Evaluation of Extraction Methods to Detect Noroviruses in Ready-to-Eat Raw Milk Minas Artisanal Cheese

Cantelli, Carina Pacheco; Silva, Marcio Roberto; Pimenta, Laís Marques; Tavares, Guilherme Caetano Lanzieri; Baduy, Gabriel Assad; Duch, André Almeida Santos; Menezes, Liliane Denize Miranda; Fialho, Alexandre Madi; Maranhão, Adriana Gonçalves; Fumian, Tulio Machado; Miagostovich, Marize Pereira; Leite, José Paulo Gagliardi

doi:10.1007/s12560-024-09588-1

Cited by 3 publications

(1 citation statement)

References 50 publications

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“…The mengovirus (MgV vMC 0 ), a non-virulent murine mutant strain lacking the poly(C) tract and a member of the Picornaviridae family, was used as an internal extraction process control. Previously, MgV vMC 0 was propagated in sub-confluent VERO cells (ATCC-CCL-81), as described by Cantelli et al [ 35 ]. Ten microliters of MgV vMC 0 (7.7 × 10 6 genome copies (GC)/µL) were spiked in each bivalve sample (a pool of 12–15 digestive glands was dissected) and homogenized for 3 min in a vortex with 2 mL of a proteinase K solution (100 µg/mL).…”

Section: Methodsmentioning

confidence: 99%

Assessment of Gastroenteric Viruses in Marketed Bivalve Mollusks in the Tourist Cities of Rio de Janeiro, Brazil, 2022

Cantelli,

Tavares,

Sarmento

et al. 2024

Viruses

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This study investigated the prevalence and genetic diversity of gastroenteric viruses in mussels and oysters in Rio de Janeiro, Brazil. One hundred and thirty-four marketed bivalve samples were obtained between January and December 2022. The viral analysis was performed according to ISO/TS 15216, and the screening revealed the detection of norovirus GII/GI (40.3%), sapovirus (SaV; 12.7%), human mastadenovirus (7.5%), and rotavirus A (RVA; 5.9%). In total, 44.8% (60) of shellfish samples tested positive for one or more viruses, 46.7% (28/60) of the positive samples tested positive for a single viral agent, 26.7% (16) tested positive for two viral agents, 8.3% (5) for three viral agents, and 13.3% (8) for four viral agents. Additionally, three mussel samples were contaminated with the five investigated viruses (5%, 3/60). Norovirus GII showed the highest mean viral load (3.4 × 105 GC/g), followed by SaV (1.4 × 104 GC/g), RVA (1.1 × 104 GC/g), human mastadenovirus (3.9 × 103 GC/g), and norovirus GI (6.7 × 102 GC/g). Molecular characterization revealed that the recovered norovirus strains belonged to genotypes GII.2, GII.6, GII.9, GII.17, and GII.27; SaV belonged to genotypes GI.1 and GIV.1; RVA to genotypes G6, G8, P [8]-III, and human mastadenovirus to types F40 and F41. The GII.27 norovirus characterized in this study is the only strain of this genotype reported in Brazil. This study highlights the dissemination and diversity of gastroenteric viruses present in commercialized bivalves in a touristic area, indicating the potential risk to human health and the contribution of bivalves in the propagation of emerging pathogens.

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Section: Methodsmentioning

confidence: 99%

Assessment of Gastroenteric Viruses in Marketed Bivalve Mollusks in the Tourist Cities of Rio de Janeiro, Brazil, 2022

Cantelli,

Tavares,

Sarmento

et al. 2024

Viruses

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Testing of Retail Cheese, Butter, Ice Cream, and Other Dairy Products for Highly Pathogenic Avian Influenza in the US

Suarez,

Goraichuk,

Killmaster

et al. 2025

Journal of Food Protection

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Testing of retail cheese, butter, ice cream and other dairy products for highly pathogenic avian influenza in the US

Suarez,

Goraichuk,

Killmaster

et al. 2024

Preprint

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The recent outbreak of highly pathogenic avian influenza (HPAI) in dairy cows has created public health concerns about the potential of consumers being exposed to live virus from commercial dairy products. Previous studies support that pasteurization effectively inactivates avian influenza in milk and an earlier retail milk survey showed viral RNA, but no live virus could be detected in the dairy products tested. Because of the variety of products and processing methods in which milk is used, additional product testing was conducted to determine if HPAI viral RNA could be detected in retail dairy samples, and for positive quantitative real-time RT-PCR (qRT-PCR) samples to be tested for presence of live virus. Revised protocols were developed to extract RNA from solid dairy products including cheese and butter. The solid dairy product was mechanically liquified with garnet and zirconium beads in a bead beater diluted 1 to 4 with BHI media. This pre-processing step was suitable in allowing efficient RNA extraction with standard methods. Trial studies were conducted with different cheese types with spiked in avian influenza virus to show that inoculation of the liquified cheese into embryonating chicken eggs was not toxic to the embryos and allowed virus replication. A total of 167 retail dairy samples, including a variety of cheeses, butter, ice cream, and fluid milk were collected as part of nationwide survey. A total of 17.4% (29/167) of the samples had detectable viral RNA by qRT-PCR targeting the matrix gene, but all samples were negative for live virus after testing with embryonating egg inoculation. The viral RNA was also evaluated by sequencing part of the hemagglutinin gene using a revised protocol optimized to deal with the fragmented viral RNA. The sequence analysis showed all viral RNA positive samples were highly similar to previously reported HPAI dairy cow isolates. Using the revised protocols, it was determined that HPAI viral RNA could be detected in a variety of dairy products, but existing pasteurizations methods effectively inactivate virus assuring consumer safety.

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Evaluation of Extraction Methods to Detect Noroviruses in Ready-to-Eat Raw Milk Minas Artisanal Cheese

Cited by 3 publications

References 50 publications

Assessment of Gastroenteric Viruses in Marketed Bivalve Mollusks in the Tourist Cities of Rio de Janeiro, Brazil, 2022

Assessment of Gastroenteric Viruses in Marketed Bivalve Mollusks in the Tourist Cities of Rio de Janeiro, Brazil, 2022

Testing of Retail Cheese, Butter, Ice Cream, and Other Dairy Products for Highly Pathogenic Avian Influenza in the US

Testing of retail cheese, butter, ice cream and other dairy products for highly pathogenic avian influenza in the US

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