Toxoplasmagondii oocysts, Giardia cysts and Cryptosporidium oocysts in outdoor swimming pools in Brazil

• Published in: Zoonoses and public health 2020-11, Vol.67 (7), p.785-795
• Main Authors: Pineda, Carolina Ortiz, Leal, Diego Averaldo Guiguet, Fiuza, Vagner Ricardo da Silva, Jose, Juliana, Borelli, Guilherme, Durigan, Mauricio, Pena, Hilda Fátima Jesus, Bueno Franco, Regina Maura
• Format: Article
• Language: English
• Subjects: Amplification; Animals; B1 gene; Backwash; Brazil; Chlorine; Cryptosporidium; Cryptosporidium - isolation & purification; Cysts; Deoxyribonucleic acid; DNA; Fluorescent Antibody Technique; Giardia; Giardia - isolation & purification; Health risks; Humans; Immunofluorescence; Immunomagnetic separation; Ingestion; Membrane filtration; Oocysts; Oocysts - isolation & purification; Pathogens; Phylogenetics; Phylogeny; Protozoa; Purification; Recreation; Recreational swimming; Recreational waters; Risk Factors; rRNA 18S; Swimming; Swimming Pools; Toxoplasma - isolation & purification; Toxoplasma gondii; Water - parasitology; Water purification; Water treatment; waterborne protozoa
• ISSN: 1863-1959; 1863-2378
• DOI: 10.1111/zph.12757

The accidental ingestion of treated recreational water is an important transmission route of waterborne protozoa worldwide. The present study aimed to provide the first evaluation of swimming pools in Brazil, analysing the presence of pathogenic protozoa (Toxoplasma gondii, Cryptosporidium spp. and Giardia spp.) by parasitological and molecular methods. A total of 57 samples were collected from 21 public swimming pools, either directly from the pool or filter backwash water and concentrated using the membrane filtration technique. Giardia cysts and Cryptosporidium oocysts were visualized by direct immunofluorescence assay after purification by immunomagnetic separation. Toxoplasma gondii oocysts were detected by autofluorescence visualization using the supernatant discarded during the purification step as a sample. Positive samples were submitted to molecular analysis. The molecular markers were used: SSU‐rRNA, tpi, gdh and bg, for Giardia DNA amplification, and 18S rRNA gene fragment amplification was used for the Cryptosporidium oocysts. The 529‐bp repeat element (REP529) fragment and the 35‐fold repetitive B1 gene were employed as a target for T. gondii. Amplified products were submitted to sequencing and phylogenetic analysis. Giardia cysts were detected in 19.0% and Cryptosporidium oocysts in 9.5% of swimming pools. In one swimming pool (4.7%), both protozoa were detected on at least one occasion. Structures similar to T. gondii oocysts were detected in 33.3% of the samples, ranging from one to 23 per slide. Giardia was confirmed by DNA amplification in three swimming pools; Giardia duodenalis Assemblage A was identified by the phylogenetic positioning of the β‐giardin gene. Toxoplasma gondii DNA was detected in 14.2% of swimming pools. The present study represents the first report of the occurrence of T. gondii oocysts in swimming pools. Recreational activity in swimming pools contaminated by chlorine‐resistant protozoa can represent a high risk of infection for bathers and swimmers.