Giardia duodenalis and Cryptosporidium spp. as contaminant protozoa of the main rivers of western Romania

genetic characterization and public health potential of the isolates

Kálmán Imre, Claudia Sala, Adriana Morar, Marius S. Ilie, J. Plutzer, Mirela Imre, Florin Hora, Corina Badea, Mihai V. Herbei, Gheorghe Dărăbuș

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The objective of this study was to establish the prevalence, contamination level, and public health significance of Giardia duodenalis and Cryptosporidium spp. in the primary rivers of western Romania. A total of 53 sampling points in the 24 most important western Romanian rivers in four counties (Arad, Bihor, Caraș-Severin, and Timiș) were investigated from March to September 2016. Surface water samples were collected by microfiber filtration. Cryptosporidium and Giardia (oo)cysts were isolated using immunomagnetic separation (IMS) according to the USEPA 1623 method and, after staining with fluorescently labeled (FITC) monoclonal antibodies, were identified and counted under a microscope. The Cryptosporidium and Giardia (oo)cysts were identified to species and assemblage/sub-assemblage level through the nested PCR–RFLP procedure targeting the 18S ribosomal RNA and gdh genes, respectively. PCR-based techniques were utilized for all water samples. Overall, 22 samples (41.5%) were determined to be positive for Giardia cysts (ranging from 0.05 to 300 cysts per liter), and four samples (7.5%) tested positive for Cryptosporidium oocysts (0.17–48 oocysts/l). G. duodenalis was molecularly identified in 13 water samples (24.5%), indicating the presence of the sub-assemblage A-II (n = 12) and assemblage E (n = 1). PCR–RFLP showed that two samples (3.8%) contained Cryptosporidium DNA, and the identified species were Cryptosporidium parvum and Cryptosporidium canis. All positive results were successfully confirmed by DNA sequencing. Subtyping of the zoonotic C. parvum isolate based on sequence analysis of the GP60 gene revealed the occurrence of the IIaA16G1R1 subtype. The results of this study highlight considerable contamination of river waters with pathogenic Giardia spp. and Cryptosporidium spp., suggesting a potential risk for the public and animal health. This report presents the first extended published description of the presence of Giardia spp. and Cryptosporidium spp. in the aquatic environment in Romania.

Original languageEnglish
Pages (from-to)18672-18679
Number of pages8
JournalEnvironmental Science and Pollution Research
Volume24
Issue number22
DOIs
Publication statusPublished - Aug 1 2017

Fingerprint

Protozoa
Giardia lamblia
Romania
Cryptosporidium
Public health
cyst
Rivers
public health
Giardia
Public Health
Impurities
pollutant
Water
Contamination
DNA
Genes
18S Ribosomal RNA
river
Cysts
Monoclonal antibodies

Keywords

  • Cryptosporidium
  • Giardia
  • Public health
  • River water
  • Western Romania

ASJC Scopus subject areas

  • Environmental Chemistry
  • Pollution
  • Health, Toxicology and Mutagenesis

Cite this

Giardia duodenalis and Cryptosporidium spp. as contaminant protozoa of the main rivers of western Romania : genetic characterization and public health potential of the isolates. / Imre, Kálmán; Sala, Claudia; Morar, Adriana; Ilie, Marius S.; Plutzer, J.; Imre, Mirela; Hora, Florin; Badea, Corina; Herbei, Mihai V.; Dărăbuș, Gheorghe.

In: Environmental Science and Pollution Research, Vol. 24, No. 22, 01.08.2017, p. 18672-18679.

