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BioMed Research International
Volume 2013 (2013), Article ID 875048, 9 pages
http://dx.doi.org/10.1155/2013/875048
Research Article

Performance of Glutamate Dehydrogenase and Triose Phosphate Isomerase Genes in the Analysis of Genotypic Variability of Isolates of Giardia duodenalis from Livestocks

1Laboratório de Parasitologia, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, 1720 Avenida Pará, Umuarama Campus, 38400-902 Uberlândia, MG, Brazil
2Departamento de Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Avenida Prof. Dr. Orlando Marques de Paiva, 87-Cidade Universitária, 05508-270 São Paulo, SP, Brazil
3Laboratório de Biotecnologia e Marcadores Moleculares, Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de MG, 6627 Avenida Antônio Carlos, Pampulha Campus, 31270-901, Belo Horizonte, MG, Brazil
4Médico Veterinário—Instituto de Ciências Biomédicas, 1720 Avenida Pará, Umuarama Campus, 38400-902 Uberlândia, MG, Brazil

Received 16 April 2013; Revised 13 August 2013; Accepted 31 August 2013

Academic Editor: Jacques Cabaret

Copyright © 2013 Natália M. N. Fava et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Giardia duodenalis is a small intestinal protozoan parasite of several terrestrial vertebrates. This work aims to assess the genotypic variability of Giardia duodenalis isolates from cattle, sheep and pigs in the Southeast of Brazil, by comparing the standard characterization between glutamate dehydrogenase (gdh) and triose phosphate isomerase (tpi) primers. Fecal samples from the three groups of animals were analyzed using the zinc sulphate centrifugal flotation technique. Out of 59 positive samples, 30 were from cattle, 26 from sheep and 3 from pigs. Cyst pellets were stored and submitted to PCR and nested-PCR reactions with gdh and tpi primers. Fragment amplification of gdh and tpi genes was observed in 25 (42.4%) and 36 (61.0%) samples, respectively. Regarding the sequencing, 24 sequences were obtained with gdh and 20 with tpi. For both genes, there was a prevalence of E specific species assemblage, although some isolates have been identified as A and B, by the tpi sequencing. This has also shown a larger number of heterogeneous sequences, which have been attribute to mixed infections between assemblages B and E. The largest variability of inter-assemblage associated to the frequency of heterogeneity provided by tpi sequencing reinforces the polymorphic nature of this gene and makes it an excellent target for studies on molecular epidemiology.