Effect of mixing and feed batch sequencing on the prevalence and distribution of African swine fever virus in swine feed

It is critical to have methods that can detect and mitigate the risk of African swine fever virus (ASFV) in potentially contaminated feed or ingredients bound for the United States. The purpose of this work was to evaluate feed batch sequencing as a mitigation technique for ASFV contamination in a feed mill, and to determine if a feed sampling method could identify ASFV following experimental inoculation. Batches of feed were manufactured in a BSL-3Ag room at Kansas State University's Biosafety Research Institute in Manhattan, Kansas. First, the pilot feed manufacturing system mixed, conveyed, and discharged an ASFV-free diet. Next, a diet was manufactured using the same equipment, but contained feed inoculated with ASFV for final concentration of 5.6 × 10⁴ TCID₅₀/g. Then, four subsequent ASFV-free batches of feed were manufactured. After discharging each batch into a collection container, 10 samples were collected in a double ‘X’ pattern. Samples were analysed using a qPCR assay for ASFV p72 gene then the cycle threshold (Ct) and Log₁₀ genomic copy number (CN)/g of feed were determined. The qPCR Ct values (p < .0001) and the Log₁₀ genomic CN/g (p < .0001) content of feed samples were impacted based on the batch of feed. Feed samples obtained after manufacturing the ASFV-contaminated diet contained the greatest amounts of ASFV p72 DNA across all criteria (p < .05). Quantity of ASFV p72 DNA decreased sequentially as additional batches of feed were manufactured, but was still detectable after batch sequence 4. This subsampling method was able to identify ASFV genetic material in feed samples using p72 qPCR. In summary, sequencing batches of feed decreases concentration of ASFV contamination in feed, but does not eliminate it. Bulk ingredients can be accurately evaluated for ASFV contamination by collecting 10 subsamples using the sampling method described herein. Future research is needed to evaluate if different mitigation techniques can reduce ASFV feed contamination.     

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.     

Evidence of viral survival in representative volumes of feed and feed ingredients during long-distance commercial transport across the continental United States

The hypothesis that feed ingredients could serve as vehicles for the transport and transmission of viral pathogens was first validated under laboratory conditions. To bridge the gap from the laboratory to the field, this current project tested whether three significant viruses of swine could survive in feed ingredients during long-distance commercial transport across the continental US. One-metric tonne totes of soybean meal (organic and conventional) and complete feed were spiked with a 10 ml mixture of PRRSV 174, PEDV and SVA and transported for 23 days in a commercial semi-trailer truck, crossing 29 states, and 10,183 km. Samples were tested for the presence of viral RNA by PCR, and for viable virus in soy-based samples by swine bioassay and in complete feed samples by natural feeding. Viable PRRSV, PEDV and SVA were detected in both soy products and viable PEDV and SVA in complete feed. These results provide the first evidence that viral pathogens of pigs can survive in representative volumes of feed and feed ingredients during long-distance commercial transport across the continental United States.     

Detection of African swine fever virus in feed dust collected from experimentally inoculated complete feed using quantitative PCR and virus titration assays

African swine fever virus (ASFV) is a current threat to global pork production due to its high case fatality rate, lack of efficacious vaccine and recent transboundary spread into new regions of the world. Preventing introduction and further spread of ASFV is critical for countries currently negative for the virus. ASFV is stable in feed ingredients subjected to transoceanic conditions and transmission occurs through the natural consumption of contaminated feed. In this study, we investigated the use of feed dust collected from experimentally inoculated feed as a novel diagnostic sample type for ASFV detection. Moist swabs were used to collect dust from creep feeders after natural consumption of feed inoculated with 3.1–5.4 log₁₀ TCID₅₀/g ASFV Georgia 2007 in the presence and absence of antimicrobial feed additives. Results validate the potential use of feed dust swabs as a novel diagnostic surveillance tool for detection and quantification of viral nucleic acid and infectious virus titre in ASFV-contaminated feed.     

