An analysis of select swine feed ingredients and pork products imported into the United States from African swine fever virus affected countries.

The potential for feed ingredients to serve as a vehicle for African swine fever virus (ASFV) introduction to the United States remains a significant concern. It is therefore imperative that channels through which high-risk livestock feeds and feed ingredients are imported into the United States from ASFV-positive countries are identified and considered into the USDA's ASF National Response Framework. The purpose of this study is to demonstrate the use of a novel analytical tool to categorically quantify pork products and potential high-risk feed ingredients that have entered the United States from ASFV-positive countries over a 5-year period (2016-2020). Data for this study were obtained at the United States International Trade Commission Harmonized Tariff Schedule (HTS) website (www.hs.usitc.gov), a publicly available website that provides transaction information on specific trade commodities between the United States and its international trading partners. A total of 29 high-risk pork products or feed ingredients with the potential to be fed to pigs were analysed. High risk products and ingredients were defined as those that previous research has shown to facilitate extended viral survivability, and includes products such as soybean oilcake, distillers grains, pet food, and pork sausage casings. Data were exported into Microsoft Excel and organized into pivot tables to describe the quantity of each product by country of origin and Port of Entry (POE). The analysis focused on the 60 ASFV-positive countries as currently reported by the World Organization for Animal Health (OIE). In 2020, a total of 486,902 metric tons (MT) of these high-risk products were imported into the United States from a total of 19 of the 60 foreign countries currently listed as ASFV positive by the OIE. A majority of imported animal feed ingredients came from India in 2020 (85.8%; 392,243 MT), whereas the majority of pork products and by-products were imported from Poland (21,191 MT, 70.6%). Soybean oilcake from India entered the United States through a total of 15 ports of entry (POEs) in 2020. Of these POEs, a total of five POEs received greater than 91% of all of soy oilcake originating from India, including Baltimore, MD (37.7%); San Francisco, CA (30.1%); Seattle, WA (12.7%); New Orleans, LA (5.9%); and Detroit, MI (5.1%). When examining trends over a 5-year period, a few noteworthy findings include the dramatic rise in soybean oilcake imported from India when compared to China, and a dramatic decrease in the total volume of pork sausage casings imported from China into the United States. To support the risk management of feed imports, our novel approach focuses on seaport of highest risk and quantity of product received. Data provided in this report represent an initial listing of suspect pork products and feed ingredients entering the United States, much of which is destined for animal feed. Specific detailed examples are provided in order to demonstrate the tool's flexible interface, which can be quickly modified to zero-in on specific feed ingredients, countries, or POEs depending on what the user's question may be. To the authors' knowledge, the use of and application of such a tool has not been previously utilized to support ongoing risk mitigation efforts. Potential outlets for future use of the analytical tool will include a more user-friendly and interactive interface providing an inclusive analysis of global livestock feed ingredient sourcing.

Presence of hepatitis E virus in commercially available pork products.

An increasing number of hepatitis E virus (HEV) infections in industrialized countries have been foodborne and linked to the consumption of undercooked pork products. To date, data on the prevalence of HEV in pork products sold in the United States is limited and no standard processing method exists for the detection of HEV in foods. In order to develop a processing method for the detection of HEV in pork products, ground pork and pork liver were selected for method development. Murine norovirus (MNV) was used as a process control. A filtration step prior to RNA detection was shown to reduce the level of PCR inhibitors in ground pork and an additional ultracentrifugation process was successful in removing PCR inhibitors in pork liver. MNV RNA was detected in ground pork and liver samples inoculated with 4.7 log10 PFU/g and 3.0 log10 PFU/g, respectively. Using the developed method for viral RNA detection in ground pork and pork liver, 20 packages of ground pork (six 1 g sub-samples per package) and 14 pork livers (four 1 g sub-samples per liver) were screened for the presence of HEV RNA. Fifteen out of 119 (12.6%) ground pork samples tested positive for HEV RNA and 13 out of 20 packages (65%) contained at least one positive sample. Twenty-five of 56 (45%) of pork liver samples were positive for HEV RNA and 6 of 14 livers (43%) had all sub-samples test positive for HEV RNA. Overall, the results indicate ground pork and pig liver as a potential source of HEV.