Research output: Contribution to journalArticle

Imre, Kálmán ; Sala, Claudia ; Morar, Adriana ; Ilie, Marius S. ; Plutzer, J. ; Imre, Mirela ; Hora, Florin ; Badea, Corina ; Herbei, Mihai V. ; Dărăbuș, Gheorghe. / Giardia duodenalis and Cryptosporidium spp. as contaminant protozoa of the main rivers of western Romania : genetic characterization and public health potential of the isolates. In: Environmental Science and Pollution Research. 2017 ; Vol. 24, No. 22. pp. 18672-18679.
@article{42d9f5071c334603a176899863b89ded,
title = "Giardia duodenalis and Cryptosporidium spp. as contaminant protozoa of the main rivers of western Romania: genetic characterization and public health potential of the isolates",
abstract = "The objective of this study was to establish the prevalence, contamination level, and public health significance of Giardia duodenalis and Cryptosporidium spp. in the primary rivers of western Romania. A total of 53 sampling points in the 24 most important western Romanian rivers in four counties (Arad, Bihor, Caraș-Severin, and Timiș) were investigated from March to September 2016. Surface water samples were collected by microfiber filtration. Cryptosporidium and Giardia (oo)cysts were isolated using immunomagnetic separation (IMS) according to the USEPA 1623 method and, after staining with fluorescently labeled (FITC) monoclonal antibodies, were identified and counted under a microscope. The Cryptosporidium and Giardia (oo)cysts were identified to species and assemblage/sub-assemblage level through the nested PCR–RFLP procedure targeting the 18S ribosomal RNA and gdh genes, respectively. PCR-based techniques were utilized for all water samples. Overall, 22 samples (41.5{\%}) were determined to be positive for Giardia cysts (ranging from 0.05 to 300 cysts per liter), and four samples (7.5{\%}) tested positive for Cryptosporidium oocysts (0.17–48 oocysts/l). G. duodenalis was molecularly identified in 13 water samples (24.5{\%}), indicating the presence of the sub-assemblage A-II (n = 12) and assemblage E (n = 1). PCR–RFLP showed that two samples (3.8{\%}) contained Cryptosporidium DNA, and the identified species were Cryptosporidium parvum and Cryptosporidium canis. All positive results were successfully confirmed by DNA sequencing. Subtyping of the zoonotic C. parvum isolate based on sequence analysis of the GP60 gene revealed the occurrence of the IIaA16G1R1 subtype. The results of this study highlight considerable contamination of river waters with pathogenic Giardia spp. and Cryptosporidium spp., suggesting a potential risk for the public and animal health. This report presents the first extended published description of the presence of Giardia spp. and Cryptosporidium spp. in the aquatic environment in Romania.",
keywords = "Cryptosporidium, Giardia, Public health, River water, Western Romania",
author = "K{\'a}lm{\'a}n Imre and Claudia Sala and Adriana Morar and Ilie, {Marius S.} and J. Plutzer and Mirela Imre and Florin Hora and Corina Badea and Herbei, {Mihai V.} and Gheorghe Dărăbuș",
year = "2017",
month = "8",
day = "1",
doi = "10.1007/s11356-017-9543-y",
language = "English",
volume = "24",
pages = "18672--18679",
journal = "Environmental Science and Pollution Research",
issn = "0944-1344",
publisher = "Springer Science + Business Media",
number = "22",

}

TY - JOUR

T1 - Giardia duodenalis and Cryptosporidium spp. as contaminant protozoa of the main rivers of western Romania

T2 - genetic characterization and public health potential of the isolates

AU - Imre, Kálmán

AU - Sala, Claudia

AU - Morar, Adriana

AU - Ilie, Marius S.

AU - Plutzer, J.

AU - Imre, Mirela

AU - Hora, Florin

AU - Badea, Corina

AU - Herbei, Mihai V.