Stability of classical swine fever virus and pseudorabies virus in animal feed ingredients exposed to transpacific shipping conditions

Classical swine fever virus (CSFV) and pseudorabies virus (PRV) are two of the most significant trade‐limiting pathogens affecting swine worldwide. Both viruses are endemic to China where millions of kilograms of feed ingredients are manufactured and subsequently imported into the United States. Although stability and oral transmission of both viruses through contaminated pork products has been demonstrated as a risk factor for transboundary spread, stability in animal feed ingredients had yet to be investigated. The objective of this study was to determine the survival of CSFV and variant PRV in 12 animal feeds and ingredients exposed to environmental conditions simulating a 37‐day transpacific shipment. Virus was detected by PCR, virus isolation and nursery pig bioassay. CSFV and PRV nucleic acids were stable throughout the 37‐day period in all feed matrices. Infectious CSFV was detected in two ingredients (conventional soybean meal and pork sausage casings) at 37 days post‐contamination, whereas infectious PRV was detected in nine ingredients (conventional and organic soybean meal, lysine, choline, vitamin D, moist cat and dog food, dry dog food and pork sausage casings). This study demonstrates the relative stability of CSFV and PRV in different feed ingredients under shipment conditions and provides evidence that feed ingredients may represent important risk factors for the transboundary spread of these viruses.     

New perspectives for evaluating relative risks of African swine fever virus contamination in global feed ingredient supply chains

There are no published reports indicating that the African swine fever virus (ASFV) has been detected in feed ingredients or complete feed. This is primarily because there are only a few laboratories in the world that have the biosecurity and analytical capabilities of detecting ASFV in feed. Several in vitro studies have been conducted to evaluate ASFV concentration, viability and inactivation when ASFV was added to various feed ingredients and complete feed. These inoculation studies have shown that some feed matrices support virus survival longer than others and the reasons for this are unknown. Current analytical methodologies have significant limitations in sensitivity, repeatability, ability to detect viable virus particles and association with infectivity. As a result, interpretation of findings using various measures may lead to misleading conclusions. Because of analytical and technical challenges, as well as the lack of ASFV contamination data in feed supply chains, quantitative risk assessments have not been conducted. A few qualitative risk assessments have been conducted, but they have not considered differences in potential scenarios for ASFV contamination between various types of feed ingredient supply chains. Therefore, the purpose of this review is to provide a more holistic understanding of the relative potential risks of ASFV contamination in various global feed ingredient supply chains and provide recommendations for addressing the challenges identified.     

The risk and mitigation of foot-and-mouth disease virus infection of pigs through consumption of contaminated feed

Transboundary movement of animal feed and feed ingredients has been identified as a route for pathogen incursions. While imports of animals and animal-derived products are highly regulated for the purpose of infectious disease prevention, there has been less consideration of the viability of infectious agents in inanimate products, such as feed. This study investigated the ability of foot-and-mouth disease virus (FMDV) to remain infectious as a contaminant of commercial whole pig feed and select pig feed ingredients, and to establish the minimum infectious dose (MID_F) required to cause foot-and-mouth disease (FMD) in pigs that consumed contaminated feed. FMDV viability in vitro varied depending on virus strain, feed product, and storage temperature, with increased duration of infectivity in soybean meal compared to pelleted whole feed. Specifically, both strains of FMDV evaluated remained viable through to the end of the 37 day observation period in experimentally contaminated soybean meal stored at 4 or 20°C . The MID_F for pigs consuming contaminated feed varied across virus strains and exposure duration in the range of 10^6.2 to 10⁷ TCID₅₀. The ability of FMDV to cause infection in exposed pigs was mitigated by pre-treatment of feed with two commercially available feed additives, based on either formaldehyde (SalCURB®) or lactic acid (Guardian™). Our findings demonstrate that FMDV may remain infectious in pig feed ingredients for durations compatible with transoceanic transport. Although the observed MID_F was relatively high, variations in feeding conditions and biophysical characteristics of different virus strains may alter the probability of infection. These findings may be used to parameterize modelling of the risk of FMDV incursions and to regulate feed importation to minimize the risk of inadvertent importation.     

