survey of Toxacara canis in normal and hospitalized eye patients from Baghdad, Iraq. Online Journal of Veterinary Research, 21, 45-50.

Holland CV. 2017. ‘Knowledge gaps in the epidemiology of Toxocara: the enigma remains’, Parasitology, 144, 81-94. https://doi.org/10.1017/S0031182015001407.

Holsinger KE, Weir S. 2009. ‘Genetics in geographically structured populations: defining, estimating and interpreting F ST’, Nature Reviews Genetics, 10, 639.

Jacobs DE, Zhu X, Robin BG, Chilton NB. 1997. ‘PCR-based methods for identification of potentially zoonotic ascaridoid parasites of the dog, fox and cat’, Acta tropica, 68, 191-200. https://doi.org/10.1016/S0001-706X(97)00093-4

Jex AR, Andrea W, Littlewood DT, Hu M, Gasser RB. 2008. ‘The mitochondrial genome of Toxocara canis’, PLoS neglected tropical diseases, 2,  e273. https://doi.org/10.1371/journal.pntd.0000273.

Johnstone IL. 2000. ‘Cuticle collagen genes: expression in Caenorhabditis elegans’, Trends in Genetics16, 21-27. https://doi.org/10.1016/S0168-9525(99)01857-0.

Khademvatan S, Fakher R, Mahdi T, Rahman A, Mahmoud H. 2013. ‘PCR-based molecular characterization of Toxocara spp. using feces of stray cats: a study from Southwest Iran’, PLoS ONE, 8, e65293. https://doi.org/10.1371/journal.pone.0065293.

Klis FM, Jong M, Brul S, Groot PW. 2007. ‘Extraction of cell surface‐associated proteins from living yeast cells’, Yeast, 24,253-58. https://onlinelibrary.wiley.com/doi/full/10.1002/yea.1476.

Krücken J, Julia B, Denny M, Janina D, Sabrina R, Daniela A, Roland S, Georg SH. 2017. ‘Small rodents as paratenic or intermediate hosts of carnivore parasites in Berlin, Germany’, PLoS ONE,12, e0172829. https://doi.org/10.1371/journal.pone.0172829.

Li MW, Lin RQ, Song HQ, Wu XY, Zhu XQ. 2008. ‘The complete mitochondrial genomes for three Toxocara species of human and animal health significance’, BMC genomics,9, 224. https://doi.org/10.1186/1471-2164-9-224.

Ma GX, Zhou RQ, Hu L, Luo YL, Luo YF, Zhu HH. 2018. ‘Molecular characterization and transcriptional analysis of the female-enriched chondroitin proteoglycan 2 of Toxocara canis’, Journal of helminthology, 92: 154-60. https://doi.org/10.1017/S0022149X17000359.

Maniatis T, Fritsch EF, Sambrook J. 1982. Molecular cloning: a laboratory manual (Cold Spring harbor laboratory Cold Spring Harbor, NY).The First edition.

Mikaeili F, Kia EB, Sharbatkhori M, Sharifdini M, Jalalizand N, Heidari Z, Zarei Z,  Stensvold CR, Mirhendi H. 2013. ‘Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes’, Experimental parasitology, 134, 155-59. https://doi.org/10.1016/j.exppara.2013.02.008.

Mikaeili F, Mirhendi H, Mohebali M, Hosseini M, Sharbatkhori M, Zarei Z, Kia KB. 2015. ‘Sequence variation in mitochondrial cox1 and nad1 genes of ascaridoid nematodes in cats and dogs from Iran’, Journal of helminthology, 89, 496-501. https://doi.org/10.1017/S0022149X14000133.

Morgan JAT, Blair D. 1995. ‘Nuclear rDNA ITS sequence variation in the trematode genus Echinostoma: an aid to establishing relationships within the 37-collar-spine group’, Parasitology, 111, 609-15. https://doi.org/10.1017/S003118200007709X.

Mughini-Gras L, Margriet H, Wilfrid P, Elena P, Titia K. 2016. ‘Seroepidemiology of human Toxocara and Ascaris infections in the Netherlands’, Parasitology research, 115: 3779-94. https://link.springer.com/article/10.1007/s00436-016-5139-6.

Nolan MJ, Cribb TH. 2005. ‘The use and implications of ribosomal DNA sequencing for the discrimination of digenean species’, Advances in parasitology, 60, 101-63 https://doi.org/10.1016/S0065-308X(05)60002-4.

Oliart-Guzmán H, Breno MD, Antonio CM, Saulo ASM, Athos MB, Thasciany MP, Branco F, Ramalho AA, Campos RG, Fontoura PS. 2014. ‘Epidemiology and control of child toxocariasis in the Western Brazilian Amazon–a population-based study’, The American journal of tropical medicine and hygiene, 90, 670-81. https://doi.org/10.4269/ajtmh.13-0506.

Owen SP. 2017. ‘The first epidemiological study on the prevalence of cardiopulmonary and gastrointestinal parasites in cats and dogs from the Algarve region of Portugal using the FLOTAC technique’, Universidade de Lisboa, Faculdade de Medicina Veterinária. http://hdl.handle.net/10400.5/14521.

Sambrook J, Russell DW. 2001. ‘Molecular cloning: a laboratory manual 3rd edition’, Coldspring-Harbour Laboratory Press, UK.

Santos SV, Pedro PC, Fabiana MP, Ronaldo CBG, Susana AZL. 2017. ‘Experimental toxocariasis in BALB/c mice: relationship between parasite inoculum and the IgG immune response’, Memórias do Instituto Oswaldo Cruz: 0-0. http://dx.doi.org/10.1590/0074-02760160341 .

Sarvi S, Ahmad D, Mehdi S, Mohammad TR, Mohammad HK, Siavash M, Abolghasem S, Seyed-Abdollah H, Shirzad G. 2018. ‘Zoonotic intestinal parasites of carnivores: A systematic review in Iran’, Veterinary world,11: 58. https://doi.org/10.14202/vetworld.2018.58-65.

Wang ZQ, Li ZL, Peng J, Li NL, Jing C. 2012. ‘Molecular identification and phylogenetic analysis of Trichinella isolates from different provinces in mainland China’, Parasitology research, 110, 753-57.

Wickramasinghe S, Lalani Y, Rajapakse RP, Takeshi A. 2009. ‘Toxocara canis and Toxocara vitulorum: molecular characterization,discrimination, and phylogenetic analysis based on mitochondrial (ATP synthase subunit 6 and 12S) and nuclear ribosomal (ITS-2 and 28S) genes’, Parasitology research 104, 1425. http://dx.doi.org/10.1007/s00436-009-1345-9.

Zhu XQ, Ameli So, Gasser RB, Yang TB, Paggi L, He F, Lin RQ, Song HQ, Ai L, LiAX. 2007. ‘Practical PCR tools for the delineation of Contracaecum rudolphii A and Contracaecum rudolphii B (Ascaridoidea: Anisakidae) using genetic markers in nuclear ribosomal DNA’, Molecular and Cellular Probes21, 97-102. http://dx.doi.org/10.1016/j.mcp.2006.08.004

Ziętkiewicz E, Magdalena W, Patrycja D, Jadwiga ZG, Mariusz G, Barbara J, Michał W. 2012. ‘Current genetic methodologies in the identification of disaster victims and in forensic analysis’, Journal of applied genetics53, 41-60. http://dx.doi.org/10.1007/s13353-011-0068-7.