Molecular diversity and frequency of the diarrheagenic enteric protozoan Giardia duodenalis and Cryptosporidium spp. in a hospital setting in Northern Spain

• Published in: PloS one 2017-06, Vol.12 (6), p.e0178575-e0178575
• Main Authors: Azcona-Gutiérrez, José Manuel, de Lucio, Aida, Hernández-de-Mingo, Marta, García-García, Concepción, Soria-Blanco, Luis Miguel, Morales, Lucía, Aguilera, María, Fuentes, Isabel, Carmena, David
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Annual variations; Assaying; Biology and Life Sciences; Care and treatment; Child; Child, Preschool; Children; Cryptosporidiosis; Cryptosporidiosis - diagnosis; Cryptosporidium; Cryptosporidium - classification; Cryptosporidium - genetics; Cryptosporidium - isolation & purification; Dehydrogenase; Developed countries; DNA, Protozoan - genetics; Epidemiology; Female; Flow; Gastrointestinal diseases; Genes; Genetic aspects; Genotypes; Genotyping; Geographical distribution; Giardia; Giardia lamblia - classification; Giardia lamblia - genetics; Giardia lamblia - isolation & purification; Giardiasis; Giardiasis - diagnosis; Glutamate dehydrogenase; Glutamic acid; Glycoproteins; Health aspects; Health care; Hospitals; Humans; Infant; Infection; Infections; Interannual variability; Longitudinal Studies; Male; Medicine and Health Sciences; Methods; Microscopy; Middle Aged; Multilocus Sequence Typing; Parasites; Parasitology; Pathogens; Patients; People and Places; Polymerase chain reaction; Population; Population studies; Prevalence; Prevention; Protozoa; Public health; Real time; Research and Analysis Methods; Ribonucleic acid; RNA; rRNA; Seasonal variations; Spain; Surveys; Transcription; Young Adult; Zoonoses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0178575

Human giardiosis and cryptosporidiosis are caused by the enteric protozoan parasites Giardia duodenalis and Cryptosporidium spp. Both pathogens are major contributors to the global burden of diarrhoeal disease, affecting primarily children and immunodebilitated individuals in resource-poor settings. Giardiosis and cryptosporidiosis also represent an important, often underestimate, public health threat in developed countries. In Spain only limited information is currently available on the epidemiology of these infections. Molecular data on the diversity, frequency, geographical distribution, and seasonality of G. duodenalis assemblages/sub-assemblages and Cryptosporidium species/sub-genotypes are particularly scarce. A longitudinal molecular epidemiological survey was conducted between July 2015 to September 2016 in patients referred to or attended at the Hospital San Pedro (La Rioja, Northern Spain) that tested positive for G. duodenalis (N = 106) or Cryptosporidium spp. (N = 103) by direct microscopy and/or a rapid lateral flow immunochromatographic assay. G. duodenalis infections were subsequently confirmed by real-time PCR and positive isolates assessed by multi-locus sequence genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. Cryptosporidium species and sub-genotypes were investigated at the 60 kDa glycoprotein or the small subunit ribosomal RNA genes of the parasite. Sociodemographic and clinical parameters of infected patients were also gathered and analysed. Out of 90 G. duodenalis-positive isolates by real-time PCR a total of 16 isolates were successfully typed. AII (44%, 7/16) was the most prevalent sub-assemblage found, followed by BIV (31%, 5/16) and BIII (19%, 3/16). A discordant genotype result AII/AIII was identified in an additional (6%, 1/16) isolate. No mixed infections A+B were detected. Similarly, a total of 81 Cryptosporidium spp. isolates were successfully typed, revealing the presence of C. hominis (81%, 66/81) and C. parvum (19%, 15/81). Obtained GP60 sequences were assigned to sub-type families Ib (73%, 59/81) within C. hominis, and IIa (7%, 6/81) and IId (2%, 2/81) within C. parvum. A marked inter-annual variation in Cryptosporidium cases was observed. Human giardiasis and cryptosporidiosis are commonly identified in patients seeking medical care in Northern Spain and represent a more important health concern than initially thought. Assemblage A within G. duodenalis and sub-genotype IbA10G2 within C. hom