Detection of Giardia intestinalis in water samples
collected from natural water reservoirs and wells in
northern and north-eastern Poland using LAMP, real-time
PCR and nested PCR
Anna Lass, Beata Szostakowska, Krzysztof Korzeniewski
and Panagiotis Karanis
ABSTRACT
Giardia intestinalis is a protozoan parasite, transmitted to humans and animals by the faecal–oral
route, mainly through contaminated water and food. Knowledge about the distribution of this parasite
in surface water in Poland is fragmentary and incomplete. Accordingly, 36 environmental water
samples taken from surface water reservoirs and wells were collected in Pomerania and Warmia-
Masuria provinces, Poland. The 50 L samples were filtered and subsequently analysed with three
molecular detection methods: loop-mediated isothermal amplification (LAMP), real-time polymerase
chain reaction (real-time PCR) and nested PCR. Of the samples examined, Giardia DNA was found in 15
(42%) samples with the use of LAMP; in 12 (33%) of these samples, Giardia DNA from this parasite was
also detected using real-time PCR; and in 9 (25%) using nested PCR. Sequencing of selected positive
samples confirmed that the PCR products were fragments of the Giardia intestinalis small subunit rRNA
gene. Genotyping using multiplex real-time PCR indicated the presence of assemblages A and B, with
the latter predominating. The results indicate that surface water in Poland, as well as water taken from
surface wells, may be a source of Giardia strains which are potentially pathogenic for humans. It was
also demonstrated that LAMP assay is more sensitive than the other two molecular assays.
Anna Lass
Beata Szostakowska
Department of Tropical Parasitology, Institute of
Maritime and Tropical Medicine in Gdynia,
Medical University of Gdansk,
9b Powstania Styczniowego Str.,
Gdynia 81-519,
Poland
Anna Lass
Panagiotis Karanis (corresponding author)
State Key Laboratory of Plateau Ecology and
Agriculture, Center for Biomedicine and
Infectious Disease, Qinghai Academy of Animal
Sciences and Veterinary Medicine,
Medical School of Qinghai University Xining,
1#Wei’er Road, Qinghai Biological Scientific Estate
Garden,
Xining 810016,
P. R. China
E-mail: panagiotis.karanis@uk-koeln.de
Krzysztof Korzeniewski
Epidemiology and Tropical Medicine Department in
Gdynia,
Military Institute of Medicine in Warsaw,
Grudzinskiego St. 4,
Gdynia 81-103,
Poland
Key words | environment, genotype, Giardia, LAMP, nested PCR, real-time PCR
INTRODUCTION
Giardia intestinalis is a protozoan parasite causing gastroin-
testinal diseases in humans and animals worldwide (Karanis
& Ey ; Smith et al. ; Reynolds et al. ; Ryan &
Cacciò ). It has been estimated that ingestion of as few
as ten cysts can lead to the development of infection in
humans and animals (Rendtorff ). The clinical manifes-
tation of giardiasis includes severe diarrhoea, abdominal
cramps, nausea, weight loss, and an acute phase which
usually lasts a few weeks and which may develop into a
chronic recurring disease (Wolfe ; Farthing ;
Adam ; Lebwohl et al. ). The prevalence of Giardia
infections in humans was previously estimated at 2‒5% in
industrialised countries and 20‒30% in developing countries
(Thompson & Monis ). However, regarding Europe, the
epidemiology report from ECDPC in 2014 confirmed 17,278
giardiasis cases, reported by 23 countries in the EU/EEA,
with the highest number of cases reported by Germany
(n ¼ 4,011), followed by the United Kingdom (n ¼ 3,628),
which accounted for 44% of cases (ECDPC ).
This parasite is transmitted by the faecal–oral route,
through contaminated water and food (Karanis et al. ;
Yoshida et al. ; Baldursson & Karanis ; Efstratiou
et al. ). The presence of Giardia cysts in water is a
problem throughout the world. They are resistant to
775 © IWA Publishing 2017 Journal of Water and Health | 15.5 | 2017
doi: 10.2166/wh.2017.039
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