First molecular characterization of Giardia duodenalis from goats in Malaysia

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Abstract

In the present study, 310 faecal samples from goats from eight different farms in Malaysia were tested for the presence of Giardia using a PCR-coupled approach. The nested PCR for SSU amplified products of the expected size (∼200 bp) from 21 of 310 (6.8%) samples. Sixteen of these 21 products could be sequenced successfully and represented six distinct sequence types. Phylogenetic analysis of the SSU sequence data using Bayesian Inference (BI) identified Giardia assemblages A, B and E. The identification of the ‘zoonotic’ assemblages A and B suggests that Giardia-infected goats represent a possible reservoir for human giardiasis in Malaysia.

Introduction

Giardia duodenalis (Protista) represents an important group of enteric pathogens of mammals, including humans and ruminants [1], [2]. Giardiasis causes intestinal disease in cattle, particularly young calves [3]. Animals that are affected by giardiasis can exhibit clinical signs of diarrhoea, weight loss and a failure to thrive, often resulting in significant production losses and sometimes death [4]. The relatively high prevalence of G. duodenalis in young ruminants (often >30%) [5] is a public health concern, because of the possibility of zoonotic transmission from infected animals to humans (e.g., farmers and their families) and, on a larger scale, through the contamination of the environmental or water with viable cysts, which have the potential to infect large human populations [6].

Based on molecular evidence, G. duodenalis is currently divided into eight genotypes or assemblages (A to H) [3], [7], with assemblages A, B, and, occasionally E, being found to infect humans [8]. In ruminants, G. duodenalis assemblage E is most frequently being recorded in calves [9], sheep [10], [11] and goats [12]. However, the “zoonotic” assemblages A and B are also common (with prevalences varying from 2.9% to 14%, depending on study) [13], [14], [15], suggesting that ruminant hosts could be significant reservoirs of G. duodenalis infection to humans.

Presently, there is considerable information on the epidemiology of Giardia of ruminants (e.g., [11], [16], [17]), particularly goats [5], [12], [18]. However, molecular studies have not been widely conducted on Giardia from animals in Malaysia. In this country, goat farming is a key agricultural sector, and goats represent a major food source. Goats constitute ∼70% of the total small ruminant livestock industry in this country, being second after cattle and involving ∼12,500 farms (Department of Veterinary Services, 2006; http://agrolink.moa.my/jph/). Except for a study of the prevalence of Giardia in cattle using a conventional microscopic detection method [6], there has been no study of Giardia of any ruminant species in Malaysia using molecular methods. The present study genetically characterized, for the first time, Giardia from goats in this country to assess whether they harbour genotypes with the potential to infect humans.

Section snippets

Materials and methods

A total of 310 faecal samples were collected from goats (Jamnapuri breed; both sexes; 3 months to 7 years of age) from eight farms in four states of peninsular Malaysia (i.e., Selangor, Perak, Terengganu and Sarawak). The farms had similar farm management strategies, in accordance with guidelines for goat breeding (Department of Veterinary Services, Malaysia; http://www.dvs.gov.my/web/guest/penerbitan). Goats were kept in a semi-intensive system in raised wooden houses with slated flooring. The

Results and discussion

All 310 genomic DNA samples derived from the goat faecal samples from eight farms in four different states in Malaysia were subjected to the molecular detection of Giardia DNA. The nested PCR for SSU amplified products of the expected size (∼200 bp) from 21 of 310 (6.8%) samples. Sixteen of these 21 amplicons could be sequenced successfully and represented six distinct sequence types [GenBank accession numbers HQ283227 (isolate GS5), HQ283228 (isolate GT8), HQ283229 (isolate GT9), HQ283230

Acknowledgements

The authors thank the farmers for their collaborative efforts and Bakhtiar Affendi Bukari for his technical assistance. We acknowledge the funding support from the Ministry of Higher Education, Malaysia, University of Malaya [projects RG221-10HTM (YLAL and RBG) and PV024/2011B (YLAL and TKT)]. Funding support from the Australian Research Council (ARC) and Melbourne Water Corporation are gratefully acknowledged (RBG and ARJ).

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    Note: The nucleotide sequences determined in the present study are available from the GenBank database under accession numbers HQ283227 to HQ283232

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