Detection of Hepatitis E Virus in the Pig Livers and Retail Pork Samples Collected in Selected Cities in China.

Hepatitis E virus (HEV) is a biological hazard that must be controlled and is a recognized etiological agent in viral hepatitis. This is a zoonotic virus and can be transmitted through the fecal-oral route. The pig is an important reservoir host of HEV, and is a source of contamination for the consumer after the consumption of raw or undercooked pork products. When detected, the most prevalent genotype of HEV in China is genotype 4 (denoted as HEV-4). To ensure the safety of this food of animal origin, we undertook a survey of HEV contamination in pig livers and pork samples available for sale, in retail outlets in selected cities in China. Viral RNA was purified from samples collected by lysing in Trizol followed by purification using trichloromethane and virus RNA extract kit. An additional step was applied to improve the recovery rate by adding RNase OUT when extracting virus RNA from pig livers, and the RNA productions were washed in 75% (v/v) ethanol to remove inhibitors. In total, 158 pig livers and 80 pork samples were procured and analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). After purification of total RNA from all samples taken and analyzed by RT-qPCR, a single pig liver was positive by this method for HEV. The positive rate was calculated as 0.63%. In this study, a single positive sample was detected. Considering the dietary habits of Chinese people, pork is a popular food that on occasion may be contaminated with HEV, thereby posing a threat to consumer health. Ongoing surveillance is required to assess the risk to human health arising from HEV-contaminated pork being offered for sale, at retail outlets, especially in the areas of China where pig production is practiced.

Foodborne transmission of hepatitis A and hepatitis E viruses: A literature review.

Foodborne viruses have been recognized as a growing concern to the food industry and a serious public health problem. Hepatitis A virus (HAV) is responsible for the majority of viral outbreaks of food origin worldwide, while hepatitis E virus (HEV) has also been gaining prominence as a foodborne viral agent in the last years, due to its zoonotic transmission through the consumption of uncooked or undercooked infected meat or derivatives. However, there is a lack of scientific reports that gather all the updated information about HAV and HEV as foodborne viruses. A search of all scientific articles about HAV and HEV in food until March 2020 was carried out, using the keywords "HAV", "HEV", "foodborne", "outbreak" and "detection in food". Foodborne outbreaks due to HAV have been reported since 1956, mainly in the USA, and in Europe in recent years, where the number of outbreaks has been increasing throughout time, and nowadays it has become the continent with the highest foodborne HAV outbreak report. Investigation and detection of HAV in food is more recent, and the first detections were performed in the 1990s decade, most of them carried out on seafood, first, and frozen food, later. On the other hand, HEV has been mainly looked for and detected in food derived from reservoir animals, such as meat, sausages and pate of pigs and wild boars. For this virus, only isolated cases and small outbreaks of foodborne transmission have been recorded, most of them in industrialized countries, due to HEV genotype 3 or 4. Virus detection in food matrices requires special processing of the food matrix, followed by RNA detection by molecular techniques. For HAV, a real-time PCR has been agreed as the standard method for virus detection in food; in the case of HEV, a consensus assay for its detection in food has not been reached yet. Our investigation shows that there is still little data about HAV and HEV prevalence and frequency of contamination in food, prevalent viral strains, and sources of contamination, mainly in developing countries, where there is no research and legislation in this regard. Studies on these issues are needed to get a better understanding of foodborne viruses, their maintenance and their potential to cause diseases.

Stability of hepatitis E virus at high hydrostatic pressure processing.

