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. 2021 May 4;10(5):1005.
doi: 10.3390/foods10051005.

Survival of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and Herpes Simplex Virus 1 (HSV-1) on Foods Stored at Refrigerated Temperature

Janak Dhakal  1 , Mo Jia  2 , Jonathan D Joyce  3 , Greyson A Moore  4 , Reza Ovissipour  5 , Andrea S Bertke  2
Affiliations

Survival of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and Herpes Simplex Virus 1 (HSV-1) on Foods Stored at Refrigerated Temperature

Janak Dhakal et al. Foods. .

Abstract

Outbreaks of coronavirus infectious disease 2019 (COVID-19) in meat processing plants and media reports of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection on foods have raised concerns of a public health risk from contaminated foods. We used herpes simplex virus 1, a non-Biosafety Level 3 (non-BSL3) enveloped virus, as a surrogate to develop and validate methods before assessing the survival of infectious SARS-CoV-2 on foods. Several food types, including chicken, seafood, and produce, were held at 4 °C and assessed for infectious virus survival (herpes simplex virus 1 (HSV-1) and SARS-CoV-2) at 0 h, 1 h, and 24 h post-inoculation (hpi) by plaque assay. At all three time points, recovery of SARS-CoV-2 was similar from chicken, salmon, shrimp, and spinach, ranging from 3.4 to 4.3 log PFU/mL. However, initial (0 h) virus recovery from apples and mushrooms was significantly lower than that from poultry and seafood, and infectious virus decreased over time, with recovery from mushrooms becoming undetectable by 24 hpi. Comparing infectious virus titers with viral genome copies confirmed that PCR-based tests only indicate presence of viral nucleic acid, which does not necessarily correlate with the quantity of infectious virus. The survival and high recovery of SARS-CoV-2 on certain foods highlight the importance of safe food handling practices in mitigating any public health concerns related to potentially contaminated foods.

Keywords: COVID-19; HSV-1; RT-qPCR; SARS-CoV-2; food contamination; foodborne illness; foodborne transmission; plaque assay; qPCR.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Rinse versus massage method comparison. Herpes simplex virus (HSV-1) was recovered from chicken thigh, apple skin, and mushrooms incubated at 4 °C for 0 h, 1 h, and 24 h using (A) rinse method and (B) massage method, quantifying infectious virus titer by standard plaque assay on Vero 76 cells and shown as log PFU/mL (n = 3). Inoculum (Inoc) titer is shown as PFU/sample (log); 10 µL of a 2.3 × 108 PFU/mL stock (≈2.5 × 106 PFU/sample) were inoculated onto each food sample. The dashed line indicates the detection limit (0.7 log or 5 plaques/mL) of the viral plaque assay. a–c Means within an incubation time that have a common lowercase letter are not significantly different (p < 0.05).
Figure 2
Figure 2
Survival of HSV-1 on foods. HSV-1 was recovered from foods immediately after inoculation (0 h) and at 1 h and 24 h after incubation at 4 °C (n = 3). Infectious virus titer was quantified by plaque assay on Vero 76 cells and shown as log PFU/mL. Inoculum (Inoc) is shown as PFU/sample. Dashed line indicates detection limit of the plaque assay (0.7 log pfu/mL). a–d Means within an incubation time that have a common lowercase letter are not significantly different (p < 0.05).
Figure 3
Figure 3
Survival of SARS-CoV-2 on foods. SARS-CoV-2 was recovered from foods immediately after inoculation (0 h) and at 1 h and 24 h after incubation at 4 °C (n = 3). Infectious virus titer was quantified by plaque assay on Vero E6 cells and shown as log PFU/mL. Inoculum (Inoc) is shown as PFU/sample. Dashed line indicates detection limit of the plaque assay (0.7 log PFU/mL). a–d Means within an incubation time that have a common lowercase letter are not significantly different (p < 0.05).
Figure 4
Figure 4
Comparison of infectious virus titer and viral genome copy number. Infectious viral titers were determined by plaque assay for (A) HSV-1 on Vero 76 cells, and (C) SARS-CoV-2 on Vero E6 cells, shown as log PFU/mL. Viral genome copies were determined by qPCR/RT-PCR for (B) HSV-1 by qPCR using primers and probe specific for the HSV-1 thymidine kinase (TK) gene, and (D) SARS-CoV-2 by RT-qPCR using primers and probe specific for the SARS-CoV-2 nucleocapsid (N) gene, shown as viral genome copies/mL.
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