Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR

• Published in: Brazilian journal of microbiology Vol. 53; no. 3; pp. 1465 - 1471
• Main Authors: Pedrosa de Macena, Lorena da Graça, Pereira, Joseane Simone de Oliveira, da Silva, Jansen Couto, Ferreira, Fernando César, Maranhão, Adriana Gonçalves, Lanzarini, Natália Maria, Miagostovich, Marize Pereira
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01-09-2022; Springer Nature B.V
• Subjects: Biomedical and Life Sciences; Brackish water; Capsids; Chemical analysis; Environmental Microbiology - Research Paper; Food Microbiology; Infectivity; Life Sciences; Medical Microbiology; Microbial Ecology; Microbial Genetics and Genomics; Microbiology; Micrometers; Mycology; Photoactivation; Photolysis; Raw sewage; Reduction; Risk assessment; Room temperature; Seawater; Sewage; Viruses; Water analysis; Propidium monoazide; Raw sewage; PMAxx-qPCR; HAdV; Brackish water; Seawater
• ISSN: 1517-8382; 1678-4405
• DOI: 10.1007/s42770-022-00775-5

Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess
human mastadenovirus
(HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 μM, 50 μM, and 100 μM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 μM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 μM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 10
4
, 6.81 × 10
2
, and 2.33 × 10
2
GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water.