AU - Dărăbuș, Gheorghe

PY - 2017/8/1

Y1 - 2017/8/1

N2 - The objective of this study was to establish the prevalence, contamination level, and public health significance of Giardia duodenalis and Cryptosporidium spp. in the primary rivers of western Romania. A total of 53 sampling points in the 24 most important western Romanian rivers in four counties (Arad, Bihor, Caraș-Severin, and Timiș) were investigated from March to September 2016. Surface water samples were collected by microfiber filtration. Cryptosporidium and Giardia (oo)cysts were isolated using immunomagnetic separation (IMS) according to the USEPA 1623 method and, after staining with fluorescently labeled (FITC) monoclonal antibodies, were identified and counted under a microscope. The Cryptosporidium and Giardia (oo)cysts were identified to species and assemblage/sub-assemblage level through the nested PCR–RFLP procedure targeting the 18S ribosomal RNA and gdh genes, respectively. PCR-based techniques were utilized for all water samples. Overall, 22 samples (41.5%) were determined to be positive for Giardia cysts (ranging from 0.05 to 300 cysts per liter), and four samples (7.5%) tested positive for Cryptosporidium oocysts (0.17–48 oocysts/l). G. duodenalis was molecularly identified in 13 water samples (24.5%), indicating the presence of the sub-assemblage A-II (n = 12) and assemblage E (n = 1). PCR–RFLP showed that two samples (3.8%) contained Cryptosporidium DNA, and the identified species were Cryptosporidium parvum and Cryptosporidium canis. All positive results were successfully confirmed by DNA sequencing. Subtyping of the zoonotic C. parvum isolate based on sequence analysis of the GP60 gene revealed the occurrence of the IIaA16G1R1 subtype. The results of this study highlight considerable contamination of river waters with pathogenic Giardia spp. and Cryptosporidium spp., suggesting a potential risk for the public and animal health. This report presents the first extended published description of the presence of Giardia spp. and Cryptosporidium spp. in the aquatic environment in Romania.

AB - The objective of this study was to establish the prevalence, contamination level, and public health significance of Giardia duodenalis and Cryptosporidium spp. in the primary rivers of western Romania. A total of 53 sampling points in the 24 most important western Romanian rivers in four counties (Arad, Bihor, Caraș-Severin, and Timiș) were investigated from March to September 2016. Surface water samples were collected by microfiber filtration. Cryptosporidium and Giardia (oo)cysts were isolated using immunomagnetic separation (IMS) according to the USEPA 1623 method and, after staining with fluorescently labeled (FITC) monoclonal antibodies, were identified and counted under a microscope. The Cryptosporidium and Giardia (oo)cysts were identified to species and assemblage/sub-assemblage level through the nested PCR–RFLP procedure targeting the 18S ribosomal RNA and gdh genes, respectively. PCR-based techniques were utilized for all water samples. Overall, 22 samples (41.5%) were determined to be positive for Giardia cysts (ranging from 0.05 to 300 cysts per liter), and four samples (7.5%) tested positive for Cryptosporidium oocysts (0.17–48 oocysts/l). G. duodenalis was molecularly identified in 13 water samples (24.5%), indicating the presence of the sub-assemblage A-II (n = 12) and assemblage E (n = 1). PCR–RFLP showed that two samples (3.8%) contained Cryptosporidium DNA, and the identified species were Cryptosporidium parvum and Cryptosporidium canis. All positive results were successfully confirmed by DNA sequencing. Subtyping of the zoonotic C. parvum isolate based on sequence analysis of the GP60 gene revealed the occurrence of the IIaA16G1R1 subtype. The results of this study highlight considerable contamination of river waters with pathogenic Giardia spp. and Cryptosporidium spp., suggesting a potential risk for the public and animal health. This report presents the first extended published description of the presence of Giardia spp. and Cryptosporidium spp. in the aquatic environment in Romania.

KW - Cryptosporidium

KW - Giardia

KW - Public health

KW - River water

KW - Western Romania

UR - http://www.scopus.com/inward/record.url?scp=85021308108&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85021308108&partnerID=8YFLogxK

U2 - 10.1007/s11356-017-9543-y

DO - 10.1007/s11356-017-9543-y

M3 - Article

VL - 24

SP - 18672

EP - 18679

JO - Environmental Science and Pollution Research

JF - Environmental Science and Pollution Research

SN - 0944-1344

IS - 22

ER -