High occurrence of viable forms of Cryptosporidium and Giardia in domestic sewage from an agricultural region of Brazil

Abstract

Cryptosporidium and Giardia are the main etiologies of waterborne outbreaks caused by protozoa. These parasites are commonly detected in wastewater; however, there is little knowledge about the concentration of viable forms in treated sewage, mainly in small communities. To understand more about the presence of viable oocysts and cysts in domestic sewage, we monitored the affluent and effluent of a wastewater treatment plant (WWTP) in inner-city Brazil. Ten samplings and seven follow-ups were performed in 2020. Samples were concentrated by centrifugation, filtration and purified by fluctuation. Viability was accessed by propidium-monoazide (PMA) associated with nPCR and qPCR. Both viable protozoa were detected in all raw sewage samples (average: 438.5 viable oocysts/L). Regarding treated sewage, Cryptosporidium was detected in all of the samples (average: 92.8 viable oocysts/L) and Giardia was detected in 70% with viable cysts in 30%. Considering the follow-ups, 31.17% of Cryptosporidium viable oocysts remained in the effluent after the treatment. High amounts of Cryptosporidium and a high frequency of Giardia were detected, therefore both arrived at WWTP and were discharged into the river. These alert the presence of agro-industrial effluents into domestic sewage and demonstrated the effectiveness of the concentration technique for monitoring protozoa in wastewater. HIGHLIGHTS Viable oocysts and cysts persist after the stabilization pond treatment system.; High concentration of viable Cryptosporidium was quantified in treated sewage.; An accurate methodology to concentrate and purify (oo)cysts in wastewater was described.; PMA-PCR is a cheap and efficient methodology to detect (oo)cysts viability.; Agro-industrial effluents might influence protozoa concentration in domestic sewage.;

Keywords

• contamination
• propidium-monoazide
• qpcr
• quantification
• wastewater
• wwtp