Hepatitis E virus (HEV) is the causative agent of acute and chronic hepatitis in humans. The zoonotic HEV genotype 3 is the main genotype in Europe. The foodborne transmission via consumption of meat and meat products prepared from infected pigs or wild boars is considered the major transmission route of this genotype. High hydrostatic pressure processing (HPP) is a technique, which can be used for inactivation of pathogens in food. Here, preparations of a cell culture-adapted HEV genotype 3 strain in phosphate-buffered saline (PBS) were subjected to HPP and the remaining infectivity was titrated in cell culture by counting fluorescent foci of replicating virus. A gradual decrease in infectivity was found by application of 100 to 600 MPa for 2 min. At 20 °C, infectivity reduction of 0.5 log10 at 200 MPa and 1 log10 at 400 MPa were observed. Slightly higher infectivity reduction of 1 log10 at 200 MPa and 2 log10 at 400 MPa were found by application of the pressure at 4 °C. At both temperatures, the virus was nearly completely inactivated (>3.5 log10 infectivity decrease) at 600 MPa; however, low amounts of remaining infectious virus were observed in one of three replicates in both cases. Transmission electron microscopy showed disassembled and distorted particles in the preparations treated with 600 MPa. Time-course experiments at 400 MPa showed a continuous decline of infectivity from 30 s to 10 min, leading to a 2 log10 infectivity decrease at 20 °C and to a 2.5 log10 infectivity decrease at 4 °C for a 10 min pressure application each. Predictive models for inactivation of HEV by HPP were generated on the basis of the generated data. The results show that HPP treatment can reduce HEV infectivity, which is mainly dependent on pressure height and duration of the HPP treatment. Compared to other viruses, HEV appears to be relatively stable against HPP and high pressure/long time combinations have to be applied for significant reduction of infectivity.

Surge Response: How the Panhandle Handled One of the State's Biggest COVID-19 Outbreaks.

IN TEXAS, COVID-19 outbreaks have been especially pronounced in three types of facilities: nursing homes, jails or prisons, and meatpacking plants. The Amarillo area has plenty of all three. But it was the meatpacking plants that drew national attention to Amarillo's COVID-19 problems. The city, which straddles Potter and Randall counties, has numerous plants that employ 12,000 to 15,000 people. In early April, Amarillo's two hospitals began filling up with COVID-19 patients who worked at a plant in neighboring Moore County. Two weeks later, workers from a plant in Potter County flooded in.

Effect of Sodium Chloride, Sodium Nitrite and Sodium Nitrate on the Infectivity of Hepatitis E Virus.

Hepatitis E virus (HEV) infection can cause acute and chronic hepatitis in humans. The zoonotic HEV genotype 3, which is highly prevalent in Europe, is mainly transmitted by consumption of raw meat and raw meat products produced from infected pigs or wild boars. High salt concentrations represent an important measure to preserve meat products and to inactivate foodborne pathogens. Here, an HEV preparation in phosphate-buffered saline (PBS) was subjected to different salt concentrations and the remaining infectivity was measured in a cell culture assay. Treatments with up to 20% sodium chloride for 24 h at 23 °C, with and without addition of 0.015% sodium nitrite or 0.03% sodium nitrate, did not lead to virus inactivation as compared to PBS only. Conditions usually applied for short-term and long-term fermented raw sausages were simulated by incubation at 22 °C for up to 6 days and at 16 °C for up to 8 weeks, respectively. Only 2% sodium chloride with 0.015% sodium nitrite showed a weak (< 1 log10), but significant, infectivity reduction after 2 and 4 days as compared to PBS only. Addition of 2% sodium chloride and 0.03% sodium nitrate showed a slight, but not significant, decrease in infectivity after 2 and 8 weeks as compared to PBS only. In conclusion, HEV is highly stable at high salt concentrations and at salt conditions usually applied to preserve raw meat products.

Detection and quantification of hepatitis E virus RNA in ready to eat raw pork sausages in the Netherlands.

