An Investigation of Giardia duodenalis Subgenotype AI in Human and Cattle Isolates in Baghdad
Keywords:
Giardia duodenalis, restriction fragment length polymorphism (RFLP), Glutamate dehydrogenase gene (gdh), GenotypeAbstract
The aims of this study was designed for detection and determined the genotype of Giardia duodenalis isolates from human and cattle. To achieve this goal, thirty stool samples (5g) from patient were collected in sterile plastic cups, another 17 fecal samples from cattle were collected, all samples were examined microscopically by Saline wet-mount method for Giardia detection, Giardia duodenalis cysts were purified by using zinc flotation method, genomic DNA was successfully extracted from all samples by using AccuPrep® Stool DNA extraction kit, the gdh gene (432 bp fragment) were amplified by using specific primers of this gene, and the restriction enzyme 2U of NIaVI was used for amplified gdh gene and found the sequence of this region for 9 human and 17 cattle sample. The results indicated that the restriction pattern showed 9samples in human isolates refer to genotype A (The digested subgenotype AI yields a fragment of 150, 120, 90bp), and 21 samples refer to genotype B (Subgenotype B allele yields an intact 432 bp fragment), while all the 17 cattle samples refer to genotype A, than result of sequencing of gdh gene and alignment showed that the 99% similarity of sample with wild type of the gdh gene of Giardia intestinalis from the Gene Bank, there are one transition at position +92 T/C single nucleotide polymorphism that cause a phenylalnine (F) to phenylalnine substitution. Conclusion: The subgenotype AI of G.duodenalis was found in human and cattle isolate.
Â
References
• Aline, C. C., Nathalia, M. O., Marcus, V., Halim, A. N., Luciane, M. and Ricardo, L. Giardiasis Zoonosis: between proof of principle and paradigm in the North Western region of Sao State. Brzil. Braz J. Infect. Dis. 15(4):51-53, 2011.
• Appelbee, A. J., Frederick, L. M., Heitman, T. L. and Olson, M.E. Prevalence and genotyping of Giardia duodenalis from beef calves in Alberta. Canada.Vet. Parasitol, 112: 289-294, 2003.
• Berrilli,F., DiCave,D., D’Orazi,C., Oreechia,P., Caca,P. and Divizia,M. (2006). Prevalence and genotyping of human isolates of Giardia duodenalis from Albania. Parasitol. International, 55: 295- 297, 2006.
• Breathnacg, A.S.; Mchugh, T.D. and Butcher, P.D. Prevalence and clinical correlation of genetic subtypes of Giardia lamblia in an urban setting. Epidemiol. Infect, 138: 1459-1467, 2010.
• Fallah, E., Nahavandi, K.H., Jamali, R., Poor, B. M. and Asgharzadeh, M. Molecular Identification of Giardia duodenalis isolates from human and animal reservoirs by PCR- RFLP. J. Biol. Sci, 14: 1-6, 2008.
• Feng,Y., Ortega,Y., Cama,V., Terrel, J. and Xiao, L. High intragenotypic diversity of Giardia duodenalis in dairy cattle on three farms. Parasitol. Res, 103: 87-92, 2008.
• Guneyt, B., Ozgur, K., Naser, S., Hande, D., Asli, T., Koray, E., Kor, Y. and Seray, O. Genotyping of Giardia lamblia in a Cohort of Turkish patient:A research for a relationship between symptoms and genotypes. Kafkas Univ.Vet.FakDerg, 18: 125-131, 2012.
• Kareem, H.N., Esmaeel, F., Mohammad, A., Nasrin,M. and Behroz, M. Glutamate dehydrogenase and triose-phosphate - isomerase coding genes for detection and genetic characterization of Giardia lamblia in human feces by PCR and PCR-RFLP. Turk. J. Med. Sci, 41(2): 283-289, 2011.
• Lalle, M., Jimenez-Cardosa, E., Caccio, S. M. and Pozio, E. Genotyping O Giardia duodenalis from humans and dogs from Mexico using a beta-giardin nested polymerase Chain reaction assay. J. Parasitol; 91: 203- 205, 2005.
• Leonhard,S., Pfister,K., Beelitz,P., Wielinga,C. and Thompson, R.C.A. The molecularCharacterization of Giardia from dogs in Southern Germany. Vet. Parasitol; 150 (1-2): 33-38,2007.
• Mahdy, A.K., Surin, I., Mohd-Adnan, A., AL- Mekhlafi, M.S. and Lim, Y.A. Giardia intestinalis genotypes :risk factor and correlation with clinical symptoms. ActaTrop, 112:67- 70, 2009.
• Maria,C., Edward, R. and Alverne, P. Prevalence and associated risk factor for Giardia Lamblia infection among children hospitalized for diarrhea in Goias stat, Brazil, Rev. Inst. Med.Trop., 49(3):139-145, 2007.
• Nash,S.F., and Patel, R. The Giardia lamblia VSP gene repertoire, characteris, Genomic organization,and evolution. BMC. Genomic,11:424, 2010.
• Parima,B., Suradej.S., Mathirut,M., Peerapan,T.and Suovanee, L. Improved sensitivity of PCR amplification of gdh gene for detection and genotyping of Giardia duodenalis in Stool specimen. Research Note, 41(2):280-284, 2012.
• Read, C. M., Monis, P.T. and Thompson, R.C. Discrimination of all genotypes of Giardia duodenalis at the glutamate dehydrogenase locus using PCR - RFLP. Infect Gene Evol, 4:125- 130, 2004.
• Sprong, H., Caccio, S. and Vander, J. Identification of zoonotic genotypes of Giardia duodenalis .PLOS. Negl .Trop. Dis, 3(12) : 558, 2009.
• Trout, J. M., Greiner, E. and Fayer, R. Prevalence of Giardia duodenalis genotypes in preweaned dairy calves.Vet. Parasitol,124:179- 186,2004.
• Volotao,A.C., Costa-Macedo, L. M., Haddad, F.S., Brandao, A., Peralta, J. M. and
• Fernandes, O. Genotyping of Giardia duodenalis from Southern brown howler
• monkeys from Brazil. Vet.Parasitol, 158: 133-137, 2008.
• Zajac, A.M.; Johnson, J. and King, S.E. (2002). Evaluation of the importance of centrifugation As a component of zinc sulfate fecal flotation examinations. Journal of American Animal Hospital Association; 38: 221- 224.
Downloads
Published
Issue
Section
License
- Papers must be submitted on the understanding that they have not been published elsewhere (except in the form of an abstract or as part of a published lecture, review, or thesis) and are not currently under consideration by another journal published by any other publisher.
- It is also the authors responsibility to ensure that the articles emanating from a particular source are submitted with the necessary approval.
- The authors warrant that the paper is original and that he/she is the author of the paper, except for material that is clearly identified as to its original source, with permission notices from the copyright owners where required.
- The authors ensure that all the references carefully and they are accurate in the text as well as in the list of references (and vice versa).
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Attribution-NonCommercial 4.0 International that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
- The journal/publisher is not responsible for subsequent uses of the work. It is the author's responsibility to bring an infringement action if so desired by the author.