Feed safety collaborations: Experiences,progress and challenges

Concerns were raised regarding the role feed and feed ingredients play for risk of disease introduction and dissemination after PEDV was first identified mid-2013. Subsequently there has been a body of research and reviews completed. The results suggest a subset of contaminated feed ingredients could serve as vehicles for transboundary disease introduction into the United States. That has led to the development of biosecurity information from the pork and feed industry associations. At this time, implementation is voluntary. In 2019, representatives from pork producers, veterinarians, pork and other agriculture commodity associations and animal food industry associations formed a feed safety task force. The United States Department of Agriculture, the United States Food and Drug Administration and the Canadian Food Inspection Agency were also invited and attended. The task force operates under the premise that all participants agree there is risk of introduction of pathogens into and within the US via imported feed products. It is agreed that any actions should be achievable, are based on science and should minimize trade disruptions. The pork and feed industries have the same goal – a healthy, productive US swine herd. While our two industry sectors may have different ideas on how to prevent the introduction of diseases via imported feed ingredients, there is agreement that the general foundation for these approaches must be science based, cost effective and minimize negative impacts on market and international trade. Noncompliance with voluntary mitigation measures puts the entire pork industry at risk, all allied industries, and the US agricultural economy in general. Because of that it is essential to continue to evaluate the role of effective regulation to ensure risk of introduction is minimized through implementation of programs that will be broadly and uniformly applied.     

Could African swine fever and classical swine fever viruses enter into the United States via swine products carried in air passengers’ luggage?

On average 8,000 pork derived products are annually confiscated by Customs and Border Protection at the United States (US) ports of entry such as international airports, harbours or mail offices. These swine products with unknown sanitary status could pose a risk for foreign animal diseases introduction into the US. This study aimed at analysing the risk of African swine fever virus (ASFV) and classical swine fever virus (CSFV) being introduced into the US through prohibited swine products carried by air passengers (PSPAP) and identifying locations and time periods at higher risk where and when preventive and mitigation measures should be implemented. Our results estimated that the risk for CSFV entry was seven times higher and further spread between US airports than for ASFV. Specifically, the overall mean annual probability of ASFV entry was estimated as 0.061 at 95% confidence interval (CI) [0.007, 0.216] while the probability of CSFV entry was estimated as 0.414 (95% CI [0.074, 1]). For both diseases, July and May were the months at highest risk for entry. For ASFV, the origin countries of those PSPAP that represented the highest risk (above 70% of the total risk) were Ghana, Cape Verde, Ethiopia and the Russian Federation, while for CSFV above 90% of the risk at origin was concentrated in the Dominican Republic and Cuba, followed by India, Colombia, Peru, Ecuador and China. These results could be used to implement and feed real time surveillance systems, which could potentially help customs to increase the detection rate of smuggled products, indicating when and where to look for them. Similarly, these systems could be adapted and implemented to other diseases improving the cost‐effectiveness of the resources invested in preventing entrance of diseases via air passengers’ luggage.     

Quantitative Risk Assessment for the Introduction of African Swine Fever Virus into the European Union by Legal Import of Live Pigs

The recent incursion and spread of African swine fever virus (ASFV) in the Russian Federation and Caucasus region, close to European Union (EU) borders, have increased the concerns regarding the probability of ASFV introduction into the EU. There are many potential routes of ASFV entry into EU, but here we specifically aimed to assess the probability of ASFV introduction by legal trade of pigs, which historically has been one of the most important ways of exotic diseases introduction into the EU. A stochastic model was used to estimate the monthly probability of ASFV introduction for each country of the EU. Results of this model suggest an annual probability for ASFV introduction in the whole EU by this way of 5.22*10⁻³, which approximately corresponds with one outbreak in 192 years. The risk of ASFV introduction via live pigs was highest in Poland (69%), particularly during the months of November and December. As expected, Russian Federation is the country that most contributes to this risk, representing 68% of the overall annual risk. Methods and results presented here may be useful for informing risk‐based surveillance and control programmes and, ultimately, for prevention and control of potential ASFV incursions into the EU.     

Validation of sampling methods in bulk feed ingredients for detection of swine viruses

Animal feed can be contaminated with fomites carrying swine viruses and subsequently be a vehicle for viral transmission. This contamination may not be evenly distributed, and there is no validated sampling method for detection of viruses in animal feed or ingredients. The purpose of this experiment was to evaluate the sensitivity of ingredient sampling methods for detection of porcine epidemic diarrhoea virus (PEDV). No animals were used in this experiment, so approval from an animal ethics committee was not necessary. Thirteen kg soybean meal was used in a 2 × 2 factorial plus a control, with 2 doses of PEDV (Low: 10³ TCID₅₀/g versus High: 10⁵ TCID⁵⁰/g) and two sample types (individual probes versus composite sample). Soybean meal was confirmed PEDV negative, then loaded into individual, 1‐kg polyethylene tote bags with PEDV introduced after loading the first 100 g. There were six replicates per PEDV dose plus a control. Ten individual probes or one composite sample per bag were created and analysed for PEDV via qRT‐PCR. The interaction, dose and sample type were significant for both PEDV presence and quantity. No control samples had detectable PEDV. At the low dose, no PEDV RNA was detected in individual probes or composite samples, but was confirmed in 100% (32.4 C_t) of the inoculant samples. This is likely due to loss of sensitivity during the analysis process, which has been previously reported to cause a loss up to 10 C_t when detecting PEDV in feed or ingredients. At the high dose, only 37% (37.7 C_t) of the probes had detectable PEDV RNA. Composite samples were more sensitive (p < .05), with PEDV RNA detected in 100% of samples (35.7 C_t). In summary, sampling bulk ingredients for PEDV should include compositing at least 10 individual samples. Future research is needed to identify alternative methods that have a similar sensitivity, but require less time and effort to collect such a sample.     