The aim of the present study was to assess raw pork sausages collected on the Dutch market for the presence of hepatitis E virus (HEV) RNA. 46 of 316 (14.6%) products sampled from Dutch retail stores in 2017-2019 were positive for HEV RNA. HEV RNA was detected in 10.8% of "cervelaat" (n = 74), 18.5% of salami (n = 92), 26.1% of "metworst" (n = 46), 16.3% of "snijworst" (n = 43) samples. This was significantly more often than in other raw pork sausages like dried sausages, fuet or chorizo (3.3%, n = 61). The percentage of HEV RNA positive products was not significantly different for products sold as either sliced or unsliced deli meat. The average viral load in positive tested products was 2.76 log10 genome copies per 5 g, incidentally reaching up to 4.5 log10 genome copies per 5 g. The average HEV RNA level was significantly higher in samples collected in 2017 than those in samples collected in 2018, and most of the samples in 2019. Typing by sequence analysis was successful for 33 samples, all revealing genotype 3c. The results support recent epidemiological studies that identified specific raw pork sausages as risk factor for hepatitis E virus infection in the Netherlands. Persons at risk, including Dutch transplant recipients, have been advised to avoid the consumption of raw pork sausages. The study warrants a continuation of monitoring to follow the HEV RNA levels in pork products for use in risk assessments and risk management.

Assessment of hepatitis E virus (HEV) in the edible goat products pointed out a risk for human infection in Upper Egypt.

Hepatitis E virus (HEV) infection is endemic in developed and developing countries. Although the seroprevalence of HEV among the Egyptians is high, the sources of HEV infection in Egypt are not completely identified. Zoonotic HEV transmission among Egyptians is underestimated. Recently, we detected HEV in the milk of cows, this suggests the possibility of HEV transmission through the ingestion of contaminated milk. However, the role of small ruminants especially the goats in HEV epidemiology in Egypt remains unclear. Herein, we screened HEV markers in the edible goat products, mainly the milk and liver and we assessed the risk factor for HEV infection to the goat owners. A total of 280 goat milk samples were collected from 15 villages in the Assiut governorate. Anti-HEV IgG and HEV Ag were detected in 7.14% and 1.8% of the samples, respectively. HEV RNA was detected in 2 milk samples, cladogram analysis revealed that the isolated viruses belonged to HEV-3 subtype 3a. One viral isolate showed high homology to HEV recently isolated from the cow milk in the same geographic area. The level of anti-HEV IgG and HEV Ag were comparable in the milk and matched blood samples. While the urine and stool of HEV seropositive goats tested negative for HEV markers. HEV RNA was also detectable in the fresh goat liver samples (n = 2) derived from HEV seropositive goats. Finally, we analyzed HEV seroprevalence in households (n = 5) that owned the seropositive goats and households (n = 5) that owned the seronegative goats. Interestingly, anti-HEV IgG was recorded in 80% of households owned and frequently consumed the products of HEV seropositive goats, while HEV markers were not detectable in the owners of the seronegative goats. In conclusion: Here, we report HEV in the milk and liver of goats distributed in the villages of Assiut governorate. Higher HEV seroprevalence was recorded in the households that owned the seropositive goats. Investigation of the goat products is pivotal to assess the risk factor of HEV transmission to villagers in the Assiut governorate.

High Predictive Power of Meat Juice Serology on the Presence of Hepatitis E Virus in Slaughter Pigs.

Hepatitis E virus (HEV) as a zoonotic agent can be responsible for an acute hepatitis in humans, which is usually self-limiting. Progression toward a chronic stage is possible, especially in immunocompromised patients. In the past decade, the number of hepatitis E cases in humans in Germany has increased enormously to 3491 cases in 2018. Domestic pigs have been identified as a main animal reservoir and the consumption of raw and undercooked pork products, that is, livers or liver products, meat or meat products, is known as a potential risk of foodborne HEV infection. The aim of this study was to determine whether serological tests are appropriate to predict the occurrence of HEV in the liver and muscle of domestic pigs in Germany. In 2018, samples of meat juice, liver, and ham muscle were collected from 250 fattening pigs at an abattoir in North West Germany. Samples were analyzed for the presence of HEV antibodies using enzyme-linked immunosorbent assay respectively for the presence of HEV RNA using real-time reverse transcription-polymerase chain reaction. In total, 62% (155/250) of the meat juice samples were positive for HEV antibodies at a single animal basis. At herd level, 72% (18/25) of the herds were seropositive. The HEV prevalence in the liver was 17.2% (43/250). Each positive liver sample originated from seropositive herds respectively from HEV seropositive pigs. This study demonstrates for the first time the significant correlation between a positive HEV serology and the occurrence of HEV RNA in the liver of slaughter pigs (χ2 = 31.83; p < 0.001), highlighting the significant predictive power of positive serological results on the occurrence of HEV RNA in the liver.

