Human coronaviruses (HCoVs) are mainly associated with respiratory infections. However, there is evidence that highly pathogenic HCoVs, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Middle East Respiratory Syndrome (MERS-CoV), infect the gastrointestinal (GI) tract and are shed in the fecal matter of the infected individuals. These observations have raised questions regarding the possibility of fecal-oral route as well as foodborne transmission of SARS-CoV-2 and MERS-CoV. Studies regarding the survival of HCoVs on inanimate surfaces demonstrate that these viruses can remain infectious for hours to days, however, there is limited data regarding the viral survival on fresh produce, which is usually consumed raw or with minimal heat processing. To address this knowledge gap, we examined the persistence of HCoV-229E, as a surrogate for highly pathogenic HCoVs, on the surface of commonly consumed fresh produce, including: apples, tomatoes, cucumbers and lettuce. Herein, we demonstrated that viral infectivity declines within a few hours post-inoculation (p.i) on apples and tomatoes, and no infectious virus was detected at 24h p.i, while the virus persists in infectious form for 72h p.i on cucumbers and lettuce. The stability of viral RNA was examined by droplet-digital RT-PCR (ddRT-PCR), and it was observed that there is no considerable reduction in viral RNA within 72h p.i.

Hepatitis E virus (HEV) causes acute hepatitis with approximately 20 million cases per year globally. Based on genetic diversity, HEV is classified into different genotypes, with genotype 3 (HEV-3) being most prevalent in Europe and North America. The transmission of HEV-3 has been shown to be zoonotic and mainly associated with the consumption of raw or undercooked pork products. Herein, we investigated the efficacy of high-pressure processing (HPP) in inactivation of HEV-3 using a cell culture system. HPP has been indicated as a promising non-thermal pathogen inactivation strategy for treatment of certain high-risk food commodities, without any noticeable changes in their nature. For this purpose, we treated HEV-3 in media with different conditions of HPP: 400 MPa for 1 and 5 min, as well as 600 MPa for 1 and 5 min, at ambient temperature. All four HPP treatments of HEV in media were observed to result in a 2-log reduction in HEV load, as determined by the amounts of extracellular HEV RNA produced at 14-day post-infection, using the A549/D3 cell culture system. However, application of the same treatments to artificially contaminated pork pâté resulted in 0.5 log reduction in viral load. These results indicate that the efficacy of HPP treatment in the inactivation of HEV-3 is matrix-dependent, and independent of maximum pressure between 400 and 600 MPa and hold time between 1 and 5 min. Based on the obtained results, although the HPP treatment of pork pâté reduces the HEV-3 load, it might not be sufficient to fully mitigate the risk.

13Hepatitis E virus (HEV) causes acute hepatitis with approximately 20 million cases per year 14 globally. While HEV is endemic in certain regions of Asia, Africa and South America, it is 15 considered an emerging foodborne pathogen in developed countries. Based on genetic diversity, 16 HEV is classified into different genotypes, with genotype 3 (HEV-3) being most prevalent in 17 Europe and North America. The transmission of HEV-3 has been shown to be zoonotic and 18 mainly associated with the consumption of raw or undercooked pork products. Herein, we 19 investigated the efficacy of high-pressure processing (HPP) in the inactivation of HEV-3 using a 20 cell culture system. HPP has been indicated as a promising nonthermal pathogen inactivation 21 strategy for treatment of certain high-risk food commodities, without any noticeable changes in 22 their nature. For this purpose, we treated HEV-3 in media as well as in artificially inoculated 23 pork pâté, with different conditions of HPP: 400 MPa for 1 and 5 minutes, as well as 600 MPa 24 for 1 and 5 minutes, at ambient temperature. In general, we observed approximately a 2-log 25 reduction in HEV load by HPP treatments in media; however, similar treatment in the pork pâté 26 resulted in a much lower reduction in viral load. Therefore, the efficacy of HPP treatment in the 27 inactivation of HEV-3 is matrix-dependent. 28Importance: HEV is an emerging foodborne pathogen in industrialized countries, and its 29 transmission is associated with the consumption of contaminated undercooked pork product. In 30 this work, we employed an infectivity assay to investigate the potential of high-pressure in 31 inactivation of HEV in media and ready-to-eat pork pâté. We demonstrated that the effect of 32 HPP on inactivation of HEV depends on the surrounding matrix. 33 PCR 35 36 HEV is a single-stranded RNA virus with positive polarity belonging to the Hepeviridae family 37 (1). The HEV genome has 3 open reading frames (ORFs): ORF1 encodes a long non-structural 38 polyprotein with multiple functions; ORF2 encodes the viral capsid protein; and ORF3 encodes a 39 small phosphoprotein with structural and non-structural functions (2), (3). The Hepeviridae 40 contain two genera Orthohepevirus and Piscihepevirus, which infect a wide range of vertebrate 41 hosts (4). Four genotypes (HEV-1, HEV-2, HEV-3 and HEV-4) of the species Orthohepevirus A 42 are associated with human illness. HEV-1 and HEV-2 are restricted to humans and are prevalent 43 in regions with poor water sanitation, such as the developing countries of Asia, Africa, South and 44Central America (5, 6). On the other hand, HEV-3 and HEV-4 are considered to be zoonotic 45 pathogens as they have a much wider range of mammalian hosts including, among others, 46 domestic and wild swine and ruminants (7-9). Hepatocytes have been identified as the primary 47 sites of HEV replication, but the virus can replicate in other tissues such as epithelial cells of the 48 small intestine, placenta, and muscle (10-13). 49Clinical manifest...

Human coronaviruses (HCoVs) are mainly associated with respiratory infections. However, there is evidence that highly pathogenic HCoVs, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Middle East Respiratory Syndrome (MERS-CoV), infect the gastrointestinal (GI) tract and are shed in the fecal matter of the infected individuals. These observations have raised questions regarding the possibility of fecal-oral route as well as foodborne transmission of SARS-CoV-2 and MERS-CoV. Studies regarding the survival of HCoVs on inanimate surfaces demonstrate that these viruses can remain infectious for hours to days, however, to date, there is no data regarding the viral survival on fresh produce, which is usually consumed raw or with minimal heat processing. To address this knowledge gap, we examined the persistence of HCoV-229E, as a surrogate for highly pathogenic HCoVs, on the surface of commonly consumed fresh produce, including: apples, tomatoes and cucumbers. Herein, we demonstrated that viral infectivity declines within a few hours post-inoculation (p.i) on apples and tomatoes, and no infectious virus was detected at 24h p.i, while the virus persists in infectious form for 72h p.i on cucumbers. The stability of viral RNA was examined by droplet-digital RT-PCR (ddRT-PCR), and it was observed that there is no considerable reduction in viral RNA within 72h p.i.