title: Virus recovering from strawberries: Evaluation of a skimmed milk organic flocculation method for assessment of microbiological contamination.
abstract: Skimmed milk organic flocculation method was adapted, optimized and compared with polyethylene glycol (PEG) precipitation and filtration methods for recovering viruses from a strawberry matrix. Spiking experiments with norovirus genogroup II genotype 4 (NoV GII.4) and murine norovirus 1 (MNV-1) demonstrated that the organic flocculation method associated with a glycine elution buffer, filter bag and cetyltrimethylammonium bromide (CTAB) showed a recovery percentage of 2.5 and 32 times higher than PEG precipitation and filtration methodologies for NoV recovering. Furthermore, this method was used for investigating NoV and human adenoviruses (HAdVs) in 90 samples of fresh strawberries commercialized in Rio de Janeiro markets. NoV GI and GII were not detected in those samples and MNV-1, used as internal process control (IPC), was recovered in 95.5% (86) of them. HAdVs were detected in 18 (20.0%) samples and characterized by nucleotide sequencing as Human Mastadenovirus specie F and as type specie HAdV-2. Bacterial analysis did not detect Salmonella spp. and Listeria monocytogenes, however, 3.3% of fecal coliforms were detected in those samples. These results indicate the organic flocculation method as an alternative for recovering enteric viruses from strawberries, emphasizing a need for virus surveillance in food matrices.
authors.affiliation: Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: fabianagilmelgaco@gmail.com.
authors.forename: Fabiana Gil
authors.initials: FG
authors.lastname: Melgaço
authors.affiliation: Laboratory of Molecular Virology, Regional Norte, CENUR Noroeste, Universidad de la República, Salto, Uruguay. Electronic address: matvicmon@yahoo.com.
authors.forename: Matias
authors.initials: M
authors.lastname: Victoria
authors.affiliation: Laboratory of Virology, Fluminense Federal University, Niterói, RJ, Brazil. Electronic address: adrianacorrea@id.uff.br.
authors.forename: Adriana Abreu
authors.initials: AA
authors.lastname: Corrêa
authors.affiliation: Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: acganime@ioc.fiocruz.br.
authors.forename: Ana Carolina
authors.initials: AC
authors.lastname: Ganime
authors.affiliation: Laboratory of Microbiology and Parasitology, Faculty of Pharmacy, Rio de Janeiro Federal University, Macaé, RJ, Brazil. Electronic address: fabio.malta@macae.ufrj.br.
authors.forename: Fábio Correia
authors.initials: FC
authors.lastname: Malta
authors.affiliation: Food Sector, Laboratory of Food and Sanitizing, Department of Microbiology, National Institute of Quality Control in Health, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: marcelollb8@gmail.com.
authors.forename: Marcelo Luiz Lima
authors.initials: ML
authors.lastname: Brandão
authors.affiliation: Food Sector, Laboratory of Food and Sanitizing, Department of Microbiology, National Institute of Quality Control in Health, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: valeria.medeiros@incqs.fiocruz.br.
authors.forename: Valéria
authors.initials: V
authors.lastname: de Mello Medeiros
authors.affiliation: Food Sector, Laboratory of Food and Sanitizing, Department of Microbiology, National Institute of Quality Control in Health, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: carla.rosas@incqs.fiocruz.br.
authors.forename: Carla
authors.initials: C
authors.lastname: de Oliveira Rosas
authors.affiliation: Food Sector, Laboratory of Food and Sanitizing, Department of Microbiology, National Institute of Quality Control in Health, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: silvia.bricio@incqs.fiocruz.br.
authors.forename: Silvia Maria Lopes
authors.initials: SM
authors.lastname: Bricio
authors.affiliation: Laboratory of Comparative and Environmental Virology, Oswaldo Cruz Institute, Fiocruz, Rio de Janeiro-RJ, Brazil. Electronic address: marizepm@ioc.fiocruz.br.
authors.forename: Marize Pereira
authors.initials: MP
authors.lastname: Miagostovich
mesh_terms: D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D004770:Enterovirus; D005243:Feces / Q000382:microbiology / Q000821:virology; D005427:Flocculation; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D031985:Fragaria / Q000821:virology*; D006801:Humans; D008089:Listeria monocytogenes / Q000235:genetics / Q000302:isolation & purification; D008892:Milk / Q000737:chemistry*; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry*; D012367:RNA, Viral / Q000235:genetics; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral; D011092:Polyethylene Glycols
keywords: Enteric virus; Foodborne; Low cost; Skimmed milk flocculation method; Strawberry