Stability of hepatitis E virus at different pH values.

Infection with the hepatitis E virus (HEV) can cause acute and chronic hepatitis in humans. The zoonotic HEV genotype 3 is mainly transmitted by consumption of raw and fermented meat products prepared from infected pigs or wild boars. Lowering of pH during fermentation is one of the microbiological hurdles considered to inhibit growth of certain pathogens. However, no data are currently available on pH stability of HEV. As a reliable and reproducible measurement of HEV infectivity in meat products is not established so far, the stability of the cell culture-adapted HEV genotype 3 strain 47832c was analyzed here in phosphate-buffered saline (PBS) at different pH values. Only a minimal decrease of infectivity (up to 0.6 log10 focus forming units) was found after treatment at pH 2 to 9 for 3 h at room temperature. At pH 10, a decrease of about 3 log10 was evident, whereas no remaining virus (>3.5 log10 decrease) was detected at pH 1. The conditions usually achieved during curing of raw sausages were simulated using D/L-lactic acid added to PBS resulting in pH 4.5 to 6.5. After incubation at 4 °C for 7 days at these conditions, no significant differences as compared to a standard PBS solution at pH 7.7 were evident. At room temperature, a 0.8 log10 decrease was found at pH 4.7 after 7 days incubation compared to pH 7.7, but less at the other pH values. In conclusion, only minimal inactivating effects were found at pH conditions commonly occurring during food processing. Therefore, remaining infectious virus might be present in fermented meat products if HEV-contaminated starting material was used. Additional effects of other factors like high salt concentrations and low aw values should be investigated in future studies.

Occupational Animal Contact in Southern and Central Vietnam.

Despite the global zoonotic disease burden, the underlying exposures that drive zoonotic disease emergence are not understood. Here, we aimed to assess exposures to potential sources of zoonotic disease and investigate the demographics, attitudes, and behavior of individuals with sustained occupational animal contact in Vietnam. We recruited 581 animal workers (animal-raising farmers, slaughterers, animal health workers, and rat traders) and their families in southern and central Vietnam into a cohort. Cohort members were followed for 3 years and interviewed annually regarding (1) demography and attitudes regarding zoonotic disease, (2) medical history, (3) specific exposures to potential zoonotic infection sources, and (4) socioeconomic status. Interview information over the 3 years was combined and analyzed as cross-sectional data. Of the 297 cohort members interviewed, the majority (79.8%; 237/297) reported raising livestock; almost all (99.6%; 236/237) reported being routinely exposed to domestic animals, and more than a quarter (28.7%; 68/237) were exposed to exotic animals. Overall, 70% (208/297) reported slaughtering exotic animals; almost all (99.5%; 207/208) reported consuming such animals. The consumption of raw blood and meat was common (24.6%; 73/297 and 37%; 110/297, respectively). Over half (58.6%; 174/297) reported recent occupational animal-induced injuries that caused bleeding; the use of personal protective equipment (PPE) was limited. Our work demonstrates that individuals working with animals in Vietnam are exposed to a wide range of species, and there are limited procedures for reducing potential zoonotic disease exposures. We advocate better education, improved animal security, and enforced legislation of PPE for those with occupational animal exposure in Vietnam.

Infectious phage particles packaging antibiotic resistance genes found in meat products and chicken feces.