Quantitative risk assessment of African swine fever virus introduction to Japan via pork products brought in air passengers’ luggage

The spread of African swine fever (ASF) has reached pandemic levels over the last decade, and outbreaks of this disease in China, Mongolia, Vietnam and Cambodia in 2018 and 2019 could accelerate its transmission to neighbouring Asian territories. Thus, the risk that the ASF virus (ASFV) will be introduced to disease‐free territories increases each year. Since Japan is an island nation, the most likely way in which ASFV would be introduced is via pork products brought in air passengers’ luggage (PPAP). Therefore, in the present study, we assessed the risk of ASFV introduction to Japan via PPAP. For the purposes of this analysis, we considered 214 international commercial flights travelling from 47 origin territories to 31 destination airports as potential routes of ASFV introduction via PPAP. The risk was estimated quantitatively through a stochastic model that considered the volume of air passengers’ luggage, the amount of confiscated pork products that were carried in air passengers’ luggage and the disease status of the origin territory. The overall mean annual probability of ASFV introduction to Japan via PPAP was found to be 0.941 [95% confidence interval (CI), 0.661–1.000], which approximately corresponds to one introduction every 1.06 years. At the origin territory level, Mongolia was led as the highest risk territory, with a risk of 0.864 (95% CI, 0.434–1.000), followed by China (0.697; 0.223–0.999), Vietnam (0.662; 0.196–0.998) and the Russian Federation (0.136; 0.018–0.401). At the destination airport level, Narita International Airport had the highest risk (0.905; 0.537–1.000), followed by Kansai International Airport (0.496; 0.109–0.961), Tokyo International Airport (0.389; 0.072–0.879) and Chubu Centrair International Airport (0.338; 0.058–0.816). This information will help improve risk management activities and monitoring systems to prevent the introduction of ASFV to Japan.     

Interventions to reduce porcine epidemic diarrhea virus prevalence in feed in a Chinese swine production system: A case study

Research has shown that feed and feed ingredients can be one of the potential routes of transmitting viral pathogens into swine farms. In this short communication, we report two cases of Porcine Epidemic Diarrhea (PED) in two sow farms located in eastern China. Immediately after the outbreaks, extensive sampling and testing for genetic materials of PEDV was carried out on farms, and at the feed mill, in an effort to identify possible sources of infection based on field observations of local area viral spread and interventions already implemented to lower risk of this spread. Samples collected from personnel or supplies entering the farms were inspected and proved as low risk factors. In contrast, feed and feed ingredient samples collected at the on-farm feed bins, and at the feed mill, tested positive for PEDV RNA. Based on these data, multiple interventions to lower viral spread via feed were implemented including (1) simplification of diet formulation excluding high-risk ingredients, (2) extension of thermal treatment during pellet conditioning and (3) maximising feed quarantine on farm up to 7 days from feed delivery to consumption. Collectively, these interventions appeared to have a positive effect as the prevalence of PED-related disease and the number of PEDV-positive feed or feed ingredient samples decreased considerably following implementation.     

