Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

doi:10.1371/journal.pntd.0004822

. 2016 Jul 7;10(7):e0004822.

doi: 10.1371/journal.pntd.0004822. eCollection 2016 Jul.

Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

Catrin E Moore ^{1

2

3

4}, Kristin Elwin ^{5

6}, Nget Phot ⁷, Chanthou Seng ⁷, Saroeun Mao ⁷, Kuong Suy ⁷, Varun Kumar ⁷, Johanna Nader ⁸, Rachel Bousfield ⁹, Sanuki Perera ³, J Wendi Bailey ⁸, Nicholas J Beeching ^{8

10}, Nicholas P J Day ^{1

4}, Christopher M Parry ^{1

2

11

12}, Rachel M Chalmers ^{5

6}

Affiliations

¹ Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
² Cambodia-Oxford Medical Research Unit, Angkor Hospital for Children, Siem Reap, Cambodia.
³ Modernising Medical Microbiology Consortium, Nuffield Department of Clinical Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom.
⁴ Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
⁵ Cryptosporidium Reference Unit, Public Health Wales Microbiology, Singleton Hospital, Swansea, United Kingdom.
⁶ Swansea University Medical School, Grove Building, Swansea University, Singleton Park, Swansea, United Kingdom.
⁷ Angkor Hospital for Children, Siem Reap, Cambodia.
⁸ Clinical Sciences Group, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.
⁹ Microbiology Department, Addenbrooke's Hospital, Cambridge, United Kingdom.
¹⁰ NIHR Health Protection Research Unit (HPRU) in Gastrointestinal Infections, University of Liverpool, Liverpool, United Kingdom.
¹¹ London School of Hygiene and Tropical Medicine, London, United Kingdom.
¹² School of Tropical Medicine and Global Health, Nagasaki University, Nagasaki, Japan.

PMID: 27387755
PMCID: PMC4936737
DOI: 10.1371/journal.pntd.0004822

Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

Catrin E Moore et al. PLoS Negl Trop Dis. 2016.

. 2016 Jul 7;10(7):e0004822.

doi: 10.1371/journal.pntd.0004822. eCollection 2016 Jul.

Authors

Catrin E Moore ^{1

2

3

4}, Kristin Elwin ^{5

6}, Nget Phot ⁷, Chanthou Seng ⁷, Saroeun Mao ⁷, Kuong Suy ⁷, Varun Kumar ⁷, Johanna Nader ⁸, Rachel Bousfield ⁹, Sanuki Perera ³, J Wendi Bailey ⁸, Nicholas J Beeching ^{8

10}, Nicholas P J Day ^{1

4}, Christopher M Parry ^{1

2

11

12}, Rachel M Chalmers ^{5

6}

Affiliations

¹ Mahidol-Oxford Tropical Medicine Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
² Cambodia-Oxford Medical Research Unit, Angkor Hospital for Children, Siem Reap, Cambodia.
³ Modernising Medical Microbiology Consortium, Nuffield Department of Clinical Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom.
⁴ Centre for Tropical Medicine and Global Health, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
⁵ Cryptosporidium Reference Unit, Public Health Wales Microbiology, Singleton Hospital, Swansea, United Kingdom.
⁶ Swansea University Medical School, Grove Building, Swansea University, Singleton Park, Swansea, United Kingdom.
⁷ Angkor Hospital for Children, Siem Reap, Cambodia.
⁸ Clinical Sciences Group, Liverpool School of Tropical Medicine, Liverpool, United Kingdom.
⁹ Microbiology Department, Addenbrooke's Hospital, Cambridge, United Kingdom.
¹⁰ NIHR Health Protection Research Unit (HPRU) in Gastrointestinal Infections, University of Liverpool, Liverpool, United Kingdom.
¹¹ London School of Hygiene and Tropical Medicine, London, United Kingdom.
¹² School of Tropical Medicine and Global Health, Nagasaki University, Nagasaki, Japan.

PMID: 27387755
PMCID: PMC4936737
DOI: 10.1371/journal.pntd.0004822

Abstract

Background: In a prospective study, 498 single faecal samples from children aged under 16 years attending an outpatient clinic in the Angkor Hospital for Children, northwest Cambodia, were examined for Cryptosporidium oocysts and Giardia cysts using microscopy and molecular assays.

