Prevalence and molecular typing of Giardia spp. in captive mammals at the zoo of Zagreb, Croatia.

A total of 131 faecal samples from 57 mammalian species housed at the zoo of Zagreb, Croatia, were tested for the presence of Giardia spp. cysts using epifluorescence microscopy. The overall prevalence (29%) was high, yet all animals were asymptomatic at the time of sampling. Positive samples were characterized by PCR and sequence analysis of both conserved and variable loci, for the identification of Giardia species and G. duodenalis assemblages and genotypes. Assemblages A and C were identified in Artiodactyla, assemblage B in Primates, Rodentia and Hyracoidea, and assemblages A, B, C and D, as well as Giardia microti, in Carnivora. Genotyping at the ITS1-5.8S-ITS2 region, at the triose phosphate isomerase, glutamate dehydrogenase and beta-giardin genes revealed extensive polymorphisms, particularly among assemblage B isolates. A phylogenetic analysis of concatenated sequences showed that isolates from captive mammals housed at the zoo are genetically different from isolates of human and domestic animal origin. This is the first survey in a zoological garden to include a molecular characterization of the parasite, and provides novel sequence data of G. duodenalis from many previously uncharacterized hosts.

A large survey of Croatian wild mammals for Giardia duodenalis reveals a low prevalence and limited zoonotic potential.

Wild mammals are considered an important source of potentially zoonotic Giardia duodenalis parasites, yet surprisingly little information is available on the actual prevalence and the genetic identity of the species they harbor. A large survey was conducted in Croatia by collecting 832 fecal samples from red deer (Cervus elaphus, n = 374), roe deer (Capreolus capreolus, n = 21), wild boars (Sus scrofa, n = 144), foxes (Vulpes vulpes, n = 66), bears (Ursus arctos, n = 19), wolves (Canis lupus, n = 127), jackals (Canis aureus, n = 8), and hares (Lepus europeus, n = 73). Fecal samples were tested for the presence of Giardia cysts using fluorescent microscopy. The observed prevalence ranged from low (1% in red deer, 1.7% in wild boars, and 4.5% in foxes) to moderate (10% in wolves and 12.5% in jackals) to high (24% in roe deer). No cysts were observed in bears and hares. Polymerase chain reaction was performed on microscopically positive samples to amplify fragments of the small subunit ribosomal gene, the ribosomal 5.8S gene and the two flanking internal transcribed sequences, and the triose phosphate isomerase gene. Sequence analysis showed a predominance of G. duodenalis assemblage A in both ruminants (genotypes A1 and A3) and carnivores (genotype A1). G. duodenalis assemblages B, C, and D, as well as Giardia microti, were also detected in this study. This is the first molecular description of the parasite from the red deer, the wolf, and the jackal. The data point to a minor role of wild mammals as reservoirs of zoonotic assemblages of G. duodenalis, albeit cycling between sylvatic and domestic animals is possible.