Diagnostic specificity of the African swine fever virus antibody detection enzyme‐linked immunosorbent assay in feral and domestic pigs in the United States

African swine fever (ASF) is a highly contagious haemorrhagic disease of pigs that has the potential to cause mortality nearing 100% in naïve animals. While an outbreak of ASF in the United States’ pig population (domestic and feral) has never been reported, an introduction of the disease has the potential to cause devastation to the pork industry and food security. During the recovery phase of an outbreak, an antibody detection diagnostic assay would be required to prove freedom of disease within the previously infected zone and eventually nationwide. Animals surviving an ASF infection would be considered carriers and could be identified through the persistence of ASF viral antibodies. These antibodies would demonstrate exposure to the disease and not vaccination, as there is no ASF vaccine available. A well‐established commercial enzyme‐linked immunosorbent assay (ELISA) detects antibodies against ASF virus (ASFV), but the diagnostic specificity of the assay had not been determined using serum samples from the pig population of the United States. This study describes an evaluation of the World Organization for Animal Health (OIE)‐recommended Ingezim PPA COMPAC ELISA using a comprehensive cohort (n = 1791) of samples collected in the United States. The diagnostic specificity of the assay was determined to be 99.4% (95% confidence interval (CI): [98.9, 99.7]). The result of this study fills a gap in understanding the performance of the Ingezim PPA COMPAC ELISA in the ASF naïve pig population of the United States.     

Recombinant Protein p30 for Serological Diagnosis of African Swine Fever by Immunoblotting Assay

This article is devoted to the development and evaluation of the immunoblotting test system for serological diagnosis of African swine fever (ASF), based on the highly purified recombinant p30 of ASF virus (ASFV) strain Stavropol 01/08 (Stavropol 2008), representative of the ASFV currently circulating in the Russian Federation. The main project stages are as follows: (i) cloning of the central hydrophilic region of the ASFV gene CP204L (p30) into a prokaryotic vector; (ii) expression and chromatographic purification of the recombinant product p30 with thioredoxin and poly‐histidine site (p30e1_TrxA_6xHis); (iii) development of the immunoblotting test system (Rec p30‐IB) using the highly purified recombinant p30; and (iv) evaluation of Rec p30‐IB using sera and organ samples from domestic pigs and wild boars experimentally or naturally infected by ASFV. Testing of the Rec p30‐IB showed the diagnostic specificity and sensitivity of the assay to be 98.75% and 100.00%, respectively. High sensitivity of the Rec p30‐IB allowed the detection of ASFV‐specific antibodies in samples of organs of the immune system and blood sera, collected from domestic pigs and wild boars, starting from 6 to 8 days post‐infection, regardless of virus virulence, seroimmunotype and geographic origin of the samples (East Europe, South Europe, West Europe, Central and south‐east Africa).     

The Canadian 2014 porcine epidemic diarrhoea virus outbreak: Important risk factors that were not considered in the epidemiological investigation could change the conclusions

The introduction and spread of porcine epidemic diarrhoea virus (PEDV) in North America resulted in significant death loss in the swine industry. As the industry learned how to manage this disease, many new risks were identified, including the potential for feed and feed ingredients to become contaminated and spread PEDV. In addition, biosecurity practices were reevaluated and strengthened throughout the industry. At the time of the outbreak epidemiologists did not understand, as well as they are understood today, all the risk factors that contribute to the spread of PEDV. As a result, the epidemiological investigations into the 2014 PEDV outbreak in eastern Canada may not have investigated all risk factors as thoroughly as they would be investigated today. In retrospect, many of the Bradford Hill criteria used to determine causation were not fulfilled. This review identifies risk factors that were not included in the 2014 epidemiology. If these risk factors were included in the epidemiology, the conclusions and determination of causation may have been different.     

Cost-effective osteoporosis treatment thresholds: the United States perspective

Summary A United States-specific cost-effectiveness analysis, which incorporated the cost and health consequences of clinical fractures of the hip, spine, forearm, shoulder, rib, pelvis and lower leg, was undertaken to identify the 10-year hip fracture probability required for osteoporosis treatment to be cost-effective for cohorts defined by age, sex, and race/ethnicity. A 3% 10-year risk of hip fracture was generally required for osteoporosis treatment to cost less than $60,000 per QALY gained. Introduction Rapid growth of the elderly United States population will result in so many at risk of osteoporosis that economically efficient approaches to osteoporosis care warrant consideration. Methods A Markov-cohort model of annual United States age-specific incidence of clinical hip, spine, forearm, shoulder, rib, pelvis and lower leg fractures, costs (2005 US dollars), and quality-adjusted life years (QALYs) was used to assess the cost-effectiveness of osteoporosis treatment ($600/yr drug cost for 5 years with 35% fracture reduction) by gender and race/ethnicity groups. To determine the 10-year hip fracture probability at which treatment became cost-effective, average annual age-specific probabilities for all fractures were multiplied by a relative risk (RR) that was systematically varied from 0 to 10 until a cost of $60,000 per QALY gained was observed for treatment relative to no intervention. Results Osteoporosis treatment was cost-effective when the 10-year hip fracture probability reached approximately 3%. Although the RR at which treatment became cost-effective varied markedly between genders and by race/ethnicity, the absolute 10-year hip fracture probability at which intervention became cost-effective was similar across race/ethnicity groups, but tended to be slightly higher for men than for women. Conclusions Application of the WHO risk prediction algorithm to identify individuals with a 3% 10-year hip fracture probability may facilitate efficient osteoporosis treatment. The authors comprise the National Osteoporosis Foundation Guide Committee.     

