Evaluation of High-Pressure Processing in Inactivation of the Hepatitis E Virus

Abstract

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...

Examining the Effect of Organic Acids on Inactivation of Hepatitis E Virus

“…The protective effect of food against different inactivation strategies has been studied for various foodborne viruses and their surrogates (reviewed in Cook et al (4)(5)(6)). The protective effect of food against inactivation of HEV-3c by HPP has been previously demonstrated by our group (23). It can be speculated that food components in pork pâté, such as fat, protein, and salt, can interact with the viral capsid or organic acids, reducing the inactivation efficiency.…”

“…HEV extraction: pork pâté. The modified ISO 15216-1:2017 method for viral extraction of bivalve molluscan shellfish (13) was employed as described previously (23). Following treatments, each pâté sample was placed in a stomacher bag with 16 mL of Tris glycine beef extract and incubated on a rocking plate for 20 min at room temperature.…”

“…The RNeasy Mini Kit (Qiagen, Mississauga, Ontario, Canada) protocol "Purifying Total RNA from Animal Cells with Spin Technology" was used to isolate RNA from the A549/D3 cells. The recovered RNA was quantified using Bio-Rad ddRT-PCR technology (Bio-Rad Laboratories, Mississauga, Ontario, Canada) as previously described (22,23) for determination of intracellular HEV RNA. The One-Step ddRT-PCR Kit for Probes (Bio-Rad) was employed according to manufacturer's instructions.…”

“…HEV is resistant to drying and is stable for hours to days on steel and plastic at room temperature (37). Previously, we investigated the application of high pressure processing (HPP) for the inactivation of HEV-3 in artificially contaminated pork pâté (23). HPP treatment resulted in minimal reduction in HEV infectivity (0.5-log reduction).…”

New food safety challenges of viral contamination from a global perspective: Conventional, emerging, and novel methods of viral control

“…Nasheriet al. (2020) studied the HPP treatment of hepatitis E virus, type‐3 (HEV‐3) in cell culture medium, and pork pâté. In the cell culture medium, they found 1.6 and 1.95 log 10 reductions in the load of HEV‐3 after a treatment at 400 MPa during 1 and 5 min, respectively.…”