237 FOLIA PARASITOLOGICA 59 [3]: 237–240, 2012 ISSN 0015-5683 (print), ISSN 1803-6465 (online) © Institute of Parasitology, Biology Centre ASCR http://folia.paru.cas.cz/ Address for correspondence: P. Solarczyk, Department of Biology and Medical Parasitology, Poznań University of Medical Sciences, Faculty of Medicine I, 10 Fredry Street, 61-701 Poznań, Poland. Phone +48 61 8546080; E-mail: psolar@ump.edu.pl Abstract: A total of 181 faecal samples were collected from wild cervids in two regions of Poland. Giardia cysts were de- tected in one faecal specimen from red deer and in two samples from roe deer. Fragments of the β-giardin (bg) triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) genes were successfully amplified from the Giardia isolate obtained from red deer, whereas only amplicons of bg and gdh were obtained from Giardia isolates derived from two roe deer. The result of genotyping and phylogenetic analysis showed that the G. duo- denalis isolate from red deer belonged to sub-assemblage AIII, which has never been identified in humans, whereas isolates from roe deer clustered within zoonotic sub-assemblage AI. Fur- ther studies are necessary to explain which Giardia assemblages and/or sub-assemblages occur in wild cervids in various regions of the world. Moreover, the impact of Giardia infection on the health of wild cervids should also be elucidated. Keywords: Giardia, genotypes, molecular epidemiology, phy- logeny, zoonoses The cosmopolitan flagellate Giardia duodenalis (Lambl, 1859) (syns. G. intestinalis, G. lamblia) is one of the most com- mon intestinal protozoan parasites of humans and many spe- cies of animals (Thompson 2004, Lasek-Nesselquist et al. 2010, Yang et al. 2010). This species exhibits great genetic heterogene- ity and eight major genetically distinct assemblages (A–H) have been recognized. These assemblages differ in host specificity. Giardia duodenalis isolates belonging to assemblages A and B are found in both humans and animals, whereas the rest of the assemblages (C–H) are more host-adapted: C and D occur in ca- nids, E in livestock, F in cats, G in rodents, H in marine mammals (Sprong et al. 2009, Lasek-Nesselquist et al. 2010). However, further genotyping studies at different levels of resolution have indicated greater genetic variability among some assemblages than was previously found (Cacciò and Ryan 2008, Sprong et al. 2009). Besides that, new G. duodenalis genotypes are still being identified, mostly in wild animals (Lalle et al. 2007, Gaydos et al. 2008, Lebbad et al. 2010, Thompson et al. 2010). Such considerable genetic variability among G. duodenalis isolates obtained from different hosts and from various geo- graphic regions complicates determination of the role of animals as a source of human infection. Whereas the role of pets and livestock in Giardia transmission has been studied intensively, the role of wild animals, particularly artiodactyls, has only re- cently been taken into consideration (Trout et al. 2003, Lalle et al. 2007, Robertson et al. 2007, Kutz et al. 2008, Beck et al. 2010). Wild cervids might play a significant role in contamina- tion of the environment with Giardia cysts because large popu- lations occur worldwide, they excrete a relatively large volume of faeces, and their feeding ranges usually overlap with cattle pastures. Thus, they might be potential reservoirs of Giardia infection both for humans and livestock, and might represent a public and veterinary health interest. However, there are few genotyping studies of Giardia isolates found in wild cervid animals. So far, sub-assemblage AI (with zoonotic potential) and AIII (which is mainly cervid-adapted), as well as non-zoonotic assemblage E, have been identified in cervids in different geographic regions (Trout et al. 2003, van der Giessen et al. 2006, Lalle et al. 2007, Robertson et al. 2007, Beck et al. 2010, Lebbad et al. 2010). Moreover, recent studies showed variations in dissemination of G. duodenalis genotypes among geographic regions and be- tween farmed and wild ruminants (Feng and Xiao 2011). The aim of this study was a multilocus genotyping of Giardia iso- lates recovered from wild cervids. A total of 181 faecal samples were collected from wild cer- vids in west-central and north-eastern regions of Poland. Speci- mens were taken from 65 fallow deer (Dama dama) (Linnaeus), 61 red deer (Cervus elaphus) Linnaeus, 50 roe deer (Capreolus capreolus) (Linnaeus), and five moose (Alces alces) (Linnaeus). Most of the faecal samples (n = 174) were taken from the hunt- er-killed animals during selective shootings, whereas only seven specimens were picked up immediately after excretion next to feeding sites during animal observation. Each faecal sample was concentrated using the sucrose gra- dient centrifugation technique, with the final sediment being examined using a light microscope. Wet and trichrome stained smears were microscopically examined for the presence of Gia- rdia cysts. Total genomic DNA was directly extracted from faecal sam- ples. A FastDNA kit (BIO101, Vista, California, USA) was used for extraction of the Giardia DNA based on a protocol described previously (da Silva et al. 1999). The eluted DNA was purified with the QIAquick PCR purification kit (Qiagen, Hilden, Ger- many) according to the kit instructions. RESEARCH NOTE Multilocus genotyping of Giardia duodenalis isolates from red deer (Cervus elaphus) and roe deer (Capreolus capreolus) from Poland Piotr Solarczyk 1 , Anna C. Majewska 1 , Bożena Moskwa 2 , Władysław Cabaj 2 , Miroslawa Dabert 3 , Piotr Nowosad 1 1 Department of Biology and Medical Parasitology, Poznań University of Medical Science, Faculty of Medicine I, 10 Fredry Street, 61-701 Poznań, Poland; 2 Witold Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, Warsaw, Poland; 3 Molecular Biology Techniques Laboratory, Faculty of Biology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznań, Poland