Monoglyceride reduces viability of porcine epidemic diarrhoea virus in feed and prevents disease transmission to post-weaned piglets

Outbreaks of African swine fever virus (ASFv) and porcine epidemic diarrhoea virus (PEDv) have revealed the susceptibility of livestock to disease transmitted through feed. Several viruses, including PEDv, survive in feed and may introduce disease that causes significant morbidity and mortality. In 2013, PEDv, which causes severe diarrhoea and vomiting, reached North America after spreading for decades across Eurasia. The global exchange of ingredients has created demand for products that prevent disease transmission from feed. Formaldehyde-based products are highly effective at inactivating enveloped viruses when applied at 3.25 kg/t. Alternative products to formaldehyde, including carboxylic acids, essential oils and medium chain fatty acids (MCFAs), have exhibited mixed efficacy against PEDv and require application rates higher than formaldehyde. Amphiphilic molecules like MCFAs disrupt the bilayer-lipid membranes that protect viral nucleic acids through the formation of micelles. Monoglycerides form micelles at lower concentrations than MCFAs, which suggests they may be more potent against enveloped viruses. The potential efficacy of monoglycerides against enveloped viruses in feed led to the development and examination of an experimental monoglyceride blend. The proprietary monoglyceride blend significantly (p < .0001) reduced PEDv viability in vitro after application to feed at 1.5, 2.5 and 3.5 kg/t. The monoglyceride was tested in a natural feeding behaviour challenge model in piglets. The feed was contaminated with ice-blocks containing viable PEDv, and the piglets were exposed to PEDv through the feed bin for 20 days. At the end of the 20-day challenge period, all pigs were rectally swabbed and tested for PEDv by qPCR. In the untreated control group 54.8% of the piglets tested positive for PEDv, whereas none of the MCFA-treated feed (10 kg/t inclusion) transmitted PEDv. Strikingly, the monoglyceride-treated groups (1.5, 2.5 and 3.5 kg/t) all exhibited 100% protection from PEDv. These data support the use of this proprietary monoglyceride blend in mitigation and prevention of viral disease transmission to piglets from contaminated feed.     

Time trends in reported prevalence of kidney stones in the United States: 1976-1994

BACKGROUND: A body of evidence establishes that the occurrence of kidney stone disease has increased in some communities of industrialized countries. Information on recent temporal trends in the United States is lacking and population-based data on epidemiologic patterns are limited. Study objective was to determine whether kidney stone disease prevalence increased in the United States over a 20-year period and the influence of region, race/ethnicity, and gender on stone disease risk. METHODS: We measured the prevalence of kidney stone disease history from the United States National Health and Nutrition Examination Survey (II and III), population-based, cross-sectional studies, involving 15,364 adult United States residents in 1976 to 1980 and 16,115 adult United States residents in 1988 to 1994. RESULTS: Disease prevalence among 20- to 74-year-old United States residents was greater in 1988 to 1994 than in 1976 to 1980 (5.2% vs. 3.8%, P < 0.05), greater in males than females, and increased with age in each time period. Among 1988 to 1994 adults, non-Hispanic African Americans had reduced risk of disease compared to non-Hispanic Caucasians (1.7% vs. 5.9%, P < 0.05), and Mexican Americans (1.7% vs. 2.6%, P < 0.05). Also, age-adjusted prevalence was highest in the South (6.6%) and lowest in the West (3.3%). Findings were consistent across gender and multivariate adjusted odds ratios for stone disease history, including all demographic variables, as well as diuretic use, tea or coffee consumption, and dietary intake of calcium, protein, and fat did not materially change the results. CONCLUSION: Prevalence of kidney stone disease history in the United States population increased between 1980 and 1994. A history of stone disease was strongly associated with race/ethnicity and region of residence.