Bacteriophages can package part of their host's genetic material, including antibiotic resistance genes (ARGs), contributing to a rapid dissemination of resistances among bacteria. Phage particles containing ARGs were evaluated in meat, pork, beef and chicken minced meat, and ham and mortadella, purchased in local retailer. Ten ARGs (blaTEM, blaCTX-M-1, blaCTX-M-9, blaOXA-48, blaVIM, qnrA, qnrS, mecA, armA and sul1) were analyzed by qPCR in the phage DNA fraction. The genes were quantified, before and after propagation experiments in Escherichia coli, to evaluate the ability of ARG-carrying phage particles to infect and propagate in a bacterial host. According to microbiological parameters, all samples were acceptable for consumption. ARGs were detected in most of the samples after particle propagation indicating that at least part of the isolated phage particles were infectious, being sul1the most abundant ARG in all the matrices followed by ß-lactamase genes. ARGs were also found in the phage DNA fraction of thirty-seven archive chicken cecal samples, confirming chicken fecal microbiota as an important ARG reservoir and the plausible origin of the particles found in meat. Phages are vehicles for gene transmission in meat that should not be underestimated as a risk factor in the global crisis of antibiotic resistance.

Occurrence of HEV-RNA in Italian Regional Pork and Wild Boar Food Products.

Hepatitis E is an emerging threat in industrialized countries. The foodborne transmission linked to consumption of pork and game meat is considered the main source of autochthonous infection. In Europe, small outbreaks have been reported linked to the consumption of pork liver sausages and wild boar meat. Based on previous findings and on increasing evidence of pork and game meat as a vehicle for HEV infections, the present study investigated the occurrence of HEV in 99 pork and 63 wild boar sausages and salami sold in Southern Italy. The HEV genome was detected in four wild boar sausages. Sequencing from 2 wild boar sausages confirmed that the HEV strains detected belonged to HEV-3 genotype, not assigned to any defined subtype. Data obtained confirmed the possible occurrence of HEV in pork products and in game. Although the detection rate is low, these products are frequently consumed raw after curing, whose effect on virus viability is still unknown.

The risk of introduction of swine vesicular disease virus into Kenya via natural sausage casings imported from Italy.

Pig production in Kenya is hampered by seasonal markets. As an alternative outlet for the finished pigs, several value-added meat-processing firms have been established. Sausage, which is produced using casings derived from intestines of pigs, is one form of processed meats. Kenya imports several kgs of natural casings every year; and a recent concern is Swine vesicular disease virus (SVDV), which has never been reported in Kenya, might be introduced via natural casings imported from Italy. To determine conditions (with associated probabilities) that could lead to the introduction of SVDV, a quantitative risk assessment model was developed. Using Monte Carlo simulations at 10,000 iterations, the probability of introducing SVDV was estimated to be 1.9x10-8. Based on the suggested volume of import and mitigations used in the analysis, contaminated casings derived from an estimated 0.003 (Range = 8.1x10-8 - 0.08) infected pigs will be included in the consignment each year. The critical pathway analysis revealed that rigorous surveillance programs in Italy have a potential to dramatically reduce the risk of introducing SVDV into Kenya by this route.

Hepatitis E virus (HEV)-The Future.

Hepatitis (HEV) is widely distributed in pigs and is transmitted with increasing numbers to humans by contact with pigs, contaminated food and blood transfusion. The virus is mostly apathogenic in pigs but may enhance the pathogenicity of other pig viruses. In humans, infection can lead to acute and chronic hepatitis and extrahepatic manifestations. In order to stop the emerging infection, effective counter-measures are required. First of all, transmission by blood products can be prevented by screening all blood donations. Meat and sausages should be appropriately cooked. Elimination of the virus from the entire pork production can be achieved by sensitive testing and elimination programs including early weaning, colostrum deprivation, Caesarean delivery, embryo transfer, treatment with antivirals, protection from de novo infection, and possibly vaccination. In addition, contaminated water, shellfish, vegetables, and fruits by HEV-contaminated manure should be avoided. A special situation is given in xenotransplantation using pig cells, tissues or organs in order to alleviate the lack of human transplants. The elimination of HEV from pigs, other animals and humans is consistent with the One Health concept, preventing subclinical infections in the animals as well as preventing transmission to humans and disease.

Monitoring of pork liver and meat products on the Dutch market for the presence of HEV RNA.