Methodology/principal findings: Cryptosporidium oocysts were detected in 2.2% (11/498) of samples using microscopy and in 7.7% (38/498) with molecular tests. Giardia duodenalis cysts were detected in 18.9% (94/498) by microscopy and 27.7% (138/498) by molecular tests; 82% of the positive samples (by either method) were from children aged 1-10 years. Cryptosporidium hominis was the most common species of Cryptosporidium, detected in 13 (34.2%) samples, followed by Cryptosporidium meleagridis in 9 (23.7%), Cryptosporidium parvum in 8 (21.1%), Cryptosporidium canis in 5 (13.2%), and Cryptosporidium suis and Cryptosporidium ubiquitum in one sample each. Cryptosporidium hominis and C. parvum positive samples were subtyped by sequencing the GP60 gene: C. hominis IaA16R6 and C. parvum IIeA7G1 were the most abundant subtypes. Giardia duodenalis was typed using a multiplex real-time PCR targeting assemblages A and B. Assemblage B (106; 76.8% of all Giardia positive samples) was most common followed by A (12.3%) and mixed infections (5.1%). Risk factors associated with Cryptosporidium were malnutrition (AOR 9.63, 95% CI 1.67-55.46), chronic medical diagnoses (AOR 4.51, 95% CI 1.79-11.34) and the presence of birds in the household (AOR 2.99, 95% CI 1.16-7.73); specifically C. hominis (p = 0.03) and C. meleagridis (p<0.001) were associated with the presence of birds. The use of soap was protective against Giardia infection (OR 0.74, 95% CI 0.58-0.95).

Conclusions/significance: This is the first report to describe the different Cryptosporidium species and subtypes and Giardia duodenalis assemblages in Cambodian children. The variety of Cryptosporidium species detected indicates both anthroponotic and zoonotic transmission in this population. Interventions to improve sanitation, increase hand washing after defecation and before preparing food and promote drinking boiled water may reduce the burden of these two parasites.

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Conflict of interest statement

The authors have declared that no competing interests exist.

References

1. Moore CE, Nget P, Saroeun M, Kuong S, Chanthou S, Kumar V, et al. Intestinal parasite infections in symptomatic children attending hospital in siem reap, cambodia. PLoS One. 2015;10(5):e0123719 10.1371/journal.pone.0123719 - DOI - PMC - PubMed
1. Yoder JS, Harral C, Beach MJ, Centers for Disease C, Prevention. Giardiasis surveillance—United States, 2006–2008. MMWR Surveill Summ. 2010;59(6):15–25. . - PubMed
1. Yoder JS, Gargano JW, Wallace RM, Beach MJ, Centers for Disease C, Prevention. Giardiasis surveillance—United States, 2009–2010. MMWR Surveill Summ. 2012;61(5):13–23. . - PubMed
1. Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, et al. Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet. 2013;382(9888):209–22. 10.1016/S0140-6736(13)60844-2 . - DOI - PubMed
1. Thompson RC. The zoonotic significance and molecular epidemiology of Giardia and giardiasis. Vet Parasitol. 2004;126(1–2):15–35. 10.1016/j.vetpar.200409008 . - DOI - PubMed

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[1] Moore CE, Nget P, Saroeun M, Kuong S, Chanthou S, Kumar V, et al. Intestinal parasite infections in symptomatic children attending hospital in siem reap, cambodia. PLoS One. 2015;10(5):e0123719 10.1371/journal.pone.0123719 - DOI - PMC - PubMed

[2] Moore CE, Nget P, Saroeun M, Kuong S, Chanthou S, Kumar V, et al. Intestinal parasite infections in symptomatic children attending hospital in siem reap, cambodia. PLoS One. 2015;10(5):e0123719 10.1371/journal.pone.0123719 - DOI - PMC - PubMed

[3] Yoder JS, Harral C, Beach MJ, Centers for Disease C, Prevention. Giardiasis surveillance—United States, 2006–2008. MMWR Surveill Summ. 2010;59(6):15–25. . - PubMed

[4] Yoder JS, Harral C, Beach MJ, Centers for Disease C, Prevention. Giardiasis surveillance—United States, 2006–2008. MMWR Surveill Summ. 2010;59(6):15–25. . - PubMed

[5] Yoder JS, Gargano JW, Wallace RM, Beach MJ, Centers for Disease C, Prevention. Giardiasis surveillance—United States, 2009–2010. MMWR Surveill Summ. 2012;61(5):13–23. . - PubMed

[6] Yoder JS, Gargano JW, Wallace RM, Beach MJ, Centers for Disease C, Prevention. Giardiasis surveillance—United States, 2009–2010. MMWR Surveill Summ. 2012;61(5):13–23. . - PubMed

[7] Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, et al. Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet. 2013;382(9888):209–22. 10.1016/S0140-6736(13)60844-2 . - DOI - PubMed

[8] Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, et al. Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet. 2013;382(9888):209–22. 10.1016/S0140-6736(13)60844-2 . - DOI - PubMed

[9] Thompson RC. The zoonotic significance and molecular epidemiology of Giardia and giardiasis. Vet Parasitol. 2004;126(1–2):15–35. 10.1016/j.vetpar.200409008 . - DOI - PubMed

[10] Thompson RC. The zoonotic significance and molecular epidemiology of Giardia and giardiasis. Vet Parasitol. 2004;126(1–2):15–35. 10.1016/j.vetpar.200409008 . - DOI - PubMed

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Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

Affiliations

Molecular Characterization of Cryptosporidium Species and Giardia duodenalis from Symptomatic Cambodian Children

Authors

Affiliations

Abstract

Conflict of interest statement

References

Publication types

MeSH terms

Grants and funding

LinkOut - more resources

Full Text Sources

Other Literature Sources

Medical