The aim of the present study was to assess pork liver and meat products present on the Dutch market for the presence of hepatitis E virus (HEV) RNA. HEV RNA was detected in 27.3% of 521 products sampled from Dutch retail stores in 2016. 12.7% of livers were positive for HEV RNA (n = 79), 70.7% of liverwurst (n = 99), 68.9% of liver pate (n = 90), but in none of the pork chops (n = 98), fresh sausages (n = 103) or wild boar meat (n = 52). The highest level of HEV RNA contamination was observed in a liver (reaching up to 1 × 106 copies/g), followed by ready to eat liverwurst and liver pate (up to 3 × 104 copies/g and 7 × 104 copies/g respectively). Sequence analyses revealed mainly genotype 3c, but also some 3a, 3e and 3f strains. One strain derived from a liver sample was 100% (493 nt) identical with one isolated from a HEV case with onset of disease close in time and geography, although no direct epidemiological link could be established. Despite liverwurst and liver pate undergo heat treatment (information dd. Mid 2017) that may be sufficient to inactivate HEV, persons at risk, including Dutch transplant recipients, have been advised to avoid the consumption of raw liver as well as liverwurst and liver pate.

Determination of lumpy skin disease virus in bovine meat and offal products following experimental infection.

Lumpy skin disease (LSD) has recently expanded its range northwards to include the Balkans, Turkey and Russia. Because there was no solid evidence conclusively verifying the transmission mechanism in the field and LSDV viraemic animals with overt and asymptomatic presentation of disease and their products may represent a risk as an indirect transmission pathway. In this work, we used PCR positivity and infectivity in clinical and subclinical infection to evaluate the safety of meat and offal products from cows infected with the virulent LSDV strain Russia/Dagestan/2015. At day 21 post infection, seven of the 12 animals developed the generalized disease, and four animals became subclinically infected without apparent clinical signs. Upon examination and necropsy, the animals with the generalized disease had skin lesions; noticeably enlarged lymph nodes; and lesions in the lungs, trachea and testicles; whereas subclinically ill animals exhibited only enlarged lymph nodes and fever. For both disease presentations, testing of skeletal meat by PCR and virus isolation showed that the skeletal meat did not contain live virus or viral genome, whereas in cattle with generalized disease, meat with gross pathology physically connected under the site of a skin lesion was positive for the live virus. In subclinical infection, only enlarged lymph nodes carried the infectious virus, while the other internal organs tested in both types of disease manifestation were negative except for the testicles. Overall, our findings demonstrate that clinically and subclinically infected animals are reservoirs of live LSDV in lymph nodes and testicles, whereas deep skeletal meat in both types of infection do not carry live virus and the risk of transmission through this product seems very low. The detection of LSDV in testicular tissues in subclinically ill animals is concerning because of the potential to spread infection through contaminated semen. This aspect requires reconsideration of surveillance programmes to identify these Trojan horses of LSDV infection.

Persistent viremia and presence of hepatitis E virus RNA in pig muscle meat after experimental co-infection with porcine reproductive and respiratory syndrome virus.

Although hepatitis E virus (HEV) transmission has been demonstrated after consumption of products containing infected pig liver, human cases can be also associated with other pig meat products, such as sausages. Data on HEV viremia and dissemination in muscle meat of infected animals are still sparse, especially during long-term infection. Previously, we have shown that experimental co-infection of pigs with HEV and porcine reproductive and respiratory syndrome virus (PRRSV) lengthens HEV infection up to 49 days and increases the likelihood of the presence of HEV RNA in the liver of the pig at a later stage of infection. In the present study, we show that during experimental HEV-PRRSV co-infection, prolonged HEV viremia, up to 49 days post-inoculation (dpi), is detected. The long-term viremia observed was statistically associated with the absence of HEV seroconversion. HEV RNA was also frequently detected, at a late stage of infection (49 dpi), in the three different types of muscle tested: femoral biceps, psoas major or diaphragm pillar. The HEV RNA load could reach up to 1 ·â€¯106 genome copies per gram of muscle. Detection of HEV in muscle meat was statistically associated with high HEV loads in corresponding liver and fecal samples. The presence of HEV in pig blood, femoral biceps and major psoas, corresponding to ham and tenderloin muscles respectively, is of concern for the food industry. Hence, these results indicate new potential risks for consumers and public health regarding pork products.